Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareCOVID-19: Plasma Therapy and Stem Cell Therapy English0102Anoop KallingalEnglishEnglishhttp://ijcrr.com/abstract.php?article_id=3609http://ijcrr.com/article_html.php?did=3609

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareOccupational Awareness in Hen Poultry Farm Women Workers: A Letter to the Editor English0303Debasish Kar MahapatraEnglishEnglishhttp://ijcrr.com/abstract.php?article_id=3610http://ijcrr.com/article_html.php?did=3610

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareAntiprostatic Activity of Ferulene in Male Rats English0408Tukhtasheva Visola FarmonovnaEnglish Rejepov JumadillaEnglish Djakhangirov Farkhod NabievichEnglish Zakhidova Lola TishaevnaEnglish Halilov Ravshanjon MuratjanovichEnglish Saidkhodjayeva Dilfuza Mir-TaxirovnaEnglishEnglish Ferula finely dissected, Ferula tenuisecta, Esters of sequiterpine alcohols, Ferulen, Prostate, Chronic toxicityINTRODUCTION Prostate cancer (PC) is currently one of the most pressing diseases in urology oncology. Survival outcomes for patients with hormonal treatment are different. PC is a fairly common disease. On average, 71 cases per 100 thousand men are registered per year. Moreover, in the last decade, there has been an increase in the incidence of 3% annually. This is an alarming signal that suggests an increase in the number of registered cases by almost 2 times already by 2030. Treatment of patients with prostate cancer is an important task of modern urology due to the prevalence of the disease and the unresolved issues of its therapy. Drug therapy occupies an important place in the treatment of patients with prostate cancer and is based on the results of the latest research on the pathogenesis of this disease. For the drug treatment of patients with prostate cancer, 5-α-reductase inhibitors, α1-adrenergic receptor blockers, polyene antibiotics, antiandrogens, estrogens, and preparations of plant and biological origin are used.1,2 Prostate cancer is one of the most common diseases of men of elderly and senile age, occupying the 2-3rd place in the structure of oncological diseases of the male population and showing a clear tendency to an increase in frequency.3,4 The main functions of the gland are the partial production of semen (about 30% of the total volume), as well as participation in the process of ejaculation. A man's ability to retain urine is also related to prostate function.5,6 Because of the above, the creation of a new agent for the treatment of adenoma and prostate cancer is a scientific novelty and an urgent task. Based on the esters of ferutinin (C22H30O4), ferutin (C23H32O5), tenuferin (C23H32O6), tenuferidine (C22H30O5) and fertidine (C27H36O4) isolated from the roots of the finely dissected ferula (Ferula tenuisecta celery), a new family has been created. the drug "Ferulen" for the treatment of adenoma and prostate cancer.3 Technology has been developed for the preparation of ferulene, which consists in the extraction of plant materials with alcohol, thickening of the alcohol extract, dilution with water and extraction of esters with ethyl acetate, treatment with 5% potash solution, extraction of esters with 1% KOH solution, acidification with sulfuric acid, extraction with ethyl acetate, evaporation of ethyl acetate solution, chromatographic purification of esters on a column of silica gel (eluent - ethyl acetate-hexane 1: 3) and drying of the esters by adding microcrystalline cellulose. The yield of the ferulene substance is 18.6% by weight of the raw material, esters - 80% of the content in the raw material.4,7 The purpose of the study was to substantiate the feasibility of using the developed new drug ferulene as a means of antiprostatic action in the body. ?ATERIALS AND METHODS Animals and habitat All animals were kept in stationary vivarium conditions on a regular diet with free access to water. Experiments with them were carried out by the rules adopted by the International Convention for the Protection of Vertebrate Animals used for Experimental Purposes.6,8 Experimantal design To assess the effect of ferulene on androgen-dependent organs, the drug was administered to male rats orally using a metal probe at doses of 1-5-10 mg/kg for 10 days. Each dose of the drug was tested on 10 rats. The control group of animals under similar experimental conditions was injected with a solvent (distilled water with the addition of apricot gum). After the end of the experiment, all experimental and control animals were decapitated, the weight of the ventral prostate, coagulating gland, seminal vesicles and testes was determined; the content of testosterone in the blood serum was determined by the radiomunal method on the counter "Gamma-12" with kits for the in vitro radioimmunoassay and the immunoenzyme (Human GmbH (Germany) kit "Diagnostiksistema", Russia) by the method for the determination of testosterone in the blood serum of the Czech company "JMMUNOTECH as" Content of bioactive luteinizing hormone in the blood plasma of male rats was determined using the enzyme immunoassay analyzer MINDRAY-MR-96A. (the work was carried out at the Research Institute of Endocrinology, Ministry of Health of the Republic of Uzbekistan), histological studies were also carried out. The study of the effect of ferulene on androgen-dependent organs in comparison with surgical castration was carried out on white male rats weighing 300-320 g. the drug was administered orally using a metal probe at a dose of 10 mg/kg for 10 days. Surgical castration (orchiectomy) was performed with a longitudinal incision at the base of the scrotum, the skin and testicular membrane were dissected to the albuminous. The testicle with the epididymis and part of the spermatic cord was bluntly isolated. The rope was clamped in parts with a clamp and crossed. The proximal part of the transected cord was sutured with strong ligatures and the testicle and cord were removed. The control group of animals was injected with a solvent under similar experimental conditions. After the end of the experiment, all experimental and control animals were decapitated, the weight of the ventral prostate, coagulating gland, seminal vesicles and testes was determined; determined the content of luteinizing hormone and testosterone in the blood serum.3,4 The study of the chronic toxicity of the drug Ferulen was carried out in experiments on 40 white outbred rats - males. The drug was injected into the stomach daily at doses of 10-25-50 mg/kg for 2 months. Each dose was tested in 10 rats. The control group of animals was injected with the solvent under similar conditions. All experimental and control animals were kept in the same conditions and on the usual diet. Statistical analysis The digital material obtained during the experiments was statistically processed using the Student's t-test. RESULTS AND DISCUSSION The results of the studies showed that oral administration of ferulene at a dose of 1 mg/kg reduced the weight of the ventral prostate, coagulating gland, seminal vesicles and testicles compared with the control group of rats by 35%, 26.2%, 34% and 11%, respectively. With an increase in the dose of the drug to 5 mg/kg, ferulene reduced the mass of these organs by 56%, 39%, 41% and 23%, respectively. A further increase in the dose of ferulene to 10 mg/kg reduced the mass of the ventral prostate and the coagulating gland to values close to post-castration and compared with the control group, the mass of the ventral prostate decreased by 73%, the coagulating gland by 61%, seminal vesicles by 60% and testes by 41% (Table 1). The results of the studies carried out to assess the content of luteinizing hormone and testosterone in the blood serum showed that ferulene with 10 days of oral administration in doses of 1-5-10 mg/kg leads to a decrease in testosterone, compared with control animals by 19%, 28% and 54 % of a luteinizing hormone by 24.5%, 31% and 47.13%, respectively (Table 2). The results of a histological study showed that the introduction of ferulene at a dose of 1 mg/kg caused in some animals a slight decrease in the amount of secretion in the lumen of the prostate, a decrease in the height of epithelial cells and an increase in the connective tissue of the organ. In other animals, the reaction to the introduction of ferulene at a dose of 1 mg/kg was significantly stronger. When the dose of ferulene was increased to 5 mg/kg, the histological changes became permanent and deeper. The size of the lumens of the end sections of the gland decreased, even more, the volume of the cytoplasm of epithelial cells, their shape changed, they became low-cylindrical or cubic. Desquamation of the epithelium from a large area of the alveoli was observed. The connective tissue grew, replacing the atrophied secretory parts of the gland. The cells of the basal layer of the glandular epithelium had a pycnotic nucleus and an optically transparent cytoplasm. Hypertrophy and hyperplasia of myocytes around the secretory glands were observed.4,7-9 With a further increase in the dose of ferulene to 10 mg/kg against the background of hyperplasia of connective tissue and muscle elements, pronounced dystrophic and destructive changes in the secretory epithelium were observed. Thus, the results of the studies carried out have established that, depending on the administered dose, ferulene, to varying degrees, reduces the mass of androgen-dependent organs, causes dystrophic destructive changes in the prostate and reduces the content of luteinizing hormone and testosterone in the blood serum of male rats (Table 1 and 2). The results of the studies on the effect on the mass of androgen-dependent organs in comparison with orchiectomy are presented in Table 3. As can be seen from the table, ferulene at a dose of 10 mg/kg reduced the mass of the ventral prostate and coagulating gland at the level of surgical castration, and compared with the control animals, the mass of the ventral prostate decreased by 67%, the coagulating gland by 66%, seminal vesicles by 59%. and testicles by 35% (Tables 3, 4 and 5). The results of the studies have shown that long-term oral administration of ferulene at doses of 10-25-50 mg/kg is well tolerated by experimental animals. All experimental animals did not differ from control rats in general condition, in haematological parameters (Tables 6 and 7). The results of the studies have shown that long-term oral administration of ferulene at doses of 10-25-50 mg/kg is well tolerated by experimental animals. All experimental animals did not differ from the control rats in general condition, behaviour, and haematological parameters (Table 6 and7). On the part of biochemical parameters, a decrease in urea was noted, b- lipoproteins, cholesterol and lipids (Table 5) CONCLUSION The studies carried out in this work showed that ferulene has a pronounced antiprostatic effect when administered orally, reduces the mass of adrenogenesis-dependent organs (ventral prostate, coagulating gland, seminal vesicles, testes) and reduces the level of testosterone in the blood. In terms of antiprostatic activity, ferulene is not inferior to surgical castration. When applied again, it does not accumulate. Long-term use of ferulene is well tolerated by experimental animals and does not have a toxic effect on the part of biochemical parameters, a decrease in the mass of androgen-dependent organs (ventral prostate, coagulating gland, seminal vesicles, testes) and a decrease in the content of testosterone in the blood associated with the specific activity of the drug concerning target organs. Conflict of Interest: None Source of Funding: None Englishhttp://ijcrr.com/abstract.php?article_id=3611http://ijcrr.com/article_html.php?did=3611 Huggins C, Hodges CV. Studies on prostatic cancer: I. The effect of castration, estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate. Cancer Res 2019;1(19):293-297. Tukhtasheva VF, Rejepov J, Djakhangirov FN, Azamatov AA, Zakhidova LT. Preclinical study of safeness of the drug ferulen. Am J Med Sci Pharm Res 2020;2(6):134-143. Guryeva MV, Yu. B,  Kotov VA.  Daily monitoring of blood pressure and heart rate in the diagnosis. Russ Bull Obstet Gynecol 2013;3:4-9. Wu YW, Colford Jr JM. Chorioamnionitis as a risk factor for cerebral palsy: a meta-analysis. JAMA 2000;284(11):1417-142 Fox R, Kitt J, Leeson P, Aye C, Lewandowski AJ. Preeclampsia: Risk Factors, Diagnosis, Management, and the Cardiovascular Impact on the Offspring. J Clin Med 2019;8(10):162 Vasiliev V, Tyagunova  AV, Drozheva VV. Renal function and indicators of endogenous intoxication with gestosis.  Obstet Gynaecol 2013; 4:16-20. Mallick S. Study on the clinical profile of patients with cerebral palsy (Doctoral dissertation) 2011; 52(172):127-4 Tukhtasheva VF, Rejepov J, Djakhangirov FN, Azamatov AA, Amonov MA. Study on the antiandrogen activity of ferulen in the experiment. Eur Appl Sci 2018;1:30-32. Garg, SA, Chakravarti R, Singh NR, Masthi RC, Goyal GR. Jammy E. Dengue serotype-specific Seroprevalence among 5- to 10-Year-Old Children in India: A Community-Based Cross-Sectional Study. Int J Infer Dis 2017;54:25–30.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcarePrevalence of Premenstrual Syndrome and its Influence on Job Performance Among Nurses English0915Reham Mohammad KharabahEnglish Abdulaziz Saleh Eid AljohaniEnglish Khalid Ahmad AmaraEnglishEnglishPremenstrual syndrome (PMS), Nurses, Work related quality of life, AlMadinah, Saudi ArabiaINTRODUCTION       Premenstrual syndrome (PMS) is characterized by a wide range of emotional and physical symptoms and behavioural changes, happening before the menstruation phase of the menstrual cycle and dropping after the starting of the menstrual period, 1-6and it is classified as physical disease in the 10th revision list of the international classification of disease (ICD).6,7,8            PMS is variously defined.2 The American College of Obstetricians and Gynecologists (ACOG) defined PMS as “a clinical condition characterized by the cyclic presence of physical and emotional signs and symptoms unconnected to any organic illness that show during the 5 days before menses in each of the three previous menstrual cycles and vanish within 4 days of the onset of menses, without repetition until at least the day 13th of the cycle.4,9 Also, the American Psychiatric Association (APA) has determined criteria for the diagnosis of severe PMS or premenstrual dysphoric disorder (PMDD).4,10 Where women are diagnosed with PMDD when their lives are considerably affected by moderate to severe symptoms as defined in the Diagnostic and Statistical Manual of Mental Disorders, fifth edition.4,11            PMS disturbs the daily lives of women and can worsen their quality of life and social skills.3,12,13 The severity of PMS symptoms is associated with its duration in how it ruins the daily lives of women.3,13 Known risk factors for PMS are hormonal imbalance, fluid retention, thyroid dysfunction, genetic factors, hypoglycemia, stress and psychological factors.6,14,15 Over 200 premenstrual symptoms have been reported; however, very few are explained by changes in the menstrual cycle.5,16          Symptoms widely associated with PMS include physical symptoms such as bloating, headache, breast swelling and tenderness, nausea, weight gain and sweating, and psychological symptoms such as restlessness, anger and irritability.1-6 The incidence of PMS is very common among women of reproductive ages.3 Studies have stated that the rate of PMS with moderate and severe symptoms ranges from 4.1% to 80.2%.1-6,17-19 Where PMS prevalence rates can vary due to cultural characteristics, sample differences, and diagnostic methods.3          Few epidemiological studies have investigated the prevalence of premenstrual syndrome and its influence on job performance among nurses, particularly in Saudi Arabia. The current study aimed to  investigate the prevalence of premenstrual syndrome and its influence on job performance among nurses in Al-Madinah, Saudi Arabia, MATERIALS AND METHODS Study design This was a cross-sectional descriptive study conducted among nurses working at Madinah maternity and child hospital and affiliated Primary Health care Centers in Madinah. The calculated sample size was 300 with 95% confidence limits, 5% accepted errors, the prevalence of premenstrual syndrome =4.1% to 80.2%, 1-6and a 10% (27 nurses) was added to the sample to avoid withdrawing and refusing to participate. Procedure       A stratified random technique (proportional allocation) was applied to select the required sample size representing different clinical departments. Targeted departments were: Emergency department, Intensive care Units, medical department, surgical department, pediatric department, Oncology Physiotherapy and 7 primary healthcare centres PHCCs. A structured self –administered questionnaire. The questionnaire was taken from previous studies4,5,20,21 and modified by two experts from family medicine, it contains four main parts; A – Socio-demographic characters of the nurses. B –Menstrual and premenstrual characteristics. C –Premenstrual symptoms. D- Premenstrual syndrome scale. Variables       The dependent variable of the study was Job performance. And the independent variable were sociodemographic characteristics, menstrual and premenstrual characteristics, and premenstrual symptoms. Statistical analysis Data was entered and processed by using SPSS software version 21 for analysis and interpretation. P-Value is considered statistically significant if it is ≤ 0.05. Study approvals The following approvals were obtained: the research ethical committee, ministry of health, and the general Directors of Madinah maternity and child hospital (MMCH) and affiliated primary healthcare centres (PHCCs) in AL-Madinah. Written consent was obtained from each participant. And the collected data was handled confidentially. RESULT The Shapiro-Wilk statistic test was done for the following continuous variables (PMS score, age, children number, body mass index (BMI), absence days by week, absence days by months and age at first ministration). The result for PMS score was ( 0.991, p value= 0.745) indicating normal distribution. On the other hand the result for the rest continuous variable were (0.943, 0.851, 0.321, 0.465,0.485 and 0.963) with (pEnglishhttp://ijcrr.com/abstract.php?article_id=3612http://ijcrr.com/article_html.php?did=3612 Shameem I. Clinical Study of Mutlazima Qabl Haiz (Premenstrual Syndrome) and its Management with Unani Formulation-A Randomized Controlled Trial. Int J Curr Res Rev 2014;06(13):51-57. Erbil N. Prevalence of depressive symptoms among Turkish women experiencing premenstrual symptoms and correlated factors. Alexandria J Med 2018;54:549–553. Sut HK and  Mestogullari E. Effect of Premenstrual Syndrome on Work-Related Quality of Life in Turkish Nurses. Safety Health Work 2016;7:78-82. Buddhabunyakan N, Kaewrudee S, Chongsomchai C, Soontrapa S, Somboonporn W, Sothornwit J. Premenstrual syndrome (PMS) among high school students. Int J Women’s Health 2017;9:501–505. Erbil N, Karaca A, Kiris T. Investigation of premenstrual syndrome and contributing factors among university students. Turk J Med Sci 2010;40(4):565-573. Sahin S, Ozdemir K, Unsal A. Evaluation of premenstrual syndrome and quality of life in university students. J Pak Med Assoc 2014;64(8):915-922. Nagata C, Hirokawa K, Shimizu H. Soy, fat, and other dietary factors in relation to premenstrual symptoms in Japanese women. BJOG 2004;111:594-599. Domoney CL, Vashisht A, Studd WWJ. Premenstrual syndrome and the use of alternative therapies. Ann Ny Acad Sci 2003;997:330-343. Frank RT. The hormonal causes of premenstrual tension. Arch Neurol Psychiatr 1931;26:1052–1057. ACOG Practice Bulletin. Premenstrual syndrome. Clinical management guidelines for obstetrician – gynecologists.  J Obstet Gynecol 2001;73:183–191. American Psychiatric Association. Diagnostic and Statistical Manual of Mental Disorders. 5th ed. Washington, DC: APA Press; 2012: 465–475. Speroff L, Fritz MA. Menstrual disorder. Clinical Gynecologic Endocrinology and Infertile. 8th ed. Philadelphia, PA: Lippincott Williams & Wilkins; 2011:568–578. Direkvand-Moghadam A, Sayehmiri K, Delpisheh A, Sattar K. Epidemiology of premenstrual syndrome (pms)da systematic review and meta-analysis study.  J Clin Diagn Res 2014;8:106-109. Baker FC, Driver HS. Circadian rhythms, sleep, and the menstrual cycle. Sleep Med 2007;8:613-622. Erbil N, Bolukbas N, Tolan S, Uysal F. Determination of the premenstrual syndrome and affecting factors among married women. Int J Human Sci 2011;8:427-438. Halbreich U. The diagnosis of premenstrual syndromes and premenstrual dysphoric disorders-clinical procedures and research perspectives. Gynecol Endocrinol 2004; 19: 320-334. Demir B, Algül LY, Güvendag? Güven ES. The incidence and contributing factors of premenstrual syndrome in health working women. Turk J Obstet Gynecol 2006;3(4):262–270. Ad?güzel H, Task?n O, Danac? AE. The Symptomatology and Prevalence of Symptoms of Premenstrual Syndrome in Manisa. Turk Psikiyatri Derg 2007;18(2):215–222. Hamaideh SH, Al-Ashram SA, Al-Modallal H. Premenstrual syndrome and premenstrual dysphoric disorder among Jordanian women. J Psyhiatr Ment Health Nurs. 2014;21(1):60–68. Gencdogan B. A new instrument for premenstrual syndrome. Psychiatry in Turkey 2006;8:81-87. Padmavathi  P, Sankar  R, Kokilavani  N, Dhanapal K, Ashok B. Validity and Reliability Study of Premenstrual Syndrome Scale (PMSS). Int J Adv Nurs Manag 2014; 2(1):4-5. Marjoribanks J, Brown J, O’Brien PMS, Wyatt K. Selective serotonin reuptake inhibitors for premenstrual syndrome. Cochrane Database Syst Rev 2013;7:  CD001396. Cheng SH, Shih CC, Yang YK, Chen KT, Chang YH, Yang YC. Factors associated with premenstrual syndrome - a survey of new female university students. Kaohsiung J Med Sci 2013;29(2):100–105. ACOG Committee on Practice Bulletins--Gynecology. Premenstrual Syndrome. ACOG Practice Bulletin No. 15. Obstet Gynecol 2000;95(4):1-9. Anandha LS, Saraswathi I, Saravanan A. Prevalence of premenstrual syndrome and dysmenorhoea among female medical students and its association with college absenteeism. Int J Biol Med Res 2011;2:1011-1016. K?rcan N, Ergin F, Adana F, Arslantas H. The prevalence of premenstrual syndrome in nursery students and its relationship with quality of life. J ADÜ Med Faculty 2012;13:19-25. Kutlu R, Demirbas N, Dag?stan F. Frequency and Affecting Factors of Premenstrual Syndrome Among Turkish Female University Students. J Clin Anal Med 2017;8(5): 416-420. Direkvand-Moghadam A, Sayehmiri K, Delpisheh A, Sattar K. Epidemiology of premenstrual syndrome (pms)da systematic review and meta-analysis study.  J Clin Diagn Res 2014;8:106-109. Chayachinda C, Rattanachaiyanont M, Phattharayuttawat S, Kooptiwoot S. Premenstrual syndrome in Thai nurses. J Psychosom Obstet Gynaecol 2008;29:199-205. Demir B, Algül LY, Güven ESG. The incidence and the contributing factors of premenstrual syndrome in health working women. J Turk Obstet Gynecol 2006;3:262e70. Bertone-Johnson ER, Whitcomb BW, Missmer SA, Manson JE, Hankinson SE,  Rich-Edwards JW. Early life emotional, physical, and sexual abuse and the development of premenstrual syndrome: a longitudinal study. J Womens Health 2014;23:729-739. Abdelmoty HI, Youssef MA, Abdallah S, Abdel-Malak K, Hashish NM, Samir D, et al. Menstrual patterns and disorders among secondary school adolescents in Egypt. A cross-sectional survey. BMC Womens Health 2015;15:70. Erbil N, Karaca A, K?r?s T. Investigation of premenstrual syndrome and contributing factors among university students. Turk J Med Sci 2010; 40 (4): 565-573.  Babacan-Gumus A, Bayram N, Can N, E. Kader. Premenstrual syndrome in university students: an investigation in terms of somatization and some variables. Anato J Psychiatry 2012;13:32-38. Borenstein JE, Dean BB, Endicott J, Wong J, Brown C, Dickerson V, Yonkers KA. Health and economic impact of the premenstrual syndrome. J Reprod Med 2003;48:515-524. Ozturk S, Tanr?verdi D. Premenstrual Syndrome and Management. Journal of Anatolia Nurs Health 2010;13:3:57-61. Weisz G, Knaapen L. Diagnosing and treating premenstrual syndrome in five western nations. Soc Sci Med 2009;68:1498–1505. K?sa S, Zeyneloglu S, Güler N. Prevalence of Premenstrual Syndrome among University Students and Affecting Factors. Gümü?hane Uni J Health Sci 2012;1(4):284-297.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareTo Evaluate the Sleep Pattern in Children with and without Developmental Co-Ordination Disorder English1619Sujatha BaskaranEnglish Jagatheesan AlagesanEnglish Pooja KEnglish Thirumal SEnglishEnglishSleep pattern, DCD, School children, Questionnaire, DSMVINTRODUCTION           In general, many clinical conditions may Impact motor skills such as epileptic syndromes, autism spectrum disorders, Attention Deficit Hyperactivity Disorder (ADHD), primary headache, learning disorders, and obesity. Whereas, when motor coordination is primarily impaired, Developmental Coordination Disorder (DCD) may be identified.1           Developmental coordination disorder is characterized by significant difficulty in the development of motor skill in the absence of obvious sensory or neurological impairment. Children with DCD struggle to perform everyday movement tasks requiring fine manipulation, eye-hand coordination and balance and agility.2 Sleep disturbance is common in groups of children with other neurodevelopmental disorders and psychiatric disturbances, such as anxiety and depression.2,3            Previous studies have estimated that the prevalence of sleep disturbance in typically developing children is in the range of 20-40% conversely, the prevalence of sleep disturbance in children with developmental disorders, such as Pervasive developmental disorder (PDD) and Attention deficit hyperactivity disorder (ADHD), has been reported to the two to three-fold higher.1             Many children with DCD perform poorly at school despite average intellectual ability and may fail to achieve their academic potential. A range of socio-economical problems has also been reported, including low self-esteem, high anxiety and poor peer relations. Emotional and behavioural problems are seen in populations of children with a range of other developmental disorder, which suggests that the link impaired motor function may not be direct.2            The current study has compared the sleep pattern of typically developing children with developmental coordination disorder, evaluated rate of sleep disturbance are continuously reported in children with the range of medical and psychiatric conditions and developmental disorder.2 The children sleep habit questionnaire (CSHQ) has been used in several countries to assess children sleep patterns and sleep problems as reported by parents.1           Thus many of the adverse effects may be rectified by addressing sleep disturbances, to enhance the overall functioning of the child and family. The children sleep habit questionnaire has high validity and reliability, specifically designed for school-aged (9 years to 14 years) children, the design of children sleep habit questionnaire is based on common clinical symptom presentations of the most prevalent pediatric international classification of sleep disorders diagnoses.5           The BOT-02 tool of motor proficiency is a standardized, norm-referenced measure used by the physical therapist in the clinic and school practice settings. The BOT-02 tool is an individually administered measure of fine and gross motor skills of children 9 through 14 years of age. It is intended for use by practitioners and researchers as a discriminative and evaluative measure to characterize motor performance, specifically in the areas of fine motor control, manual coordination, body coordination, strength and agility.6 The aim is to evaluate the sleep pattern in children with and without developmental coordination disorder.6,7 MATERIALS AND METHOD           The Study designed as an Observational Study, Study setting done at Schools in the Chennai district and the sampling method is the Conventional sampling method. The sample size is 14 School going children. Boys and girls between 9-14 years, Children without musculoskeletal disorders and Children who scored 15-57 in the Developmental coordination disorder 2007 DCD 07 questionnaire were included. Subject with a history of respiratory illness, Subjects with existing cardiovascular disease, Subjects who are already in the treatment program, Subjects with neurological and sensory impairment and Subjects with any congenital deficit were excluded. Materials required are developmental coordination disorder 07 questionnaires, BOT-02 and children sleep habit questionnaire.           A total of 414 school-going children were screened after obtaining permission from the school heads, detailed study procedure was being explained to the parents, teachers and children. Written informed consent was obtained from the parent or guardian. Children were allocated in two groups. Group A consisted of 14 children with developmental coordination disorder, who scored 15-57 in the DCD-07 Questionnaire and Group B consisted of 14 children without Development coordination disorder. The outcome measure used in the study is, Diagnostic and Statistically Manual of Mental disorder (DSM V) has four criteria to rule out Development coordination disorder and they are, Criteria-A: The acquisition and execution of coordinated motor skills are substantially below the expected given the individual chronological age and opportunity for skill learning and use. Difficulties are manifested as clumsiness (e.g., dropping or bumping into objects) as well as slowness and inaccuracy of performance of motor skills (e.g., catching an object, using scissors or cutlery, handwriting, riding a bike or participating in sports.) Criteria-B: The motor skills deficit in criterion A significantly and persistently interferes with activities of daily living appropriate to chronological age (e.g., self-care and self-maintenance) and affects academic/ school productivity, prevocational and vocational activities, leisure and play. Criteria-C: The onset of symptoms is in the early developmental period. Criteria-D: The motor skills deficits are not better explained by intellectual disability (intellectual developmental disorder) or visual impairment and are not attributable to a neurologic condition affecting movement (e.g., cerebral palsy, muscular dystrophy, degenerative disorder). To fulfil the Criteria- A of DSM V, The Bruininks- Oseretsky test of motor performance 2 (BOT 2) tool was also used to assess the motor function of children with DCD. Scores greater than 25 indicates well about average (WAA) motor function, 20 to 24 indicates above average (AA) motor function, 11 to 19 indicates the average motor function, 6 to 10 indicates below average (A) motor function, and less than 5 indicates well below average (WBA) motor function respectively. To fulfil Criteria-C of DSM V, the parental interview was conducted by the researcher about the onset of developmental symptoms of other children. To fulfil Criteria- D of DSM V every child with probable DCD was assessed by the physician to rule out any other major illness and neurological disorder.  RESULTS The number of subjects screened for this study was 414 out of which 14 (4.8%) subjects were found to be under DCD criteria. In gender distribution among the 14 subjects, 9(64.3%) were boys and 5(35.7%) were girls who have DCD. According to the data collected, the DCD children have an increased sleep pattern compared to the normal children, you have a normal sleep pattern. Based on the children sleep habit questionnaire the mean values of individual subscales were calculated. DCD children showed 28.7% (16.79) at bedtime, 41.1% (24) in sleep behaviour, 5.4% (9.29) in morning waking and 8.9% (5.21) in Day time sleepiness. On comparing sleep habit in DCD children with non-DCD children, the results of the current study show a significant p-value Englishhttp://ijcrr.com/abstract.php?article_id=3613http://ijcrr.com/article_html.php?did=36131. LeBourgeois MK, Hale L, Chang AM, Akacem LD, Montgomery-Downs HE, Buxton OM. Digital media and sleep in childhood and adolescence, Pediatrics 2017;140(2):S92-6. 2. Gringras P, Green D, Wright B, Rush C, Sparrowhawk M, Pratt K, et al. Weighted blankets and sleep in autistic children - A randomized controlled trial. Pediatrics 2014;134(2):298-306. 3. Cattane N, Richetto J, Cattaneo A. Prenatal exposure to environmental insults and enhanced risk of developing Schizophrenia and Autism Spectrum Disorder: focus on biological pathways and epigenetic mechanisms. Neurosci Biobehav Rev 2018;4:123-126. 4. McLeod KR, Langevin LM, Goodyear BG, Dewey D. Functional connectivity of neural motor networks is disrupted in children with developmental coordination disorder and attention-deficit/hyperactivity disorder. NeuroImage Clin 2014;4:566-575. 5. Schoemaker MM, Niemeijer AS, Reynders K, Smits-Engelsman BC. Effectiveness of neuromotor task training for children with developmental coordination disorder: a pilot study. Neural Plast 2003;10. 6. Missiuna C, Moll S, King S, King G, Law M. A trajectory of troubles: parents impressions of the impact of developmental coordination disorder. Phys Occup Ther Pediatr 2007;27(1):81-101. 7. Polatajko HJ, fox M, Missiuna C. An international consensus on children with developmental coordination disorder. Can J Occup Ther 1995;62:3-6. 8. Henderson SE, Sugden DA, Barnett AL. Movement Assessment Battery for Children, 2nd edn. London: Pearson, 2007. 9. Schoemaker MM, flapper B, Verheij NP, Wilson BN, Reinders-Messelink HA, de Kloet A. Evaluation of the developmental coordination disorder questionnaire (DCDQ) as a screening instrument. Dev Med Child Neurol 2006;48(8):668-673. 10. Magalhães LC, Missiuna C, Wong S. Terminology used in research reports of developmental coordination disorder. Dev Med Child Neurol 2006;48(1):937-941. 11. Mandich AD, Polatajko HJ, Rodger S. Rites of passage: understanding participation of children with developmental coordination disorder. Human Movement Sci 2003;22(4-5):583-595. 12. Magalhães LC, Missiuna C, Wong S. Terminology used in research reports of developmental coordination disorder. Dev Med Child Neurol 2006;48(1):937-941. 13. Wilson BN, Kaplan BJ, Crawford SG, Campbell A, Dewey D. Reliability and validity of a parent questionnaire on childhood motor skills. Am J OccupTher 2000; 54(5):484-493. 14. Terzano M, Parrino L, Smerieri A, Carli F, Nobili L, Donadio S, et al. CAP and arousals are involved in the homeostatic and ultradian sleep processes. J Sleep Res 2005;14:359-368. 15. Biotteau M, Péran P, Vayssière N, Tallet J, Albaret JM. Neural changes associated to procedural learning and automatization process in Developmental Coordination Disorder and/or Developmental Dyslexia. Eur J Paediatr Neurol 2017;21(2):286-299.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareMother’s Education and Mortality under Age Three: An Investigation in Presence of Some Socio-Demographic Correlates English2028Brijesh P. SinghEnglish Madhusudan JVEnglish Sandeep SinghEnglishEnglish Binary logistic regression, Mortality under age three, Mother’s education Introduction The level of infant and under-five mortality is still not up to the mark in India. The infant mortality rate (IMR) is the probability of dying before one year of age expressed per1000 live-births and the under-five mortality rate (U5MR) is the probability of dying between birth andage 5 expressed per 1000 live-births have been used as measures of children’s well-being for manyyears. Previous studies in developing countries showed that education played a crucial role in improving the survival status of children.1,2 There are some pieces of literature are available that documented maternal education as a significant determinant of child health outcome.3-9 These studies discuss several reasons for the inverse association between maternal education and infant/child death. It is expected that better maternal education leads to higher use of the modern health care system10 and highly educated mothers are more likely to take decisions on self illness control11,12 which helps prevent premature deaths13-16 and increases empowerment of women about health care and decision making.10,14,17 Finally, educated mothers have higher levels of physical movements which are associated with their concern for antenatal care18 and thus treatment-seeking for a sick child.19When assessing the health benefits of increased education in less developed countries, many researchers have been concerned about the omission of important determinants of an individual’s education from the models. An increase in education would reduce mortality not only because more women would enter an educational category associated with lower mortality, but also because everyone, including those who themselves remained uneducated, would benefit from the generally higher level of education in the community. Such a community level contribution was seen in recent analyses of fertility from Africa20 and India,21 and a similar effect of literacy was shown for India.22These studies have shown that among all socio-economic factors, maternal education has the most influence on the variation of infant and child mortality at the regional level. Some other studies in the domain of education and health have largely focused on examining the role of mother’s education in reducing the morbidity and mortality of their children.23-26 Mortality under age three has special importance because the child in the developing world is vulnerable to infectious diseases such as diarrhoea, malaria, acute respiratory infections (ARI) and parasites as well as immunizable ones like neonatal tetanus, measles and pertussis.27 Children have a higher mortality rate (of 30 to 40 %) than any other age group and most of the deaths (70%) occur from infectious diseases such as diarrhoeal diseases, acute respiratory infections (pneumonia), measles and malaria.28 The above diseases are especially uncommon under six months of age due to presumably protection of breastfeeding. Thereafter, disease prevalence increases with the age of the child. After age five months they may wean from breast milk and supplementary foods may be introduced and at that moment their immune system may weaker than in older age. Several studies suggest considering special care for children betweenthe age of 7 and 36 months. Poverty usually coexists with illiteracy, unemployment, malnutrition, poor health and low status of women. Child mortality differs remarkably between urban and rural area. Mother’s education, mother’s age at birth, birth order, basic immunization, household size and sex are the important determinants of infant and child mortality.29-32 It is noted that preceding short birth intervals adversely affect infant survival and infant and child mortality.33-35 This study intends to quantify the contribution of education in explaining the gap in mortality under age three with a special focus on the effect of maternal education. The purpose of this paper is to investigate the level of a child under age three mortality in Uttar Pradesh, the most populous state of India, according to some socio-demographic characteristics. Also examines the relative importance of different sets of variables (such as socio-demographic variables) in influencing under-three mortality.36,37 MATERIALS AND METHODS We have utilized the data from the fourth round of the National Family Health Survey (NFHS)38, conducted in 2015-2016. The NFHS is a nationally representative survey firstly conducted by the International Institute for Population Science, Mumbai as the Nodal Agency under the patronage of the Ministry of Health and Family Welfare, Government of India during 1992-1993. Consecutively four National Family Health Surveys (NFHS) were completed in India. The NFHS 2015-2016 surveyed 601,509 households, 699,686 women aged 15-49 years with a response rate of 97%, and 112,122 men aged 15-54 years with a response rate of 92%. Besides women and men, this survey also provides relevant information about their children. In this study, we have restricted our sample children born three years before the survey and their mother age is between 16-30 years to reduce recall biases. A detailed description of the sampling design and survey procedure is provided in the national report of NFHS-4, 2015-2016. The births before the three years of the survey in Uttar Pradesh are considered for analysis. For this study, a total of 6482 births which occurred three years before the survey are found and 452 among these is dying before reaching their third birthday. Mortality under age three is (children below 3 years) the outcome of interest in this study. Direct estimates of the probabilities of under age three mortality were calculated for each category of independent variables. Mortality under age three of the child is in dichotomous form (if the child was dead coded as ‘1’ and otherwise ‘0’). Explanatory variables maternal education is the main predictor variable in this study. Maternal education was categorized into four groups, i.e., no education or illiterate, primary education, secondary education and higher education. Other explanatory variables included in this study are religion, wealth index, residence, maternal age at birth, preceding birth interval, sex of children, birth order, basic immunization, household size, number of eligible women (currently married women aged 15-49 living with her husband) living in the house and media exposure. Descriptive statistics are used to show sample characteristics and bivariate percentage distribution was carried out to assess the differentials in mortality under age three by selected predictor variables. Finally, a binary logistic regression is used to examine the associations between maternal education and child mortality. The regression results were estimated by unadjusted and adjusted odds ratio with a 95% confidence interval. A Brief Discussion of the Logistic Regression Regression analysis is a major subset of multivariate analysis and a very useful tool not only to predict the response variable with the help of one or more predictors but also to understand factors that are responsible to bring out the changes in the response variable. Linear regression is the simplest form of regression. In this form of the regression model, it is assumed that each predictor variable has a direct effect on the study variable. Since many predictor variables affect the response or study variable and these may affect another intervening variable and thus problem of multi-collinearity may come into the picture. In regression analysis, the study variable is generally a quantitative variable measured on the interval scale.36,37 In case the study variable is dichotomous then the linear regression model is not very effective due to the violation of various assumptions, i.e. linearity, homoscedasticity etc. In such a situation if the regression model is used, the fit of the line will be very poor having low R2 and even testing of the hypothesis is invalid. Under these circumstances (when the study variable is dichotomous and independent variable are of a different type, some are measured on an interval scale and others on the categorical scale) logistic regression model is used. Sometimes it is also known as the logit regression model. The basic form of Logistic function is   Where z is the predictor variable. Let us suppose that z instead of being a single predictor variable, is a linear combination of a set of predictor variables i.e.and therefore  This function is known as logit of P or log odds, and this is the familiar form of an ordinary multiple regression equation and therefore the interpretation of logit regression coefficient is the same as regression coefficients but the difference is that the effect is measured not on the study variables but the logit of the study variable.38,39 Results We have analysed 6482 children’s information in this paper and Table 1 shows that more than seventy % (75.6%) of the children are from rural areas and the rest of the children from urban areas. Among the child of the mother, 40.6 % are illiterate and about 11 %of mothers are well educated. About three fourth children belong to the Hindu religion. Half of the children belong to poor households and about 30 % are from rich households, only 18 %of children having middle wealth index category. The majority of mother’s age at birth (77.4%) lies between 20-30 years and very few (5.3%) mother’s age at birth below 20 years. One-third of mothers have one child and the majority (79.0%) have up to three births. More than half of the mothers have their preceding birth interval is more than 24 months and 8.1 %s havea preceding birth interval less than 12 months. As for the sex of the child is concerned, male children are more preferred than females in the sample. Two-third of deliveries of children are observed in hospital (not at home) and the majority of deliveries are normal only 9 %s are by caesarean section. One year immunization coverage (3 dose polio+3 dose DPT and Measles) is considered in this study and found about two-thirdsof children are immunized. 37 %of mothers are not exposed to any type of media (newspaper, radio and TV). We observed in the sample about 18 %of children’s household size is less than or equal to 4. About 48 %of household size is 7 or more. About 36 %of the children’s house, there are 2 or more eligible women are living.  Table 2 shows the under three child mortality rates according to some selected socio-demographic characteristics. The result exhibits that, child mortality under age three is 5 % higher among rural child than their urban counterparts. Education is inversely related to child mortality. Well educated mothers experience lower child mortality. Religion does not pose a difference in mortality under age three. Children of households with poor wealth index category experienced a higher probability of dying. Mothers belong to the age at birth category 20-30 have less probable to have mortality under age three than another comparison group. Higher birth order shows higher under age three mortality (113/1000). When the preceding birth interval is less than 12 months the mortality under age three is about 132 per thousand. Smaller household size shows higher mortality under age three (121 per thousand). Chi-square test and p-value reveal that all other predictors are significant except the place of residence, sex of the child, type of delivery and eligible women. Overall the estimate of mortality under age three is 75 per thousand for the study sample. The logistic regression analysis is employed to identify the important contribution of variables that influence mortality under age three. This analysis is also aimed at the relative importance of two sets of socio-demographic characteristics in influencing under age three mortality. In this analysis, the dependent variable is the survival status of the child. A variable is considered significantly associated with underage three mortality when its p-value is below 0.1 at least. The results of the analysis are shown in Table 3. In the first model, when all the variables were taken separately, the analysis suggests that, among the socio-demographic variables, mother’s education, wealth index, mother’ age at birth, birth order, preceding birth interval, immunization, household size and media exposure are statistically significant concerning the survival status of the mortality under age three. Discussion As expected, the child of illiterate mothers has experienced significantly higher mortality (about 3 times) as compared with the well-educated mothers. Children of households with a poor wealth index are 1.6 times more likely to die than those of households having a better economic standard. The child had born to the mothers at age less than 20 years and 30+ years experienced 1.8 and 1.5 times significantly higher risk of dying respectively, as those born to the mother’s age between 20-30 years. Children of birth order 1 is 1.35 times more likely and child with birth order 4+ are 1.98 times more likely than those of 2-3 birth order women. The child with a preceding birth interval less than equal to 12 months and 12-24 months faced significantly higher odds of dying (2.5 and 1.8 times respectively) than those with the birth interval of 24+ months and above. Male child’s experienced a 4 % higher risk of dying than female children but it is insignificant. Children who are immunized are 36 % significantly less vulnerable to die before their third birthday. Children from larger household size show a significantly lower chance of dying.40-42 In the second model, when all the socioeconomic variables are taken together except the mother’s education. This model reveals that the wealth index, mother’s education, birth order 4+, preceding birth interval, immunization status and household size are significant predictors for mortality under age three. The third model considered as a full model in this study when all the socio-demographic variables with the mother’ education are taken together in the model to examine the net effect of the variables. Though the controls for socio-demographic variables have decreased the effect of mother’s education on mortality under age three, yet only mother’s education, birth order 4+, preceding birth interval, immunization and household size continue to remain statistically significant for mortality under age three. Some model suitability criterion such as -2Log likelihood, Cox & Snell R square and NagelkerkeR Square shows that Model 3 is better than Model-2 to explain the under age three mortality.42,43 It indicates that for explaining under age three mortality mother’s education is emerging as an important predictor even in presence of socio-demographic variables. Conclusion This study demonstrates that the mother’s education emerges as an important factor apart from other socio-demographic variables to influence the child under age three mortality. Similar findings have been observed.In this study, education of mother, preceding birth interval, immunization, birth order and household size has emerged as a very important factor to influence under age three mortality. It is generally agreed that education acts as independent determinants of under age three mortality and at times it is considered as a proxy variable for other social variables. Education of the mother influence the child survival through various pathways; which are enhanced socio-economic status, greater health choice for children, including interaction with medical personnel, cleanliness, emphasis on child quality in terms of fewer children, and greater food and capital investment23,41. Findings of the study show that illiterate mothers experienced higher under age three mortality. Moreover, maternal education seems to have indirect effects through the treatment-seeking behaviours as well as family formation patterns in case of infant mortality. The economic standard of the household, which was taken as the proxy of family income, emerged as one of the most important factors influencing under age three mortality. The better economic standard enables the mother to have greater health choice for children and to have greater food and capital investment. The study reveals that the children of the households with low economic standard experienced more risk of dying than those of the households having a better economic standard. Thus steps should be taken to improve the education and economic standard of the people. In addition to that, free health services could be provided to people in remote rural areas. A mother’s age at childbirth, birth order, and preceding birth interval havea powerful effect on the survival chances of a child. These are also found important variables to influence under age three mortality. Young mothers face more risk of a child dying because they may not be physiologically and emotionally mature enough to adequately manage a pregnancy. Moreover, young mothers also have poor child care skills, which derive partly from inexperience in child-rearing. A preceding birth interval of fewer than one years is significantly associated with higher child mortality risk than those with birth intervals of more than one years. Increasing education and the age at marriage, reproductive span could be cut off to some extent, which in turn will help increase mother’s age of childbirth. People should also be encouraged to keep their number of kids small. Acknowledgements: The authorsare extremely thankful to the reviewers for their valuable suggestion for theimprovement of the paper. Competing interests: Authors have declared that no competing interests exist. Financial support: We don’t have any financial assistance from anywhere.A Ethical Issue: The data used in this study is taken from NFHS-IV and freely available for research use thus there is no need for ethical clearance. Englishhttp://ijcrr.com/abstract.php?article_id=3614http://ijcrr.com/article_html.php?did=3614 Caldwell J, McDonald P.Influence of maternal education on infant and child mortality: levels and causes. Health Policy Edu1982;2(3-4):251-267. Cochrane SH, Leslie J, O’Hara DJ. Parental education and child health: intra country evidence. Health Policy Edu 1982;2:213-250. Jejeebhoy S. Women’s education, autonomy and reproductive behaviour: experience from developing countries. Oxford: Clarendon Press, 1995. Govindasamy P, Ramesh BM. Maternal education and utilization of maternal and child health services in India. NFHS Subject Reports Number 5, Int.Institute of Population Sci: Mumbai, (1997). Hatt LE, Waters, HR. Determinants of child morbidity in Latin America: a pooled analysis of interactions between parental education and economic status. Soc Sci Med 2006;62:375-386. Pamuk, ER, Fuchs, R, Lutz, W. Comparing relative effects of education and economic resources on infant mortality in developing countries. Popul Dev Rev 2011;37:637-664. Singh A, Pathak PK, Chauhan RK, Pan W. Infant and child mortality in India in the last two decades: a geospatial analysis. PloS One 2011;6(11):e26856. Singh Brijesh P, Maheshwari, S. Infant mortality and its determinates in Uttar Pradesh, india. J Stat Appl Prob Lett 2014;1(3):47-52. Vikram K, Vanneman R, Desai S. Linkages between maternal education and childhood immunization in India. Soc Sci Med 2012;75:331-339. Frost MB, Forste R, Haas DW. Maternal education and child nutritional status in Bolivia: finding the links. Soc Sci Med 2005;60:395-407. Basu AM, Stephenson R. Low levels of maternal education and the proximate determinants of childhood mortality: a little learning is not a dangerous thing. Soc Sci Med 2005;60(9):2011-2023. Desai S, Alva S. Maternal education and child health: is there a strong causal relationship? Demography1998;35:71-78. Alemayehu AA, Huang W. Maternal education, linkages and child nutrition in the long and short-run: evidence from the Ethiopia Demographic and Health Surveys. Int J Afr Dev 2014;1(2):5-28.  Aslam M, Kingdon GG. Parental education and child health-understanding the pathways of impact in Pakistan. World Dev 2012;40:2014-2032. Glewwe P. Why does mother’s schooling raise child health in developing countries? Evidence from Morocco. J Hum Resour 1999; 124-159. Webb P, Block S. Nutrition information and formal schooling as inputs to child nutrition. Econ Dev Cult Change 2004;52:801-820. Choudhury PK. Explaining the role of parental education in the regional variations in infant mortality in India. Asia Pac Policy Stud 2015;2:544-572. Bloom SS, Wypij D, Das Gupta M. Dimensions of women’s autonomy and the influence on maternal health care utilization in a North Indian city. Demography 2001;38(1):67-78. Dasgupta M. Death clustering, mother’s education, and the determinants of child mortality in rural Punjab, India. Population Stud 1990;44(3):489-505.  Kravdal O. Education and fertility in sub-Saharan Africa: individual and community effects. Demography 2002;39(2):233-250. Moursund A,KravdalO. Individual and community effects of women’s education and autonomy on contraceptive use in India. Population Stud 2003;57(3):285-301. McNay K, Cassen R, Arokiasamy P. Why are uneducated women in India using contraception? A multilevel analysis. Population Studies 2003;57(1):21-40. Caldwell J. Education as a factor in mortality decline: an examination of Nigerian data. Population Stud1979;33(3):395-413. Gakidou E, Cowling K, Lozano, R, Murray CJL. Increased educational attainment and its effect on child mortality in 175 countries between 1970 and 2009: A systematic analysis. Lancet 2010;376(9745):959-974. Papageorgiou C, Stoytcheva P. Education inequality among women and infant mortality: A cross-country empirical investigation. November 2012. http://www.chrispapageorgiou.com/papers/PS.pdf. Saikia N, Singh A, Jasilionis D, Ram F. Explaining the rural-urban gap in infant mortality in India.DemographRes2013;29(18):473-506. Tsui AO, Declerque J, Mangani N. Maternal and socio-demographic correlates of child morbidity in Bas Zaire: the effects of maternal reporting. Soc Sci Med1988;26:701-713. Yohannes AG, Streatfield K, Bost L. Child morbidity patterns in Ethiopia. J BiosocSci1992;24:143-155. Bairagi R, Santosh Chandra Sutradhar, Nurul Alam. 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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareAntibiogram and Isolation of S. aureus from the Urinary Tract Infections: Comparison of Meca Gene Detection and Phenotypic Methods for Detection of Methicillin-Resistant S. aureus English2933Basavaraj C. MetriEnglish Jyothi PEnglishEnglish Urinary tract infections, S.aureus, MRSA, PCR, mecA geneIntroduction             Bacterial resistance to antibiotics increases mortality, the likelihood of hospitalization and the length of stay in hospital. Resistance is related to the increasing usage of antimicrobial agents; growing numbers of patients with impaired immunity; increasing instrumentation, and emphasis on cost control. Furthermore; it no longer remains the domain of Gram-negative bacteria. Antimicrobial usage may be controlled by antibiotic policies, but these can only be formulated if the antimicrobial susceptibility pattern of prevalent bacterial pathogens is known.1 Urinary tract infections (UTIs) are one of the most common infectious diseases, and nearly 10% of people will experience a UTI during their lifetime.2-4 The infections may be symptomatic or asymptomatic, and either type of infection can result in serious sequelae if left untreated. Klebsiella, Staphylococci, Enterobacter, Proteus, Pseudomonas, and Enterococci species are more often isolated from inpatients, whereas there is a greater preponderance of E. coli in an outpatient population. Corynebacterium urealyticum has been recognized as an important nosocomial pathogen. Anaerobic organisms are rarely pathogens in the urinary tract.5-8 Coagulase Negative Staphylococci are a common cause of urinary tract infection in some reports. Staphylococci saprophyticus tends to cause infection in young women of sexually active age.9,10 S. aureus is one of the most widely spread human pathogens. Considering the havoc it causes on life and subsequently on the economy, it became necessary to determine its incidence and antibiogram in our environment for adequate control and treatment.11 of infections caused by S. aureus can be one of the gratifying experiences in clinical practice. Survey of resistant patterns of microbes to drugs has shown a rise in the incidence of microbial resistance to most prescribed antibiotics.  The study aimed to detect  MRSA among S. aureus causing UTI  and to know the antibiotic sensitivity pattern of the isolates in our hospital setting. Materials and methods Study design, setting This study conducted in the Department of Microbiology, Shri B M Patil Medical College over 2 years from January 2017 to December 2018. Sample collection The samples included midstream urine specimen, catheterized urine samples, supra-pubic aspirates collected in sterile universal bottles. The urine specimens were transported to the bacteriology laboratory within 2 hours of collection.12 Statistical analysis Values were expressed in terms of Mean ± SD. Analysis was done by using SPSS software version 16.  P≤0.05 was considered statistically significant. Microbiological analysis All urine samples were examined by routine microscopic examination by the wet mount of urine sediment. All urine samples were cultured over routine culture media with a sterile standard loop. These plates were incubated at 37°C for 2 consecutive days. Culture results were interpreted according to the standard criteria.13 Cultures with more than three colonies were discarded, as contaminants 12 The isolates were identified by gram staining, colony morphology and standard biochemical tests catalase, slide and tube coagulase, mannitol salt agar test, phosphatase test.14 Antimicrobial susceptibility testing All isolates were tested for antimicrobial susceptibility on Mueller Hinton agar by the standard disc diffusion method recommended by the Clinical and Laboratory Standards Institute (CLSI).15 Detection of MRSA The Cefoxitin Disc Diffusion Test: the test was carried out on Mueller-Hinton agar by using a  30 μg cefoxitin disc. An interpretation was done using the Kirby-Bauer charts. An inhibition zone diameter of ≤ 21 mm was reported as methicillin resistant.16 The Oxacillin Disk Diffusion Method: The Oxacillin disk (1 μg) diffusion method was carried out on Mueller-Hinton agar which was supplemented with 4% NaCl to detect MRSA according to the CLSI guidelines. The isolates were considered as resistant when the diameter of inhibition was ≤10 mm.17 Genotypic detection of MRSA by PCR (mec A gene) DNA Extraction Procedure was done by Modified Proteinase-K method.18,19 MRSA strains were amplified by conventional PCR. Following a set of PCR primers were used which were specific to Methicillin-resistant S.aureus.20 Forward Primer:  5'- TGC TAT CCA CCC TCA AAC  AGG -3' Reverse Primer:  3'-AAC GTT GTA ACC ACC CCA  AGA -5' Figure 1: Results of mecA gene (left to right), Lane 1: Molecular weight marker, Lane 2: MRSA ATCC 43300, Lane 3:  MSSA ATCC 25923, Lane 4, 6, 11,12,14 and 15: MRSA isolates from clinical samples(280 BP),  Lane 5,7-10,13,16: MSSA isolates from clinical sample. We had chosen a primer set that gives an amplified product of size 280 base pair. So the well which gives a DNA band of 280 base pair is considered positive, whereas the well which does not have any DNA band is indicated as negative ( Figure 1). Results Discussion UTI(UTIs) are one of the most prevalent extra-intestinal bacterial infections. Nowadays, it represents one of the most common diseases encountered in medical practice affecting people of all ages from the neonate to the geriatric age group.21,22 In our study, the UTI was reported more among the age group of 21 to 40 with a rate of 42%. the findings are in agreement with the study conducted by Inaba et al,.23 and El-Sweih et al,.24 this  can be explained by the fact the structure of the female's urethra and vagina makes it susceptible to trauma during sexual intercourse and pregnancy and or childbirth.24,25              S. aureus  UTI more often occurs in urinary-catheterized and pregnant individuals. The majority of S. aureus UTI isolates are methicillin-resistant and S. aureus bacteriuria is associated with subsequent development of invasive infection.26-28 Like S. saprophyticus, S. aureus also encodes an active urease enzyme. Two nickel ABC-transporters (Opp2 and Opp5a) have been identified as necessary for urease activity in vitro. These, along with a third ABC-transporter that imports nickel and cobalt when zinc is depleted, are both involved in UTI colonization and virulence in a mouse model.29-32 Urinary tract infection is one of the most important causes of morbidity in the general population and is the second most common cause of hospital visits.32 Among the male patients, UTI was reported more in elderly patients with 50% of cases occurring between the age group of 40-60 years. Our  finding is in agreement with a study conducted by Das et al.33             With advancing age, the incidence of UTI increases in men due to prostate enlargement and neurogenic bladder.34 Recurrent infections are common and can lead to irreversible damage of the kidneys, resulting in renal hypertension and renal failure in severe cases.33 UTIs have been reported to be the majority caused by Gram-negative bacteria with E. coli being the most prevalent. However, there is an increasing prevalence of S. aureus as a UTIs etiological agent with an alarming rate of developing antimicrobial resistance (Table 1 and 2).35 Linezolid was found to be the most effective drug overall against S. aureus followed by vancomycin, tetracycline, gentamycin and cefuroxime. The highest percentages of resistance were found for penicillin, pefloxacin. these results are basically in agreement with other studies carried out around the world. Our findings illustrate that antimicrobial therapy needs to be selected based on actual culture findings and antimicrobial sensitivity patterns of isolates (Tables 3 and 4).             Antibiotic susceptibility pattern revealed a high resistance to routinely used antibiotics. Resistance to quinolones I,e. ciprofloxacin and pefloxacin were high in this study. This is comparable to the study done by Sanjana et al,36 in Nepal. Majumder et al,.37 also revealed that resistance to various antibiotics with methicillin-resistant strains was s higher in comparison to methicillin-sensitive isolates. Factors responsible for drug resistance in MRSA are as follows. Antibiotics are available without a prescription at drug stores or even at general stores and injudiciously used in communities, animal husbandries, and fisheries and  use  of allopathic drugs by traditional practitioners.38           In our study, the mecA gene PCR  detected  20 isolates as MRSA and the  25 isolates as MSSA. Detection of mecA gene is considered the best method  for MRSA confirmation.The accurate and early determination of methicillin resistance is of key importance in the prognosis of infections caused by S. aureus.39 This higher sensitivity to cefoxitin can be explained by the increased expression of the mecA-encoded protein PBP2a, cefoxitin being an inducer of the mecA gene.39 Our study reveals that cefoxitin disc is better than oxacillin disc for the detection of methicillin resistance. Conclusion To conclude, we found that UTIs were more among young females patients and elderly male patients. PCR was the best method for the detection of MRSA but in peripheries where it is not available Cefoxitin disc diffusion test is the best alternatives for PCR for the identification of MRSA. Antibiotic sensitivity revealed MRSA were resistant to many antibiotics but were sensitive to tetracycline, gentamicin, vancomycin and linezolid.  Acknowledgements: The authors acknowledge the immense help received from the scholars whose articles are cited and included in references to this manuscript. The authors are also grateful to authors/editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Conflict of interest: Nil Source of Funding: Nil Englishhttp://ijcrr.com/abstract.php?article_id=3615http://ijcrr.com/article_html.php?did=3615 Winstanleya  TG,  Limba DI, Eggingtona R, Hancock F. A 10 year survey of the antimicrobial susceptibility of urinary tract isolates in the UK: the Microbe Base project. J  Antimicrob Chemother 1997;40:591–594. Hoberman A, Wald ER. UTIin young febrile children. Pediatr Infect Dis J 1997;16:11-17. Delanghe J, Kouri TT, Huber AR, Hannemann-Pohl K, Guder WG, Lun A, et al,. The role of automated urine particle flow cytometry in clinical practice. Clin Chim Acta 2000;301:1-18. Hryniewicz K, Szczypa K, Sulikowska A, Jankowski K, Betlejewska K, Hryniewicz W. Antibiotic susceptibility of bacterial strains isolated from UTIin Poland. J Antimicrob Chemother 2001;47:773-780. Farajnia S,  Alikhani M Y, Ghotaslou R, Naghili B, Nakhlband A. Causative agents and antimicrobial susceptibilities of UTI in the northwest of Iran. J Infect Dis 2009;13:140-144. Bronsema DA, Adams JR, Pallares R. Secular trends in rates and aetiology of nosocomial UTI at a university hospital. J Urol 1993;150:414-416. Soriano F, Aguado JM, Ponte C. Urinary tract infection caused by Corynebacterium group D2: Report of 82 cases and review. Rev Infect Dis 1990;12:1019-1034. Jacobs LG. Fungal UTI in the elderly: Treatment guidelines. Drugs Aging 1996;8:89-96. Schneider PF, Riley TV. Staphylococcus saprophyticus urinary tract infections: Epidemiological data from Western Australia. Eur J Epidemiol 1996;12:51-54. Amin M, Mehdinejad M, Pourdangchi Z. Study of bacteria isolated from UTI and determination of their susceptibility to antibiotics. Jundishapur J  Microbiol 2009;2(3):118-123. Akortha EE, Ebadin OK. Incidence and antibiotic susceptibility pattern of Staphylococcus aureus amongst patients with urinary tract infection (UTI) in UBTH Benin City, Nigeria. Afr J Biotech 2008;7(11):1637-1640. Collee JG, Duguid JP, Fraser AG, Marmion BP, Simmons A. Laboratory strategy in diagnosis of infective syndromes. In: Collee JG, Duguid JP, Fraser AG, Marmion BP, Simmons A (editors). Mackie and McCartney Practical Medical Microbiology, 14th ed. London: Churchill Livingstone ; 1996:53-94. Cruickshank R, Duguid JP, Marmion BP. Tests for identification of bacteria. In: Medical Microbiology. 12th ed. London: Churchill Livingstone; 1975:170-89. Forbes BA. Sahm DF, Weissfeld AS. Bailey and Scott's Diagnostic microbiology, 12th edition, Mosby Elsevier; 2007:842-55. The Clinical and Laboratory Standards Institute. Performance Stan­dards for Antimicrobial Susceptibility testing, twenty-first informational supplement, M100-S21, Clinical and Laboratory Standards Institute, 2011. Isenberg  HD,  editor.  Clinical microbiology procedures handbook.  2nd ed. Washington DC: ASM Press; 2004:192-98. Brown DF, Edwards DI, Hawkey PM, Morrison D, Ridgway GL, Towner KJ, et al.Guidelines for the laboratory diagnosis and susceptibility testing of methicillin-resistant Staphyloccocus aureus. J Antimicrob Chemother 2005;56:1000-1018. Rao  Venkatakrishna  I,  Bhat  Kishore  G,  Kugaji  Manohar  S,  Manjula  PV.     Detection of methicillin resistance in  Staphylococcus aureus:  Comparison of Disc diffusion and MIC with mecA gene detection by PCR. Int J Pharm Bio Sci 2011;1:518-21. Vanpelt E, Belkum VAV, Hays JP. Principles and Technical Aspects of PCR amplification. Springer 2008;34. Boucher HW, Corey GR. Epidemiology of methicillin-resistant Staphylococcus aureus. Clin Infect Dis 2008;46(Suppl 5):S344–S9. Kunin CM. UTI in females. Clin Infect Dis 1994;18:1-10. Gupta K, Scholes D, Stamm WE. Increasing prevalence of antimicrobial resistance among uropathogens causes acute uncomplicated cystitis in women. J Am Med Assoc 1999;281:736-738. Inabo HI, Obanibi HBT. Antimicrobial susceptibility of some urinary tract clinical isolates to commonly used antibiotics. Afr J Biotech 2006;5:487-489. Al-Sweih N, Jamal W, Rotimi VO. Spectrum and antibiotic resistance of uropathogens isolated from hospital and community patients with UTI in two large Hospitals in Kuwait. Med Princ Pract 2005;14:401-407. Manikandan S, Ganesapandian S, Singh M, Kumaraguru AK. Emerging of Multidrug Resistance Human Pathogens from Urinary Tract Infections. Curr Res Bacteriol 2011;4: 9-15. Muder RR, Brennen C, Rihs JD, Wagener MM, Obman A, Stout JE, et al. Isolation of Staphylococcus aureus from the urinary tract: association of isolation with symptomatic urinary tract infection and subsequent staphylococcal bacteremia. Clin Infect Dis 2006;42:46–50. Baraboutis IG, Tsagalou EP, Lepinski JL, Papakonstantinou I, Papastamopoulos V, Skoutelis AT, et al. Primary Staphylococcus aureus urinary tract infection: the role of undetected hematogenous seeding of the urinary tract. Eur J Clin Microbiol Infect Dis 2010;29:1095-1101. Gilbert NM, O’Brien VP, Hultgren S, Macones G, Lewis WG, Lewis AL. Urinary tract infection as a preventable cause of pregnancy complications: opportunities, challenges, and a global call to action. Glob Adv Health Med 2013;2:59–69.  Hiron A, Posteraro B, Carrière M, Remy L, Delporte C, La Sorda M, et al. A nickel ABC-transporter of Staphylococcus aureus is involved in urinary tract infection. Mol Microbiol 2010;77:1246–1260. Remy L, Carrière M, Derré-Bobillot A, Martini C, Sanguinetti M, Borezée-Durant E. The Staphylococcus aureus Opp1 ABC transporter imports nickel and cobalt in zinc-depleted conditions and contributes to virulence. Mol Microbiol 2013;87:730–743. Kline KA, Lewis AL. Gram-Positive Uropathogens, Polymicrobial Urinary Tract Infection, and the Emerging Microbiota of the Urinary Tract. Microbiol Spectr 2016; 4(2). Ronald AR, Pattulo MS. The natural history of urinary infection in adults. Med Clin North Am 1991;75:299-312. Das RN, Chandrashekhar TS, Joshi HS, Gurung M, Shrestha N, Shivananda PG. Frequency and susceptibility profile of pathogens causing UTI at a tertiary care hospital in western Nepal. Singapore Med J 2006;47(4):281.  Liperky BA. Urinary tract infection in men: epidemiology, pathophysiology, diagnosis and treatment. Ann Intern Med 1989;111:138-150. Onanuga A, Awhowho GO. Antimicrobial resistance of Staphylococcus aureus strains from patients with UTI in Yenagoa, Nigeria J Pharm Bio Sci 2012;4:226-230. Sanjana RK, Shah R, Chaudhary N, Singh YI. Prevalence and antimicrobial susceptibility pattern of methicillin-resistant Staphylococcus aureus (MRSA) in CMS-teaching hospital: a preliminary report. J College Med Sci Nepal 2010;6:1-6. Majumder D, Bordoloi JS, Phukan AC, Mahanta J. Antimicrobial susceptibility pattern among methicillin-resistant staphylococcus isolates in Assam. Indian J Med Microbiol 2001;19:138-140. Metri BC, Peerapur BV, P Jyothi. Comparison of antimicrobial resistance pattern of hospital-and community-acquired Methicillin-resistant Staphylococcus aureus. J Chem Pharm Res 2014;6:201-205. Anand KB, Agrawal P, Kumar S, K Kapila K. Comparison of cefoxitin disc diffusion test, oxacillin screen agar, and PCR for mecA gene for detection of MRSA. Indian J Med Microbiol 2003;27:27-29.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareEstimation of Fluoride Levels in Drinking Water and its Association with Acquired Hypothyroidism in Children - A Prospective Observational Study English3439Srinivasan PurusothamanEnglish Lal DVEnglish Radha KumarEnglishEnglish Acquired hypothyroidism, Drinking water, Fluoride levels, Thyroid-stimulating hormone, Groundwater, Iodine deficiencyINTRODUCTION Fluorosis and Iodine Deficiency Disorders (IDD) are endemic disorders in India and co-occur in certain parts of the country.1 Endemic fluorosis resulting from high fluoride concentration in groundwater is a public health problem in India as it is located in the geographical fluoride belt extending from Turkey to China and Japan through Iraq, Iran and Afghanistan.2,3 Previous studies reveal that 15 States in India are endemic for fluorosis with fluoride level in drinking water >1.5 mg/l, leading to dental, skeletal and non-skeletal fluorosis.1,4,5 Among those affected, 6 million are children below the age of 14 years.5 Fluoride is naturally present in low concentration (0.01–0.3 ppm) in surface rain or river water. Fluoride levels in groundwater (well water) concentrations can vary, depending on the mineral composition of the local ground and in some locations, the drinking water contains dangerously high levels of fluoride leading to serious health problems.2,3 Fluoride can interfere with the function of the thyroid gland and lead to delayed development in children as well as impairment of brain function.6-8 Since fluoride is more electronegative than iodine, it easily displaces iodine, thereby affecting the functioning of the thyroid gland. Fluoride has been known to produce biochemical changes within the body of an individual, which included deranged thyroid hormonal levels. A negative feedback mechanism regulates the production of thyroid hormones in the body by releasing TSH from the anterior pituitary gland following a drop of T3 levels in circulation. TSH stimulates the thyroid gland which in turn accelerates the production of the thyroid hormone T4, now considered a “pro-hormone”. Most the circulating T3 arises from peripheral deiodination of T4 and not from thyroid secretion. The process of deiodination is catalysed by the enzyme iodothyronine deiodinases and fluoride is known to interfere with the activity of the deiodinases enzyme.  In optimally fluorinated parts of the world, the daily dietary intake of fluoride is 0.05 mg/kg/day whereas the dietary intake of fluoride is much lower in children from developing countries. The upper limit of fluoride concentration by the World Health Organization (WHO) in drinking water at 1.5 mg/litre. The maximum daily limit of fluoride intake as per Bureau of Indian Standards is 1.0 mg/l and the “lesser the better”.6 Whenever the intake of fluoride is higher than the optimum level, it can lead to dental and skeletal fluorosis.  Since there are only a few population studies in children that examine the association of thyroid disease with fluoride intake in water, our study was performed to determine if acquired hypothyroidism in the children is related to consuming water with excess fluoride content. MATERIAL AND METHODS Study design: Hospital-based prospective study Period of study: June 2016 to May 2017  Study area: Department of Paediatrics, Saveetha Medical College Hospital, Chennai. Study sample: All children with clinical symptoms and signs of acquired hypothyroidism in the age group 5 to 15 years who presented to the Paediatric outpatient department were enrolled in the study. A sample of 45 children during the study period diagnosed with acquired hypothyroidism was included in the study. Symptoms of hypothyroidism which were included were poor physical growth, poor concentration, poor scholastic performance, fatigue, weak muscles, depression, dry and itchy skin, unusual hair loss, dry hair, cracking nails, infrequent bowel movements or hard stools, unexplained weight gain, swelling at the front of the neck, difficulty in tolerating cold and menstrual disturbances in adolescent girls. Inclusion criteria:  All children aged 5 years to 15 years presenting with clinical features of acquired hypothyroidism. Exclusion criteria: Children diagnosed with congenital hypothyroidism, unwillingness to participate in study and children with any pre-existing or severe concurrent severe comorbid illness. Procedure: After obtaining a detailed history, general examination, anthropometry and examination of the neck was done.  Informed written consent about the study was obtained from the parents or guardian.  Blood samples were taken for estimation of thyroid profile.  Only those children who were confirmed as the case of acquired hypothyroidism were included in the study.  Information about the usual source of drinking water was collected. Participants were asked about the primary source of drinking water at their current residence whether municipal water supply, groundwater, bottled water, reverse osmosis water (RO) treated or others and for how long they have been consuming the water.RO is a water purification process that uses a partially permeable membrane to remove ions, unwanted molecules and larger particles from drinking water.   For those children who were confirmed as overt hypothyroidism (based on the abnormal thyroid profile), the drinking water samples (minimum 50 ml, maximum 200 ml in a  Special plastic bottle provided for the sample analysis) was collected on a follow-up visit and sent to Ekdant Enviro services Pvt limited, Chennai which is a NABH accredited lab to estimate the fluoride levels. Test to detect fluoride level was performed upon the water samples as per the World Health Organisation recommendation. Correlation between the fluoride levels in water and hypothyroidism. Sample size and sampling technique: Prevalence of acquired hypothyroidism in school children is around 3-4%, and assuming 5% precision and 95% confidence interval, statistically significant sample size came out to be 39.  Hence 45 children between 5-15 years were recruited by Consecutive sampling.  The study was conducted over one year.  International Business Machines (IBM) SPSS version 22 was used for statistical analysis. Statistical methods: The primary outcome variables were: Free T3, Free T4, TSH.Primary explanatory variable: Fluoride content in drinking water.Secondary explanatory variable: Age, gender, family history of disorders, height and weight. Inferential statistics: Quantitative outcome-The mean values were compared with the quantitative outcome and categorical variables. The differences between mean levels of FT3, FT4, TSH with mean fluoride levels with 95% confidence intervals were assessed. Statistical significance was done using an Independent sample t-test/ ANOVA. The association of variables Pearson correlation coefficient and the data was represented in a scatter diagram. Linear regression analysis was done. The regression coefficient, along with its 95% Confidence Intervals ( CI )and P values is presented.  Categorical outcome - The association between explanatory variables and categorical outcomes was assessed by cross-tabulation and comparison of percentages. Odds ratio along with 95% CI are presented and the Chi square test was used to test statistical significance. P-value > 0.05 was considered statistically insignificant whereas P-value ≤ 0.05 was considered statistically significant. RESULTS The total number of children who presented with clinical features of acquired hypothyroidism were 156 out of which 3 parents refused consent for blood sampling. A thyroid profile was performed for the remaining 153 children and 45 children had elevated TSH levels while 108 children had normal TSH levels. The 45 children with elevated TSH were included in the study. Out of the total of 45 children who were analysed, the mean age was 11.51 years with a minimum age of 6  years and maximum of 15 years of age in the study (95%  CI  10.74  -  12.28).   24.44% of children were between the age group of 6 to 9 years whereas children in the age group of 10 to 12 years and 13 to 15 years was 31.11% and 44.44% respectively. In the study, 75.56% were girls and 24.44% were boys (Table 1). Among the study population, 57.77% of children had symptoms of fatigue. Dry and itchy skin was present in 13.33%whereas 17.7 % complained of excessive hair loss and 42.2 % had dry hair. The proportion of constipation, unexplained weight gain, swelling in front of the neck, difficulty in tolerating cold, sleep disturbances, menstrual irregularities history of poor scholastic performance were 31.11%, 20%, 17 .77 %, 13.33%, 28. 88%, 17.77%, 24.44% respectively. In the study population, 33.33% had a positive family history of thyroid disorder. As seen in Table 3, among the study population, 71.11% of children had age-appropriate weight ( 5 to - 95th centile) as per the CDC growth chart. 10 (22.22 %) children and 3 ( 6.66%) children had weight for age less than < 5th centile (undernourished) and weight for age > 95 the centile as per CDC growth chart respectively. The mean TSH of the study population was 31.21 with a minimum of 5.36 and a maximum of 106 (95% CI 22.82 - 39. 61 ) as seen in table 4. The mean Free T3  was 3.38 with a minimum of 0.94 and a maximum of 8.90  (95%  CI  2.90  – 3.87 ). The mean Free T4 was 3.10 with a minimum of 0.36 and a maximum of 16.80 (95 % CI 1.83 – 4. 39) (Table 2). Among the study population, the majority of them were consuming pipeline water ( panchayat / municipal) 57.78% while groundwater (bore well/ well) and purified water (R.O/ can water) was consumed by 24 .44 % and 17.78% respectively as seen in table 5. The mean fluoride content in drinking water was 0.462, the minimum and the maximum levels were 0. 10 mg/l and 1. 20 mg/l respectively as seen in table 6. Most of the patients were consuming a permissible range of fluoride levels in drinking water. In the study all children had high TSH level as seen in table 7. 15 children (33.33 %) had low values and 30 children (60.67%) had normal levels of Free T3; whereas FT4 measurements showed low and normal values in 5 ( 11.11%) and 40 (88. 89%) children respectively. The Fluoride content in drinking water was low in  42  children  (93.3 %) and high in 3 children (6.7%). Ethical clearance number:0Z8/08/2016/KEC/SU. DISCUSSION Iodine is an important microelement required by the body for neural development and neuronal functioning. Consumption of drinking water with high Fluoride levels for long periods can lead to acquired hypothyroidism since iodine gets easily displaced by fluoride. Fluoride can inhibit TSH release from the pituitary gland, thereby reducing thyroid hormone release.4 In our study, the mean age of presentation of acquired hypothyroidism was 11.51 years. The majority of the participants were females (75.56%). A similar male to female ratio of 1:3 was reported by Shah NA et al in their study of 50 children.9 Contrastingly, the proportion of females in the study by Peckham et al was equal (49.9%) to that of males.7 In our study goitre was present in 17.77% of cases, fatigue in 57.77%, cold intolerance in 13. 33%, short stature in 20. 00% and constipation by 31.11 %. In contrast, the study by de Vries et al showed that goitre was present in 77. 2% of cases, fatigue in 21.6%, cold intolerance in 6.9 %, and constipation in  5.9% of cases.10 In a study by Al-Agha AE et al the commonest clinical presentations were short stature (32 .5 %), fatigability ( 12.1%),  constipation  (9.8%), cold intolerance (5. 3 %), goitre ( 2.6%) and menstrual irregularities 7.9%.11 Among the study population, 24.44% of patients consumed groundwater from either bore well or well. The proportion of pipeline water (panchayat/ municipal) was 57 .78 % and the proportion of purified water (RO/  can water)  was 17.78%. In our study, the mean Fluoride content in drinking water was 0.462ppm the minimum level was 0.10ppm and the maximum level was 1.20 ppm in the study population. The recommended level of fluoride in drinking water in India is 0.5 to 0. 8 mg/l.7 In their study, Singh N et al reported a  higher concentration of fluoride in drinking water of both cases (1. 6 - 5.5ppm) and controls (0. 98 -1ppm) with acquired hypothyroidism.12 Therefore it is important to test fluoride levels of drinking water in children with acquired hypothyroidism. In their hospital-based study of 275 patients, Sachdeva et al. categorised the patients into 3 groups based on the source of drinking water.14 They found that the mean fluoride level group A (consuming groundwater), were 2.50±1.27 mg/dl, in group B (municipal water) it was 0.82±0.07 mg/ dl and in group C ( reverse osmosis water) it was 0.76 ±0. 11 mg/ dl. The mean TSH levels were 9.66 ± 4 .02, 7. 48 ± 3 .21 and 8.38 ± 4.28 respectively. The mean FT3 levels were 3. 76 ± 1. 41, 4.13 ± 2. 13 and 4.02 ± 2.30 respectively. In our study, the mean TSH in the study population was  31.21 m IU/L with a minimum of 5. 36 and a maximum of 106 in the study population ( 95% CI 22.82 - 44.65). The mean Free T3 was 3. 38 with a minimum of 0. 94 and maximum 8 .90 (95% CI 2. 90 – 3.87). The mean Free T4 was 3.10 with a minimum of 0.36 and a maximum of 16. 80 (95% CI 1. 83 – 4.39). In  their case-control study, Singh N et al found significantly higher mean TSH levels between the groups ( 3.7+/ - 1. 9), while the mean FT3 levels (3 . 07+/- 1. 1) and mean FT4 levels ( 1.19+/- 0.21) was non-significant.12 Though children with low free T3 levels were consuming relatively higher fluoride content water (0.502 ppm) compared to those with normal free T3  levels having water with a  fluoride level of 0.442 ppm, the difference was not statistically significant. 37 Children who consumed water with low fluoride levels had normal FT4 (88 .09 %) and 5 of them had low FT4(11.9%). Out of 3 children who drank water with high Fluoride content, all had a normal level of FT4. In our study, there was a non-significant correlation between the either with FT3 (R=0.23: P=0. 12) levels or FT4 (R=0.21: P=0. 17) levels with fluoride levels in the drinking water.  A similar insignificant correlation with the values of FT3 and FT4  was observed by Singh et al.12 Previous evidence suggests that intake of the high level of fluoride can interfere with thyroid gland function with derangements of thyroid hormone levels. 12,13Excess of fluoride in drinking water, especially groundwater has been shown to demonstrate a linear correlation with raised TSH levels, most of which are subclinical. Also, fluorosis is known to compound its detrimental effects in areas previously deficient  in  their  iodine status.14 Susheela et al compared thyroid hormone status of 90 children with dental fluorosis (drinking water fluoride ranging from 1.1 to 14.3 mg/L) and 21  children without dental fluorosis (0.14?0.81 mg/L fluoride in drinking water) in areas where iodine supplementation was considered adequate.15 They reported that 49 (54.4%) children had “well?defined hormonal derangements; findings were borderline in the remaining 41 children. In another study conducted among school children in the Nalgonda district, the prevalence  of dental caries was more in  below optimal  fluoride  area  followed  by  areas  where fluoride concentration was 5 ppm and above.16 Limitations of the study were that sample size was small, other source of fluoride consumption were not included in the study, fluoride level measurements were not done in serum and body fluids and serum auto-antibodies levels were not measured. CONCLUSION In our study, there was no significant correlation between TSH levels, FT3 levels and FT4 levels with fluoride levels in the drinking water. Fluoride levels in the drinking water of most of the study participants were within the permissible range. Only 3 children had fluoride level of more than  1 ppm. Though permissible drinking water fluoride content in India is 0.7  mg/l can have an effect on the thyroid gland causing hormone derangement. Acquired hypothyroidism is a common endocrine disorder and few population studies examine the association of this disease with fluoride intake.  More studies with a large sample size have to be conducted to assess for any positive correlation between high fluoride and thyroid dysfunction. ACKNOWLEDGEMENT The authors acknowledge the immense help received from the scholars whose articles are cited and included in references to this manuscript. The authors are also grateful to authors/editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed CONFLICT OF INTEREST: NIL SOURCE OF FUNDING: NIL Englishhttp://ijcrr.com/abstract.php?article_id=3616http://ijcrr.com/article_html.php?did=3616 Hetzel BS PB, Dulberg EM. The iodine deficiency disorders: nature, pathogenesis and epidemiology. World Rev Nutr Diet 1990;62:59–119. Kotecha PS, Bhalani KD, Shah D, Shah VS, Mehta KG. Prevalence of dental fluorosis & dental caries in association with high levels of drinking water fluoride content in a district of Gujarat, India. Indian J Med Res 2012;135(6):873–877. Saravanan S, Vijayarani M, Jayakodi P,  Felix A,  Nagarajan S. Prevalence of dental  fluorosis  among primary school children in rural areas of Chidambaram Taluk Cuddalore District, Tamil Nadu. India. Indian J Commun Med 2008;33:146-150. NRC. Fluoride in drinking water: a scientific review of EPA’s standards. Washington DC; 2006. AK S. Fluorosis: Indian scienario: A treatise on fluorosis. New Delhi, India: Fluorosis Research and Rural Development Foundation; 2001 Ha H CZ, Liu XM .The influence of fluoride on human embryo. Chin J Ctrl Endem Dis 1989;4:136–137. Peckham S, Lowery D, Spencer S. Are fluoride levels in drinking water associated with hypothyroidism prevalence in England? A large observational study of GP practice data and  fluoride levels  in    drinking    water.    J    Epidemiol    Community    Health 2015;69(7):619-624 . Newton JN, Young N, Verne J, Morris J. Water fluoridation and hypothyroidism: results of this study need much more cautious interpretation. J Epidemiol Community Health 2015;69(7):617-618. Shah NA MP, Bhalodia JN, Desai NJ. Evaluation of thyroid diseases by hormonal analysis in pediatric age group. Natl J Med Res 2013;3(4):367-70. de Vries L, Bulvik S, Phillip M. Chronic autoimmune thyroiditis in children and adolescents: At presentation and during long-term follow-up. Arch Dis Child 2009;94:33-37.  Al-Agha AE, Alshugair RM, Aljunedi WA, Badakhan BA. Clinical Presentation of  Acquired  Hypothyroidism and Associated Disorders in Children and  Adolescents at  King Abdul- Aziz University Hospital in a Western Region of Saudi Arabia. J Pat Care 2016;2:3. Singh N VK, Verma P, Sidhu GK, Sachdeva S.  A  comparative study of fluoride ingestion levels, serum thyroid hormone & TSH level derangements, dental fluorosis status among school children from endemic and non-endemic fluorosis areas. Springerplus 2014;3:7. Xiang Q CL, Liang Y, Wu M. Fluoride and thyroid function in children in two villages in China. J Toxicol Environ Health Sci 2009;1:54–59. Sachdeva S, Ahmed J, Singh B. Thyroid dysfunction associated with excess fluoride intakes: Scope for primary prevention. Thyroid Res Pract 2015;12(2). Susheela AK, Bhatnagar M, Vig K, Mondal NK.Excess fluoride ingestion and thyroid hormone derangements in children living in Delhi. Fluoride 2005;38:98-108. Desai VC,  Manjula M, Reddy RE, Shaik H. Prevalence of dental caries at different levels of fluoride ion concentrations among the school children in Nalgonda district.  Int J Curr Res Rev 2013;5(5):135-139. 

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareResuscitation in Ayurveda with Reference to Care of Newborn English4045Shinde Rekha VEnglish Patil Sunanda CEnglish Uke PunamEnglishEnglishPranapratyagamana, Resuscitation, Newborn Care, Najaat Shishu paricharya Introduction Resuscitation (rise again) is defined as the act of reviving a newborn from unconsciousness, administering emergency measures to support adaptation with airway, breathing and circulation of the newborn to extrauterine life.1 Immediately after birth (the moment the baby come out of maternal passages); the question that arise is about evidence of baby breathing or crying. When the answer to the above question is YES, the baby needs routine care but when the answer is NO, the baby is provided resuscitation. In Ayurveda, the word resuscitation is replaced by Prana pratyagaman. The word meaning of resuscitation is the procedure used for apparently lifting the baby from death-like condition. The meaning of Prana pratyagaman is Re-establishing the vital Prana back in the body2. Both words carry equal meaning. The respirations which commenced, and enfeebled by the labour pains thus become vigorous. Stimuli for the first breath may be multifactorial. The first functional breath after birth is produced by the integrated activity.2 The environmental changes that occur with birth, is tactile and thermal, increased noise and light; activate several sensory receptors that may help initiate and maintain breathing. The carotid bodies and peripheral chemoreceptors located at the bifurcation of the common carotids are stimulated during hypoxia to increase minute ventilation. Soon after birth, the foetal respiratory activity must transition to normal spontaneous breathing. To overcome the viscosity and resistance of fluid-filled lungs, the recoil and resistance of the chest wall, the lungs and airways of the infants must generate negative pressure so that air moves from an area of higher pressure to one of lower pressure. In asphyxiated infants who cannot increase minute ventilation, profound bradycardia may resulted.3                 Foetal breathing or chest wall and diaphragmatic movements begin at approximately 11 weeks of gestation and increases in strength and frequency throughout gestation. Neonatal transition requires spontaneous breathing and successful cardiopulmonary changes as well as other changes to independent organ system functions. The first minute of life (Golden minute) is considered as crucial for the integrity and survival of a newborn baby. The first functional breath after birth is produced by the integrated activity of several stimuli, hypoxia, acidosis, cord occlusion and thermal changes. During intrauterine life, the foetal lungs are filled with fluid and they do not serve any ventilatory purpose since the placenta supplies oxygen to the foetus. During vaginal delivery, one-third of foetal lung fluid is removed as the chest is squeezed and lungs fluid comes out of the nose and mouth. The first new breaths of most newborn babies are extremely powerful to inflate the alveoli and replace the lung fluid with air.4 Infant who is apnoeic at birth and those having weak respiratory efforts cannot achieve this function. If a fetus is exposed to hypoxia due to any reason, the fetus behave like a strangulated individual and make desperate movements that end with reduced or absent movements. At birth the lungs must transit (change) rapidly to become the sight for gas exchange, otherwise, cyanosis and hypoxia develops rapidly.4 Most babies have a smooth transition from foetal to neonatal life and establish spontaneous breathing at birth without any active assistance. About 5% to 10% of babies are likely to have difficulty initiating spontaneous breathing at birth and need active resuscitation. In such conditions, resuscitative measures should be promptly followed. It requires excellent assessment skills, having a resuscitation team and a thorough understanding of fetal and neonatal physiology.5 Newborn infants who need extensive resuscitation should be rapidly identified. Term infants with clear amniotic fluid, adequate respiratory efforts and good muscle tone should receive routine care. The goals of resuscitation are to assist the baby with the initiation and maintenance of adequate ventilation and oxygenation. Along with this adequate cardiac output, tissue perfusion, normal coral temperature and serum glucose are also maintained. These goals may be attained more rapidly when risk factors are identified early, neonatal problems are anticipated, equipment is available, personnel are qualified and trained and a care plan is formulated. Infants do not meet the criteria for routine care need additional steps in their resuscitation. This includes the initial stabilization (provide warmth, position, clear the airway, dry stimulate and reposition). It may also include ventilation, chest compression and medication.6 Materials and Methods The methodology includes a detailed description of Ayurvedic procedures of Resuscitation and its scientific approach. Every birth must be considered as a medical emergency. In the uterus, the Placenta serves to transfer nutrition and oxygen from the mother. After separation from the mother, the baby must breathe immediately to safeguard against anoxic damage to the brain and other vital organs. In the labour room, the newly born baby should help to establish independent breathing without delay to avoid asphyxiated morbidity and quality of life among survivors. Conditions required resuscitation are; Poster babies have more risk of asphyxia Preterm babies have more risk due to lungs immaturity IUGR babies Problems related to uterine contractions Abnormal presentations Chromosomal anomalies Maternal disorders Drugs and procedure during pregnancy Congenital problems with the baby Meconium aspiration 7 In Ayurveda, the word resuscitation is replaced by Prana pratyagaman. The word meaning of resuscitation is a procedure used for apparently lifting the baby from death-like condition. The meaning of Prana pratyagaman is Re-establishing the vital Prana back in the body. Both word carry equal meaning. ???????????  ?????????? ??????????? ?????? ????????? ??????????? ??????? ??????????? ?????????? , ????????????????? ?? ?????????? , ??? ?????????????? ?????? ?????? ; ?????? -?? ???????? ?????????    ? ??????????????????? ????? ????????????????? ??? ?????? ????????? ????????? ???????????  ?                                                                                  ?. ?? . ? / ?? Charaka mentions different procedures to be conducted immediately after birth in case of Achestha Shishu -baby showing no movements (chestha) or any signs and symptoms of life at birth. The asphyxiated baby does not cry, is unconscious or restless due to instability of dhatus and has hyper-aesthesia; to revive the child do the necessary measures. Word Achesta can be correlated with asphyxiated state, terminal or secondary apnoea like conditions with APAGAR score less than 2 at one 1minute or 5 minutes. This Achesta might have occurred due to prenatal events, antenatal events, aspirations, primary apnoea, terminal apnoea etc. resulting in the unconscious, none crying, Limp, flaccid baby at birth. Such baby should be resuscitated with the procedures mentioned, till it regains Chestha or till there is the re-entry of vital Vayu Prana inside the body is evidenced. This clearly shows Charaka has faced situations of the floppy baby without spontaneous respiration giving rise to the impression that Prana has left out of the body. In such cases, it should be treated till its recovery and vital Prana should be re-established in the body. This explains the resuscitation of the baby in Ayurveda.7, Following procedures were done to stimulate respiration All these procedures are aimed to revive the unconscious baby by initiating resuscitation, pulse, heart rate like vital functions. When we compare modern methods of resuscitation, principles adopted behind these procedures explore scientific knowledge. ??????? ??????????? ??????????;                                             This is the first procedure that should be done when the baby is not crying or Achestha. Two stone pieces are taken and stroked together to produce a dull sound near the base of the ear (mastoid process). This leads to the stimulation of respiration. How this will help to revive the baby? Sensory stimulation: Receptors for different sensations are present in the skin, like the receptors for pain, heat, vibration, pressure, crude touch, fine touch etc. As we know all the sensations from the peripheral parts of the body are carried through the sensory tract. These sensations are carried through the sensory tract in the posterior lobe of the spinal cord. These fibers are riches to the hypothalamus and then cross to the opposite side. When it crosses, it also gives a branch to the respiratory Centre therefore, an extreme degree of sensory stimulation is always associated with stimulation of respiration this is also true in higher sensory functions like hearing, vision etc. which are connected with the respiratory Centre through respective cranial nerves. Hence stimulation of the sensory pathway is one of the easiest indirect methods to stimulate the respiratory centre. Modern approach The modern method of resuscitation also believes in the stimulation of respiration by the sensory stimulus. Methods of sensory stimulation are cleaning and drying of the baby along with the face, striking the palms and soles, pressure and pain sensation. Rubbing at the back (spinal cord) area is direct stimulation of the sensory pathway.9 Examination of reflex There is reflex by name startling reflex. By striking stones near the ear base, we are eliciting startling reflex-like stimulation which helps to identify the alertness of the baby. Stimulation at the base of the ear – is direct stimulation of the respiratory centre. It has been told that sounds should be produced at the base of the ear which corresponds to the mastoid area where the vestibule-cochlear nerve passes through. While testing conductive deafness also keep tuning the fork at the level of the mastoid area only. Why stones are selected rather than metal as we can produce a louder sound by striking metal rods instead of stone intentions may be as follows stones are easily available, sharp sound of metals may damage the Labyrinth, loud sound may cause reflex bradycardia in baby, stones produce a dull sound which is comfortable.10 ????????????????? ?? ?????????; This is the second procedure in the sequence of resuscitation, hot and cold water should be sprinkled over the face of the baby alternatively. Sensory receptors which are abundantly present on the facial skin are stimulated by hot and cold sensation. Sensation will be carried through the sensory segment of the facial nerve and the Trigeminal nerve to stimulate the respiratory centre. In the hot season, cold water and in winter season hot water can be used. Hot and cold water can be sprinkled alternatively. Sensory stimulation by hot and cold sensation for initiation of breathing- This looks to be the second method of sensory stimulation by heat- cold sensation so that the respiratory centre is stimulated. In the first step, they have tried with the Shabda Guna of the Akash Mahabhoot, a failure to which necessitates simulation of Vayu mahabhut with its Sparsha Guna(touch which is also a second-most perceptive Guna by the body). An alternate sprinkling of hot and cold water also stimulates the sympathetic nervous system, which in turn stimulates the respiratory centre. A baby with apnoea can be easily awakened by this method. A sprinkling of cold water to a person during the state of unconsciousness/semi-consciousness is generally practised in community practice, which also works on the same principle. Minor cases will be revived by this method. This should not be done continuously. If the baby is not responding after doing it three to four-time we have to go for the next step of Prana pratyagaman modern science also believes in the same that one should not waste time on giving sensory stimulation if the baby is not responding rather, quickly shift to bag and mask ventilation. Prolonged sprinkling also carries the risk of hypothermia.11 Thus s Ashmno Sangathana looks to be the stimulation of respiration by sensation carrying in through the vestibule-cochlear pathway guided by sound stimulus, while in the case of touch stimulus, in this procedure drying causes stimulation of the trigeminal nerve pathway. Sudden cooling after birth when the human newborn baby is delivered may lose up to 600-kilocalorie per minute at room temperature provides respiratory drive by operating through trigeminal cold receptors located on the facial skin.12 The sudden sprinkling of cold water over the facial region may serve the purpose. The further facial area is the Adhistana of maximum Indriyas like shabda, Rasa, Ghrana etc. Hence it is easy to stimulate all Indriyas by Mukha parisheka. Blood circulation is maximum on the facial area and the face is supplied by external carotid arteries which are connected to the internal carotid artery containing baroreceptors. The rule of baroreceptors in the stimulation of respiration is well known. If these two methods of sensory- stimulation fail to revive the baby immediately shift to the next method. ??????????????????? ????? ????????????????? ??? ?????? ????????? ????????? ???????????; When both the above procedures have failed, this method of stimulation is indicated. This is a method, where fresh air or oxygenated air is provided to the baby by continuous fanning over the area where the baby is kept. For fanning, a black colour earthen broken pots and winnowing basket made out of broad leaves are used. This method is continued till the baby start breathing spontaneously. This looks like a method of ventilating the baby by using room air which contains 21% of Oxygen and which is sufficient to ventilate the baby in emergency conditions. Many times bag and mask ventilation is also carried out by using room air during an emergency. Fanning the spot where the baby has been kept, replaces the impure air and creates a negative pressure on the spot which results in fresh air being sucked into the spot. This provides a high concentration of O2 to the baby. Winnowing basket can be made into a square shape structure where the baby&#39;s head will be placed and meanwhile fanning will be continued. This method provides steel higher connection of O2 to the square area. This method is almost similar to the oxygen Hood box method, where the baby will be provided with a higher concentration of O2. This method is almost similar to free-flow oxygen therapy or nursing the baby under the oxygenated box. Of course, an advanced method like bag and mask (positive pressure ventilation) where mechanically air is pushed inside to initiate respiration has not been mentioned in Ayurveda. Modification of atmospheric air or oxygen- Although the method of collection of oxygen was unknown to them, they will well be versed in the modification of oxygen present/ available air that is existing in the atmosphere. Haritha gives the concept of the modification of the Year he explains five variety of fanning air for the baby. Here atmospheric air while passing through a particular fanning material gets modified. This is just like a hundred per cent O2 collected in O2 cylinder when made to pass through water gets converted into humidified oxygen which is comfortable for the body and is devoid of the bad effects of dry oxygen.13 Ayurveda explains the modification of air as follows. Fanning with the leaves of Bamboo air becomes- dry and induces sleep. Fanning with the Kamsya Patra absorbed the sweat, suppresses Vata. Fanning with the red cloth harmful and it&#39;s contraindicated. Fanning with a palm tree or Banana leaves is very good as it relieves the fatigue, the air become humid brings down daha, pitta and induces sleep. When air passes through a banana leaf, gets moistened and humidified due to sheeta guna of Kadhi Patra. This is almost similar to humidified oxygen. Why Krishna kapalika is used- Krishna kapalika is commonly available in those days or light for use. The black colour absorbs heat imparts coldness. Black colour helps to protect from physiological photophobia. Alternate black and white Fanning may initiate a light reflex that is Rupa Guna Teja. This helps in the initiation of breathing by stimulating visual sensation.14 ???? ???? Pichu dharan is well explained by Sushruta. A Kavalika (cotton pad) has to be dipped in the oil medium like Bala tail and kept in the Brahma Randra pradesha. This point where Pichu has to be placed refers to anterior fontanels, the soft area over the skull which is devoid of ossification and prone to injuries. Hence it protects this highly sensitive area. This may be aimed to prevent and hyperthermia during resuscitation as maximum heat loss take place through the scalp and forehead area. Hence Kavalika covers this area. This procedure may provide an opportunity for examination of anterior fontanels for its elevation of intracranial pressure, haemorrhage, for depression, shock, fluid loss or injuries, so that early management will be insured. Tail which is dedicated to dipping Kavalika is Bala taila which is very good for suppression of Vata. This is proved for its beneficial effects in neurological disorders.15, 16 ????? ??????? After striking the stones near the base of the ear and by doing Bala taila Parisechana if the baby is not revived then the last step is chanting Mantra near the base of the right ear. The mantra which is explained by Astanga Hridaya has to be chanted by the father on the ear of the baby if the baby has not revived by doing all the procedures of resuscitation. Now the concept of Mantra which is mentioned in this context does not tell about the type of mantra to be chanted. This may be one method of auditory stimulation as the father whispers in the ear of the baby or as a last attempt/ hope to revive the baby if it&#39;s Atma Bala and Daiva Bala is good.17,18 Effectiveness of the Mantra chikitsa in reviving the baby- Vedic rhymes or Veda Mantras have a specific way of chanting by maintaining Rhythm, frequency, amplitude. When this is chanted for a long time this may stimulate electromagnetic waves of the brain and in turn, stimulate respiration. Mantras too act as an auditory method of stimulation. Sama Veda Mantra can be considered as the great musical rhymes that can be considered under music therapy and the role of music therapy to reduce the stress factor is a topic of hot discussion nowadays. Mantras are considered as the last option when there is no hope of saving the baby sometime by luck or Atma bala, Daiva bala of the baby it may work, hence this method might have been tried such similar explanations are seen in other disorders also. In asphyxiated infants who cannot increase minute ventilation, profound bradycardia may result.19 TABC of resuscitation in Ayurveda T - Temperature is maintained as Construction of Sutikagara by heat resistant materials. Continuously burning stems of medical drugs. Lighting a lamp of Sarshapa taila. Construction of Sutikagara by keeping a view of airflow- sunlight (south and east facing). Closely packed from all sides. A – Airway Mukha Shodhana by the index finger. Cleaning of Kantha Pradesha. Keeping a baby on a flat surface. Garbhodaka Vaman by Saindhava and Sarpi B – Breathing Fanning the baby. Different sensory stimulations to initiate breathing. Fanning with different leaves to moisten the air. C – Circulation Pichu dharan Raksha karma Balataila abyanga Temperature maintainance Jatkarma samskara Stanyapanayana Suwarna prash atipooj Namakarana samskara Karna vyadhana Chooda karma. Complications and their management The affected child may develop deep unconsciousness or coma, generalized increase in body temperature, unable to cry excessively according to bodyache, rasa, rakta mamsa etc dhatus are unstable during this condition. Dhatus do not attend maturity state or in other words, dhatus do not reach up to their final normal metabolite which keeps the body in normal state., touch by hand cloth or bed gives the pain as it is being cut by the saw; that is in hyperesthesia stage other parameters are continuous involuntary movements of body parts being unconsciousness the baby frequently seems dead that is apnea beings developed due to obstruction of the foetus and compressed by the uterine muscles. In such conditions to revive the baby, Bala oil irrigation should be done along with striking stones near to the ear. The Bala oil is prepared with many drugs which may be useful to review the baby as well as in combating many complications by providing drugs through the skin. The above-said complications seem similar to the feature of HIE(Hypoxic Ischemic Encephalopathy) that appears in its different stages.20 Discussion The ancestral or conventional child care practices are by and large, based on core knowledge and wisdom although some of them may have emerged purely from institutional superstitions and unfounded beliefs. Ayurveda means knowledge of life and art of science of living.17,18 Ayurveda adds not only years to life but also life to years. Most of our health care practices have their origin in our traditions based on the core knowledge and wisdom of our ancestor. These practices are easy and a part of our lifestyle. They are available in the footstep of the people and they are readily acceptable to society. To promote these practices of indigenous medicines, the government of India provided the medicines called AYUSH medicine to treat common day to day illness .there is a need to do further research to ascertain the efficacy and safety of these procedures comes from the Indian system of medicine. In Ayurveda the word resuscitation is replaced by Prana pratyagaman. The word meaning of resuscitation is the procedure used for apparently lifting the baby from death-like condition. The meaning of Prana pratyagaman is Re-establishing the vital Prana back in the body. Both words carry equal meaning. The procedures described in Ayurvedic classic regarding resuscitation of a newborn child are time-honoured and easily available in the surroundings of human being.20 Conclusion The procedures mentioned in Ayurveda to pranapratyagamana (resuscitate) are defined as the total of the knowledge, skills and practices based on the theories of the indigenous system. Though most of these procedures are harmless and having a scientific base but due to lack of their utility they may lead to seeking medical aid with resultant deterioration in the condition of the newborn child. Acknowledgement: The author’s acknowledge the immense help received from Datta Meghe Ayurvedic Medical College, Hospital and Research Centre Nagpur for concluding the present study. Conflict of interest: None Source of funding: Nil Englishhttp://ijcrr.com/abstract.php?article_id=3617http://ijcrr.com/article_html.php?did=3617 Singh M. Medical Emergencies in Children. Revised 5th edition, CBS Publishers and Disributors Pvt Ltd 2016; 1:25. Acharya’s Text book of Kaumarbritya Vol.1. Srinidhi Kumar Acharya, Chaukhamba Orientalia, Varanasi. Chapt 28:p348. Singh B, Text Book of Balaroga Kaumarbhritya, Chaukhamba Orientalia, Varanasi 2015. Singh M, Care of Newborn, Revised 8th edition, CBS Publishers and Distributors Pvt Ltd 2017. Chapt 1:p Singh M, Medical Emergencies in Children, revised 5th edition, CBS Publishers and Distributors Pvt Ltd 2016. Section 1:60. Chheda M, Practical Aspects of Pediatrics 7th edition, CBS Publishers and Distributors Pvt Ltd 2017. Chapt 2:p53. Acharya’s Text book of Kaumarbritya vol.1. Srinidhi Kumar Acharya, Chaukhamba Orientalia, Varanasi., Chapt 28:p512. Vd. Joshi YG. Editor, Charaka Samhita, Sharirsthana 8,42, Reprint 2003. Vaidyamitra Publication. 2003. Dr. B. M. Singh, Text Book of BALAROGA Kaumarbhritya, Chaukhamba Orientalia, Varanasi 2015;2:223. Deepshikha, Amit Kumar Rai, Critical Analysis of Neonatal Care (Navjat Shishu Paricharya) in Brihat Trai. Ayurpharm Int J Ayur Alli Sci 2014;3(10):306-313. Lakshmanaswamy A. Clinical Paediatrics, 4th edition, Published by Wolters Kluwer (India) Pvt. Ltd., New Delhi, 2017. Singh M. Care of Newborn, Revised 8th edition, CBS Publishers and Distributors Pvt Ltd 2017;9:169. Singh M. Pediatric Clinical Methods, 4th edition, CBS Publishers and Distributors Pvt Ltd 2017;12:184. Acharya SK. Acharya’s Text book of Kaumarbritya. Chaukhamba Orientalia, Varanasi. Chapt 2012;28:567. Kaviraj AS, Ed Sushruta Smhita, Hindi commentary,sharirsthana 10, 12 Reprint 2013,  Chaukhamba Sanskrita SansthanaVaranasi 2013. Brahmhananda T. Ashtanga Hridaya, Published by Chaukhambha Sanskrita Pratishtana Delhi 2012, Uttartantra 1/17. Brahmhananda T, Ashtanga Hridaya, Published by Chaukhambha Sanskrita Pratishtana Delhi 2012, Uttartantra 1/1-2. Dr Shivaprasad S, Ashtanga Sangraha with Shashilekha Sanskrita commentary by Indu, Published by Chaukhamba Sanskrita Sansthana Varanasi 2006, Uttaratantra 1/ 3-4. Srinidhi KA,Chaukhamba O .Acharya’s Text book of Kaumarbritya . 2011;28: 673. Ghai OP, Gupta P, Paul VK. Essential Paediatrics, 8th Edition, Published by CBC Publishers, New Delhi,  2013;2:74.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareA Review on the Clinical Aspect of Guda (jaggery) in Brihatrayi and laghutrayi English5163Prashant UmateEnglish Pramod KhobragadeEnglish Harsha ThanviEnglish Sarju ZilateEnglishEnglishGuda, Jaggery, Brihatrayi, Laghutrayi, Medicinal properties and usesINTRODUCTION Ayurveda is called Veda for ayu (Life, longevity, health and wealth). For thousands of year, it was played a vital role in health problems i.e. Prevention (Swastha rakshana) and the cure of diseases (Vyadhiparimoksha)[i] of not only human being but also another living thing in the world. It has mentioned very details and specific about ahara (diet) and aushadi(medicine). All Samhita has given importance to Sahara before medicine and also stated the first stage towards the disease cure process. Some of the product mentioned in Ayurveda Samhita plays an important role as diet and medicine also. Jaggery is one of them and the most useful and beneficial product in traditional Indian kitchen since ancient time. There are many types of research on jaggery that has been published but most of them were specifically concerned about marketing and focused on the nutritional value and colour product (jaggery). Very few study concern about the medicinal uses of jaggery butAyurvedasamhita has given details about its various types, medicinal properties and uses. There are three types viz. Naveen, Purana and PrapuranaGuda mentioned in Samhita but the clear demarcation is not mentioned. It has been specified in Nighantu. Properties of all types of Guda are different hence the medicinal uses also changed accordingly. General properties are described in various Samhita are Na-atishleshmakar, Mala-Mutra shuddhikar, Krimi-Majja-shyonit-Meda-Mamsavardhaka, Agnimandya Hridya and Pathya.[ii]-4 To date, the medicinal properties and uses of guda in bruhatrayi (Charaka, Sushruta, Vagbhata) and laghutrayi (Sharngadhara, Bhavprakasha, Madhavnidan) are not compiled in one rooftop. Hence the topic is selected for study and organised clinical aspect of guda in brief with the tabular form which has given in Brihatrayi and Laghutrayi?. Which will be more helpful for clinician and scholar to find the appropriate reference in one place. Madhavnidan is a specific concern about diagnosis hence it has not mentioned any information about jaggery and its medicinal uses. REVIEW Charaka has given guna of Guda, i.e. The Krimi-Majja-shyonit-Meda-Mamsavardhak, Annapanavidhiadhyaya in Sutrasthana. Alpa doshkar is the final prepared shudhha Guda. In Chikitsa sthana for chikitsa, where Guda is one of the ingredients, binding agent, as anupana with a particular Kalpa according to disease, hetu of disease.  chikitsa Charak Samhita listed various Kalpa. All data summarized in Table 1 for quick reference.[iii] Sushruta has mentioned Guda and its properties in sutrasthana adhyaya 45, Dravdravya vidhyaninya shlok no.160. Guna karma of Naveen and Puran Guda was separately mentioned by shushurta. As the Sushruta was mainly concern with shalyatantra, Kalpa of Guda found less in numbers than in Charak Samhita but pathya-apthya specifically given in sutrasthana which are summarized in Table 2.[iv] Vagbhat mentioned properties of guda in Sutrasthana, dravdravyavdhyniya, shlok 48, 49 (AH.Su 5/48, 49). NavvenGuda is kaphakarak and create agnimandya while Puran Guda is Hridya, Pathya. There are very scattered references in Samhita about use of guda in chikitsa of urdhvajatrugata roga, grahani roga, arsha, skin diseases etc are summarised in Table 3.[v] Asthangsangrahkara mentioned Guda in Sutrasthana, dravdravyavdhyniya, shlok 87 (As.Su 6/87). He explained more or same uses as Astangahriday like in jwara, swasa, Kasa, hikka, swarabheda, aruchi, arsha, Grahani, pandu, udara, pleeha and rasayana in combination with other drug or solitary which was concluded in Table 4.[vi] Sharangdhar has given more detailed information about matra (proportion) of Guda in Panchavidh Kasay kalpana. Guda is used in various diseased condition as kalpa form or as anupana. There are several Kalpa like Hemagarbha pottali rasa, Shital Jwaradi Rasa,  Pathyadi kwatha, Shadanga Kwatha and used in many formulations especially in asava- aristha kalpana and many more these are summarised in Table 5.[vii] Bhavaprakasha Samhita written by Acharya Bhava Mishra. Guda is described at Nighantu part of samhita in Ikshuvarga. NaveeenGuda properties are mentioned as Vrushya, Guru, Snigdha, Vatanashak, Mutrashodhan, Na-ati-pittakara, Medakara, Kaphakara, Krimikara, Balakara, Swasakara and Puran Guda is Laghu, Pathya, Anbhishandi, Agnijanaka, Pusthikruta, Pittaghna, Madhur, Vrushya, Vataghna, Raktaprasadaka. Bhavprakasha has mentioned specific guna with particular anupan like Guda with Ardraka- quickly reduces the enhanced kapha, with Haritaki it acts on Pitta and when used with Sunthi it acts as Vatahara. The scattered reference of guda used in chikitsa is summarised in Table 6.[viii] Modern view According to modern parameter, jaggery is far complex than sugar hence get digested slowly than sugar and releases energy accordingly slow and not spontaneously thus provides energy for a longer time and is become beneficial to the body. As it is prepared in iron vessels Jaggery hence rich in ferrous salts (iron) and popularly used in anaemia for a long time. It also contains traces of mineral salts which are very helpful for the body. Jaggery act as a very good cleansing agent. It acts as a purifying agent for lungs, respiratory tracts, stomach, intestines and oesophagus. It is highly recommended to take jaggery daily who workplaces are full of dust as it can keep them safe from asthma, cough & cold, congestion in chest etc. It is rich in significant minerals required for the human body (viz., Potassium-1056 mg, Calcium-40-100 mg, Magnesium-70-90 mg, Phosphorus-20-90 mg, Sodium-19-30 mg, Iron-10-13 mg, Manganese-0.2-0.5 mg, Zinc-0.2- 0.4 mg, Copper-0.1-0.9 mg, and Chloride-5.3 mg per 100 g of jaggery), vitamins (viz., Vitamin A-3.8 mg, Vitamin B1-0.01 mg, Vitamin B2- 0.06 mg, Vitamin B5-0.01 mg, Vitamin B6-0.01 mg, Vitamin C-7.00 mg, Vitamin D2-6.50 mg, Vitamin E-111.30 mg, Vitamin PP-7.00 mg), and protein-280 mg per 100 g of jaggery, which can be made available to the masses of malnutrition and undernutrition to lessen the health issues. It has been reported micronutrients present in the jaggery plays a better role in antitoxic and anti-carcinogenic properties. All features of Guda shown in Table 1 to Table 6 according to various Ayurved books.[ix]     Discussion Properties of Guda described by various Acharya area approximately similar. Uses in various diseases through many formulations are different and peculiar.  There are few references where Guda used solely in treatment like virechana, ajirna, shola etc and abundant reference found in combination with other medications. There are some refrances where guda is considered as hetu of some diseases like krimi or in some karma (Procedure). It is specifically called apathya in apya, anupa mamsa, Vrana[i] and virudhha (incompatible) when taken with kakamachi (Solanum nigrum), Varaha Mamsa and Guda is Ssanyog viruddha.11,12 It is a very useful product of sugar cane which is almost useful in all systems of the body. According to modern research, it will help in anaemia, jaundice etc diseases which are already mentioned in samhita. It is one of the examples and proof that whatever matter mentioned in samhita will be used in today’s era also. Conclusion The compiled references of medicinal properties and uses of Guda (Jaggery) which is a very common ahar (Dietary) as well as aushadhi (Medicinal) dravya in Ayurveda will simplify the search in Brihatrayi and Laghutari. Conflict of interest: Nil Source of funding: Nil Englishhttp://ijcrr.com/abstract.php?article_id=3618http://ijcrr.com/article_html.php?did=3618 [i]. Thakaral K, Dalhana & Nyaychandrika comentri of Shushruta, Sutrasthana 1/14, vol-1 Reprint 2016;9. [ii]. Agnivesha krit, charak pratisansrit, Edited by Pandit Kashinath-Charak Samhita-Purvardha, Reprint yr 2008. [iii]. Agnivesha krit, charak pratisansrit, Edited by Pandit Kashinath-Charak Samhita-Purvardha, Reprint yr 2008 4. Ambikadatta Shastri ed, Sushruta samhita of Sushruta, Chikitsasthan 2/10, Chaukhamba Sanskut Sansthan, Varanasi. 2005;1: 2/10 [v]. Vagbhatkrit Ashtangahridaya, Sartha Vgbhata, edited by Dr. Ganesh Krishna Garde Anmol Prakshan. Reprint 2008;5/48-49:21. [vi].Vagbhatkrit Astangasangraha, edited by Kaviraj Atridev Gupta, Chaukhamba Krisnadas Academic, Varanasi, Reprint 2016;6/87:55. [vii]. Pandit Parsurama sashtri,Vidyasagar, Sharangdhar samhita with commentary Adhmalla’s dipika Madhyam and uttar khand,Choukhmba orientalia , Varanasi,7th edition 2008 [viii]. Bhavmishra, Bhavprakash, edited by Pandit Shri Brahma Sankara Mishra with vidyotani hindi commentary, Choukhamba Sanskrita Bhavan, Varanasi, Vol 1 & 2. Reprint 2016. [ix]. Nath DD. Review on Recent Advances in Value Addition of Jaggery based Products. J Food Process 2015;6(4);1-4.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareReview of Viparit lajjalu (Biophytum sensitivum Linn.) and its Effect on Sperms English6469Dhiraj ZadeEnglish Swanand PathakEnglish Pramod KhobragadeEnglish Sonali WairagadeEnglish Prafulla FadanvisEnglishEnglishViparit Lajjalu, Sperm, VajikaranIntroduction Folklore herbal medicines are getting more attention now a day. Folklore medicines are a more important source of knowledge for research and development.1 The traditional therapist more often carries the traditional philosophies, civilisations, and stories of a community, which they receive from the previous generations.2 There is scope to encourage the new idea, manner and treatment modalities from older healing procedure.3 The traditional medical practitioner provides health maintenance by using herbal plants, inorganic and organic materials and certain other approaches based on the societal, ethnic and spiritual backgrounds as well as the fundamental knowledge.4,5 Traditional practitioner used different medicinal formulas from various natural substances (herbal plants, inorganic and organic materials). They have wide knowledge based on herbal plants for treatment and dietary purposes. Currently, traditional therapists have their area of expertise. There are different types of medicinal plants that are known to traditional therapists, Biophytum sensitive Linn is one of the tremendous effect, variety of uses even though not into the lumen of research. There are so many therapeutic myths on different types of traditional medicine but lacking scientific study on them.6 Most of the folklore medicines are given without doing any ethical, well-mannered research even in this era of science. Single herbs have so many qualities, activity on different diseases in so many types of permutation and combinations. Infertility is emerging as one of the embarrassing problems in the present world. But now the male infertility of a day is unexpectedly raising & putting up a major concern for both patients and doctors. Infertility affects around 1 in 7 reproductive-age couples, frequently causing significant psychological distress. In men, infertility may results from impaired sperm quality, quantity and abnormality. Azoospermia or Oligospermia is usually idiopathic but may be a consequence of hypogonadism.1 Male infertility is commonly due to deficiency in the semen and semen quality is used as a surrogate measure of male fecundity.2 It may cause due to y-chromosomal microdeletions, varicocele, hypogonadism, previous vasectomy, previous sexually transmitted infection and others.3 To verify the number, shape and movement of sperm in the ejaculate, the diagnosis should require a medical history of the individual and a physical examination along with a semen analysis. To verify the levels of hormones that regulate the development of sperm, blood tests may also be performed. Genetic investigations and testicular biopsies are sometimes done. For this there are some treatments, today doctors will advise that the couple take Reproductive Assisted Technologies (ART), such as IVFF (in vitro fertilisation). The cause of infertility is not healed or treated by ART but may help couples achieve pregnancy even if the man&#39;s sperm count is very low.4,6,7 All these treatments may be expensive, prolonged which cannot possible for every couple and there is no reassurance about fertility after these treatments. It ultimately results that some men have to deal with the reality that nothing can be done about their infertility. In Bhavaprakash about Laxmana as Putrajanani means ability to produce healthy progeny. Viparit Lajjalu (Biophytum sensitive) is one drug under the category of Laxmana in Bhavprakash5. Laxmanamula Strivandhtyatvanashana and Vandhyatva in Shushrutasamhita, which improves the consistency, quantity and abnormality of the female reproductive system, have also been described by Acharya Shushruta.6 B. sensitive Linn. has been liberally used all over India. Even in Folklore in West Africa and Mali region it is used for Aphrodisiac activity, wound healing and malaria.7 B. sensitive Linn the whole plant is also eaten to induce sterility in man.8 Due to different opinion on the activity of sperm of B. sensitive Linn.,  the proper scientific study should be done for an accurate conclusion of the effect on sperms. Ayurvedic review Shukra is considered as the last of the seven Dhatus which are responsible for maintaining and providing strength to the human body. Ingested food material after digestion “Rasa Dhatu” is produced which after the action of Agnis of different Dhatus produces Rakta, Mamsa, Meda, Asthi, Majja while these Dhatus are formed different types of waste products are produced and are thrown out of the Dhatu resulting into the Mala Rahit (devoid of waste products) Last Dhatu i.e. Shukra.9 As the function described to Shukra Dhatu is to produce “Garbha” (Fetus) it must be the cleanest Dhatu. Being cleanest of the Dhatus it is also one of the “Ten Pranayatanani” i.e. one of the ten body parts where the “Jiva” i.e. life resides10. Any harm to these ten parts costs the life of the person who is hurt at those places. Shukra Dhatu is formed or produced in the body from the Rasa Dhatu i.e. an absorbed form of the digested food material from the Gut. Rasa Dhatu then produced Rakta and then Mansa& so on till the Shukra the last of the Seven Dhatus is produced.  Acharyas consider that it requires 3 day nights to produce the next Dhatu thus it takes 21 days for producing Shukra. Some Acharyas believe that after one month Shukra Dhatu is produced. There are interesting theories to explain the formation of these Dhatus one of which is denoted by giving an example of a canal that irrigates the farm turn by turn, this theory is known as “Kedar kullya nyay”. The second theory indicates that as the milk is turned into curd and so on likewise Rasa turns to Rakta and Rakta to Mansa and so on, this theory is known as “ksheer dadhi nyay”.11,12 The third theory explains that as the pigeon collects grain from the field where grains are extracted, pigeons whose nest is nearby will reach earlier whereas whose nest is faraway will take more time to reach likewise Rakta being the next Dhatu will be formed earlier; Mansa being third in line will take more time. Shukra being the farthest will be the last to be formed. This theory is known as “khale kapot nyay.”11 When these theories are seen collectively it becomes clear that rasa contents raw material to produce different Dhatus. It flows in the body continuously providing raw material to different manufacturing systems of the Dhatus, which after receiving the suitable raw material produces particular Dhatu with help of Dhatawagnis which converts them properly and the waste material ready to be thrown out of the body. There are many opinions regarding the production of Dhatus particularly that of Shukra. Some Acharyas consider that it takes one day and one night (i.e. 24 Hrs) to produce the next Dhatu likewise at the end of the seventh day after ingestion of food Shukra is produced. The digested and absorbed Rasa contents the raw material required for the formation of different Dhatus which are processed by different Agnis and next Dhatu have formed including Shukra and oja there after. When these processes go wrong due to various reasons including heredity, to dietary deficiencies, the previous history of the diseases, deficient Dhatawagnis, abnormalities of the system concerned whether congenital or acquired, the Dhatu produced is not normal or optimum. Such abnormal or deficient Dhatu is also deficient in its performance. The same is the case of ShukraDhatu; if the production of the ShukraDhatu is deficient due to any reasons mentioned above or any other reason it cannot produce “Garbha”. The production of “Garbha” is the prime function of ShukraDhatu.13,14,15 According to Kashyap Samhita, though Shukra is present in the body it remains dormant and becomes vyakta at the age of 16 years. As it becomes Sampurna i.e. Complete hence becomes visible. Of these, the SarvdehikShukra produces secondary sex characters and the SthanikShukra is ready for ejaculation and producing Garbha. The “Garbhotpatti” is possible only if a good “Shonit” i.e. Stribija and good healthy Shukra unite to form a Garbha” i.e. Zygote. This Zygote when reaches the healthy uterus it’s implanted and grows normally in a well-nourished mother. But, if any of these four factors are deficient the formation of “Garbha” i.e. Zygote or its implantation is hampered causing ApatyaKamana or “Vandhyatwa” i.e Infertility. Ayurvedic literature describes the lakshan of “ShuddhaSukra” and “AshuddhaSukra”. Facilities of microscopic examinations were lacking for ancient physicians who were aware of the fact that the “Vikrities” abnormalities of the semen cause infertility.16-18 Modern view With the advancement of Teratozoospermia, the physics and tools offered by it for medical usage the microscopic examination of the semen reveal abnormalities of the semen particularly of the sperms and their viability. The sperm with abnormalities in its anatomy and physiology is unable to fertilize the ovum. In cases where the number of such abnormal sperms is high ovum cannot be fertilized causing infertility. Many such abnormalities of the sperm ranging from semen without sperm (Azoospermia), Dead sperms (Necrospermia), Fewer sperms (Oligozoospermia), sperms with pus cells (Pyospermia) & abnormalities of its head, body, tail are now encountered. Amongst these abnormalities, Teratozoospermia is one such abnormality where the head, body or tail of the sperm is abnormal separately or collectively.19,20 Modern drug review There is a generic medicine that affected sperm quality and quantity. Clomiphene is the licensed medicine for female infertility, but clomiphene can also be administered to males by physicians.12 It will decrease hyperandrogenic (low testosterone) symptoms, increase sperm count, and potentially help boost non-obstructive azoospermia, a blockage that prevents semen from entering the sperm.13 Originally designed for breast cancer, Anastrozole or Arimidex has become a viable off-label treatment for particular groups of infertile men. Anastrazole inhibits the enzyme, aromatase, which prevents testosterone from transforming into estradiol, unlike most fertility medications.14 This process helps increase the level of testosterone and reduces the level of estrogen. Anastrozole was found to decrease symptoms of male hypoandrogenism, such as low energy, decreased muscle mass, low libido, and erectile dysfunction. It has also been shown in males who are azoospermic or have a low sperm count to increase sperm production and sperm recovery rates.15 By directly stimulating the testes, human chorionic gonadotropin acts to produce its testosterone and increase sperm production. In hypogonadism men, research has found HCG to improve energy, libido, strength, and mood. This was observed in men who are hypogonadism due to unknown reasons, as well as in men with lower production of testosterone due to long-term replacement therapy with testosterone. The improvement in testosterone output from HCG can also increase the chance of successful sperm retrieval in men with non-obstructive azoospermia, according to another study.16 One of the declines in sperm motility and seminal vesicle secretions, irregular sperm morphology, is also methadone hydrochloride.17,18 Spermatogenic arrest/decreased sperm count by nitrofurantoin (in high doses).19,20 Ayurvedic drug review Taxonomical Description Botanical name –  Biophytum sensitive Linn. Synonyms Sanskrit:   Jhullipuspa, Lajjaluka, Panktipatra, Pitapushpa, Vipareetalajjaalu20 Common name:  Life plant, little tree plant, sensitive plant21 Hindi: Lajalu, Lajjaalu, Lakshmana, Zarer22 Marathi: Jharera, Lajwanti, Lahanmulaka23 Family: Oxalidaceae Genus: Biophytum Species: sensitivum24 It has been used in traditional medicine, especially in Indian medicine, for different ailments. 25,26 In the tradition and culture of the state of Kerala in India, the flower of this plant is regarded as one of the ten sacred plants called Dasapushpam27 Botanical description It is an annual herb that looks like a miniature palm, with 2.5 to 25 cm of unbranched, erect, glabrous or hairy stems. Leaves, with 6-12 pairs of leaflets, are sensitive, pinnately compound, crowded into rosette at the top of the stem, and 5-12 cm long. The leaflets are opposite, the petiole is short, and the size increases upwards steadily, being 1.5 cm long, oblong and apiculate at the apex. Flowers are dimorphic, numerous, and up to 10 cm long, 8 mm across the yellow peduncle. With parallel nerves, sepals are 5, lanceolate, imbricate, and acute. Petals are 5, red-marked gold, connate into a salver-shaped corolla and rounded lobes far exceeding the sepals. The fruit is a capsule that is apiculate, ellipsoid, slightly exceeding the sepals. Seeds are ovoid and striate transversely. There are 10 stamens, free of separate filaments, five inner ones are longer, and five styles.28 Movement of plant leaves B.sensitivum displays nastic movement in the leaves in response to touch, contact with the foreign body, rainfall, wind, vibration, heat, and closing of its leaflets, and the movement is independent of the direction of the stimulus. Depending on the stimulus intensity applied, the degree of motion varies. When the sensitive plant&#39;s leaf apex is gently touched, only a few pairs of leaflets close up, and when roughly touched, all the leaflets respond in the same way from the apex downward. A motor structure formed by a rod of collenchyma surrounded by sclerenchyma is the pulvinus. In their extended location, the cells of the entire collenchyma are distended with water, and the cells in the lower half of the pulvinus react by expelling potassium and chlorine ions and taking up calcium ions upon receipt by contact of the action potential signal. This results in an osmotic gradient that pulls water out of the affected cells. The lower pulvinus cells temporarily shrink due to water depletion. In the manner of a fan, this causes the entire structure to bend down. The pulvinus no longer acts as a support in this contracted role and the petiole droops.29 Furthermore, botanists have found a signalling molecule called turgorin, which is known to be a new phytohormone class that regulates all leaf motions by regulating the turgor of plant cells. A highly acidic, free sulphuric acid group is included in the turgorin molecule and it was found that diluted sulphuric acid-induced leaf closing as strongly as turgorin. Therefore it was concluded that turgorin&#39;s leaf-closing activity is due to its sulphuric acid group&#39;s high acidity.30 Properties of Vipareeta lajjaalu(Biophytum sensitivum Linn) - Guna: Laghu Rasa - Tikta, Madhur, Kashaya, Veerya – Sheeta Vipak - Katu Gana:-Guduchyaadivraga B.H.P. Chemical composition: There are different chemical components in the whole plant, such as phenolic and polyphenolic compounds, saponin, essential oil, polysaccharides and pectin. The key constituent was found to be amentoflavone in methanolic extract of roots, stems and leaves.31 Other therapeutic utilities of the plants:- 1.  In Urine calculus Quath of the root of B. sensitivum administered. 2. In piles Roots powder paste applied. 3. The root of B. sensitivum is tied on Hydrocele patient on locally.32,33 Safety profile Acute toxicity for B. Sensitivum extracts has been tested in rodents. Aqueous plant leaf extract was tested and found to be non-toxic at 100, 200 and 300 mg/kg dose levels by oral route in mice 33 Methanol extract B. Sensitivum whole plant is well tolerated up to an oral dose of 4000 mg/kg as no mortality was observed within 24 h34 The median dose (LD50) of plant in hexane, chloroform, ester, N-butanol and ethanol extracts found to be greater than 1 g/kg when intraperitoneally administered to rats.35 Clinical Studies B. sensitivum is used for the treatment of diabetes traditionally said to have an insulin-like compound. The mechanism of action is not well understood, but tropical insulin properties appear to be present. Clinical studies on the B. Sensitivum containing formulation have been published36 Since 2002, DB14201 has been sold under an Ayurvedic license granted under the trade name Diabedrink by the Drug Controller of the State of Kerala. In Ayurveda, it is a mixture of 16 herbs used including B. sensitivum. Blinded, placebo-controlled, randomized clinical trials have been reported in 30 patients with type 2 diabetes aged 29-71 with single oral diabetes in both sexes. It is confirmed that when administered along with glibenclamide, the herbal formulation DB14201 is safe in T2DM patients and enhances the efficacy of glibenclamide in providing improved glycemic control. Compared to the addition of placebo, it also offers a substantial increase in fasting and post-prandial blood sugar levels and also significantly decreases HbA1c levels.36 Herbal emulsifying creams and gels are prepared by adding the dry methanolic extract of the whole plant B. Sensitivum has been assessed for in-vitro antibacterial efficacy against four distinct bacterial strains (Salmonella typhi, Staphylococcus aureus, Escherichia coli and Bacillus subtillis). The findings revealed that when formulated as cream and gel for topical use B. sensitivum has a high potential as an antibacterial agent.37 B. sensitivum was prepared with an optimized tablet formulation by using the methanolic extract from the dried whole plant. Their antioxidant properties were evaluated in vitro based on their total flavonoid content (TFC) against the regular flavonoid, Quercetin, and also in vivo for the antidiabetic activity of Streptozotocin (STZ)-induced diabetic rats against the anti-diabetic drug Glibenclimide.38-39 The drug with the formulations showed antioxidant and anti-diabetic properties37,40 anti-inflammatory anti-cancerous,41-43  Immunomodulatory effect and Radioprotective. Conclusion It has historically been recognized that parts of the plant have a broad range of medicinal properties, including antiseptic properties, including beneficial effects on a variety of skin infections and diabetes care. Scientific study has shown that anti-bacterial, anti-fungal and anti-diabetic practices have been carried out. The plant has been extensively assessed and confirmed to have anti-inflammatory, antipyretic, antimicrobial, anti-obesity, antioxidant, anti-diabetic, anti-fungal, anti-cancer, larvicidal, anti-obesity, anti-hypertensive, anti-epileptic, wound healing and anti-fertility activity for various pharmacological activities. In the treatment of different diseases, the entire plant is often traditionally used. B. sensitivum has been traditionally recommended for Diabetes treatment in the Ayurvedic Philosophy of Medicine. After critical review, no clinical work has been done previously and the conclusions of existing studies are contradictory to each other in therapeutic uses. One article is suggesting Aphrodisiac activity and the other one sterility activity of B. sensitivum. Whereasthe drug is used for infertility by tribal people. Previous evidence search on B. sensitivum no fact has been generated to prove the exact mechanisms on sperm by B. sensitivum.To generate the evidence regarding specific action B. sensitivum, it is necessary to overcome the confusion about the activity of B. sensitivum on sperm in a scientific manner. Conflict of interest: Nil Source of funding: Nil Englishhttp://ijcrr.com/abstract.php?article_id=3619http://ijcrr.com/article_html.php?did=36191. Walker B. Ralstan S. Davidso principle and practice of medicine; 21st edi.British Library Cataloguing in Publication Data. 2019;758-60. 2.  Cooper TG, Noonan E, Von Eckardstein S, Auger J, Baker HW, Behre HM, Haugen TB, Kruger T, Wang A. World Health Organization reference values for human semen characteristics. Human Reprod Update 2009;16(3):231–245. 3.  Colledge N, Walker B, Ralstan S. Davidson’s principle and practice of medicine; 21st edi. British Library Cataloguing in Publication Data 2019;758-760 4. AndrologyAustrelia. Male infertility Andrology Australia, School of Public Health & Preventive Medicine, Monash University; Melbourne; (Last modified: March 23, 2016) cited 2017 feb.10 5.  Mishra B. Vaishya R. Bhavaprakash of Bhavamishra. Vol.I Parishishta 2; chaukhamba Sanskritbhavan, Varanasi. Bhavaprakash Nighantu Madhyam Khanda, Guduchyadi Varga. 2017;147-148,357-359. 6.  Shushrutasamhita, ChiktsaSthana, 37/16. 7. Tom EG. Ethnopharmacological survey of six medicinal plants from Mali, West-Africa. J Ethn Ethnomed 2008;4(26). 8.    Charak Samhita, Sutrasthana, Dirghajivitam Adhya-1/42-43. 9. Charakchiktsa, chakrapaanitika, Grahanidoshachkitsa, 15/16- 19. 10. Moskovic D, Katz D, Akhavan A, Park K, Mulhall J. Clomiphene citrate is safe and effective for long-term management of hypogonadism. BJU Internat 2012;110(10):1524-1528. 11. Hussein A, Ozgok Y, Ross L, Niederberger C. Clomiphene administration for cases of nonobstructive azoospermia: a multicenter study. J Androl 2005;26(6):787-791. 12. Kim E, Crosnoe L, Bar-Chama N, Khera M, Lipshultz L. The treatment of hypogonadism in men of reproductive age. J Fert Ster 2012;99(3):678-79. 13. Kim E, Crosnoe L, Bar-Charma N, Khera M, Lipshultz L. The treatment of hypogonadism in men of reproductive age. J Fert Ster 2013;99(3):718-24. 14. Shinjo E, Shiraishi K, Matsuyama H. The effect of human chorionic gonadotropin-based hormonal therapy on intratesticular testosterone levels and spermatogonial DNA synthesis in men with non-obstructive azospermia. J Andr 2013;1(6):929-935. 15.Cicero TJ, Bell RD, Wiest WG, Allison JH, Polakoski K, Robins E. Function of the male sex organs in heroin and methadone users. N Engl J Med 1975(292):882–887. 16. Ragni G, De Lauretis L, Gambaro V, Di Pietro R, Bestetti O, Recalcati F, et al. Semen evaluation in heroin and methadone addicts. Acta Eur Fertil 1985;16:245–249. 17. Albert PS, Mininberg DT, Davis JE. The nitrofurans as sperm immobilising agents: their tissue toxicity and their clinical application. Br J Urol 1975;47:459–462. 18. Editorial Committee of the Flora of Taiwan; Flora of Taiwan, 2nd ed. 1993. Biophytum sensitivum (Oxalidaceae) at the culture sheet. http://culturesheet.org/doku.php?id=oxalidaceae :Biophytum:Sensitivum 19. Pawar AT and Vyawahare NS: Phytochemical apd Pharmacological profile of BiophytumSensitivum (L.) DC. Int J Pharm  Pharmac Sci 2014;6(11):18-22. 20. Kala SC, Mallikarjuna K. Short review on callus studies of Biophytumsensitivum Linn. World J Microb Res 2015;4(4):985-991. 21. Kakade RT, Sandu N and Senthilkumar KL: In-vitro anthelmintic activity of leaves and stems extract of BiophytumSensitivum Linn. Asian J Plant Sci  Res 2013;3(6):64-68. 22. Biophytumsensitivum var. assamicum (life plant). Available from: http://zipcodezoo.com/Plants/Biophytum_sensitivum. 23. Jirovetz L, Buchbauer G, Wobus A, Shafi MP, Jose B. Medicinal used plants from India: Analysis of the essential oil of air- dried Biophytumsensitivum (L.) DC. Sci Pharm 2004;72:87–96. 24. Inngjerdingen KT, Colibaly A, Diallo D, Michaelsen TE, Paulsen BS. A Complement fixing polysaccharide from Biophytumpetersianum Klotzch, a medicinal plant from Mali, West Africa. Biomacrom 2006;7:48–53. 25. Varghese KJ, Anila J, Nagalekshmi R, Resiya S, Dasapushpam JS. The traditional uses and the therapeutic potential of ten sacred plants of Kerala state in India. Int J Pharma Sci  Res 2010;34:50–59. 26. Metcalfe CR, Chattaway MM, Hare CL, Richardson FR, Slatter EM, Chalk L. Anatomy of the dicotyledons: Leaves, stem, and wood in relation to taxonomy, with notes on economic uses. Int J Pharma Sci Res 1950;1:724. 27. Ueda M, Sugimoto T, Sawai Y, Ohnuki T, Yamamura S. Chemical studies on plant leaf movement controlled by a biological clock. Pure Appl Chem 2003;75:353-358. 28. Ravishankara MN, Pillai DA, Padh H and Rajani M: A Sensitive HPTLC method for estimation of amentoflavone, a bioactive principle from Biophytum sensitivum (Linn.) DC. and Putranjiva roxburghii Wall. J Med Clin 2003;16(3):36. 29. Mishra B. Vaishya R. Bhavaprakash of Bhavamishra; guduchyadi varg. Vol I; Achaukhamba Sanskrit Bhavan, Varanasi. 2018; 357-359. 30. Bhaskar VH, Rajalakshmi V. Anti-tumour activity of aqueous extract of Biophytumsensitivum Linn. Biol Res 2010;3(1):76-80. 31. Chatterjee TK, Mishra M, Pramanik KC, Bandyopadhyay D. Evaluation of anti-inflammatory, antipyretic and analgesic properties of Biophytumsensitivum. J Ind Drug 2008; 45(2): 123-131. 32. Johnson DB, Kumar DC, Arunkanth KR, Giles D, Gopal M, Hubert VG. Antifertility activity of Biophytumsensitivum. J Ind Drugs 2003; 40(9): 523-525. 33. Krishnan G, Gopalakrishna P. Pharmacological Integration: Adjunct effect of Db14201, A new herbal formulation developed based on ayurvedic principles, when co-administered with Glibenclamide: Results of a placebo-controlled trial. J Ind Drugs  2015; 2(1): 2-10.. 34. Pal TK, Dutta D, Banerjee R, Maity S. Formulation and evaluation of antimicrobial topical semisolid dosage form containing whole plant extract of B. sensitivum. J Pharm Res 2013; 6(7): 641. 35. Pal TK, Kalita P, Burman TK, Chatterjee TK, Maity S. Formulation and evaluation of antidiabetic tablet containing whole plant extract of Biophytum sensitivum on the basis of total flavonoid content. World J Pharm Res 2007;2(4):986-1007. 36. Bucar F, Jachak SM, Noreem Y, Karting T, Perera P, Bohlin L, et al. Catalysed prostaglandin M. Amentoflavone from Biophytum sensitivum and its effect on COX-1/COX-2biosynthesis. Planta Med 1998;64:373-374. 37. Puri D. The insulintropic activity of a Nepalese medicinal plant Biophytum sensitivum: Preliminary experimental study. J Ethnopharm 2001;78:89-93. 38. Guruvayoorappan C, Kuttan G. Amentoflavone stimulates apoptosis in B16F-10 melanoma cells by regulating bcl-2, p53 as well as caspase-3 genes and regulates the nitric oxide as well as proinflammatory cytokine production in B16F-10 melanoma cells, tumour associated macrophages and peritoneal macrophages. J Exp Ther Oncol 2008;7:207-18. 39. Guruvayoorappan C, Kuttan G. Amentoflavone Inhibits Experimental Tumor Metastasis Through a Regulatory Mechanism Involving MMP-2, MMP-9, Prolyl Hydroxylase, Lysyl Oxidase, VEGF, ERK-1, ERK-2, STAT-1, nm23 and Cytokines in Lung Tissues of C57BL/6 mice. Immunopharm Immunot 2008;30:711-727. 40. Guruvayoorappan C, Kuttan, G. 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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareOccupational Therapy and Yoga for Children with Autism Spectrum Disorder for Rehabilitation Professional English7073K. KalaichandranEnglish P. SwarnakumariEnglish R. SankarEnglishEnglishINTRODUCTION Autism is a neurodevelopment syndrome that is defined by deficits in social reciprocity and communication, and by unusual restricted, repetitive behaviours (American Psychiatric Association 2000). Autism is a disorder that usually begins in infancy, at the latest, in the first three years of life. Autism is caused by a combination of genetic and environmental factors. Risk factors include certain infections during pregnancy such as rubella as well as valproic acid, alcohol, or cocaine use during pregnancy.1-3 Controversies surround other proposed environmental causes; for example, the vaccine hypotheses, which have been disproven. Autism affects information processing in the brain by altering how nerve cells and their synapses connect and organize; how this occurs is not well understood. In the DSM V, autism is included within the autism spectrum (ASDs), along with Asperger syndrome which is less severe, and a pervasive developmental disorder, not otherwise specified (PDD-NOS).4-6 Autism symptoms range in presentation and severity. Autism children commonly have an intellectual disability, but in some cases, autistic children will have average intelligence. Children with autism may also have motor abnormalities, fear, anxiety, seizures, sleep cycle disturbances, gastrointestinal problems, immune dysfunction and sensory disturbances. Not all the features are present in every child. Instead, a subset of features underlies autism in each individual.7,8             Gross Motor Functional Measure scale (GMFMs) and Modified Berg Balance Scale (BBS) / Pediatric Balance Scale as a Standardized screening tool to assess gross motor function and fall in children with Autism Spectrum Disorder, the score of less than 0-20 are indicative of a high fall risk that may have limited activities of daily living (ADL) skills and demonstrates increased risk of fall.9 The ability to maintain balance during activities of daily living is essential functional independence and safety of these children with Autism Spectrum Disorder.10          Yoga is a popular and now common form of exercise for children with Autism Spectrum Disorder. It helps teach calming techniques, building muscle, and increasing flexibility, balance, and more. Occupational Therapy helps students organizing sensory systems and increase body awareness. Occupational therapy and yoga have a natural affinity for one another. Both disciplines promote improved physical and functional outcomes and used together create a dynamic state of improved wellness. Yoga has emerged as a potential and valuable tool to boost a child’s ability to focus, as well as control anxiety and sensory-related ecological by Radha krishna , Rosen blatt. Yoga as an effective behavioural treatment for children diagnosed with an Autism Spectrum Disorder by triggers, Porter and Jennifer.11,12          Literature suggests that gross motor and balance training of both yoga and occupational therapy helps to improve gross motor skills and to prevent falls in children with Autism Spectrum Disorder, only a few studies have mentioned the fall prevention training programme and children education for Autism Spectrum Disorder.9-13 Therefore, in this study, evaluation is done on the effectiveness of gross motor skills and fall prevention training programme for children with Autism Spectrum Disorder. MATERIALS AND METHODS Participants          Participants of this study were a convenience sample of children diagnosed with Autism Spectrum Disorder. All children were between ages 4-8 [Mean] age 5.97) and attended 22-week Occupational Therapy and yoga program. A total of 9 patients included, 5 Male Children and 4 Female Children diagnosed with Autism Spectrum Disorder participated in this study. All children attended the intervention phase of the study, for both Occupational Therapy and Yoga program. This study was ethically approved by the Institutional Ethical Committee (IEC) with Ref No: 000113/ 0207/2018 Dated 4/07/2018. Procedure The participants were randomly assigned for occupational therapy and yoga program, pre and post-test were done in front of parents/caregivers. Standardized assessment tool which includes Gross Motor Functional Measure scale (GMFMs) and Modified Berg Balance Scale (Pediatric Balance Scale) to assess Gross Motor function and Fall in children with Autism Spectrum Disorder and intervention was provided for 22 weeks training program with appropriate occupational therapy management such as peg-board activities, dexterity, puzzles, balancing, ball play and yoga program such as for children with Autism Spectrum Disorder. The entire children received the intervention of 45 min each over 22 weeks, for both occupational therapy and yoga intervention. Criteria for selection Children with Autism Spectrum Disorder for both male and female those who are prone to fall and poor gross motor skills were selected for this study. RESULTS           The statistical analysis of Gross Motor Functional Measure scale (GMFMs) mean value is 86.7777, SD 7.89905 and post-treatment mean value is 121.444, S.D 7.5881, t-test value of GMFM is -8.95182 and p-value is < .00001. The Statistical analysis of Berg Balance Scale between pre-treatment mean values is 24.777, S.D is 3.18948 and post-treatment mean value is 40.222, S.D 4.7088, t-test value of BBS is 7.68085 and p-value is < .00001.  Table 1 shows the Mean and SD value of Pre and Post-therapy Management of the Gross Motor Functional Measure scale (GMFMs) for Children with Autism Spectrum Disorder. Table 2 shows the Mean and SD value of the Pre and Post-therapy Management of Berg Balance Scale (BBS) for Children with Autism Spectrum Disorder.  This statistical analysis shows that there is a significant difference between pre and post-therapy values of both GMFM and BBS score. Hence, it is suggested that the gross motor skills were improved as well the rate of fall was comparatively reduced for children with Autism Spectrum Disorder.       It is observed that the t value is greater than the table values hence, the null hypothesis is rejected. It implies that there is a significant difference in the mean values before and after treatment. Hence, it is concluded that the treatment significantly shows improvements in children with Autism Spectrum Disorder. We can recommend that the period of treatment might increase and further studies are warranted to increase the sample size. CONCLUSION Throughout this study, I have concluded that Occupational Therapy and Yoga can be used effectively as one of the techniques to improve gross motor skills and to prevent fall in children with Autism Spectrum Disorder. Therapists may consider these interventions when treating children with Autism Spectrum Disorder. Acknowledgement: I Thankful to my Most bellowed Research Supervisor Dr.P.SwarnaKumari Madam and my Family Members. Conflict of Interest and: - This work has done my interest. Source of Funding: - Done, my own cost Patient informed consent: Parent consent form, parental permission for children participation in research were approved by the institutional ethical committee (IEC), Post Graduate & Research, Department of Rehabilitation Science, Holy Cross College, (000113), which was obtained from all the participants of this study. Ethical committee approval and consent to participate:  We would like to inform you that, our research  has been approved by institutional ethical committee (IEC), Post Graduate & Research, Department of Rehabilitation Science, Holy Cross College, (000113), which was obtained from all the participants of this study and which is enclosed Financial supports and sponsorship / Funding:  We did not receive any specific grant from founding agencies in the commercial or public sectors. Conflict of interest: There are no conflicts of interest to declare. Author Contribution: We (KK, PS & RS) have spent lot of time and lit bit of our own money. Kalaichandran Kothandapani :  https://orcid.org/0000-0002-4682-7842 Dr. P. Swarnakumari:                 https://orcid.org/0000-0002-6683-5743 Dr. R. Sankar:                          https://orcid.org/0000-0002-1045-9507 Englishhttp://ijcrr.com/abstract.php?article_id=3620http://ijcrr.com/article_html.php?did=3620 Amaral DG, Geschwind DH. Autism Spectrum Disorder. Oxford University Press. 2011. Minjarez MB, Williams SE, Mercier EM, Hardan AY. Pivotal response group treatment program for parents of children with autism. J Autism Dev Disord 2011;41(1):92-101. Porter J. Yoga as an Effective Behavioral Intervention for Children Diagnosed with an Autism Spectrum Disorder. Graduate Annual 2013;9. Bharah S. Yoga India Teaching Manual Level 1 and 2. One Month Yoga Teacher Training Course In Mumbai, India (theyogainstitute.org)  Cox A, Hopkinson K, Rutter M. Psychiatric interviewing techniques: II. Naturalistic study: Eliciting factual information. Br J Psychiatry 1981;138:283-291. Cox A, Rutter, M, Holbrook, D. Psychiatric interviewing techniques: Experimental study: Eliciting factual information. Br J Psychiatry 1981;139:29-37. Volkmar FR, Cicchetti DV, Bregman J, Cohen DJ. Childhood disintegrative disorder: Issues for DSM-IV. J Autism Dev Disord 1992;22:483-492. Fombonne E. Is there an epidemic of autism? Pediatrics 2001;107:411-413. Autism and Developmental Disabilities Monitoring (ADDM) Network, Available at website Fong L, Wilgosh L, Sobsey D. The experience of parenting an adolescent with autism. Int J Disabil Dev Edu 1993;40(2):105-113. Ron L, John M, Eachin JD. Work in Progress: Behavior Management Strategies and a Curriculum for Intensive Behavioral Treatment of Autism. 1999. 0966526600 (ISBN13: 9780966526608) Rahmanova S, Sabirovna S. To Study the Effects of Viral Diseases on the Human Body and Their Effective Treatment. Int J Curr Res Rev 2021;13(01):52-55. Priyanka C. Efficacy of Fumigation in Management of Rodents in Rice Shellers in District Kaithal, India. Int J Curr Res Rev 2020;12(24):130-134.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareImpact of Task-Related Games on Cognitive Flexibility of Low-Functioning Autism Spectrum Disorder Children: A Tool for Improving Quality of Life in Indian Culture English7479Kanwaljit KaurEnglish Sesadeba PanyEnglishEnglish Autism Spectrum disorder, Cognitive flexibility, Colour LED board game, Engagement, Geometry shape LED board game, Multidimensional LED board game, Music multidimensional LED board, Working MemoryINTRODUCTION Restricted, repetitive behaviour and interests, social and communicative problems dominate in autism spectrum disorder children.1 ASD is a type of neurodevelopmental disorder and children with ASD deficits in executive functioning,2 especially cognitive flexibility. ASD children show a deficit in restricted and repetitive behaviour, which is related to the impairment of cognitive flexibility.3 Till now there is no conclusive research on interventions for ASD children, still many studies indicate that individuals with ASD perform better on computerized interventions than on face to face administered tasks,4-6 but still, no conclusive data is available for both types of tasks.7 Studies on cognitive flexibility in ASD are not only inconsistent but contradictory.8 This inconsistency in result, might be due to the heterogeneity in ASD, the high level of degree comorbidity i.e. ASD with ADHD, and the overlapping in different domain of executive function i.e. working memory, inhibition, planning & organization.4,9 People with ASD find it difficult to fulfil the requirements to the changing environment, for instance, if the place of bed is changed in the room it might make them restless. However, some studies have found that cognitive flexibility deficits in ASD children.3,10-12 while others do not find any deficit.13,14 To measure cognitive flexibility among ASD children tools are used, such as the Wisconsin Card Sorting Task (WCST), Trail Making Test (TMT), or Dellis-Kaplan executive function system (D-KEFS) colour-word task; hybrid neuropsychological/experimental measures, such as the intra-dimensional/extra-dimensional (ID/ED) set shift task (Cambridge Cognition 1996), experimental task-switch paradigms, for instance, switch tasks.8,9 Children with ASD reflect deficit on the WCST15,16 and the D-KEFS colour-word task.7 Geurts et al.8 found that the failure to detect cognitive flexibility deficits in ASD is due to the predictability of the cognitive flexibility measures (such as switch tasks). Other alternative explanations are that many of the studies focus on minimizing and maximizing the WM load by varying the stimuli from simple to complex i.e. Simple geometry figure,3-D figure and3,7,11,17 and providing task cue continuously.18 The present study focuses on improving cognitive flexibility through games that initially started with minimum WM load, leads to maximum WM load to children with ASD. In this paper following questions are addressed; Do children with autism spectrum disorder (low functioning) show improvement in cognitive flexibility when exposed to task-related games? What elements of cognitive task-related games engage the children with ASD? MATERIALS AND METHODS Participant The participants were selected from Darpan academy, which is a special school for autistic children. Family members of the ASD children were contacted through the school teachers and administrators. 44 children were screened through (a) Clinical ASD diagnosis tool (Diagnostic and Statistical Manual of Mental Disorders-V) used by experienced psychiatric and psychologist in Dayanand Medical College (DMC), Ludhiana, Punjab (b) Age group 5-12 years, (c) IQ below 85. IQ was tested with Wechsler Intelligence Scale for Children (WISC-III: Kort et al.,2002; Sattler, 2001). Out of 44, eighteen children were excluded, after their screening (student’s diagnosis not confirmed n=6; age=7; students IQ Criteria does not meet, n=2; personal reasons, n=3). Seven children dropped out during the experimentation process due to their reasons. Finally, 15 potential children were selected by using inclusion criteria, for task-related games. Four children did not complete 23 cognitive sessions. three children used medication during the whole experimentation process. Pilot Test of Cognitive Flexibility Intervention Executive function performance-based test (EFPT) is meant to assess the executive function of Autistic Spectrum Disorder (ASD) children. The test consists of four major activities and each activity has two sub-activities. These activities are based on four major components i.e. working memory, inhibition, planning and organization, and cognitive flexibility. The task-related games were initially piloted with 10 students of ASD in Uddhan School for Special Children in Jalandhar, Punjab. The participants selected were between 5-10 years old. The purpose was to confirm if the interventions are appropriate to the target and to find out, what type of problem they are facing i.e. in pushing the switch, identifying the colour of the toys Intervention Intervention Description It is an individualized task where the teacher gives instructions and the child has to follow to execute the task. Different colours (Red, Green and Blue) and different shapes (Triangle, rectangle, circles and hexagonal) are inserted on the wooden board which emits different colours when switched on (attach vertically below to LED strips). Procedure Colour LED board game The participant is exposed to a wooden board, attached with the LED&#39;s (Light-emitting diodes) strips of different colours (Red, Green and Blue). First, the teacher shows them red, blue and green simple toys (i.e. Red, green and blue bucket) so the child could recall the primary colours. Then the investigator picks a toy out of a bucket and asks the participant to match the colour of the toy with the strips on the board if required cues are provided to the child. If the participant, push the correct button then he/she is rewarded, on emerging the same colour light (as the teacher instructed to match). The teacher’s instruction changes, if answered correctly by the participant (by matching the bucket colour with the one on board) as shown in figure 1. Geometry LED board game In this game, the participant is exposed to the wooden board, inscribed with the three triangles (made by using LED) of different colours (Red, Green and Blue).In this task, the teacher points a particular colour triangle i.e. red triangle (place nearby wooden block) and asks the participant to match that colour of the triangle with the wooden colour triangle (the emerging colour comes from LED stripes). The Illuminated colour of LED helps the participant to enhance the attention span and engagement in the task. The teacher will change the instructions if the answer is correct as shown in figure 2. Multidimensional LED board game A wooden board was inscribed with different shapes (triangle, circle and rectangle) made by using LED strips. The teacher points to different shapes and asks the participant to put the desired shape on the target (wooden block), in such a way that both shapes symmetrical to each other. When congruence exists between the desired and with the target shape lights emits, which reflect the symmetry between the target and wooden shape as shown in figure 3. The teacher changes the instruction according to the adaptability of the participant. Music Multidimensional LED board game A wooden block with different shapes (Circle, Rectangle and Hexagonal) was made by using LED strips along with a bird’s voice box. The teacher points towards the different shapes (desired shape place nearby wooden block) and asks the participant to put the desired shape on the target shape (shape designated on the wooden block), in such a way that both shapes symmetrical to each other as congruence exists between the desired and target shape by the participants light and birds voice are emerging as shown in figure 4. The teacher changes the instructions according to the adaptability of the participant. Procedure The study followed the time series design due to the inconsistent nature of ASD children which is difficult to generalize just only from the one test and the non-availability of children in good number to form two groups. In the present study, a single group consisting of 11 ASD category children, were taken as the subjects of the experiment. These ASD children were given intervention on task-related games. Before the treatment, they were exposed to repeated (three times) pre-tests on executive function on the dependent variables. The reason behind such repeated pre-tests was that the ASD category children’s social skills, as well as their activities either mental or behavioural, are quite inconsistent and through one pre-test it will be highly unrealistic to draw any conclusion regarding the dependent variables of the study. They were also repeatedly (three times) post-tested with the same dependent variable. RESULTS  Quantitative Analysis Research Question 1. Do children with ASD (low functioning) show improvement in cognitive flexibility when exposed to task-related games? The Means and Standard Deviation for the experimental group are presented in Table 1. The subjects of the experimental group were subjected to the cognitive flexibility test as per the time-series design and the mean values of both the pre and post-test are found to be 5.88 and 10.36 respectively. Further, when the scores were subjected to the testing of their level of significance the t ratio was reported to be 2.437 which is significant at a 0.05 level of significance (Figure 5). This result indicates that after exposure to the task-related games the ASD child showed significantly better cognitive flexibility as compared to before exposure. As earlier mentioned out of the 15 ASD children four children dropped out due to their reasons just after the pre-test and joined the group after the intervention and participated in the post-test process. As such their pre-test and post-test measures are given in table 2. In the case of the drop out ASD children, the cognitive flexibility scores were also calculated and it was observed that the pre and post-test mean scores as 2.6 and 3.3 respectively with the t ratio of 1.3 i.e. not significant (Figure 6). It indicates that the exposure of ASD children to task-related games substantially improves their cognitive flexibility. Comparison of Experimental Group with Drop-Out Group Table 3 reveals the difference between the experimental group and the drop out group. The current study shows a large effect size (0.81). It means the score of the average person in the experiment group is 0.8 standard deviation above the average person in the drop out-group and hence exceeds the score of 79% of the drop out-group. The scores varied for the two groups (Experimental Group-M=10.36, SD=4.72; Drop out Group-M=3.32, SD=0.47). At an alpha of .05, the analysis indicated a statistically significant difference among the group t =2.90 and effect size=0.81. So, we confidently (at 0.05 level) conclude that task-related games help in the improvement of cognitive flexibility among autism spectrum disorder children. Qualitative Analysis Research Question 2. What elements of cognitive task-related games engage the children with ASD? The above questions can be answered through an individual case study which discussed in detail (appendix-1). Engagement is an external measure that could be estimated by observing patterns of execution of the task. From the case studies (shown in Appendix-1), it was found that five prominent elements responsible for engaging in cognitive flexibility task and building the relationship with the ASD children are: (1) Acceptance (2) Variety of Stimulus (LED bulb, Music) (3) Motivation (4) Support system (providing cues when needed) (5) individualized instructions (Figure 7).    Children with ASD completed the games on average six to nine trials (as shown in table 4). These games completed by the children in the small number of trials that show participant pay attention to these games during the experiment which helps them to build effective interaction with the investigator as well with games. DISCUSSION The study aimed to improve the cognitive flexibility among the low functioning ASD children who reported a deficit in everyday life situations.8 In the present study, the performance of 11 ASD children was compared with drop-out students who were not exposed to the intervention due to their reasons. They were compared on basis of performance on the task mentioned in the executive function performance-based test. The task-related games based on two principles i.e. (1) Minimal Working Memory Load (2) Unpredictable and complex stimulus i.e. Different geometric shapes (difficulty level increased as proceed from game 1 to 4). This study, addressed two questions first, task-related games improve cognitive flexibility. Second, what is the pattern of behaviour of ASD student when exposed to the task-related game? Through quantitative analysis, by using a t-test it was found that there is a significant change in cognitive flexibility of ASD children as compared to those ASD children who did not get the intervention and the same trend was also observed through the case studies i.e. qualitative results. The present findings synchronized with the study findings of Kenworthy et al.4 There are at least five reasons for the positive current findings. Firstly, the Cognitive flexibility based task used in this study, unpredictable with continuous cues presented by the teacher with effective interaction. Secondly, task-related games not entirely focus on cognitive flexibility but also the far effects i.e. attention span, inhibition. Thirdly, this study focused on the individual differences approach which depicted in the variation of instructions according to the attributes of the child i.e. visual, audio with variation in the pitch. Fourth, children exposed to multiple games elements which motivated the child and increase the attention rates which may improve the training effect. Fifth, overall improvement might be due to output measure through performance task not to be measured through observation in daily life activities. Conclusion The current study indicates that the cognitive flexibility of low functioning ASD children can be improved through task-related interventions including certain factors like unpredictable stimuli, effective interaction, indirect cues (LED lights, bird voice), individual differences approach, a variety of games and output measures. This study found limited evidence for the far effect. The present study can also be extended to the population of ADHD and comorbid ASD children. Acknowledgement. The authors acknowledge the immense help received from the scholars whose articles are cited and included in the references of the manuscript. The authors are also grateful to the authors/editors. Publishers of all those articles and journals from where the literature for this article has been reviewed and discussed Conflict of Interest: The authors declare no conflict of interest associated with this work  Source of funding: There is no external funding agency associated with this article Ethical clearance: The study was approved by the departmental institutional ethics committee of CUPB ( CUPB/EDU/20-21/141). Englishhttp://ijcrr.com/abstract.php?article_id=3621http://ijcrr.com/article_html.php?did=36211. Anbarasi L, Prassanna J, Sarobin V, Rajarajeswari S, Prabhakaran R, Manikandan R. Autistic Disorder Analysis Among Children and Adult. Int J Curr Res Rev. 2020;12 (21):48-51. 2. Manggai TM, Manoj J, Muhammad Ehsan R. Analysis of Autistic Spectrum Disorder Screening Data for Adolescents. Int J Curr Res Rev 2020;12(19):23-30. 3. Yerys BE, Wallace GL, Harrison B, Celano MJ, Giedd JN, Kenworthy LE. Set-shifting in children with autism spectrum disorders: reversal shifting deficits on the Interdimensional/Extradimensional Shift Test correlate with repetitive behaviours. Autism 2009;13(5):523-538. 4. Kenworthy L, Yerys BE, Anthony LG, Wallace GL. Understanding executive control in autism spectrum disorders in the lab and in the real world. Neuropsychol Rev 2008;18(4):320-338. 5. Benyakorn S, Calub CA, Riley SJ, Schneider A, Iosif AM, Solomon M, Hessl D, Schweitzer JB. Computerized Cognitive Training in children with autism and intellectual disabilities: feasibility and satisfaction study. JMIR Mental Health 2018;5(2):e40. 6. Almeida TS, Lamb ME, Weisblatt EJ. Effects of delay, question type, and socioemotional support on episodic memory retrieval by children with autism spectrum disorder. J Autism Dev Disord 201949(3):1111-1130. 7. Van Eylen L, Boets B, Steyaert J, Evers K, Wagemans J, Noens I. Cognitive flexibility in autism spectrum disorder: Explaining the inconsistencies? Res Autism Spectr Disord 2011;5(4):1390-1401. 8.  Geurts HM, Corbett B, Solomon M. The paradox of cognitive flexibility in autism. Trends Cogn Sci 2009;13(2):74-82. 9. Monsell S. Task switching. Trends Cogn Sci 2003;7(3):134-40. 10. Hughes C, Russell J, Robbins TW. Evidence for executive dysfunction in autism.   Neuropsychologia 1994;32(4):477-492. 11. Reed P, McCarthy J. Cross-modal attention-switching is impaired in autism spectrum disorders. J Autism Dev Disord 2012;42(6):947-953. 12. Reed P, Watts H, Truzoli R. Flexibility in young people with autism spectrum disorders on a card sort task. Autism 2013;17(2):162-171. 13. Corbett BA, Constantine LJ, Hendren R, Rocke D, Ozonoff S. Examining executive functioning in children with autism spectrum disorder, attention deficit hyperactivity disorder and typical development. Psychiatry Res 2009;166(2-3):210-222. 14. Goldberg MC, Mostofsky SH, Cutting LE, Mahone EM, Astor BC, Denckla MB, et al. Subtle executive impairment in children with autism and children with ADHD. J Autism Dev Disord 2005;35(3):279-293. 15. Gillberg IC, Billstedt E, Wentz E, Anckarsäter H, Råstam M, Gillberg C. Attention, executive functions, and mentalizing in anorexia nervosa eighteen years after onset of eating disorder. J Clin Exp Neuropsychol 2010;32(4):358-365. 16. Liss M, Fein D, Allen D, Dunn M, Feinstein C, Morris R, Waterhouse L, Rapin I. Executive functioning in high-functioning children with autism. J Child Psychol Psychiatry Allied Disc 2001;42(2):261-270. 17. Robinson S, Goddard L, Dritschel B, Wisley M, Howlin P. Executive functions in children with autism spectrum disorders. Brain Cogn 2009;71(3):362-368. 18. Schmitz N, Rubia K, Daly E, Smith A, Williams S, Murphy DG. Neural correlates of executive function in autistic spectrum disorders. Biol Psychia 2006;59(1):7-16.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareHistomorphological Spectrum of Neoplastic Lesions of Kidney with a Brief Review of Literature English8085Nalini ModepalliEnglish Jyothi AnantharajEnglish Naveen ShivappaEnglish Shwetha BasavarajEnglish Sakshi BarveEnglishEnglish Kidney, Neoplasms, Morphology, Benign, MalignantMAIN  ARTICLE Introduction The kidneys are paired, bean-shaped organs located on either sides of the vertebral column in the retroperitoneal space. They  are dynamic organs serving as the main osmoregulatory system (?uid –electrolyte balance) in humans. In addition to maintaining fluid homeostasis they also act as an endocrine organ (secrete hormones - prostaglandins, and regulating vitamin D metabolism) control Red blood cell production by secreting hormone erythropoeitin and regulate blood pressure through enzyme renin. 1[D1]  The renal parenchyma though subjected to repeated trauma/insults of the noxious environment, are the last to respond . They can be involved in various pathological processes, which can be diagnosed on biopsies and nephrectomy specimens. Various disease processes affect the kidneys, some resulting in permanent damage leading to surgical removal of the organ. Nephrectomy is a common procedure in surgical practice and is of many types; partial, total and radical nephrectomy. A standard radical nephrectomy specimen consists of the entire kidney including the calyces, pelvis, variable length of ureter ,entire perirenal fatty tissue to the level of Gerota’s fascia and variable lengths of the major renal vessels at the hilus. The adrenal gland is usually removed en bloc with the kidney. Regional lymphadenectomy is not generally performed even with a radial nephrectomy. 2 A partial nephrectomy specimen may vary from a simple enucleation of the tumor to part of a kidney containing variable portions of calyceal or renal pelvic collecting system. The perirenal fat immediately overlying the resected portion of the kidney but not to a level of Gerota’s fascia is usually included. 2 Radical nephrectomy indicated in end stage renal disease, treatment of renovascular hypertension from non-correctable renal artery disease, or in severe unilateral parenchymal damage resulting from nephrosclerosis, pyelonephritis, Xanthogranulomatous pyelonephritis , vesicoureteric reflux, and congenital dysplasia. It is the treatment of choice in renal cell carcinomas. 3 Percutaneous image guided  biopsy of renal masses is a safe and accurate procedure,  being increasingly used to differentiate between benign and malignant entities to avoid unneccesary surgery. 4  In the last few years, there has been a growing interest on nephron-sparing surgery or partial nephrectomy to treat the selected cases of localized renal cell carcinoma (RCC) by open or laparoscopic approach. Renal cancer is the 9th most common cancer in men accounting for 2.5% of total malignancies.WHO (2018). 5 The histological classification of RCCs is extremely important, considering the significant implications of  the subtypes in the prognosis and treatment of these tumors. 6 In 2004 World Health Organization (WHO) published histological classification of renal tumors. 7 . In 2012, the International Society of Urological Pathology (ISUP) organized a conference in Vancouver, Canada which gave a new classification, referred to as “2012 ISUP Vancouver classification” (2012 ISUP).On the foundation of 2012 ISUP classification,2016 World Health Organization (WHO) renal tumour classification was published after considering new knowledge about pathology, epidemiology, and genetics. 3 Molecular and morphologic interrogation has driven a much-needed reexamination of renal cell carcinoma (RCC). Latest  2016 World Health Organization classification now recognizes 12 distinct RCC subtypes, as well as several other emerging/provisional RCC entities.( expanded from 4 subtypes in the 1997 Heidelberg classification to 12 recognized subtypes in 2016) 8 Important changes from previously classified  renal tumour types , to the new 2016 WHO classification refers to subtypes that have been named on the basis of predominant cytoplasmic features (eg, clear cell and chromophobe renal cell carcinomas [RCCs]), architectural features (eg, papillary RCC), anatomic location of tumours (eg, collecting duct and renal medullary carcinomas), and correlation with a specific renal disease  background (eg, acquired cystic disease– associated RCCs) as well as molecular alterations pathognomonic for RCC subtypes (eg, MiT family translocation carcinomas and succinate dehydrogenase [SDH]–deficient renal carcinomas) or familial predisposition syndromes (eg, hereditary leiomyomatosis and RCC [HLRCC] syndrome– associated RCC). 9 Emerging/provisional entities are TCEB1-mutated RCC/RCC with angioleiomyoma-like stroma/RCC with leiomyomatous stroma, RCC associated with ALK gene rearrangement, Thyroidlike follicular RCC and RCC in survivors of NB. 8 The Fuhrman system is the most frequently used grading system but has its limitations. The four-tiered WHO/ISUP grading system is recommended by the WHO . 9 However, this too has been validated as a prognostic indicator only for clear cell and papillary RCC and not for other morphotypes of renal cell neoplasms.  2016 WHO classification describes a number of new and emerging entities along with the previously recognized and also the provisional RCC types. This mandates the surgical pathologists to integrate clinical, radiologic, gross, and microscopic findings to successfully navigate challenging differential diagnoses in RCC classification. Unusual morphologic features ( clear cytoplasm, papillary architecture, and eosinophilic (oncocytic) cytoplasm) seen in various subtypes need to be kept in mind in our approach to various  RCC’s. Hence we intended to study the morphological subtypes of renal tumor using the new WHO classification. Materials and methods We conducted a cross sectional / observational study on all the  nephrectomy specimens received in Central laboratory, Department of Pathology (Histopathology division) Rajarajeswari Medical College and Hospital, Bangalore, India from Jan 2017 to Dec 2019.   Inclusion criteria- Nephrectomy Specimens received in central lab, department of Pathology, Rajarajeswari Medical College and Hospital which on histopathological examination was diagnosed as Renal tumor listed under the WHO classification of tumors of kidney were included in this study.  Exclusion criteria- The non neoplastic lesions and metastatic tumors of  kidney were excluded from the study.  Study Design - Specimens were received in 10% formalin. Essential clinical details including patient’s identification, age, sex, clinical data, investigations such as CT scan, USG, other relevant investigations were noted from the test request form. After proper fixation,  weight and dimensions were recorded  and the gross findings including examination of capsule, external surface and features on cut section were noted. Sections for histopathological examination from a lesion suspicious of tumor were taken from Tumor proper (multiple bits),  Tumor with renal capsule,  Tumor with Gerota’s fascia, hilar structures, Renal sinus, Renal pelvis , vessels , ureter and surgical  margins. Lymph nodes(if any) and sections from adjacent kidney are taken to look for associated features in the adjacent renal parenchyma. Tissue was processed, embedded, sections were cut and stained by Haematoxylin and Eosin stain and submitted for histopathological examination.  Microscopic features were noted and final reports were signed out as per the CAP protocol. RESULTS A total of 39 nephrectomy specimens were received during the study period,  out of which 15 cases(38.4%) were neoplastic and 24cases (61.6%)were non neoplastic. Among neoplastic, 4 (26.6%) cases were benign and 11cases(73.3%) were malignant tumors. Flank pain (12 cases) was the most common clinical presentation in Renal tumors followed by abdominal mass(11 cases). The highest percentage of patients belong to 61-70years age group (46.6%) with female preponderance.  Tumors were more common on the left than the right side with a predilection to the upper pole (40%). Clear cell renal cell carcinoma was more common malignant tumor and angiomyolipoma was the common one among the benign ones. Of the 10 Renal cell carcinomas, Fuhrman nuclear grading 2 was marginally more common(4 cases). Discussion Kidney are by far very resilient organs capable of withstanding the repeated stimuli by various disease processes. Once a permanent irreversible damage sets in, nephrectomy is indicated.   In our 2 year study , 15 primary renal neoplastic lesions were identified among the 39 nephrectomies  received in that period. Malignant tumors accounted for 73.3% cases which was concurrent with the studies done by Vinay KS et al 10 , Reddy et al 11 , Basir et al 12 and Gafoor A et al 13. Mean age in present study is 42.7yrs with majority of patients in the age group of 60- 70yrs , youngest being 1year old with Wilms tumour. These findings were consistent with  study conducted by Narang et al. 14 There was  female preponderance in our study with with male:female ratio of 1:1.2.This finding was in correlation with studies by Aiman et al 3 and Shifa et al 15 but most of the other studies showed a male preponderance. Left kidney showed a higher incidence of neoplastic lesions in our study as was in the studies by Chaitra.B et al 16 and Swarnalata Ajmera et al. 17 Most of the patients presented with flank pain(12 cases) and abdominal mass(11 cases) , a finding similar to study conducted by Bharti et al. 18 Grossly, most of tumors were situated in upper pole(40%) in our study comparable to Popat et al. 19 The most common malignant tumor was RCC (46.6%) , among which clear cell RCC was the most common histologic variant, consistent with the available literature. This was followed by chromophobe type RCC (2/11malignant tumors) followed by one case each of  Papillary RCC, Clear cell papillary RCC (with low grade clear cells in tubules and papillae), Multicystic RCC. Clear cell papillary renal cell carcinoma (CCP-RCC) is a recently proposed, distinctive, uncommon and indolent renal epithelial neoplasm, thought to arise in end-stage renal disease. 20 They are often small, well- circumscribed, and encapsulated, accounting for 1-4% of all resected renal tumours. These tumors have immunophenotype and molecular profiles distinct from those of clear cell and papillary RCCs. 21 CCP-RCC is composed of bland clear epithelial cells arranged in tubules and papillae, with linear nuclear alignment away from basement membrane and low Fuhrman nuclear grade. Most tumours are WHO I International Society of Urological Pathology (ISUP) grade 1 or 2. 21 Clear cell RCC rarely has papillary architecture while papillary RCC only rarely contains clear cells. Primary RCCs with both papillary architecture and cells with clear cytoplasm are diagnostically challenging for pathologists. Clues to the diagnosis of CCP-RCC include unique subnuclear cytoplasmic clearing, low nuclear grade, prominent smooth muscle stroma, and in some cases an end-stage background kidney. The morphology and typical IHC pattern of positive CK7 and CAIX but negative CD10 and AMACR should lead to accurate classification. Absence of 3p loss or trisomy of chromosomes 7 and 17 support the above diagnosis. Despite these tools, some RCCs with papillary architecture and clear cells do not fit into any of the described entities and currently remain unclassified. 22 One case of Clear cell RCC showed sarcomatoid features with 50-60% of cells arranged in solid sheets of spindle cells with high pleomorphism and high nuclear grade (Fuhrman grade 4) . Sarcomatoid RCC (sRCC) is no longer considered a separate tumor type as it can occur with all histologic subtypes. 23 It is now recognized as a high-grade undifferentiated/ dedifferentiated component of RCC, characterized by both epithelial and mesenchymal differentiation. These components share a common cell of origin as suggested by their genomic features. 24 The sarcomatoid pattern may result from activation of a distinct sarcomatoid stem cell within the tumor. The average incidence is 8% among all RCCs. 24 These tumors contain features similar to sarcomas, with spindle-like cells, high cellularity, and cellular atypia. These rare tumors show variable amounts of carcinoma elements and barely any classic RCC areas. Hence IHC using epithelial and mesenchymal   is mandatory to distinguish sarcomas from sRCC - sarcomatoid areas express cytokeratin AE1/AE3 and are negative for mesenchymal markers, such as desmin and actin. 23 These tumors are aggressive, progress rapidly and carry poor prognosis, irrespective of the underlying RCC subtype. These patients have a reported median survival time of 4-9 months after diagnosis. Major prognostic factors include proportion of sarcomatoid components, pathological stage, tumor necrosis, tumor size and genetic factors. Distant metastases are common in the lungs and bones. 24 One case of Wilms tumor in a one year old was reported in the study.  In present study Furhman grade –II was seen  more commonly among the Renal cell carcinomas, which was comparable with results of the study by Aiman et al.  3 Among the 4  benign tumors encountered in our study, 3 were Angiomyolipomas  and one oncocytoma ,these findings were similar to the results of studies by Abdul gafoor et al 13 &Aiman et al. 3 9 cases showed associated findings in the adjacent renal parenchyma which included cortical cysts, chronic glomerulonephritis and chronic pyelonephritis predominantly indicating an ongoing  chronic process. Conclusion  Histological classification of renal neoplasia has undergone significant changes with various new entities with distinct clinical , pathological and molecular characteristics.  Histologic subtype of RCC strongly correlates with prognosis, clinical implications and therapeutic strategies, underscoring the importance of accurate diagnosis. Attention to morphology and targeted immunohistochemical panel allow accurate classification of most RCCs. In others, cytogenetic and molecular findings, although expensive can establish the diagnosis. Our study provides an insight into various commonly encountered histomorphological patterns of kidney tumors and their associated features. [D2] Acknowledgement :  "Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed." [D3]  Conflict of Interest statement: The authors have no conflicts of interest to declare. Source of Funding : NIL [D4]  Authors and their contribution: Nalini M, Jyothi A Raj, and Naveen S have contributed to the conception and design of the study and have authored the main manuscript. Shwetha B and Sakshi Barve have collected the clinical and pathological data. All authors have contributed to the literature review, editing, and approval of the final draft of the manuscript. All authors agree to be accountable for the content of this manuscript and its submission to the International Journal of Current Research and review. Remove parenthesis of all the reference number in the main body of the manuscriptMention here acknowledgementMention here conflict of interest and source of funding. If there is no conflict of interest and source of funding then write NIL but mention itMention here individual author’s contribution. TABLES Table 1 : Distribution of neoplastic and non-neoplastic lesions. Englishhttp://ijcrr.com/abstract.php?article_id=3622http://ijcrr.com/article_html.php?did=3622 Alpers CE, Chang A. Robbins and Cotran Pathologic basis of disease. 9e South Asian edition. India: Reed Elsevier: 2014. Chapter 20, the Kidney: p 897-898. Srigley JR,  Zhou M, Allan R,  Amin MB, Campbell SC, Chang A et al.  Protocol for the Examination of Specimens from Patients with Invasive Carcinoma of Renal Tubular Origin. College of American Pathologists; June 2017. 15p. available from www.cap.org/cancerprotocols version : kidney 4.0.1.0 Aiman A, Singh K, Yasir M. Histopathological spectrum of lesions in nephrectomy specimens: A five-year experience in a tertiary care hospital. J Sci Soc 2013;40:148-54. Sahni VA, Silverman SG. Biopsy of renal masses: when and why. Cancer Imaging. 2009;9(1):44. Bray F, Ferlay J, Soerjomataram I, Siegal RL, Torre LA, Jemal A. Global Cancer Statistics 2018: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. Cancer J Clin. 2018;68:394-424 Muglia VF, Prando A. Renal cell carcinoma: histological classification and correlation with imaging findings. Radiol Bras. 2015 Mai/Jun;48(3):166–174. Hes O, Compérat EM, Rioux-Leclercq N, Kuroda N. The 2012 ISUP Vancouver and 2016 WHO classification of adult renal tumors: changes for common renal tumors. Diagnostic histopathology. 2016 Feb 1;22(2):41-6. Udager AM, Mehra R.  Morphologic, Molecular, and Taxonomic Evolution of Renal Cell Carcinoma - A Conceptual Perspective With Emphasis on Updates to the 2016 World Health Organization Classification. Arch Pathol Lab Med. 2016;140:1026–1037 Moch H, Cubilla AL, Humphrey PA, Reuter VE, Ulbright UM. The 2016 WHO Classification of Tumours of the Urinary System and Male Genital Organs—Part A: Renal, Penile, and Testicular Tumours  EUROPEAN UROLOGY70(2016)93–105 Vinay KS, Sujatha S. Histopathological Spectrum of Nephrectomy Specimens: Single Center Experience. Biomed J Sci&Tech Res 6(3)- 2018.  Reddy KD, Gollapalli SL, Chougani S, Sidagam S, Mohmmed AK, Bommanna A. (2016) A clinic-morphological spectrum of nephrectomy specimens-an experience from a tertiary care hospital. Int J Health Sci Res 6(11): 67-72 Bashir N, Bashir Y, Shah P, Bhat N, Salim O, Samoon N et al. Histopathological study of renal tumors in resected Nephrectomy specimens-An experience from teritary care centre. Nat J Med Res. 2015; 5(1):25-29. Ghafoor AS. Kareem A, Bashar A. Hassawi, Ziyad Ahmed. Nephrectomy. A clinicopathological study. J Am Sci 2015;11(8):97-101 Narang VI, Garg BH, Walia AS, Sood NE, Malhotra VI. Histomorphological Spectrum of Nephrectomy Specimens-A Tertairy Care Centre Experience. National Journal of Laboratory Medicine. 2016 Apr;5(2):51-4. Ibrahim SS, Dhanabalan RT, Sankari S, Shanmuganathan LK, Thandavarayan P, Ramalingam S et al. A histopathological review of nephrectomy specimens received in a referral center in South Tamil Nadu. Age. 2016;1(10). Chaitra B, Prema LP, Tejeswini V, Haritha O, Anusha M. Histopathology of nephrectomy specimens: A ten year south Indian tertiary hospital-based study. J Diagn Pathol Oncol. 2018;4(3):232-236. Ajmera S, Ajmera R (2017). Histopathological spectrum of lesions in nephrectomies- A five year study. Int J Sci Res 6(7): 44-46 Thaker BD, Singh K (2017). A histopathological review of Nephrectomy specimens Received in a Tertiary care hospital-A retrospective study. J Med Sci Cl Res 5(6): 23807-23810 Popat VC, Kumar MP, Udani D, Mundra MP, Vora DN, Porecha MM. A study on culprit factors ultimately demanding nephrectomy. Internet J Urol 2010;7(1):1-8. Kuroda N, Ohe C, Kawakami F, Mikami S, Furuya M, Matsuura K et al. Clear cell papillary renal cell carcinoma: A review. Int J Clin Exp Pathol 2014;7(11):7312-7318 Srigley JR, Cheng L, Grignon DJ, Tickoo SK. Clear cell papillary renal cell carcinoma in chapter 1: Tumors of the kidney. Moch H, Humphrey PA, Ulbright TM, Reuter VE (Eds): WHO Classification of Tumours of the Urinary System and Male Genital Organs (4th edition) IARC: Lyon 2016. pp 40-41 Ross H, Martignoni G, Argani P.  Renal Cell Carcinoma with Clear Cell and Papillary Features. Arch Pathol Lab Med. 2012;136:391–399 Shuch B, Bratslavsky G, Linehan WM, Srinivasan R. Sarcomatoid Renal Cell Carcinoma: A Comprehensive Review of the Biology and Current Treatment Strategies. The Oncologist 2012; 17: 46- 54 Liang X, Liu Y, Ran P, Tang M, Xu C, Zhu Y. Sarcomatoid renal cell carcinoma: a case report and literature review. BMC Nephrology (2018) 19: 84

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareMicro-Algal Diet for Copepod Culture with Reference to Their Nutritive Value – A Review English8696Altaff KEnglish Vijayaraj REnglishEnglishCopepods, Micro-algae, Larviculture, Aquaculture, Nutrients, FinfishIntroduction Hatchery rearing of commercially important finfish and shellfish larvae is important for successful farming of these species and live feed plays a vital role in this regard.1-3Live feeds are phytoplankton and zooplankton. They constitute the main component of the diet for finfish and shellfish larvae especially marine finfish. They are referred to as living capsules of nutrition because they contain essential nutrients such as proteins, lipids, carbohydrates, vitamins, minerals, amino acids and fatty acids.4In marine aquaculture, the use of live feed can’t replace by formulated diets in some cases. Artemia nauplii and rotifers (Brachionus plications) are still the most commonly used live feed. A good alternative is the use of copepods for rearing finfish larvae with small Gabe which could lead to the cultivation of new finfish species.5,6 Based on the above rationale, the present review article discusses the utilization of a micro-algae diet for copepods larviculture. Most research on copepods have focused on their nutrition, reproduction or physiological aspects, and culture techniques using micro-algae culture have primarily been developed at the laboratory scale. Need for alternative live feeds Past few decades, the marine hatcheries are largely dependent on the production of rotifers and brine shrimp as they could be cultured successfully and offered as traditional live feeds for fish and crustacean larvae.7-10 Rotifers are relatively easy to culture and can rapidly achieve high culture densities.11 Artemia can be obtained commercially in the form of dry cysts and their nauplii exhibit a good tolerance to culture conditions and handling.12 Research focusing on the identification of alternative feed sources that overcome the inadequacies of traditional rotifer and Artemia live feeds is critical to increase the variety and survival of species that can be successfully cultivated, and ultimately enhance the growth, sustainability and economic performance of the aquaculture industry.13-16 Copepods appear to be the most valuable candidate for this role as they are the most important natural prey for a vast majority of marine finfish larvae16-19 and their inclusion as live prey in larviculture may increase the number of fish species that can be successfully reared. Table 1 summarize the size range and main nutritional characteristics of copepods compared to the rotifers and Artemialive feed. Copepod nauplii are generally smaller than the smallest strain of rotifers and Artemia nauplii, and higher levels of the major nutrient also indicated when compared to enriched rotifers and Artemia nauplii.20-23 For the past three decades, there has been continuing interest in the development of mass culture techniques for copepods to be used as live food in aquaculture.3,7 Copepods are the most common metazoans in the marine environment, with approximately 11,500 described species. Unlike traditional live feed, copepod nutritional profiles are rich in essential fatty acids, free amino acids and other essential micronutrients.13 Moreover, the small size of copepod nauplii is vital for the first larval feeding of various fish species (Table 2). The copepod cultivation methods establishment with cost-effective protocols for mass production is still a challenge. Copepods have mostly been used at a pilot-scale or in locations where the abundant collection of natural zooplankton is possible. Still, the use of copepods as live feeds for marine fish larvae has generally led to considerably better results in terms of larval performance and quality, when compared with rotifers and Artemia nauplii.9The copepod nauplii may be produced in the rearing tanks or separate tanks and ponds. It has also been proposed that mass production of copepod resting eggs could facilitate the availability of copepod nauplii for aquaculture. However, research is needed on storage conditions of resting eggs concerning the survival and nutritional value of nauplii.18,36-38 Utilization of micro-algal diet for copepods larviculture In aquaculture, micro-algae are used as a direct food source for various filter-feeding larval stages of organisms. They are also used as an indirect food source, in the production of copepod culture which in turn is used as food for the carnivorous larvae of many of the marine finfish species presently farmed.39-41 The intensive rearing of bivalves has so far relied on the production of live micro-algae, which comprises on average 30% of the operating costs in a bivalve hatchery. For culture of marine finfish larvae according to the green water technique the micro-algae are used directly in the larval tanks.42This technique is nowadays a normal procedure in marine larviculture. It has been widely reported to improve fish larval growth, survival and feed ingestion which showed that micro-algae seemed to provide nutrients directly to the larvae that contribute to nutritional quality as it plays an important role in the microflora diversification of both the tank and the larval gut.22 Whenever micro-algae are used as a direct food source or indirect food source in the production of copepods, the growth of the animals is usually superior when a mixture of several microalgal species is used. The micro-algae diets of copepods culture are listed in Table 2. Nutritional importance of micro-algae Increasing the needs for protein and the high cost of fish meal in recent years has led to the search for new alternatives, as animal and plant sources of protein for sustainable aquaculture.42 One such accessible and relatively inexpensive food component that could respond successfully to the challenging question raised by aquaculture is algae. Cultivation of microalgae is mandatory in hatchery as it is a basic and nutritious diet for live feed organisms, specifically the zooplankton.91 However, factors such as manpower requirement, infrastructure facilities and other related costs involved in the high-density culture escalate the price of the production. For the cost-effective production of micro-algae, new approaches have to be adopted with an improvised culture environment. Micro-algal species can vary significantly in their nutritional value, and this may also change under different cultural conditions.92,93  In the laboratory-scale production of micro-algae, light plays a fundamental role in the development of microalgae through photosynthesis.94,95 It is one of the major environmental factors which control the performance of micro-algae through phototrophic growth and productivity. However, the extreme light intensity may result in photoinhibition which reduces the photosynthetic rates and growth. In the indoor culture based on the light-dark cycle periodicity duration of L D 12: 12 illumination, it could be modified to 14:10 L D or a maximum of 16:8 LD for optimizing photosynthesis of micro-algae For some microalgae and diatoms it has been reported that changes in the frequency of light-dark cycle enhance exponentially rate of photosynthesis. Some aquaculture hatcheries adopt a longer dark period compared to the light period for enhancing photosynthetic competence.96-99  Many algae have been found to contain good nutritional properties. Several factors can contribute to the nutritional value of a micro-alga, which include their size and shape, digestibility, biochemical composition, nutrients profile and enzymes. These factors satisfy the nutritional requirements of the larvae feeding on the algae. Many studies have attempted to correlate the nutritional value of micro-algae with their biochemical and nutritional profile.100-101 In the primary growth phase of the micro-algae, they contain 30 to 40% protein, 10 to 20% lipid and 5 to 15% carbohydrate51 and the high dietary protein provided the best growth for Oithona davisae.102 The major nutritional composition (Carbohydrates, Protein and Lipid) of some commercially important micro-algal species is comparable with the available feed ingredients which are used in the aquafeed. For the supply of sufficient nutrient for primary and secondary consumers, micro-algae should have the nutrient profile of protein (6–52%), carbohydrate (5–23%) and lipid (7–23%). Most microalgal species have a similar amino acid composition with enriched essential amino acids and high concentrations of ascorbic acid (1–16 mg g−1 dry weight) and riboflavin (20–40 μg g−1).103 Further high percentage of nutrient contents, carbohydrates, proteins and lipids were recorded in common micro-algae.104-109 Microalgal protein could be a plausible alternative to fishmeal protein because of its desired quality and amino acid profiles comparable with that of other reference protein sources.109 There are several studies on the utilization of biomass of Arthrospira sp.110, Chlorella sp.111 Scenedesmus sp.112 Nanofrustulum sp.113 and Tetraselmissuecica 114as valuable supplementary protein sources. Lipids have a role as high energy storage molecules. Oil content in micro-algae can exceed 60% by weight of dry biomass, while levels of 20 to 50% are common. PUFAs derived from micro-algae such as docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and arachidonic acid (AA) are known to be essential bioactive molecules for various larval cultures.115  Dunstan et al.,116 reported the most micro-algal species to have moderate to high percentages of EPA (7 to 34%) as in the case of major micro-algae species such as Pavlova spp. and Isochrysis sp. A high amount of DHA (0.2 to 11%) is present inNannochloropsis spp. and diatoms consisting of the highest percentages of AA (4%). Vitamins are essential micronutrients and their content can vary in different micro-algae. Brown and Miller117 reported that ascorbic acid shows the greatest variation (1 to 16 mg g-1) in different micro-algae. The β-carotene content ranged from 0.5 to 1.1 mg g-1. The vitamin B complex is rich in micro-algae and amount of different compounds of this vitamin showed variation in their levels (thiamin 29 to 109 μg g-1; riboflavin 25 to 50 μg g-1; niacin (0.11 to 0.47 mg g-1; pantothenic acid 14 to 38 μg g-1; pyridoxine 3.6 to 17 μg g-1; biotin 1.1 to 1.9 μg g-1; folates 17 to 24 μg g-1; cobalamin 1.8 to 7.4 μg g-1).  The α-tocopherol content of micro-algae ranged between 0.07 and 0.29 mg g-1. Nutritional importance of copepods Copepods can consume different unicellular micro-algae and also capable of feeding on filamentous algae which might provide all necessary nutrients. Several studies have demonstrated correlations between zooplankton productions and some amino acids and fatty acids content in dietary algae.118 The free-living copepod nauplii more particularly those of harpacticoid are highly suitable starter feed for marine finfish larvae due to their small size and nutritive value. The larvae of dolphinfish(Coryphaenahippurus) fed with harpacticoid copepod (Euterpinaacutifrons) nauplii showed desirable growth, survival and biochemical profile in the hatchery rearing. The mass culture system using micro-algae for the production of benthic marine harpacticoid copepod described by Sun and Fleeger could be an ideal starter feed for rearing larvae of many marine finfish with small mouth size.119 The high content of lipid is reported from many marine copepods and docosahexaenoic acid (DHA; 22:6n-3), eicosapentaenoic acid (EPA; 20:5n-3) and the saturated fatty acid (16:0) constituted predominant fatty acids. In Temoralongicornis andEurytemora sp. total lipid content reported between 7% and 14% of the dry weight of which DHA accounted 26- 42%, EPA 15-24% and Palmitic acid (16:0) 8-12%. Comparable lipid content of DHA, EPA and Palmiticacid ranging between 21% and 32.5%,15- 21% and 9-15% of total fatty acids, respectively is also reported from another calanoid copepod, Calanus finmarchicus.7 The calanoid copepods especially Acartiatonsa are a rich source of essential highly unsaturated fatty acids (HUFAs) such as docosahexaenoic acid and eicosapentaenoic acid.32 The fatty acid composition of Acartiatonsacontain high amount of monounsaturated fatty acid (411 mg g−1), and polyunsaturated fatty acids (360 mg g−1). A large amount of DHA and EPA (170 and 96 mg g−1, respectively) reported in this species.120 Several marine copepods have a high content of both docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) (60% of total fatty acids).7 Also, the copepods are natural sources of the antioxidant, astaxanthin and vitamins C and E.32 The powerful antioxidants found in copepods can protect the HUFA’s against peroxidation and are also considered beneficial to the health of fish larvae.69 An exogenous supply of free amino acids (FAA) is necessary to support growth and survival in first feeding finfish larvae.121 Copepods contain higher levels of free amino acids when compared to rotifers and Artemia nauplii.32 Copepods generally contain more vitamins and trace minerals than traditional live feeds.122 Copepods are rich in vitamin E and ascorbic acid, suggesting a high antioxidative capacity and making them particularly suitable for larvae with potential for high growth rates.32 Furthermore, copepods are rich in pigment content, particularly astaxanthin, which may be an important source of retinoids for larval fish.32 A copepod diet has been reported to promote correct pigmentation in Atlantic halibut larvae, with 55% of the copepod fed larvae exhibiting correct pigmentation of the ocular and blindsides as compared to only 13% in those fed Artemia nauplii.123 Thus it is generally accepted that many copepods are the valuable nutrient source of live feed for marine larval finfish rearing. With varying food and feeding habits and reproductive strategies among even in closely related copepod species it necessary to venture in to establish their mass production protocols.  Further such research is needed to identify the copepod species which matches nutritionally and in size spectrum with the requirement of specific marine finfish larval rearing. Conclusion This review reveals the importance of micro-algal for the culture of copepods, nevertheless, such studies are mostly of small scale level and to reach cost-effective commercial-scale production, attempts should have made to enumerating new species for mass production. They have to be mass cultured with the knowledge of their reproductive potentials and reproductive strategies. Considering copepod suitability for successful rearing of many finfish larvae further research is urgently needed to assess the culture potential of more candidate copepods as prey for the rearing of early stages of marine finfish larvae which in turn might greatly benefit the mariculture industry. Acknowledgement             The authors express their gratitude to the Management of AMET University for providing research facilities to carry out this work. The authors are thankful to the Department of Biotechnology, Govt. of India for funding a project (BT/PR30019/AAQ/3/929/2018) for Copepod culture studies. The authors acknowledge the immense help received from the scholars whose articles are cited and included in references to this manuscript. The authors are also grateful to authors/editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. 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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcarePrevalence of Post-Partum Depression and the Associated Risk Factors Among Materials in Al-Madinah City 2019 English97105Khalid Ahmad AmaraEnglish Somaya Mohammad Mahfouz AlshereifEnglish Reham Mohammad KharabahEnglishEnglishPostpartum depression (PPD), risk factors, Madinah, Saudi ArabiaINTRODUCTION Postpartum depression (PPD), a type of mood disorder, is the most frequently noted morbidity during the postpartum period.1 PPD is defined as " in the Diagnostic and Statistical Manual for Mental Disorders as major depression with postpartum onset with episodes of depression beginning within 4 weeks of giving birth"2 It is a non-psychotic depressive state that begins in the post-partum period, after the childbirth, it is a mood disorder that can occur at any time during the first year after delivery.3            PPD affects the health of both the mother and her children, especially mother-child bonding and the relationships among family members.3,4 Several studies have found that risk factors for depressive symptoms are clustered into five main groups: 1.biological, consisting of changes in hormone levels and the age of mother; 2.physical, consisting of chronic health problems and antenatal depression; 3.psychological, consisting of prenatal anxiety, stress, lack of social support and stressful life; 4.obstetrics/paediatrics, consisting of unwanted pregnancy, history of loss of pregnancy and severely ill infants; 5.and socio-cultural, consisting of the status of mother and poverty.3-8 PPD has been associated with catastrophic consequences, such as mother suicide and infanticide.3 Financial shortage, infant gender, domestic violence, hunger, and smoking during pregnancy have been reported as risk factors of PPD.3,4,9          Although PPD is a major health issue for many women from diverse cultures, this condition often remains undiagnosed.10 The use of screening scales is an easy, simple and cost-effective way to identify women who are at risk of depression.1,11 The Edinburgh Postnatal Depression Scale (EPDS) is the most widely used screening tool in identifying PPD,1,12-14 and it has been validated and used in many countries, including Saudi Arabia.15        PPD is the most common childbirth complication1 which affects 10%–15% of women in high-resource countries.16 However, the prevalence is also considerably high in developing countries, including India (23%)17, Pakistan (44%)18 and Vietnam (33%).19 In Saudi Arabia studies the prevalence was 33.0%-49.5%.15,20          Post-partum depression symptom represents a common problem among post-partum mother due to several factors. PPD symptoms result from a combination of biochemical, physical and emotional factors. The main biochemical factor causing PPD symptoms is reduced hormonal levels which lead to chemical changes in the brain that may activate mood swings. The researcher has a special interest in depression, particularly among post-partum mothers. This study aimed to investigate the prevalence and assess risk factors of postpartum depression symptom among post-partum mothers in Al-Madinah, Saudi Arabia. MATERIALS AND METHODS This study was across sectional descriptive study among post-partum mothers delivered in the hospitals in Al-Madinah city. Post-partum mothers with substances abuse or bipolar major depressive disorder were excluded. The calculated sample size was 360 with 95% confidence limits, 5% accepted errors, the prevalence of PPD syndrome =30%, 3and with 10% (30 nurses) was added to the sample to avoid withdrawing and refusing to participate.        The required sample size was divided equally to cover the three hospitals (National guard, Ohod, and MMCH hospitals) (120 post-partum mothers/ hospital). Valid structured self –administered questionnaire. The questionnaire is to obtain from previous study 15and it contains three main parts: A – Socio-demographic characters. B – pregnancy and delivery characteristics. C – The Edinburgh Postnatal Depression Scale The dependent variable of the study was the post-partum depression scale. And the independent variables were sociodemographic characteristics and pregnancy and delivery characteristics. Data was entered and processed by using SPSS software version 21 for analysis and interpretation. P-Value is considered statistically significant if it is ≤ 0.05.      The following approvals were obtained: research ethical committee, the supervisor of the general training program, primary healthcare centre PHCC director. Written consent was obtained from each participant. And the collected data was handled confidentially. RESULT Normality The Shapiro-Wilk statistic test was done for the following continuous variables Edinburgh Postnatal Depression Scale (EPDs score, age, children number, labour duration, and gravidity).The result were (0.906, 0.983, 0.925, 0.583, and 0.95) with (pEnglishhttp://ijcrr.com/abstract.php?article_id=3624http://ijcrr.com/article_html.php?did=3624 Bhusal BR, Bhandari N. Identifying the factors associated with depressive symptoms among postpartum mothers in Kathmandu, Nepal. Int J Nurs Sci 2018;5:268-274 Suhitharan T, Pham TP, Chen H, Assam PN, Sultana R, Han NL, et al. Investigating analgesic and psychological factors associated with risk of postpartum depression development: a case-control study. Neuropsychiatr Dis Treat 2016;12:1333-9. Giri RK, Khatri RB, Mishra SR, Khanal V, Sharma VD, Gartoula RP. Prevalence and factors associated with depressive symptoms among post-partum mothers in Nepal. BMC Res Notes 2015;8:111. Kalinin P, Arthur DG. Postpartum depression in Asian cultures: a literature review. Int J Nurs Stud 2009;46(10):1355–73. Mehta S, Mehta N. An overview of risk factors associated with postpartum depression in Asia. Ment Illness 2014;6(1):5370. Wisner KL, Chambers C, Sit DK. Postpartum depression: a major public health problem. JAMA 2006;296(21):2616–8. Fisher J, Mello MCD, Patel V, Rahman A, Tran T, Holton S, et al. Prevalence and determinants of common perinatal mental disorders in women in low-and lower-middle-income countries: a systematic review. Bull World Health Organ. 2012;90(2):139–49. Chaaya M, Campbell OMR, El Kak F, Shaar D, Harb H, Kaddour A. Postpartum depression: prevalence and determinants in Lebanon. Arch Women&#39;s Ment Health 2002;5(2):65–72. DØRheim Ho†Yen S, Tschudi Bondevik G, Eberhard†Gran M, Bjorvatn BR. Factors associated with depressive symptoms among postnatal women in Nepal. Acta Obstet Gynecol Scand 2007;86(3):291–7. Stewart DE, Robertson E, Dennis CL, Grace SL, Wallington T. Postpartum depression: a literature review of risk factors and interventions. https://www.who.int/mental_health/prevention/suicide/lit_review_postpartum_depression.pdf 2003. Cox JL, Chapman G, Murray D, Jones P. Validation of the Edinburgh postnatal depression scale (EPDS) in non-postnatal women. J Affect Disord 1996;39(3):185-9. Teissedre F, Chabrol H. A study of the Edinburgh Postnatal Depression Scale (EPDS) on 859 mothers: detection of mothers at risk for postpartum depression. Encephale 2004;30(4):376-81. Alvardo-Esquivel C, Sifuentes-Alvarez A, Salas-Martinez C, Martinez-Garcia S.  Validation of the Edinburgh postpartum depression scale in a population of puerperal women in Mexico. Clin Pract Epidemiol Ment Health 2006;2(33). Pitanupong J, Liabsuetrakul T, Vittayanont A. Validation of the Thai Edinburgh postnatal depression scale for screening postpartum depression. Psychiatry Res 2007;149(1e3): 253-9 Alamoudi DH, Almrstani AMS, Bukhari A, Alamoudi LH, Alsubaie AM, Alrasheed RK and Bajouh O. Prevalence and factors associated with depressive symptoms among post-partum mothers in jeddah. Int J Adv Res 2017;5(2):1542-1550. TaW Northumberland. Postnatal Depression: a self-help guide. Northumberland: Tyne and Wear (NHS Trust); 2006:2003. Patel VDN, Rodrigues M, DeSouza N. Gender, poverty, and postnatal depression: a study of mothers in Goa. Indi., Am J Psychiatr 2002;159:43-47. Khooharo YMT, Das C, Majeed N, Majeed N, Choudhry AM. Associated risk factors for postpartum depression presenting at a teaching hospital. ANNALS 2009;16(2):87-90 O&#39;Hara MW, Swain AM. Rates and risk of postpartum depression metaanalysis. Int Rev Psychiatr 1996;8:37-54 Alharbi AA1, Abdulghani HM. Risk factors associated with postpartum depression in the Saudi population. Neuropsychiatr Dis Treat 2014;10:311-6. Markhus MW, Skotheim S, Graff IE, Frøyland L, Braarud HC, Stormark KM,  et al. Low omega-3 index in pregnancy is a possible biological risk factor for postpartum depression. PLoS One. 2013;8(7):e67617. Zauderer C. Postpartum depression: how childbirth educators can help break the silence. J Perinat Educ 2009;18(2):23–31. Shah S. Frequency of postpartum depression and its association with breastfeeding: A cross-sectional survey at immunization clinics in Islamabad. J Pak Med Assoc 2017;67(8). Prakash R, Ranadip RP,  Chowdhury R, Salehi A, Sarkar K, Singh SK, et al. Postpartum depression in India: a systematic review and meta-analysis. Bull World Health Organ 2017;95(10):706–717. Nepal M, Sharma V, Koirala N, Khalid A, Shresta P. Validation of the Nepalese version of Edinburgh Postnatal Depression Scale in tertiary health care facilities in Nepal. Nep J Psychiatry 1999;1:46–50. Regmi S, Sligl W, Carter D, Grut W, Seear M. A controlled study of postpartum depression among Nepalese women: validation of the Edinburgh Postpartum Depression Scale in Kathmandu. Trop Med Int Health 2002;7(4):378–382. Turkcapar AF1, Kad?o?lu N2, Aslan E3, Tunc S4, Zay?fo?lu M, Mollamahmuto?lu L. Sociodemographic and clinical features of postpartum depression among Turkish women: a prospective study. BMC Pregn Childbirth. 2015;15:108. DØRheimHo†Yen S, TschudiBondevik G, Eberhard†Gran M, Bjorvatn BR. Factors associated with depressive symptoms among postnatal women in Nepal. Acta Obstet Gynecol Scand 2007;86(3):291–297. Muraca GM, Joseph KS. The association between maternal age and depression. J Obstet Gynecol Can 2014;36(9):803–810. Daoud N, Shoham-Vardi I, Urquia ML, O’Campo P. Polygamy and poor mental health among Arab Bedouin women: do socioeconomic position and social support matter? Ethn Health 2014;19(4):385-405. Shepard LD. The impact of polygamy on women’s mental health: a systematic review. Epidemiol Psychiatr Sci 2013;(1):47–62. Vigod S, Villegas L, Dennis C-L, Ross L. Prevalence and risk factors for postpartum depression among women with preterm and low-birth-weight infants: a systematic review. BJOG 2010;117:540–550. Ege E, Timur S, Zincir H, Geckil E, Sunar-Reeder B. Social support and symptoms of postpartum depression among new mothers in Eastern Turkey. J Obstet Gynaecol Res 2008;34(4):585-593.  Kheirabadi GR, Maracy MR, Barekatain M, Salehi M, Sadri GH, Kelishadi M, et al. Risk factors of postpartum depression in rural areas of Isfahan Province, Iran. Arch Iran Med. 2009;12(5):461-467. Savarimuthu RJ, Ezhilarasu P, Charles H, Antonisamy B, Kurian S, Jacob KS. Post-partum depression in the community: a qualitative study from rural South India. Int J Soc Psychiatry 2010 56(1):94-102. Stewart RC, Bunn J, Vokhiwa M, Umar E, Kauye F, Fitzgerald M, et al. Common mental disorder and associated factors amongst women with young infants in rural Malawi. Soc Psychiatry Psychiatr Epidemiol 2009;45(5):551-559. Clarke K, Saville N, Shrestha B, Costello A, King M, Manandhar D, et al. Predictors of psychological distress among postnatal mothers in rural Nepal: a cross-sectional community-based study. J Affect Disord 2014;156:76–86. O’hara MW, Swain AM. Rates and risk of postpartum depression-a meta-analysis. Int Rev Psychiatry 1996;8(1):37–54. Lee DT, Yip AS, Leung TY, Chung TK. Identifying women at risk of postnatal depression: prospective longitudinal study. Hong Kong Med J 2000;6(4):349–354. Goker A, Yanikkerem E, Demet MM, Dikayak S, Yildirim Y, Koyuncu FM. Postpartum depression: is mode of delivery a risk factor? ISRN Obstet Gynecol 2012;2012:616759. Saleh El-S, El-Bahei W, El-Hadidy MA, Zayed A. Predictors of postpartum depression in a sample of Egyptian women. Neuro Psychiatr Dis Treat 2013;9:15–24. Segre LS, O’Hara MW, Arndt S, Stuart S. The prevalence of postpartum depression: the relative significance of three social status indices. Soc Psychiatry Psychiatr Epidemiol 2007;42:316–321. Da-Silva VA, Moraes-Santos AR, Carvalho MS, Martins MLP, Teixeira NA. Prenatal and postnatal depression among low income Brazilian women. Braz J Med Biol Res 1998;31:799-804. Ross LE, Dennis CL. The prevalence of postpartum depression among women with substance use, an abuse history, or chronic illness: a systematic review. J Womens Health (Larchmt) 2009;18(4):475-486. Chaaya M, Campbell OM, El Kak F, Shaar D, Harb H, Kaddour A. Postpartum depression: Prevalence and determinants in Lebanon. Arch Womens Ment Health 2002;5:65–72.  Fo¨ rger F, Ostensen M, Schumacher A, Villiger PM. Impact of pregnancy on health-related quality of life evaluated prospectively in pregnant women with rheumatic diseases by the SF-36 health survey. Ann Rheum Dis 2005;64:1494–1499. Robertson E, Grace S, Wallington T, Stewart DE. Antenatal risk factors for postpartum depression: a synthesis of recent literature. Gen Hosp Psychiatry 2004;26(4):289–295. Chee CY, Chong YS, Ng TP, Lee DT, Tan LK, Fones CS. The association between maternal depression and frequent non-routine visits to the infant’s doctor – a cohort study. J Affect Disord 2008;107(1–3):247–253. Gonidakis F, Rabavilas AD, Varsou E, Kreatsas G, Christodoulou GN. A 6-month study of postpartum depression and related factors in Athens Greece. Compr Psychiatry. 2008;49(3):275–82. O’Hara MW. Postpartum depression: what we know. J Clin Psychol 2009;65:1258–1269. Orsel S, Karadag H, Turkcapar H, Kahilogullari AK. Diagnosis and classification subtyping of depressive disorders: comparison of three methods. Bull Clin Psychopharmacol 2010;20:57–65. Hamdan A1, Tamim H. The relationship between postpartum depression and breastfeeding. Int J Psychiatry Med 2012;43(3):243-259. Cristina Borra,corresponding author Maria Iacovou, and Almudena Sevilla. New Evidence on Breastfeeding and Postpartum Depression: The Importance of Understanding Women’s Intentions. Matern Child Health J 2015;19(4):897–907.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareRp-Hplc Method for Simultaneous Estimation of Vildagliptin and Metformin in Bulk and Pharmaceutical Formulations English112117V. NagalakshmiEnglish G. Srinivas RaoEnglish N. Gayathri DeviEnglish S. MohanEnglishEnglishINTRODUCTION Chemically known as 3-[diaminomethylidene]-1, 1-dimethylguanidine HCl, metformin HCl is an oral pharmaceutical product used to treat type 2 diabetes mellitus. Metformin is considered an antihyperglycemic drug because, without inducing hypoglycaemia, it reduces blood glucose concentrations in type II diabetes. Metformin is widely known as an insulin sensitizer that leads to a decrease in insulin resistance and a clinically important decrease in insulin levels from plasma fasting. Modest weight loss is another well-known advantage of this drug. For obese type II diabetes patients, Metformin is the medication of choice.1,2 Chemically, Vildagliptin is known as pyrrolidine-2-carbonitrile, (2S)-1-{2-[(3-hydroxyadamantan-1-yl) amino] acetyl}. Vildagliptin, previously known as LAF237, is a novel oral antidiabetic drug in the drug class of the new dipeptidyl peptidase-4 (DPP-4) inhibitor. Vildagliptin subsequently works by inhibiting glucagon-like peptide-1 (GLP-1) and gastric polypeptide inhibitor (GIP) inactivation by DPP-4.3,4 Literature review reveals that for estimation of metformin and vildagliptin in combination and individual dosage form5-9 and validated with parameters,10-16 various analytical methods such as UV-Vis spectroscopy, HPLC and LCMS / MS methods are available. Without using any buffers, the procedure with new composition was developed in the estimation of these compounds using RP-HPLC methods and also the development and validation of a simple, precise, fast and specific method for the determination of metformin and vildagliptin in pure form and its pharmaceutical dosage form were considered of interest. MATERIALS AND METHODS Chromatographic conditions Column: ODS (4.6×250mm, 5µm, Hypersil) Mobile phase: ACN: Methanol: Water (15:60:25) Flow rate: 1ml/min Column temperature: 35oC The volume of injection: 20µl λmax: 278nm Degradation studies The forced degradation was carried under acidic, basic, oxidative conditions. Both the drugs were separately exposed to stress conditions. After exposing stress conditions, drugs were diluted to the standard concentration of Metformin (30µg/ml & 160µg/ml) and Vildagliptin (30µg/ml & 160µg/ml). Equal volume of both drugs were mixed and analyzed in the chromatographic conditions. The acid stress condition was carried out using 1N HCl, when mixed with the drug solution and kept for 48hours and then injected into the chromatographic column, degradants evaluated using chromatograms. The alkali hydrolysis was carried out using 1N NaOH, when mixed with the drug solution and kept for 48hours and then injected into the chromatographic column to evaluate the degradedness. Hydrogen peroxide is a strong oxidant when 3% H2O2 was added to the 1000µg/ml drug and kept for 48hours and injected into the chromatographic column to check the degradation. RESULTS Validation by Method The method defined has been validated, including parameters such as suitability of the system, linearity, accuracy, precision, robustness, LOD and LOQ. System suitability Results of the system suitability study are summarized in the above table 1. Six consecutive injections of the standard solution showed uniform retention time, theoretical plate count, tailing factor and resolution for both the drugs which indicate a good system for analysis. Specificity The analytical peak was evaluated as per the methodology and observed the interference of blank, placebo with the analyte peak was there or not. Metformin and vildagliptin peaks were observed at their respective retention times of analyte peaks in figure 3 and 4.When the blank solution was injected, no peak was found in figure 2. The forced degradation study showed the method was highly specific; the chromatographic peak does not interfere with any other impurities. This proves that excipients do not affect the analytical method. On the other hand, the blank peak did not overlap the drug peak. So the method is highly selective. Accuracy Validated the accuracy in this method, accurately quantify metformin and vildagliptin tablets content at 50%, 100%, 150% and performed assay in triplicate. The mean per cent recovery of metformin and vildagliptin at each spike level should be not less than 98% and not more than 102%. Results of the accuracy study are presented in table 2. The measured value was obtained by the recovery test. The spiked amount of both the drugs were compared against the recovery amount. Precision The precision of the test method by preparing six test preparations using the product blend by mixing the active ingredient with excipients as per the manufacturing formula was determined. Repeatability of Standard metformin and vildagliptin solution was injected six times and peak areas were measured and metformin &vildagliptin per cent RSD was found to be 0.566 & 1.903. Results are given in table 3. Linearity A linear relationship between peak areas versus concentrations was observed for metformin and vildagliptin in the range of 50% to 150% of normal concentration. The correlation coefficient was 0.982 for metformin and0.988 for vildagliptin. This proves that the method is linear in the range of 50% to 150 %. Results are given in Table 4. Robustness The results of the robustness of the present method had shown by changes in the flow rate and wavelength did not produce significant changes in analytical results which were presented in table 5. As the changes are not significant we can say that the method is robust. The Detection Limit LOD for Hydrochloride of metformin = 0.6177 LOD = 0.1544 for vildagliptin Limit of Quantification LOQ for Hydrochloride of metformin = 1.8711 LOQ = 0.4688 for vildagliptin The LOD and LOQ values indicate that the method developed was sensitive, precise and reliable. Studies of stability Comparative studies were performed on drugs called metformin and vildagliptin before and after degradation, and acid, base and oxidative degradation results were reported in Table 6. Research on Stress Degradation The properties of stress degradation were analysed using a validated chromatographic method for metformin and vildagliptin. Studies of forced degradation are listed in the table. Reports show that the validated approach effectively isolated and separately classified the degradation products. From the reports, it is very clear that drugs were responsive to acidic environments where there was less degradation. DISCUSSION The proposed method for the simultaneous determination of metformin and vildagliptin in pharmaceutical dosage form was found to be precise, selective, rapid and economical.5 The interaction study between the two drugs in the standard solution was carried out by comparing peaks of each drug individually with peaks obtained in drug mixture indicating that the analytes did not interact with each other and data were within the acceptance level of ±2.0%.1 The linearity for detector response was observed in the concentration range of 50 to 150% of test concentration and the correlation coefficient (r) for the calibration curve was found to be 1.0. Per cent recovery was found to be within the range of 98.0 % to 102.0% indicating the accuracy of the method.6 The per cent RSD for the tablet analysis and recovery studied is less than 2 which is indicating a high degree of precision.5 The results of recovery studies were found to be linear in 50 % to 150 % of the final assay concentration range indicating linearity and range of the proposed method. The results of the robustness study indicate that the method is robust and is unaffected by small variations in the wavelength and flow rate trails.7 Hence, it can be concluded that the developed RP-HPLC method is accurate, precise, rapid and selective and can be employed successfully for the estimation of metformin and vildagliptin in bulk and pharmaceutical dosage forms.8 Degradation was not observed in metformin and vildagliptin stressed samples that were subjected to acid hydrolysis and oxidative conditions. However, degradation was observed under base hydrolysis. This indicates that the method is specific and stability-indicating.1 CONCLUSION In pharmaceutical formulations, the author sheds light on the enhancement of HPLC methods for estimating metformin hydrochloride and vildagliptin. It is usually important to design methods in which a very large number of samples are to be analysed with appropriate precision and accuracy in a very short period. It is possible to obtain qualitative results via the HPLC process and can thus be used in analytical analysis. This is an effective technique that provides good results for validation parameters. This approach explicitly performs well on Metformin hydrochloride and Vildagliptin. ACKNOWLEDGMENT: The authors thanks to the Department of Chemistry, Ch.S.D.St.Theresa’s College for Women (A), Eluru for providing workspace and the authors are grateful to Icon Pharmaceutical Lab for providing instrumental support. CONFLICT OF INTEREST: The authors declare that no conflict of interest for this research. No funding support for this research   Englishhttp://ijcrr.com/abstract.php?article_id=3625http://ijcrr.com/article_html.php?did=3625 Alaa SA, Soha F, Mohamad MM, Abo T. Simultaneous for the Estimation of Metformin and Empagliflozin in Pharmaceutical Dosage Form by HPLC Method. IOSR J Pharm Bio Sci 2019;14(1):75-80. Thornberry NA Gallwitz B. Mechanism of action of inhibitors of dipeptidyl-peptidase-4 (DPP-4). Best Pract Res Clin Endocrinol Metab 2009;23(4):479-486. Ahren B, Landin Olsson, Jansson M. Inhibition of dipeptidyl peptidase-4 reduces glycemia, sustains insulin levels, and reduces glucagon levels in type 2 diabetes. J Clin Endocrinol Metab 2004; 89(5):2078-2084. Mentlein R, Gallwitz B,Schmidt. Dipeptidyl-peptidase IV hydrolyses gastric inhibitory polypeptide, glucagon-like peptide-1(7-36) amide, peptide histidine methionine and is responsible for their degradation in human serum. Eur J Biochem 1993;214(3):829–835. Shirode1 AR, Maduskar PD, Deodhar MS, Kadam VJ. RP-HPLC and HPTLC methods for simultaneous estimation of Metformin Hydrochloride and Vildagliptin from bulk and their marketed formulation: Development and Validation. Br J Pharm Sci 2014;4(20):2370-2386. Abu DW, Hamad M, Mallah E, Abu DA, Awad R, Zakaria Z,  et.al. Method Development and Validation of Vildagliptin and Metformin HCl in pharmaceutical dosage form by Reverse phase-High performance liquid chromatography (RP-HPLC). Int J Pharm Sci Res 2018; 9(7):2965-2972. Subhakar N, Krishna Reddy V, Ravindranadh Reddy T. Development and Validation of RP-HPLC method for simultaneous determination of Vildagliptin and Metformin in bulk and formulation dosage. Int Res J Pharm Appl Sci 2012;2(3):44-50. Srinivas Ch, AnilkumarCh, Nagaraju B, Nagajyothi J.  Method development and Validation by RP-HPLC for the simultaneous estimation of Metformin and Vildagliptin in pharmaceutical dosage form. Eur J Pharma Med Res 2017;4(6):01-16. Priyanka DP, Saurabh D, Pandya S. The aim of this review to focus on a comprehensive update of different analytical methods for determination of oral anti-diabetic drugs for the treatment of type 2 diabetes mellitus (T2DM). World J Pharm Sci 2018;6(1):29-39. International Conference on Harmonization (ICH); Q2 (R1), Validation of analytical procedures: Text and methodology, Geneva, Switzerland, (2005).  United state Pharmacopoeia, USP 32, NF27, Page No. 2906, 2007. Mohammad A, Azim Md, Elkady EF, Fouad A. Development and validation of a reversed phase column liquid chromatographic method for simultaneous determination of two novel gliptins in their binary mixtures with Metformin. Eur J Chem 2012;3(2):152-155. Shaikh KA, Patil SD, Devkhile AB. Development and validation of a reversed-phase HPLC method for simultaneous estimation of ambroxol hydrochloride and azithromycin in tablet dosage form. J Pharm Biomed Anal 2008;48(5):1481–1484. Santhosha B, Ravindranath A, Sundari Ch. Validated method for the simultaneous estimation of Metformin Hydrochloride and VIDA by RP-HPLC in bulk and the pharmaceutical dosage form. Int Res J Pharm App Sci 2012;2(3):22-28. De AK, Ashokkumar B, Pal B. Development and Validation of Same RP-HPLC Method for Separate Estimation of Theophylline and Doxofylline in Tablet Dosage Forms. J Curr Pharm Res 2012;9(1):55-58. Devika GS, Sudhakar M.  RP HPLC method for simultaneous estimation of metolazone and ramipril in oral solid dosage form. Int J Pharm Bio Sci 2012;3(4):193–200.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareBio-conversion of Lignin Extracted from Sugarcane Bagasse and Coconut Husk to Vanillin by Bacillus sp. English118123Harpreet KaurEnglish P V PavithraEnglish Sriya Das and M KavithaEnglishEnglishVanillin, Lignin, Sugarcane bagasse, Coconut husk, Microbial bioconversionIntroduction Vanillin chemically known as 4-hydroxy 3-hydroxybenzaldehyde is a white crystalline phenolic aldehyde and it is a commonest flavouring and perfuming agent. It is naturally extracted from Vanilla planifolia which is an orchid, also known as a vanilla bean.1 Its fragrance and taste are from its fruit, vanilla pod, by an enzymatic process called curing post-harvest. After the curing process, only 2% of the vanillin is extracted. It is also a labour-intensive process leading to its high cost.2 Due to the shortage and high cost, synthetic vanillin production was researched and adopted. The methods of commercial production of vanillin were first started by using natural and readily available, eugenol also known as clove oil (4-allyl-2-methoxyphenyl).3 Later other methods of the artificial production of vanillin were followed using lignin or guaiacol. 85-90% of the production depends on guaiacol, which is a phenolic ether also known as 1-Hydroxy-2-methoxybenzene and only 10-15% is from lignin. These synthetic methods produce “naturally similar” vanillin but not natural vanillin. Lignin is the second most abundant natural polymer with cellulose being number one, making up to 10–25% of lignocellulosic biomass.4,5 The natural vanillin extract meets the global vanillin demand by around only 1%.1 It is used as a flavouring agent mainly in ice creams and chocolate manufacturing (accounting for 75%), followed by the pharmaceutical industry to mask the undesirable flavour, and food and beverages.6 Vanillin is also useful for visualization of alcohols, ketones and aldehydes separated using thin layer chromatography.7 Vanillin-HCl staining is used for tannins histochemical testing.8 Vanillin is also used as a food preservative as it contains anti-bacterial and anti-oxidant activities.9 Due to its high demand, in the recent past researcher are exploring alternative methods for the production of vanillin. To meet the expanding demand for vanillin, biotechnological approaches such as bioconversion of ferulic acid, isoeugenol, eugenol, lignin with the help of bacteria, fungi and genetically modified microorganisms are being researched to explore its full potential.1,10,11 Microbes such as white-rot fungi, bacteria such Bacillus sp. are used to degrade the complex lignin polymer to vanillin, ferulic acid and vanillic acid.12 The present study is aimed to explore the potential of Bacillus sp. in the bioconversion of lignin extracted from coconut husk and sugarcane bagasse to vanillin. Materials and Methods Materials Vanillin and catechin were procured from Sigma. Mineral Salts Medium (MSM) and other culture media ingredients were from Hi-Media, Mumbai. All other chemicals used in the study were of analytical grade obtained locally. Growth conditions for Bacillus sp. The Bacillus sp. procured from ATCC and available for regular work in the Microbiology laboratory was used in the study. It was cultured in Minimal salts medium (MSM) at 37°C for 24 hours. Stock cultures were maintained at 4°Cfor for further use. Gram’s staining was done to confirm its characteristic morphology and endospores.  Extraction of lignin from sugarcane bagasse and coconut husk Sugarcane bagasse and coconut husk were collected from local juice shop and farmland in Vellore respectively. (Figure 1-2) Lignin from sugarcane bagasse was extracted by acid hydrolysis. It was shredded into smaller pieces, mixed with formic acid and acetic acid mixture (70:30) and boiled for 2 hours. The residues were filtered, treated with peroxyformic/ peroxyacetic acid which was prepared using 35% H2O2 and boiled at 80oC for 2 hours. The lignin present in the residues was precipitated with distilled water.13 Lignin from coconut husk was extracted by alkali hydrolysis. Coconut husk was cut into smaller pieces, oven-dried and processed with 1M aqueous NaOH solution at 100oC for 1 h. It was cooled and then filtered. The filtered residue was precipitated with 20% phosphoric acid and kept overnight.14 The lignin precipitate was separated using filter paper. The separated lignin was placed in a Petri plate and dried in an oven at 55oC for 48 h till the lignin was dried completely. Characterization of Lignin by Fourier-Transform Infrared Spectroscopy analysis The extracted lignin was structurally characterized using Fourier transform infrared spectroscopy (Shimadzu). The scan was performed in the range of 500-4000 cm-1.15 Bioconversion of lignin to vanillin The bioconversion of extracted lignin to vanillin was carried out in MSM media supplemented with 0.5% and 1% lignin using Bacillus sp. The culture media were prepared, sterilised and inoculated with 1% inoculum of Bacillus sp. The culture flasks were incubated at 37oC for 48 h under shaking condition. After incubation, the culture fluid was centrifuged at 8000 rpm for 10 min at 4oC to obtain the supernatant containing vanillin for further analysis.  Vanillin assay Thiobarbituric acid assay Thiobarbituric acid reacts with vanillin to form a yellow coloured product with maximum absorbance at 434 nm which can be measured using a spectrophotometer. To 1 ml of the culture supernatant, 5 ml of 24% HCl and 2 ml of 2% thiobarbituric acid were added and mixed. The test tubes were incubated at 55oC for 10 min in a water bath followed by 20 min at room temperature. The absorbance was measured at 434 nm and compared with vanillin standard to determine the concentration of vanillin present in the supernatant.1                         Catechin assay Aromatic aldehydes when reacts with meta substituted flavanols gives a red adduct with maximum absorbance at 500 nm. 8% Hydrochloric acid (HCl) was prepared in methanol. 0.3% catechin was prepared and stored in a dark bottle at 4oC. To 1 ml of the culture supernatant, 5ml HCl reagent and 5 ml catechin were added and incubated at 50oC in the water bath for 20 min to form a red coloured adduct. The absorbance was measured at 500 nm and compared with vanillin standard to determine the concentration of vanillin present in the supernatant.16 Characterization of vanillin by High-Performance Liquid Chromatography (HPLC) analysis The vanillin was analysed using WATERS High-Performance Liquid Chromatography. The mobile phase was acetonitrile and 0.2% acetic acid in the ratio of 60:40. 20 microliter of vanillin was injected into the column and the flow rate was adjusted 1ml/minute and the graph was obtained at 280nm and compared with vanillin standard.1,16 Results and Discussion Morphology of Bacillus sp. Bacillus sp. presented with grey-white, round, opaque and medium-sized colonies on both nutrient agar and Lignin-MSM plates (Figure 3-4).  Gram staining revealed the presence of Gram-positive bacilli, arranged in chains with endospores (Figure 5). Extraction of lignin from sugarcane bagasse and coconut husk                                 In both acid hydrolysis of sugarcane bagasse and alkali hydrolysis of coconut husk, the dark brown semisolid precipitate was obtained on Whatman No.1 filter paper. (Figures 6-8) This precipitate was dried and used in further study. The yield of lignin was 6% from sugarcane bagasse and 7% from coconut husk. Characterization of lignin by FT-IR analysis The FTIR analysis of the given sample (Figure 9) was carried out and compared with the reference wave numbers of lignin reported in the literature.15 It could be observed that the wavenumbers fairly agreed with the values reported in the literature proving the presence of lignin in the extract (Table 1). In FT-IR analysis the stretching patterns indicate the presence of compounds like phenols, hydroxides, methyl groups, which are present in lignin. The bands in the region of 1600 and 1500 cm-1 indicate the presence of aromatic compounds such as phenol (Figure 9). Bioconversion of lignin to vanillin After the bioconversion reaction, the light brown supernatant obtained (Figure 10) was subjected to vanillin assay. Thiobarbituric acid assay Upon the addition of thiobarbituric acid to the tests and standards, the yellow coloured product was observed in the test sample. The absorbance was read at 434 nm and recorded and a standard graph was plotted. (Figure 13) The standard curve was calibrated from 0.2- 1 mg/ml. The regression equation obtained was y= 0.635x + 0.049. The r2 value obtained was 0.9871. From the graph, it could be interpreted that the vanillin present in the broth is 0.7 mg/ml. Catechin assay The tests and standards reacted with catechin and resulted in the formation of a light red coloured product. The absorbance was recorded at 500nm. The standard curve was calibrated from 0.2- 1 mg/mL. The regression equation obtained was y= 0.55x + 0.232. The r2 value obtained was 0.93. From the graph, it is interpreted that the vanillin present in the sample is estimated to be 0.65 mg/mL (Figure 14). Characterization of vanillin by HPLC analysis The extracted vanillin was analysed using WATERS High-performance liquid chromatography. As seen in the HPLC graph (Figure 11), the retention time of vanillin was 2.361. Few other smaller peaks were observed due to the tendency of vanillin to rapidly oxidize to vanillic acid and structural similarity between them. The formation of vanillic acid hence may have interfered in the specific detection of vanillin by HPLC. The other reason for various peaks might be a small number of impurities present in solvent or standard vanillin. Observing one of the peak retention time 2.369 of the test sample (Figure 12), we could conclude that vanillin was present in the sample. Most of the reports on lignocellulosic biomass have been on its lignification, structural properties and degradation.17,18 The setbacks in the commercial production of vanillin lies in its chemical reactivity and it is rapidly converted to vanillic acid. 19 Current research on the elucidation of the biosynthetic pathway of vanillin in microorganisms opened up new opportunities in the biotechnological production of vanillin.20 Conclusion This study demonstrated the capability of Bacillus sp. to adapt in the lignin- MSM media and the bioconversion of lignin to more valuable product vanillin. The lignin was retrieved by acidic and alkaline methods and H2O2 accelerated the process. Lignocellulosic biomass has been explored more on its lignification, structural properties and degradation. The shortcoming in the biotechnological production of vanillin includes its chemical reactivity. Vanillin gets rapidly converted to vanillic acid. To make this process industrially and economically viable it is important to inhibit the conversion of vanillin, make it more stable and develop strategies to recover it more efficiently. The increasing knowledge regarding enzymes that are involved in pathways for the biosynthesis and metabolism, identification and characterization of the corresponding genes, offers new opportunities for metabolic engineering and the construction of genetically engineered production strains to advance the production. Acknowledgement The authors thank the Vellore Institute of Technology for providing all necessary equipment and chemicals to carry out this project work.  Conflicts of Interest NIL Funding NIL Authors Contribution All the authors were involved in the conception and design of the work, experimental works, data collection, data analysis and drafting of the article. The corresponding author revised and approved the final version of the manuscript. Englishhttp://ijcrr.com/abstract.php?article_id=3626http://ijcrr.com/article_html.php?did=3626 Rana R, Mathur A, Jain CK, Sharma SK, Mathur G. Microbial production of vanillin. Int J Biotechnol Bioengi Res 2013;4 (3):227-234. Dignum MJ, Kerler J, Verpoorte R. Vanilla production: technological, chemical, and biosynthetic aspects. Food Rev Int 2001;17(2):119-120. Kaur B, Chakraborty D. Biotechnological and molecular approaches for vanillin production: a review. Appl Biochem Biotechnol 2013;169(4):1353-1372. Pothiraj C, Kanmani P, Balaji P. Bioconversion of lignocellulose materials. Mycobiology 2006;34(4):159-165. Pearl IA. Vanillin from lignin materials. J Am Chem Soc 1942;64(6):1429-1431. Vanillin: Physiochemical Properties, Production and Uses. UKEssays November 2018. Pirrung MC. Appendix 3: Recipes for TLC stains. The Synthetic Organic Chemist’s Companion. 2006:171-2. Tai A, Sawano T, Yazama F, Ito H. Evaluation of antioxidant activity of vanillin by using multiple antioxidant assays. Biochimica et Biophysica Acta 2011;1810(2):170-177. Gardner RO. Vanillin-hydrochloric acid as a histochemical test for tannin. Stain Technol 1975;50(5):315-317. Karode B, Patil U, Jobanputra A. Biotransformation of low cost lignocellulosic substrates into vanillin by white rot fungus. Phanerochaete chrysosporium NCIM. Indian J Biotechnol 2013;12:281-283. Muheim A, Lerch K. Towards a high-yield bioconversion of ferulic acid to vanillin. Appl Microbiol Biotechnol 1999;51(4):456-461. Chen P, Yan L, Wu Z, Li S, Bai Z, Yan X, et al. A microbial transformation using Bacillus subtilis B7-S to produce natural vanillin from ferulic acid. Sci Rep 2016;6:20400. Watkins D, Nuruddin M, Hosur M, Tcherbi-Narteh A, Jeelani S. Extraction and characterization of lignin from different biomass resources. J Materials Res Technol 2015;4(1):26-32. Forss KG, Talka ET, Fremer KE, inventors; Forss Kaj G, Talka Esko T, Fremer Kaj Erik, assignee. Method for the isolation of vanillin from lignin in alkaline solutions. United States patent US 4,277,626. 1981 Jul 7. Lu Y, Lu YC, Hu HQ, Xie FJ, Wei XY, Fan X. Structural characterization of lignin and its degradation products with spectroscopic methods. J Spectrosc 2017;2017: 8951658. Graf N, Altenbuchner J. Genetic engineering of Pseudomonas putida KT2440 for rapid and high-yield production of vanillin from ferulic acid. Appl Microbiol Biotechnol 2014;98(1):137-149. Kantharaj P, Boobalan B, Sooriamuthu S, Mani R. Lignocellulose degrading enzymes from fungi and their industrial applications. Int J Curr Res Rev 2017;9(21):1-7. Glasser WG. About making lignin great again-some lessons from the past. Front Chem 2019;7:565. Converti A, Aliakbarian B, Domínguez JM, Vázquez GB, Perego P. Microbial production of biovanillin. Braz J Microbiol 2010;41(3):519-530. Zheng L, Zheng P, Sun Z, Bai Y, Wang J, Guo X. Production of vanillin from waste residue of rice bran oil by Aspergillus niger and Pycnoporus cinnabarinus. Bioreso Technol 2007;98(5):1115-1119.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareTransactional Model of eGovernance: Empirical Evidence from the Healthcare Sector of India English124131Sumeet GillEnglish Priya VijEnglishIntroduction: A vast array of ICT based eGovernance initiatives are being put into practice in the healthcare sector in India to facilitate the populace and other stakeholders. However, a structural and strong healthcare management system postulates the development of an updated governance model. Objectives: The paper proposes to develop a Transactional Model governance model for the healthcare sector to augment the efficiency and social influence of eGovernance web portals. Methods: Twelve dimensions, extracted based on extensive literature review, models, NeSDA and UNDESA methodology and through Exploratory and Confirmatory Factor Analysis (EFA and CFA) have been used. Responses of 600 doctors, healthcare staff and patients have been collected through an in-depth interview and observation checklist. Structural Equation Modeling (SEM) has been applied to develop and propose a transactional model of governance after statistical testing. Results: The outcome depicts a significant positive impact of identified dimensions on Intention to Use and Service Quality construct of healthcare-related eGovernance services which ultimately improves efficiency thereby leading to social influence. Conclusion: The paper proposes an integrated and efficient governance model framework to enhance the social influence of eGovernance services in the healthcare sector. The promising outcome evinced that the proposed framework may serve as a roadmap for policy and decision-makers English eGovernance, Efficiency, Corruption, Mismanagement, Resources, Social influenceINTRODUCTION The coherent elevation and adoption of Information Communication Technology (ICT) driven eGovernance applications in the healthcare sector has transformed the mode of contact, connect and transact between hospitals, administrators, doctors, healthcare supporting staff and patients. The synthesis of the utility of ICT based eGovernance in the healthcare sector in India started with Mission Mode Projects (MMPs) of the National e-Governance Plan (NeGP) initially via using traditional communication technology viz. Electronic Media (Community Radio, and telephone), and gradually elevated to use of state-of-the-art technology viz. Video and Teleconferencing (Telehealth, Tele-medicine and Tele-referrals, online video consultations); Internet?enabled Web-based centres (Information kiosks, common service centres, eOffice, eHospital, voice web etc.);  and Cloud-based eHealth and hospital administrative services. The NeGP 2.0, eKranti, and Digital India movement facilitates services like Aadhar based online registration, appointments, payments, access to diagnostic reports, real-time tracking of data and registrations, efficient information management, patient monitoring, medical education and communication, Vaccine Tracker, eRakt Kosh); Online Medical Counseling and Admissions, online continuous medical education courses through webinars and live streaming; Swasth Bharat (Disease, Lifestyle, First Aid), NHP Indradhanush; eHMIS, Ayushman Bharat – Health and Wellness Centre (HWC) portal, Integrated Disease Surveillance Programme (IDSP) Portal, Mera Aspataal – patient Feedback mechanism and many more.1 The thrust of the National Health Policy (NAP) supported by the Digital India initiative is on augmenting the eGovernance revolution by offering medical facilities through the web, mobile, and cloud (MeghRaj) accessible to the populace equitably round the clock.2 Ministry of Health and Family Welfare (MoHFW), Government of India is endorsing eHealth to develop an efficient, evidence-based, ethical healthcare system that shall encourage, educate, and empower patients and other stakeholders through information dissemination thereby improving public healthcare delivery.1 However, the current COVID 19 pandemic situation raised a serious question about our health machinery.  Over the years governments and bureaucrats considered healthcare only as a medical subject and ignored the acute need for a structured health management system for the second largest populated country. India ranked 145 as per Global Healthcare Access and Quality (HAQ) index, ranks lower than Bangladesh, Sri Lanka, Bhutan and Sudan and also slipped to 150th rank in healthcare and survival as per World Economic Forum (WEF).3,4 Health staff always remain under stress due to scarcity of infrastructural and human resource i.e. one government doctor on 10,189 residents, one government bed on 2,046 residents and one government hospital on 90,343 residents.5 On the other hand, the Supreme Court of India has termed patient care as &#39;&#39;deplorable” and &#39;&#39;pathetic”, as patients are running from pillar to post to get admission to hospitals, whereas a large number of beds are lying vacant in the hospitals.6 The problem is not merely inadequate resources i.e. infrastructure and human, but also mismanagement of health institutions. Furthermore, not much attention is being paid to ethical issues related to the adoption of technology and the safety of personal and sensitive information related to patients.8 Besides, the most serious ethical crisis i.e. corruption in the healthcare sector is poorly addressed and doctors and hospital staff are key actors behind the same.7,8 20% of health costs in the country is due to corruption and there is corruption in healthcare education as well.9 The Organisation for Economic Co-operation and Development (OECD) estimates that 45 % of global citizens believe that the health sector is corrupt or very corrupt. Insufficient regulatory oversight and lack of transparency in governance breed corruption and reduce the quality of healthcare services.10 Apart from issues related to infrastructural, human resources, mismanagement, poor GDP allocation4 and inadequate attention towards governance11 are responsible for the pathetic situation of healthcare in India.  The outcome of various research studies has established a significant correlation between poor governance and mismanaged public healthcare services across countries.12 The gaps are visible between the ground realities and the policies framed to address them. Many strategies exist for making the healthcare environment conducive, viz. adequate funding to the public healthcare system, remunerating health care workers adequately, ensuring social accountability, etc. but eGovernance is the most vital and cost-effective strategy for the national healthcare system to work optimally.10 The induction of web, mobile and cloud-based portals in native languages has enhanced the popularity of healthcare-related eGovernance services among all and the healthcare sector steadily advanced from the eGovernance transactional phase to transformational phase i.e. participation of end-users in decision and policymaking and implementation.10 With the proliferation of information channels, the expectations of the patients related to the accessibility of public health services in a convenient, integrated, transparent, and cost-effective manner have been boosted. However, the complexity of the eGovernance in the healthcare sector has enhanced with time. The contrary research outcomes and amplified implementation challenges encouraged researchers, governing institutions and research labs towards the development of an eGovernance model viz. “DeLone and McLean IS Success Model (DandM Model), Technology Acceptance (TAM) model, Theory of Planned Behavior (TPB), Gartner&#39;s Four Stage Model, Layne and Lee Four-Stage Model, Heeks Design Reality Gap Model, Theory of Reasoned Action (TRA), Structuration Model of Technology (SMT), United Nations Five-Stage Web Presence Measurement Model, Diffusion of Innovation (DOI), Unified Theory of Acceptance and Use of Technology (UTAUT), and Theory of Planned Behavior (TPB), Heeks Onion Ring Model, EGOVSAT model” and many more to enhance the competence of the eGovernance initiatives. However, a significant gap in the expansion of existing theoretical and technology-oriented models conceptually and practically is evident in research outcomes.13 Further, the enormous failure rate of eGovernance initiatives supported by research gaps necessitates rapid and consistent horizontal and vertical assessment from the stakeholders’ perspective. National e-Governance Service Delivery Assessment 2019 (NeSDA) assessed eGovernance services portals of six sectors (Finance, Labour & Employment, Education, Local Government & Utilities, Social Welfare, Agriculture, Health and Environment) based on seven key constructs i.e. Accessibility, Ease of use, End service delivery, Integrated service delivery, Content Availability, Information security & privacy,  and Status & Request tracking, based upon Online Service Index (OSI) of UNDESA eGovernment Survey in 2019 and published survey outcome in February 2020.14 The low level of stakeholders’ satisfaction evinced in the survey particularly related to health sector demands for development of an integrated model and with this in view, the present paper proposes to develop a transactional model of eGovernance for healthcare sector from stakeholders’ perspective, with emphasis on enhancing efficiency and social influence of eGovernance web portal.  MATERIALS AND METHODS The process of model development encompasses various phases, exploring the dimensions of eGovernance using EFA by collecting responses from 600 doctors, healthcare staff and patients through in-depth interview related to twelve dimensions i.e. “Performance Expectancy, Compatibility, Job Fit, Facilitating Conditions, Intention to Use, Ease of Use, Competence, Reliability, Usefulness, Responsiveness, Product Portfolio, and Security dimensions” extracted based on extensive literature review, models, NeSDA and UNDESA survey methodology. The extracted dimensions and their relationship were affirmed using CFA.  The healthcare staff involved in governance projects serving at Central and State-funded Post Graduate Institutions of Medical Sciences (PGIMS) and patients were considered as sample unit and 600 respondents were selected based on Judgment sampling. The doctors and healthcare staff having more than three years of experience in eGovernance activities and patients possessing Smartphone with hands-on knowledge, ready to interact and participate were included in the study.  The respondents were contacted through Snowball Approach and responses were recorded on Likert Seven Point Summative Scale (SD (1) to SA (7)) through an in-depth interview and observation checklist. The data were tabulated and statistically analyzed using open source software “Jamovi” Version 0.9.5.12 and EFA, CFA, SEM was applied. The respondents were apprised regarding the objectives of the research before starting the interview. Further, the respondents were assured that their identity shall not be revealed.  RESULTS AND DISCUSSION The value of the Kaiser-Meyer-Olkin (KMO) test of Sampling Adequacy was 0.907 and the P Value of Bartlett’s Test of Sphericity was less than 0.05, which endorse the application of EFA (Tables 1 and 2). The principle Components Analysis (PCA) method was applied to execute EFA. All extracted factors having Eigenvalue >1 were retained for further analysis. Tables 3 and 4 explained extraction of 28 statements due to communality value less than 0.50, data adequacy and existence of relationship among the 21 remaining statements on which final EFA was applied. The cumulative percentage of variance was 70.546 that inferring 70 per cent of eGovernance portal efficiency is explained by these twelve dimensions. Further, CFA was applied for overall construct validation and statements falling in each of the factors derived from EFA were tabulated in Table 5. CFA was applied on twelve dimensions and model fit indices/summary (Table 6) and GFI, CFI, NFI and RMSEA values tabulated in Table 7. CFI value of 0.9 and above testifies strong unidimensionality; GFI value of more than 0.9 shows the best fit of the model, and value of RMSEA 0.079 also makes the model acceptable. The relationships between the statements and factors were strong and significant (Figure 1). All the relevant assumptions of SEM were satisfied and the transactional model of eGovernance for the healthcare sector proposed in Figure 2. CONCLUSIONS The outcome depicts the significant positive impact of “Performance Expectancy”, “Compatibility”, “Job Fit”, “Facilitating Conditions”, “Ease of Use”, “Competence”, “Reliability”, “Usefulness”, “Responsiveness”, “Product Portfolio”, and “Security” on Intention to Use and Service Quality constructs of eGovernance which ultimately improves the efficiency of the eGovernance in the healthcare sector and efficiency leads to positive social influence. The proposed transactional model of governance will guide the policymakers to focus on identified constructs to enhance and maintain efficiency and social influence. The suggested model may be tested after integrating it with new constructs based on empirical outcomes for an update. However, the masses will not be benefited unless administrative, social and political measures ensure equitable access to all. ACKNOWLEDGEMENT The authors acknowledge the help of the open source society for providing software’s access required for statistical analysis.  The authors acknowledge the immense help received from the scholars whose articles are cited and included in references to this manuscript. The authors are also grateful to authors/editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Source of Funding: Self Funded Conflict of Interest: Non Englishhttp://ijcrr.com/abstract.php?article_id=3627http://ijcrr.com/article_html.php?did=3627 Phanish C. e-Governance in Indian Healthcare Building a Stronger Nation. eHealth The Enterprise of Healthcare. 2017; https://ehealth.eletsonline.com/2017/07/e-governance-in-indian-healthcare-building-a-stronger-nation/ e-Health India, NHP Admin. 2017. https://www.nhp.gov.in/e-health-india_mty Vinay A. COVID-19 – Mismanagement and urgent need for Indian Medical Services. The Northline [Internet]. 2020 July. http://www.thenorthlines.com/covid-19-mismanagement-and-urgent-need-for-indian-medical-services/ Medical Dialogues Bureau. India Slips To 150th Rank In Healthcare: World Economic Forum. Medical Dialogues [Internet]. 2019 Dec 17. Available from: https://medicaldialogues.in/india-slips-to-150th-rank-in-healthcare-world-economic-forum?infinitescroll=1 Sanchita S. India’s public health system in crisis: Too many patients, not enough doctors. Hindustan Times [Internet] 2017 August 29. Available from: https://www.hindustantimes.com/india-news/public-health-system-in-crisis-too-many-patients-not-enough-doctors/story-39XAtFSWGfO0e4qRKcd8fO.html Sumit S. Delhi govt hospitals a spectacle of mismanagement (IANS Investigation) (Delhi Health Emergency-II). Outlook [Internet]. 2020 June 13.  https://www.outlookindia.com/newsscroll/delhi-govt-hospitals-a-spectacle-of-mismanagement-ians-investigation-delhi-health-emergencyii/1865342 Sanjay K. Health care is among the most corrupt services in India. BMJ 2003;4(10):326-37. Subrata C. Corruption in healthcare and medicine: Why should physicians and bioethicists care and what should they do? Indian J Med Ethics 2013;10(3):153-159. Corruption blamed for healthcare sector woes. The Hindu [Internet]. 2019. https://www.thehindu.com/news/cities/kozhikode/corruption-blamed-for-healthcare-sector-woes/article26361494.ece Crossing the Global Quality Chasm: Improving Health Care Worldwide. National Academies of Sciences, Engineering, and Medicine; Health and Medicine Division; Board on Health Care Services; Board on Global Health; Committee on Improving the Quality of Health Care Globally. Washington (DC): National Academies Press (US); 2018. https://www.ncbi.nlm.nih.gov/books/NBK535646/ Chandrasekhar CP, Jayati G. Covid-19: Why is India faring worse than its neighbours?. Business Line [Internet]. 2020 August 11. https://www.thehindubusinessline.com/opinion/columns/c-p-chandrasekhar/covid-19-why-is-india-faring-worse-than-its-neighbours/article32318486.ece Maureen L. Governance and Corruption in Public Health Care Systems. Center for Global Development. 2006. Working Paper Number 78. http://www1.worldbank.org/publicsector/anticorrupt/Corruption%20WP_78.pdf Anita V, Smriti S, Deboshree G. Impact of E-Governance Practices on Agriculture Sector of India. CASS Studies. 2019; 3(1), 294-307. http://heb-nic.in/cass/admin/freePDF/kji8rwvj50kbxuba8lwc.pdf Alawneh A., Al-Refai H., Batiha K. Measuring user satisfaction from eGovernment services: lessons from Jordan. Government Information Quarterly. 2013; 30(3), 277–288. DOI:https://doi.org/10.1016/j.giq.2013.03.001 Department of administrative reforms and public grievances (India). National e-Governance Service Delivery Assessment 2019, e-Governance Landscape – India’s Transformative Journey. New Delhi: Department of administrative reforms and public grievances, Ministry of personal grievances and pensions, Government of India; 2020.  https://www.nesdaportal.in/NeSDA_2019_Final_Report.pdf

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareEfficacy of Maitland Mobilization and Conventional Treatment in Patients of Osteoarthritis of Knee English132137Subrat SamalEnglish Shweta PanchbudheEnglish Snehal SamalEnglish Mohini DixitEnglish Vasant GawandeEnglishEnglishOsteoarthritis, Maitland Mobilization, Conventional treatment, Knee jointINTRODUCTION             Osteoarthritis (OA)is the most common type of arthritis and the leading cause of disability that impacts the elderly and middle-aged worldwide.1 OA is a multifactorial entity that includes several causative factors such as Trauma, Mechanical forces, Inflammation, Biochemical responses and Metabolic disturbances.2,3 The incidence of OA knee in rural and urban India is 3.9% and 5.5%.4 It is recorded to be 5.78% and 10.20% respectively in Bangladesh and India. According to research in Pakistan, 28% of the metropolitan population and 25% of the rural population have knee arthritis.5 The meta-analysis also revealed that women tend to be graphically evaluated with more serious knee OA radiographically than men, and sex differences boost with age > 55 years.6                     Osteoarthritis is a complex disease whose pathogenesis involves the contribution of biomechanical and metabolic variables that alters articular cartilage and subchondral bone homeostasis of the tissue and determine the predominance of destructive over productive procedures. The end products of cartilage emerge out and the cell lining the joint will attempt tore move them. The small bony formation “spur” will be formed in the lining of the articular surface. The physical findings of Osteoarthritis of the knee include: bony enlargement, crepitus, decreased range of motion, joint-line tenderness, and pain on passive range of motion.7 The cause of osteoarthritis is repetitive mechanical loads and aging.Decreased strength in the muscle group involving the joints which it causes significant progressive loss of function.There are two terms of Mobilization and Manipulation, but they have the same meaning and can be changed. The variable speeds and amplitude can vary from a small- amplitude force applied at quick velocity to a big amplitude force applied at slow velocity, a continuum of intensities and velocity could be applied to the method.8,9 Gentle Joint mobilization can be used by stimulating neurophysiological and mechanical effects to treat pain and muscle guarding. In neurophysiological effects, small oscillatory amplitude and distraction motion are used to boost mechanoreceptors which can prevent the transmission of nociceptive stimuli at the level of the spinal cord or brainstem.10-13 This study is carried out to find out that which treatment is effective i.e.Maitl and Mobilization or Conventional treatment in reducing Pain, ROM, functional impairment in OA knee patients. MATERIALS AND METHODS Materials used are treatment table, universal half-circle goniometer, Hydro collateral packs, stellium, cotton and talcum powder. Collection of data Seventy-two (72 subjects) of 40 to 60 years of age with a history of pain, tenderness and diagnosed with knee osteoarthritis were taken and screened by the inclusion criteria in the study and assessed for pain, range of motion and functional activity using the visual analogue scale, universal Goniometer, WOMAC Scale. Patients were randomly divided into two groups. Group A(36 subjects will receive Maitland Mobilization with conventional therapy) and Group B(36 subjects will receive conventional therapy alone).14,15 72 healthy subjects(16 male and 56 female) with osteoarthritis of the knee participated in the study. The subjects were recruited from the Department of Musculoskeletal Sciences, Ravi Nair Physiotherapy College, Sawangi (Meghe), Wardha after approval from the Institutional Ethics Committee of Datta Meghe Institute of Medical Sciences, Deemed to be University. Study Design: Interventional study. Study Setting: OPD of Musculoskeletal Sciences, Ravi Nair Physiotherapy College, Sawangi, Wardha. Sample size: Total of 72 subjects. Study Duration: 6 weeks Sampling Technique: Subjects were selected Randomly Allocated by chit method and assignedin2groupssuchasGroupA(Experimental group)andGroupB(Control group) in equal number (36 subjects in each groups). Inclusion Criteria Kellegren and Lawrence grade 1 and 2 radiographic evidence of osteoarthritis Both Genders Age between 40 to 60years Knee osteoarthritis with duration ≥ 1 year. Average knee pain ≥3 on Visual Analog Scale. Subjects with Unilateral knee joint. Tibiofemoral and Patellofemoral knee joint Tenderness over kneejoint Patient with tightness in Quadriceps, Hamstring, and Iliotibial band. Exclusion Criteria Acute exacerbation in or around kneejoints. Traumatic injury to knee joints within 6 months of study. Any surgical intervention to the kneejoints. Intra-articular steroid injection in knee joints within 3months. Subject with Psychiatric disorders/Illness. Peripheral vascular disease. Tumours/malignancies/infections associated with kneejoint. lower limb metallic implants. Impaired thermal sensation over the knee. Procedure The institutional ethics committee clearance DMIMS(DU)/IEC/2018-19/7193was obtained to conduct the study. Initially the patient was thoroughly evaluated. After satisfying inclusion and exclusion criteria the patient was allowed to participate in the study. The patients who willingly volunteered for the study were included and the purpose was explained. Total 76 OA knee subjects were screened for the inclusion and exclusion criteria of the study, out of which 4 subjects were not willing to continue treatment. Therefore 72subjects were included in this study. The procedure was well explained to all the eligible subjects and they were given informed consent (n=72) before allocating them into two groups. After randomization, they were divided into two groups- Group A (n=36) and GroupB(n=36)The preassessment were take non(0Day)and post-assessment was taken at the end of 6thweek for all outcome measures such as the Visual Analog Scale (VAS) for pain, Goniometer for ROM, WOMAC Scale for functional activities test. In Group A subjects (Experimental) received Maitland mobilization along with conventional treatment. In Group B subjects (Control) received Conventional treatment alone.  Patients were instructed to perform a home exercise program which was taught to them on the first assessment day. Group A subjects received Grade I, II and III Maitland Mobilization. Maitland mobilization for tibio femoral joint     Tibio femoral Distraction Tibio femoral Posterior Glide (To increase flexion) Tibio femoral Anterior Glide (To increase the extension) Patello femoral Joint, DistalGlide To maintain patellar mobility for normal knee flexion Group B- Conventional Therapy In conventional physiotherapy, subjects were receiving physical modalities such as Hydro collateral packs to reduce pain. StaticQuadriceps Static Hamstringcontraction VMO (strengthening of vastus medialis) Dynamic quadriceps Wall slides Partial lunges One leg standing16,17 Home Exercise Programme Self-stretching of Quadriceps muscle Self- stretching of Hamstring muscle Self -stretching of Calf muscle Outcome Measures Visual analogue scale Knee flexion range of motion on universal Goniometer WOMAC scale for functional disability. 18,19 RESULTS The data was coded and entered into a Microsoft Excel spreadsheet. Descriptive statistics included computation of means and standard deviation. Inferential statistics using student’s paired ‘t’ test (for the quantitative data to comparepre and post observation)and unpaired t-test (for quantitative data to compare within two groups) were used for comparis on of all clinical indicators.SoftwareusedintheanalysiswasSPSS22.0version. The results were concluded to be statistically significant with p < 0.05, very significant < 0.001 and highly significant p < 0.0001 (Table 1).             Table 1 and Graph 1 shows age wise distribution of subjects. 10(27.78 %) in group A and 10(27.78%)ingroupBsubjectswereintheagegroupof40-49years,24(66.67%)ingroup A and 23(63.89%) subjects in group B were in the age group of 50-59 yrs. Less than 60 years,2(5.56%) in group A and 3(8.33%) in Group B .The mean age of the subjects in groupAwas52.30 ±4.42andin groupBitwas52.58±5.74. Byusingchi-squaretestno significant difference is found in the ages of subjects of in all threegroups Table 2 Shows gender-wise distribution of subjects. There were 9(25%) in group A and 7(19.44%) male subjects in group B, 27 (75%) in Group A and 29(80.56%)female subjects in Group B. By using chi-square test no significant differences in found among the gender of both the group. Table 3 shows the comparison of pain on VAS in both group A and group B.Mean pain on VAS in Group A was 4.86±1.22 and in Group B was 2.22±0.79. By using student’s unpaired t-test showing a significant difference in both groups but Group A shows more reduction of pain than Group B. Hence, Group A shows a significant improvement in pain reduction than Group B. Table 4 shows the comparison of pain ROM on the Goniometer in both groups A and group B. Mean ROM on the goniometer in Group A was 16.38±8.34 and in Group B was 12.25±6.64. By using student’s unpaired t-test showing a significant difference in both groups.(t=2.32,P= 0.023) but Group A shows a mean increase in range of motion as compared to Group B. Hence Group A is showed a more significant result in knee flexion as compared to Group B. Table 5 shows the comparison of the functional disability scale on the WOMAC scale score in both groups. Mean of functional disability scale on WOMAC scale score Group A was 13.77±3.61and in Group B was 12.19±3.46. By using students unpaired t-test showing a significant decrease in functional disability scale on WOMAC scale (t=1.89, P=0.062) but Group A shows a significant decrease in functional disability than Group B. Hence, Group A shows significant improvement than Group B. DISCUSSION The current research was conducted to determine whether Maitland Mobilization and Conventional treatment are efficient in knee osteoarthritis subject in the Department of Musculoskeletal Physiotherapy.20,21 The outcome of this research shows that Maitland Mobilization with conventional therapy is more efficient in knee osteoarthritis. The mean data values from this research indicate that group A treated with Maitland Mobilization showed better improvement in Visual Analog Scale Pain relief, increased range of motion, WOMAC scale Physical functional ability.22,23 Table 1. showed age-wise distribution of topics in the current research. No important difference is discovered in the age of participants in both the group by using the chi-square test. The same number of subjects were affected by both the group and the maximum age group was 50-59 years in the study. Mathur et al.15 revealed that 15-40% of people over 40 years of age affected by degenerative joint disease, so it can be predicted that the most prevalent age group with the degenerative joint disease between 40 and 60 years of age. Table 2 illustrates the wise gender distribution of subjects. By using the chi-square test, there is no significant difference between the two groups. In this research, Zakir et al.24 found that both male and female OA knee subjects but observed that the percentage of male subjects (40.7 per cent) was small than female subject (59.3percent).Because of multiple factors,owing to its greater prevalence in the female population, this study had a big amount offemalesubjects (59.3).the female bony structures around the knee are different from the malecounter parts. Analysis of pain relief was ended by subjective visual analogue scale (VAS) by statistical mean. When Intragroup comparison was done there is a significant difference in both groups i.e. Group A and Group B. When intergroup comparison was done within both groups there was a significant difference, where group A showed significant improvement in terms of VAS.25,26 Mathur et al.15 studied the effectiveness of Maitland mobilizationin decreasing the knee joint pain.In earlier research,pain-decrease following passive joint mobilization has been already been stabilized.Mobilization may trigger local physiological processes and may also involve extra main procedures. These key mechanism could include activating the local inhibitory pathway in the spinal cord or lowering the inhibitory pathway from the brainstem. Analysis of knee flexion was completed by subjective Goniometer by statistical mean. When Intragroup comparison was performed there is a significant difference in both groups i.e. Group A and Group B. When intergroup comparison was done within both groups there was a significant difference, where group A showed significant improvement in terms of the range of motion for knee flexion. Analysis of Functional Disability was conducted by subjective WOMAC scale by statistical mean. When Intragroup comparison was done there is a significant difference in both groups i.e.groupAandgroupB.Whenintergroupcomparisonwasdone within both groups there was a significant difference, where group A showed significant improvement in terms of the WOMAC scale. Syed et al. 201427 showed a 56 per cent increase in complete WOMAC results in disability after administering manual physical therapy and knee osteoarthritis. However, in this study, the myofascial mobilization group and Conventional treatment group respectively noted 56.35% and 52% improvement in WOMAC scores and reported the effectiveness of Myofascial mobilization at the knee joint in reducing knee OA pain and disability.Dueto mechanical power during mobilization, including breaking Adhesions, Realigning collagen, or growing fibre glide, the efficacy of two therapy protocols can be reasonable when a particular motion stresses the particular component of the capsule. Ahmad et al.5 researched that manual techniques and exercises generated an average 56 per cent rise in self-reporting functional capacity of 54 per cent, stiffness of 54 per cent, and pain of 60 per cent as measured by the (WOMAC) scales. Knee osteoarthritis presents a serious health issue and a huge burden on society. Simple, safe, physical treatment procedures like Maitland Mobilization and conventional exercises could be of great value. This provides low-cost, easy means of treatment in subjects with knee osteoarthritis. CONCLUSION In conclusion, the present study provided evidence to support the use of physical therapy regimen in the form of Maitl and Mobilization with Conventional therapy is more effective than conventional treatment alone in relieving pain, improving range of motion and functional well-being in subjects with knee osteoarthritis. Conflict of Interest: Nil Source of Funding: Nil   Englishhttp://ijcrr.com/abstract.php?article_id=3628http://ijcrr.com/article_html.php?did=3628 Xu Q, Pang J, Zheng Y, Zhan H, Cao Y, Ding C. The effectiveness of manual therapy for relieving pain, stiffness and dysfunction in knee osteoarthritis: A systematic review and meta-analysis. Osteoarthr Cartil 2015;23:A387. Mora JC, Przkora R, Cruz-Almeida Y. Knee osteoarthritis: pathophysiology and current treatment modalities. J Pain Res 2018;11:2189–96. Kiran A, Ijaz M, Qamar M, Basharat A, Rasul A, Ahmed W. Comparison of the efficacy of mulligan’s mobilization with movement with Maitland mobilization along with conventional therapy in the patients with knee osteoarthritis: A randomized clinical trial. Libyan Int Med Univ J 2018;3(1):26. Rangey PS, Sheth MS, Vyas NJ. Comparison of Immediate Effect of Two Different Maitland Mobilization Protocols on Pain and Range Of Motion in Subjects with Osteoarthritis of Knee. Int J Med Health Res 2019; 3:124-127. Ahmad A. A Comparative Study between Joint Mobilization and Conventional Physiotherapy in Knee Osteoarthritis. Int J Physiother 2016;3(2):542-546. Hafez AR, Mohammed A, Kachanathu SJ, Alroumi M, Mohamed ES. Knee Osteoarthritis: A Review of Literature. Int J Physiother 2018;5(2):81-84. Arshad S, Rizvi SHA, Nisar M. To Evaluate the Effectiveness of Maitland Technique in Treatment of Knee Osteoarthritis in Female Patients. Int J Health Sci Res 2017;5(2):30. BD Chaurasia’s Human Anatomy, Volume 2 – Lower Limb, Abdomen and Pelvis, 6th Edition.pdf. Levangie PK, Norkin CC. Joint structure and function: a comprehensive analysis. 4th ed. Philadelphia. 2005;4:588. Kisner C, Colby LA. Therapeutic exercise: foundations and techniques. 5th ed. Philadelphia.2007;2: 928. Bennett R. Myofascial pain syndromes and their evaluation. Best Pract Res Clin Rheum 2007 Jun;21(3):427–445. Rickards LD. The effectiveness of non-invasive treatments for active myofascial trigger point pain: A systematic review of the literature. Int J Osteopath Med 2006;9(4):120–136. Mathur DMK. Comparison Between Passive Joint Mobilization And Manual Therapy Knee Protocol On Pain, Function And Quality Of Life In Patient With Chronic Osteoarthritis of Knee. Int J Osteopath Med 2018;8(2):58-62. Shukla M,  Goyal M, Effect of maitland mobilization and conventional physiotherapy exercises in osteoarthritis knee: a comparative study. Int J Physiother Res 2018;6(6):2952–2958. Jung J, Choi W, Lee Y, Kim J, Kim H, Lee K, et al. Immediate effect of self-myofascial release on hamstring flexibility. Phys Ther Rehabil Sci 2017;6(1):45–51. Vaishnavi KS, Rajeeva A. A comparative study of Maitland&#39;s mobilization along with ultrasound versus proprioceptive exercises along with ultrasound in stage ii and iii osteoarthritides of the knee joint. Int J Physical Edu.2017; 4(6):211-215. Muhammad N, Noreen A. GRADE 1-2 Osteoarthritis Of Knee Joint; Outcome Of Combination Of Grade 1-2 Knee Joint Mobilization With Quadriceps Isometrics In Patients.  Profess Med J 2017;24(7):986-991. Ganesh B. Short term effects of instrument-assisted soft tissue mobilization on pain and activities of daily living in subjects with patellofemoral joint osteoarthritis – A randomized controlled trial. Int J Curr Res Med Sci. 2017 Nov 30;4(11):55–63. Shah JP, Thaker N, Heimur J, Aredo JV, Sikdar S, Gerber L. Myofascial Trigger Points Then and Now: A Historical and Scientific Perspective. Int J Curr Res Med Sci 2015;7(7):746–761. Zakir A. Effectiveness of Manual Therapy Versus Exercise Therapy for the Management of Knee Osteoarthritis in Karachi Pakistan. Int J Physiotherapy 2016;3(1):57-62. Hhabar, P. Sathya HKC. Effect of Conventional Exercises with Balance Training & amp; Only Conventional Exercises in Patients with Osteoarthritis of Knee. Int J Innov Res Sci Eng Tech 2015;04(07):5048–5056. Ebtessam Fawzy G, Lilian Albert Z. Effect of iliotibial band myofascial release on flexibility and patellar alignment in patients with knee osteoarthritis. Int J Adv Res 2015;3(4): 399-410. Syed S, Wani S. Effect of Two Different Manual Therapy Protocols On Osteoarthritic Knee Pain &amp; Functional Disability: A Comparative Study. Int J Adv Res 2014;20(34):10. Jansen MJ, Viechtbauer W, Lenssen AF, Hendriks EJM, de Bie RA. Strength training alone, exercise therapy alone, and exercise therapy with passive manual mobilisation each reduce pain and disability in people with knee osteoarthritis: a systematic review. J Physiotherapy. 2011;57(1):11–20. Swati K, Subhash K. Effectiveness between supervised clinical exercise with Maitland manual therapy and home exercise program in treating osteoarthritis of the knee: a comparative study. Ind J Basic and App Med. 2013;3(1): 105-112. Harish S, Kashif R. Effect of Maitland Mobilization and Myofascial Release Technique in Patients with Knee Osteoarthritis. Indian J Physiother Occup Ther Int J 2013;7(4):181. Maher S, Creighton D, Kondratek M, Krauss J, Qu X. The effect of tibio-femoral traction mobilization on passive knee flexion motion impairment and pain: a case series. J Man Manip Ther 2010 Mar;18(1):29–36.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareA Study to Assess the Effectiveness of Near Peer Learning on Knowledge and Habit Making on Prevention of Water-Borne Diseases Among Primary School Children Near Peer Learning Among School Children – The Protocol English138141Rupa VermaEnglish Archana MauryaEnglishEnglish Near peer learning, Waterborne diseases, Hand washing, Personal hygiene.INTRODUCTION Learning is a key process in one’s behaviour. Our behaviour and actions are influenced by what we learn and how we learn it. We learn when we come in contact with the environment. We learn with our day to day direct or indirect experiences. For example, learning to read, write, sing, play etc. learning situations are most natural and common in our life. In most simple words, changes that are brought about by experience are known as learning.1 Near peer learning is an educational practice in which students interact with other students to attain educational objectives. It is becoming increasingly popular as a learning methodology in schools and colleges. It involves students exposed to similar circumstances, from the same educational programme, but at different levels.2,3 Near peer learning has long been a popular approach in providing mentoring to children and youth, it also comes with the additional impact of providing leadership and development opportunities to the older or near-peer youth who serve in the mentoring role.4 In simpler terms students involved in this type of learning are Seniors and juniors of the same school or college. Near Peer learning can be considered as a mode of learning for everyone, by everyone, and about any selected topic.  Near-peer learning can take place in a formal or informal teaching-learning environment. It can take place in small groups or even online. Learners are more comfortable and relaxed, and ready to learn when the teacher is their peer, whom they know and understand. At the same time, it develops communication skill, leadership skills and confidence in public dealings in the teacher peer.  In this study Near peer learning means a senior student is teaching regarding prevention of waterborne diseases to junior students. A study to assess the effectiveness of Near Peer learning on knowledge and habit making on prevention of water-borne diseases among primary school children. Operational definition Assess According to the oxford dictionary, Assess means Evaluate or Estimate the nature, ability, or quality of something. In this study, it refers to deciding the worth of child to child health education program by the significant difference in pre and post-test scores of children. Effectiveness According to the oxford dictionary, Effectiveness means the degree to which something is successful in producing the desired result. This study, it indicates the gain in knowledge as determined by the significant difference in pre and post-test scores of the children. Near Peer learning IT is an instructional method in which senior students temporarily assume the role of coach or instructor. It has been referred to as tiered or pyramidal or hierarchical learning. Senior learners solidify learning & develop additional skills by teaching peers. In this study, it refers to the elder child (13-14 years) teaching  younger children ( 9-12 years)  regarding prevention of selected water-borne diseases with the help of a pictorial booklet, songs, stories and demonstrations Prevention According to the oxford dictionary prevention means the action of stopping something from happening. Preventive aspects like hand washing and personal hygiene in the prevention of waterborne diseases will be part of this study. Waterborne diseases According to the oxford dictionary, water-borne diseases means diseases that are Conveyed by, travelling on, or involving travel or transport on water. In this study water-borne diseases refers to diarrhoea, and worm infestation. Knowledge According to the oxford dictionary, knowledge means facts, information, and skills acquired through experience or education. This study refers to the correct responses of the children to the questions related to waterborne diseases. Habit making According to Wikipedia, A Habit is a routine of behaviour that is repeated regularly and tends to occur subconsciously. New behaviours can become automatic through the process of habit making or habit formation. In this study, habit making refers to the initiation of habit formation related to practices of handwashing and personal hygiene. Primary school Children According to Wikipedia, the primary school offers primary education which is typically the first stage of compulsory education. The children of age group 6-15 years generally enrolled in these schools. This study refers to children of age group 9-12 years studying in Primary school in rural areas of Wardha district. OBJECTIVES OF THE STUDY To assess the existing knowledge regarding the prevention of water-borne diseases among primary school children. To assess the existing practices regarding the prevention of water-borne diseases among primary school children. To assess the effectiveness of Near-peer learning on knowledge regarding the prevention of water-borne diseases among primary school children. To assess the effectiveness of near-peer learning on practices related to the prevention of water-borne diseases, among primary school children. To find out the association between pre-test knowledge scores of children regarding the prevention of water-borne diseases with selected demographic variables among primary school children. To find out the association between pre-test practice scores of children regarding the prevention of water-borne diseases and selected demographic variables among primary school children. HYPOTHESIS H1- There is a significant difference in pre-test and post-test knowledge scores of children receiving Near-Peer learning. H0 – There is no significant difference in pretest and post-test scores of children receiving Near-Peer learning. H2- There is a significant difference in pretest and post-test practice scores of children receiving Near-Peer learning. H01 – There is no significant difference between pre-test and post-test practice scores of children receiving Near-Peer learning. CONCEPTUAL FRAMEWORK Conceptual framework, conceptual models or conceptual scheme ( We use the terms interchangeably here ) represent a less formal attempt at organizing phenomena than theories, As the name implies, conceptual framework deals with abstract concepts that are assembled by their relevance to a common theme. It refers to concepts that offer a framework of proposition for conducting research. The conceptual framework set up for the study is modified stuffle beam’s evaluation model of the planned programme. Stuffle beam’s “ CIPP model” prescribes four types of evaluation, context, Input, process, and product. It provides a comprehensive, systematic and continuously ongoing framework for programme evaluation.  The model is adopted in a modified form for the present study.  According to model content identifies discrepancies between intended and actual programme outcome and evaluators can develop a causal explanation for the discrepancies. Step I: context evaluation, step II: input evaluation, Step III: process evaluation, step IV Product evaluation. The core value for the present study is enhancing knowledge regarding the prevention of waterborne diseases among primary school children. MATERIALS AND METHODS Research approach- Evaluative research approach will be used in this study Research Design - one group pretest-posttest design will be used in this study. Near Peer learning: It will be composed of two phases. Phase 1 – (selection and training of elder children who will act as inducing agents in the study) Selection of 13-14 years children via purposive sampling, who are attending the school in a rural community, according to their academic abilities, group activities, and communication as reported by their teacher. Prevention of selected water-borne diseases will be taught to them by the researcher, (with the help of pictorial booklet, stories, songs and demonstration). The teaching will be repeated till they learn properly. (I.e. till they score adequate knowledge and practice scores). Inducing agents assessed by making them do rehearsals to the investigator and the same questionnaire was given to them to determine their adequate level of knowledge on the next day. If the knowledge and demonstration score were moderate or inadequate, inducing agents encouraged to do the rehearsals again until they score adequately. Once they score adequate, they were ready to teach younger children. One selected topic at a time was taught to inducing agents, after which they will teach the same to younger children. Phase 2 (trained elder children taught younger children, pretest and post-test done by the researcher) O1 X O2 O1— pretest regarding knowledge and practice of prevention of selected water-borne diseases among primary school children. (One topic at a time) X- Near – peer learning. (As per plan given below) (Elder child will teach younger children, with the help of charts, flashcards, songs and demonstrations.) One elder student will teach 10 younger students about one selected topic at a time for 30 minutes a day for 3 alternate days a week. O2 – post-test to assess the effectiveness of Near-peer learning. It will be done on the 7th day after the completion of the teaching of one selected topic DETAILED METHODOLOGY First step -Training of inducing agents (elder children) regarding prevention of Diarrhoea. Second step- These inducing agents will teach primary school children regarding the prevention of diarrhoea. Third step- Training of inducing agents (elder children) regarding prevention of worm Infestations. Fourth step- These inducing agents will teach primary school children regarding the prevention of worm Infestations. Fifth step- Training of inducing agents (elder children) regarding personal hygiene and Handwashing technique. Sixth step- Inducing agents will teach primary school children regarding personal hygiene and Handwashing technique. The pre-test is done before starting the teaching of primary school children regarding each selected topic and Post-test done on the 7th day after the completion of the teaching of the selected topic. Observation of study participants done 6 times at an interval of 15 days to evaluate whether they are practising correct hand washing technique and maintaining personal hygiene. Sample Students of age group 9-12 years, studying in primary school in rural areas of Wardha district. Sample selection criteria Inclusion Criteria Both male and female students of the age group 9-12 years at selected school. Exclusion Criteria Students who were absent on the day of data collection. Students who attended a similar type of health education program. Sampling technique- non-probability convenience sampling technique used to select samples. Sample size- 400 Data collection tool Structured questionnaire to assess demographic variables of samples. A structured questionnaire will be used to assess knowledge and practices related to the prevention of childhood diseases i.e. Diarrhea, and worm infestation. The structured observational checklist will be used to assess the practice of Handwashing and Personal Hygiene. Concealed observation technique with an observational checklist as a tool will be used to do follow up observation. Protection of human subjects The written permission will be obtained from the Principal of selected schools. Informed consent will be obtained from the Parents of school children, regarding their ward’s participation in this study. RESULT AND DISCUSSION In community paediatrics, Near Peer learning and teaching is an approach towards disease prevention and health promotion. Health-promoting change in the behaviour of children can be brought through this initiative. Children can be the change agent for bringing positive behavioural changes among their peers. Peer education is the teaching or sharing of health information, Values or behaviour in educating others who may share similar social background or experiences.  This type of teaching and learning is cost-effective and feasible in community settings.  It is believed that peers are in the best position to encourage healthy behaviour in each other, as they have a larger influence on each other. The proximity of age and recent similar experiences of peer tutors provides an added benefit as near-peer teachers have a better appreciation of the knowledge held by junior peers and can therefore target teaching at an appropriate level. This paper reflected on the design, implementation, and evaluation requirements of near-peer learning at schools to create awareness regarding healthy lifestyle and behaviour to prevent waterborne diseases. Participation in the near-peer learning programme will help students to recognise and develop their future health behaviours.5 The skill of teaching is best acquired through a sequence of training, practice and feedback.6 And so in this study special attention is given to teaching elder students by teaching them regarding prevention of waterborne diseases and then by evaluating their knowledge and practice before they will be allowed to teach the younger children. CONCLUSION Near peer learning is helpful for students to learn effectively. Health education can also be imparted using a near-peer learning strategy in all settings, and especially among school students. At a time when health resources are stretched and demands upon health personnel are increasing, it offers students the opportunity to learn from each other. It gives them considerably more practice than traditional teaching and learning methods in taking responsibility for their health.3 Developing near peer learning related to health awareness in the classrooms, neighbourhood, families and community can be instrumental in the attainment of optimum health for all. This study emphasizes utilizing elder children to impart health education regarding the prevention of waterborne diseases to younger children. Apart from increasing knowledge of children regarding the prevention of waterborne diseases, the study also aims to improve the health practices of children, specifically practices related to handwashing and personal hygiene. Conflict of Interest: None Source of Funding: None Englishhttp://ijcrr.com/abstract.php?article_id=3629http://ijcrr.com/article_html.php?did=3629 Aarti J, Communication and education technology.1st ed., lotus publisher, Jalandhar, 2019; 93-96. Lisa M, Bret W, the hidden curriculum in Near peer learning. Nur Educ Tod 2017;50:77-81. Boud D. what is peer learning and why it is important? Tomorrow&#39;s Teaching and Learning, 2002. What is Peer Learning and Why is it Important? | Tomorrow&#39;s Professor Postings (stanford.edu) Willis P, Bland R, Make L. The ABC of peer mentoring, educational research and evaluation. Int J Theo Pract 2012;18(2):231.   Annette B, Christie VD. Planning pear assisted learning in clinical schools. BMC Med Educ 2020;20(2):23-27. Marton GE, McCullough B, Ramnanan CJ, A review of teaching skills development programmes for medical students. Med Educ 2014;49:149–160.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareAnti-inflammatory Activity of Aerial and Root Extracts of Withania somnifera: A Comparative Experimental Evidence English142148Satyajyoti KanjilalEnglish Ashok Kumar GuptaEnglish Ranjana Saksena PatnaikEnglish Amitabha DeyEnglishEnglishAshwagandha, Inflammation, Interleukin, TNF alpha, Withania somniferaIntroduction Cytokines are small-secreted proteins referred inconsistently as interleukins, growth factors or chemokines. Physiologically, they activate the inflammatory mechanisms and help repair damaged tissue. During the inflammatory process, a sudden release of cytokines (cytokine storm) occurs via a cascade of activation process.1,2 The uncontrolled expression of these macrophage-derived cytokines (inflammatory mediators) tumour necrosis factor (TNF), interleukins (IL-1, IL-6, IL-8), colony-stimulating factors (CSFs) and growth factors govern the pro-inflammatory signalling pathways in the pathogenesis of rheumatoid arthritis (RA).2,3 The activated macrophages produce TNF-α in the inflamed synovial membrane tissue. This is having the capacity of induction and production of other pro-inflammatory cytokines, including Interleukin-1 beta (IL-1β) and IL-6. Combined facilitation of signalling pathways and cytokines, in turn, release chemokines that attract leukocytes to the inflamed site from the blood. The induction of proteolytic and metalloproteinase enzymes further results in the destruction of the underlying articular cartilage and bone tissue.4-10 The mainstay of the treatment of RA is to control these pro-inflammatory mediators. At therapeutics of Disease-Modifying Antirheumatic Drugs (DMARDs) such as Sulfasalazine, Methotrexate, Leflunomide, Nonsteroidal Anti-Inflammatory Drugs (NSAIDs) and biologics such as Infliximab, Adalimumab (TNF –blocker), Anakinra (IL-1 blocker), Tocilizumab (IL-6 blocker) are been looked upon for the management.4,11 The drug regimen offers efficient management, however, the chance of suboptimal control, the inability to complete remission and emerging adverse effects in long-term usage often lead to a search for more alternatives.12-16 The need-gap analysis and increasing popularity of plant or herb-based medicine often push for innovations and research which lead to the discovery of many new phytomedicines or enriched extracts or drugs.17 Many plants and formulations are reported to have anti-inflammatory and analgesic properties.18-20 Withania somnifera Dunal (Ashwagandha) or Indian ginseng has been recognized as one of the medicinally important herbs having anti-inflammatory and Rasayana (adaptogen and rejuvenator) properties. Among the other phytoconstituents, the biologically active steroidal lactones Withaferin A and Withanolides were reported to have anti-inflammatory and immunomodulatory properties contributing to their use in painful arthritic conditions.21 Roots are researched for their potent inhibitory effect on inflammatory markers and used in traditional system of medicine.21-23 The aerial part especially the leaves of the plant are also studied for its role as anticancer, antimicrobial, diabetes etc. However, a comparative study on both parts especially the commercially viable extracts are not been studied in the anti-inflammatory perspectives. Therefore, the present study was conducted to evaluate the comparative anti-inflammatory activity of standardized methanolic and hydro-methanolic extract of both the roots and the aerial part of the plant in in-vitro models of TNF-α, Interleukin-1 beta (IL-1β) and IL-6. Materials and Methods Plant Material Withania somnifera raw material of aerial part Lot No.:ASHW/PLPLR30/NOV19 and Aswagandha aerial part hydroalcoholic extract 5%, Lot No: ASHW/PLPL30/NOV were procured from Phyto Life sciences Pvt Ltd, Ahmedabad, Gujarat. The root powder was obtained from Emami Ltd, Research and Development Centre, Kolkata. Both the raw material of aerial and root were further used to get the methanolic and hydroalcoholic extracts in the phytochemistry laboratory of Emami Research and Development Centre. Chemical and Reagents, Kits, Drugs and Cell line Reagents and Kits: The TNF alpha Human ELISA Kit (Abcam; Cat. No. ab100654), ab46052 IL-1 beta Human ELISA Kit and ab46042 IL-6 (Interleukin-6) High Sensitivity Human ELISA Kit of Abcam. Drugs: The Anti-arthritic drugs were purchased from the local pharmacist - Methotrexate 25 mg (Folitrax) Batch No. At 071110, Sulfasalazine (Saaz) Batch No. ECL079007AS marketed by IPCA Laboratories Ltd, Mumbai and Diclofenac (Voveran 50) Batch No. 195009MB of Novartis India Ltd were also used. Cell culture mediums and Solutions: Dulbecco&#39;s Modified Eagle&#39;s medium (DMEM), Fetal Bovine Serum (FBS) & 1% Antibiotic/anti-mitotic solution. MTT (3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) reagent (HiMedia). Dulbecco&#39;s phosphate-buffered saline (DPBS - HiMedia). Dimethyl sulfoxide (DMSO). All the chemical and drugs are procured from Local vendor. Cell line: MG 62 and Hep was procured from National Centre for Cell Science, Pune, India. Preparation of W somnifera extracts Alcoholic and hydroalcoholic extracts of roots and leaf part of Withania somnifera were evaporated using a rotary vacuum evaporator, whereas aqueous extracts were dried by lyophilization. HPTLC (high-performance thin-layer chromatography) qualitative studies showed that the aqueous-methanolic extracts of both plant parts contain higher amounts of withanolides than all other extracts concerning withaferin A and withanolide A. Accurately weighed 1.0 g of leaf and root powder was soaked separately in 50 ml conical flask with 20 ml of 50% aq-methanol. Both the samples were sonicated at 60 C for 30 minutes and samples were filtered.  For preparing the reference standard solution, 10.0 mg of withaferin A and withanolide A were dissolved with methanol in 5 ml volumetric flasks which were further made 10-fold dilution. Silica gel aluminium sheet plate 60 F254 (Germany) was used to perform the chromatographic estimation. Linear ascending development was carried out in a twin-trough glass chamber (CAMAG) equilibrated with the mobile phase. The length of the chromatogram run was 6.0 cm. Concentrations of the compound were determined from the intensity of absorbance. The evaluation was made via peak areas versus withanolides amount in linear regression. The withanolides present in the extracts were identified by comparing the HPTLC spectrum of the standards Cell Line and Culture Condition The monolayer of the cells was maintained at sub-confluent conditions in growth media containing DMEM with 0.045 g/ml glucose, 1mM sodium pyruvate, L-glutamine, 1.5 g/L sodium bicarbonate, 100 U/ml penicillin, 100 μg/ml streptomycin, and 10% fetal bovine serum (FBS). Cells were maintained in a humidified incubator with ambient oxygen and 5% CO2 at 37°C. Cells from passages 3–15 were used in the experiment, and cells were not allowed to grow to more than 60%–70% confluence. Prepare the cell culture 100 μl of differentiated cell suspension (1 × 106 cells/ml) were seeded in a 96-well culture plate with 100 μl of DMEM – differentiated cell culture medium. The cells were incubated in an FBS-free medium for 4 to 18 hr before the examination. Effect of test samples on cell viability and proliferation: MTT assay24 Measurement of cell viability and proliferation forms the basis for numerous in vitro assays of a cell population’s response to external factors. The yellow tetrazolium MTT is reduced by metabolically active cells, in part by the action of dehydrogenase enzymes, to generate reducing equivalents such as NADH (nicotinamide adenine dinucleotide hydrogen) and NADPH (nicotinamide adenine dinucleotide phosphate hydrogen). This reduction takes place only when mitochondrial reductase enzymes are active, and therefore conversion can be directly related to the number of viable (living) cells.  Sample extracts were prepared in the concentration of 100 to 1000 µg/ml. MTT reagent was prepared aseptically by adding 6 ml of cell-based assay buffer (DPBS) in a vial containing 30 mg of MTT reagent and dissolved completely by vigorous vortexing. Assay Procedure The wells for control, samples and blank were designated in 96 well plates. The culture medium from each well was removed and washed the cells twice with DPBS. The test samples were added to the 96 well plate containing cells and incubated in a 5% CO2 incubator at 37°C for 18-20 hrs. (The volume of the test sample solution is 100 μl). After the incubation period, the plates from the incubator were removed and 10 µl of MTT reagent (5 mg/ml) were added to all the wells except blank wells. The culture plates were wrapped with aluminium foil to avoid exposure to light.  The plates were returned to the CO2 incubator and incubated for 3 to 4 hours. The culture medium was carefully removed from each well after the process of incubation was over and 100 µl of DMSO were added to each well to solubilize the Formazan crystals.  Absorbance was noted on Multimode Microplate reader: SpectraMax i3X [Molecular Devices] at 570nm. The average 570 nm absorbance values of the control wells were subtracted from the average 570 nm absorbance values of corresponding experimental wells. For optimization of the process, the cell suspension was serially diluted from 1 × 106 to 1 × 103 cells/ml using the appropriate culture medium. 100 µl of each dilution were seeded in a 96-well plate and a curve of absorbance against cell density was plotted. The optimal number of viable cells after treatment with samples are measured from the linear graph. Estimation of Cytokines – TNF-α TNF alpha Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human TNF alpha in serum, plasma and cell culture supernatants. This assay employs an antibody specific for Human TNF alpha coated on a 96-well plate. All reagents, samples and standards are prepared as per the instructions on the manual of the kit. Standards and samples 100 μL each are pipetted into the wells and TNF alpha present in a sample is bound to the wells by the immobilized antibody. At each well, the following sequence is followed with incubation at room temperature in between after each addition. The wells are washed and 100 μL of 1X Biotinylated TNF alpha Detection Antibody is added. After washing away the unbound biotinylated antibody, 100 μL of 1X HRP-Streptavidin solution is pipetted to the wells. The wells are again washed, and 100 μL of TMB One-Step Substrate Reagent is added to the wells and colour develops in proportion to the amount of TNF alpha bound. 50 μL of Stop Solution is added to each well which changes the colour from blue to yellow, and the intensity of the colour is measured at 450 nm on Multimode Microplate reader: SpectraMax i3X [Molecular Devices, USA]. The calculations were performed as per the Kit protocol. Estimation of Cytokines - IL-1β The ELISA (Enzyme-linked Immunosorbent Assay) kit for estimation of human IL 1β is intended to quantify the enzyme in blood components or culture medium. For experimentation, samples 100µL each is pipetted into the specific antibody-coated wells of microtiter strip microplates. To begin the experimental process, samples, as well as standards and 50 µL of 1X Biotinylated monoclonal anti-IL-1 beta antibody, are simultaneously incubated. After washing, the enzyme 100 μL of 1X Streptavidin- HRP, that binds the biotinylated antibody is added, incubated and washed. A 100 μL of Chromogen TMB substrate solution is added which acts on the bound enzyme to induce a coloured reaction product. Direct exposure to light is avoided by wrapping the plate in aluminium foil. 100 μL of Stop Reagent is added into each well and results are taken immediately after the addition of Stop Reagent, or within one hour, if the microplate is stored at 2-8°C in the dark. The absorbance of each well is read on a spectrophotometer (SpectraMax i3X) using 450 nm as the primary wavelength and optionally 620 nm (610 nm to 650 nm is acceptable) as the reference wavelength. The more is the concentration of the interleukin beta in the sample, the more is the strength of the coloured product. The standardized protocol mentioned in the kit is followed to calculate during experimentation and analysis. Estimation of Cytokines - IL-6 ELISA kit for Interleukin-6 in-vitro testing is intended to quantify the enzyme in different blood components, body fluids and experimental solutions. The assay method identifies natural as well as recombinant human IL-6. The coating of interleukin-6 monoclonal antibody was done before the experiment in the specified number of microtiter strips well-plates. Each well was filled with standards and control samples in the quantity of 100 μL respectively. The experiment was started with the incubation of 50 μL of 1 X Biotinylated monoclonal anti-IL-6 antibody along with either samples or standard. After washing, 100 μL of 1X Streptavidin-HRP solution, that binds the biotinylated antibody is added into all wells, incubated at room temperature for 30 minutes and washed. 100 μL of Chromogen TMB (3,3&#39;,5,5&#39;-tetramethylbenzidine) substrate solution is added which acts on the bound enzyme to induce a coloured reaction product and incubated in the dark for 12-15 minutes at room temperature. The intensity of this coloured product is directly proportional to the concentration of IL-6 present in the samples 100 μL of Stop Reagent is added into each well and results are taken immediately after the addition of Stop Reagent, The absorbance of each well was noted on a spectrophotometer (SpectraMax i3X ) using 450 nm as the primary wavelength and optionally 620 nm (610 nm to 650 nm is acceptable) as the reference wavelength. The calculations were performed as per the Kit protocol. Statistical Analysis Mean ± standard error of the mean (SEM) were calculated from the observed individual values Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Student-Newman-Keuls multiple comparison test. GraphPad Prism- 5 (GraphPad Software Inc., La Jolla, California, USA) was used for statistical analysis. P-values less than 0.05 were considered statistically significant. Results and Discussion The phytochemical analysis of W. somnifera has revealed several groups of bioactive compounds such as flavonoids, tannin, alkaloids, sitoindosides, glycosides, withanicil, steroidal lactones, and alkaloids.25,26 The most therapeutically important chemicals are withanolides especially withaferin A and withanolide D for its antiarthritic effects.27,28 Compounds in sample chromatograms were determined from the intensity of absorbance at 232 nm and identified invalidate High-performance thin-layer chromatography (HPTLC) analysis, which is an efficient tool for quantitative analysis of compounds and yields superior separation efficiency. The withanolides present in the extracts were identified by comparing the HPTLC spectrum of the standards. The quantities of withanolides were present more in 50% aq-methanolic extracts of the root part of Ashwagandha. The inflammatory mediator Tumour Necrosis Factor-alpha (TNF-α) may lead to the release of other inflammatory cytokines such as IL-1β and IL-629, which promotes synovitis causing cartilage destruction and bone erosion in RA.30,31 W. somnifera is a herb known to have multiple benefits to keep the body healthy. The roots of the plant have known usage for having anti-inflammatory properties.  Both the Leaf and root extracts of W. somnifera has been studied for analysis and standardization of Phyto-constituents and in efficacy models.32-34 The present study gave a preliminary research lead on the most effective part extract in inflammations in-vitro cytokine models. Many researchers in experimental (in-vitro and in-vivo) anti-inflammatory models have studied the leaf part. W. somnifera leaf aqueous extract and one of its active chloroform fractions studied in microglial cell lines was found to inhibit the microglial activation and migration by attenuating the pro-inflammatory markers of TNF-α, IL-1β, IL-6 and others suggesting its role in suppression of neuroinflammation.35 New withanolide glycosides and withanolide isolated from the leaves were studied for their inhibition of cycloxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) enzymes and lipid peroxidation. The majority of compounds showed selective COX-2 enzyme inhibition suggesting their anti-inflammatory activity.36 The hydro-methanolic extract in stainless steel implant induced inflammation in zebrafish in-vivo model has shown an inhibitory effect of TNF α.37 The root part of W. somnifera finds its usage more in experimental models as well as in clinical practice. Hydroalcoholic root extract showed anti-inflammatory effect by inhibition of protein (albumin) denaturation in the in-vitro model.38 Ethanolic extract was effective in acute and chronic in-vivo carrageenan-induced anti-inflammatory model.39 Aqueous extracts in gel dosage form has shown anti-inflammatory activity in trinitrobenzene sulfonic acid-induced in-vivo models for inflammatory bowel disease.40 Root powder41 and aqueous extract 42 decreased the arthritis effects in collagen-induced arthritis in-vivo model suggestive its anti-inflammatory activity. Attenuation of the pro-inflammatory markers (TNF-α, IL-1β, IL-6 and IL-10) were observed in the study by khan et.al. Root powder showed inhibitory effect on pro-inflammatory markers, proteinuria, nephritis in pristane-induced Lupus model.43 Root power exhibited anti-inflammatory activity by decreasing the pro-inflammatory markers TNF-α and IL-6 in fructose-fed rats44 and showed potent analgesic and antipyretic effect by retarding amplification and propagation of the inflammatory response in monosodium urate crystal-induced (Gout) model.45 Root powder had potent inhibitory activity towards the complement system, mitogen-induced lymphocyte proliferation and delayed-type hypersensitivity reaction suggestive usefulness in immunosuppression for the inflammatory diseases.46 Aqueous root extract showed anti?inflammatory activity on human osteoarthritic cartilage47 and in the human keratinocyte cell line by inhibiting expression of inflammatory cytokines IL?8, IL?6, TNF?α, IL?1β and IL?12, and promoting the expression of the anti?inflammatory cytokine transforming growth factor (TGF)?β1.48 The MTT method of cell determination is useful in the measurement of cell growth in response to mitogens, antigenic stimuli, growth factors and other cell growth-promoting reagents, cytotoxicity studies, and in the derivation of cell growth curves (Sigma). In the present study, the MTT assay was performed in MG 63 and Hep G2 cell line. Methotrexate confirmed the authenticity of the study by showing a decreasing trend in cell viability. All the four extracts from the concentration of 100 µg/ml up to 1000 µg/ml have shown a considerable cell survival count (Cells/ml) contributing towards its safety profile (Figures 1 & 2). Levels of inflammatory cytokine, TNF-α were measured as a marker of inflammation. Both the methanolic and hydroalcoholic extracts from the aerial part was found to have more potential compared to that of roots in inhibiting the TNF –alpha cytokines (Figure 3).  Cytokine IL-1β is reported to play critical roles in the pathogenesis of RA. The inflammatory cytokine IL-6 was measured as a marker of inflammation. In the Interleukin 6 inhibition assay, the extracts of the aerial part were better compared to their respective roots extracts. Root Hydroalcoholic extract was better than aerial methanolic extract. However, among the four, the aerial hydro-alcoholic part is the most potential. In the interleukin beta inhibition assay, both root extracts were better than aerial extract (Figure 4 and 5). Conclusion Both parts of W. somnifera possess anti-inflammatory properties however, the aerial part has shown a promising activity vis-à-vis the roots. Therefore, this can be an alternative renewable resource as raw material for preparing medicines for arthritis. Moreover, both the methanolic and hydroalcoholic extracts can be considered for Phyto-pharmaceutical drug development. Acknowledgements: The authors would like to express their gratitude to the Research and Development centre of Emami Limited, Kolkata, India for providing support for this work. The authors acknowledge the immense help and the knowledge received from the scholars whose articles are cited and included in references to this manuscript. The authors are also grateful to authors/editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Conflict of interests: Authors declare no conflict of interests. Funding Source: Nil Author’s Contribution: SJK and AB have experimented and analysed the data. SJK has prepared the initial manuscript, AKG and RKP have reviewed and modified the manuscript. The manuscript was finalized by SJK and AB after the incorporation of the comments. AKG and RKP has approved the final manuscript. Englishhttp://ijcrr.com/abstract.php?article_id=3630http://ijcrr.com/article_html.php?did=3630 Kay S, Vollrath JT, Relja B. Cytokines in Inflammatory Disease. Int J Mol Sci 2019;20(23):6008. Duff GW. Cytokines and Acute Phase Proteins in Rheumatoid Arthritis. 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New Delhi, India: Department of Indian System of Medicine and Homeopathy, Ministry of Health and Family Welfare, Government of India;2003. Chunekar KC. Bhavprakash Nighantu (Indian Meteria Medica) of Sri Bhavamishra, Pandey GS (Ed). Varanasi: Chaukhamba Bharati Academy;2015. Freshney RI. Culture of animal cells: a manual of basic techniques. In: Cytotoxicity. 5th edition. New Jersey: Wiley-Liss;2005. p. 359-74. Mirjalili MH, Moyano E, Bonfill M, Cusido RM, Palazón J. Steroidal lactones from Withania somnifera, an ancient plant for novel medicine. Molecules 2009;14(7):2373-93. Kalra R, Kaushik N. Withania somnifera (Linn.) Dunal: a review of chemical and pharmacological diversity. Phytoche Rev 2017;16:953-987. Ganzera M, Choudhary MI, Khan IA. Quantitative HPLC analysis of withanolides in Withania somnifera. Fitoterapia 2003;74(1-2):68-76. Kumar V, Dey A,Hadimani MB, Markovi? T, Emerald M. Chemistry and pharmacology of Withania somnifera: An update. Humanitas Med 2015;5:1-13. Filippin LI Vercelino R, Marroni NP, Xavier RM. Redox signalling and the inflammatory response in rheumatoid arthritis. Clin Exp Immunol 2008 Jun;152(3):415-422. Hashizume M, Mihara M. The roles of interleukin-6 in the pathogenesis of rheumatoid arthritis. Arthritis 2011;2011:765624. Morel J, Berenbaum F. Signal transduction pathways: New targets for treating rheumatoid arthritis. Joint Bone Spine 2004;71(6):503-510. Chatterjee S, Srivastava S, Khalid A, Singh N, Sangwan RS, Sidhu OP, et al. Comprehensive metabolic fingerprinting of Withania somnifera leaf and root extracts. Phytochemistry 2010;71:1085–1094.  Chaurasiya ND, Uniyal GC, Lal P, Misra L, Sangwan NS, Tuli R, et al. Analysis of Withanolides in Root and Leaf of Withania somnifera by HPLC with Photodiode Array and Evaporative Light Scattering Detection. Phytochem Anal 2008;19(2):148–54. Udayakumar R, Kasthurirengan S, Mariashibu TS, Rajesh M, Anbazhagan VR, Kim SC, et al. 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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareFormulation and Evaluation of Tocopheryl Polyethylene Glycol (TPGS) Stabilised Nanoemulsion of Curcumin for Topical Application English149155Jasjeet Kaur NarangEnglish Anmoldeep KaurEnglish R.S. NarangEnglish Ravika NandaEnglish MehakEnglish Karanbir SinghEnglishEnglishCurcumin, Nanoemulsion, Histopathological emulsion, Topical applicationINTRODUCTION Curcumin, a poorly water-soluble phytoconstituent, is limited superficially to stratum corneum after its topical administration.1 Therefore, it necessitates the formulation of lipophilic carriers that release the drug in a controlled manner. Among the different lipophilic carriers investigated nanoemulsions have gained immense importance for the delivery of drugs. Reports in the literature suggest that nanoemulsions are a promising strategy for the delivery of drugs as they can penetrate through the subcutaneous barrier into the skin.2,3 Nanoemulsions are thermodynamically stable formulations made by using oils, surfactants, cosurfactants and water.4 As compared to the conventional formulations meant for topical administration nanoemulsions facilitate an increased amount of drug entrapment. In the present study, a nanoemulsion of curcumin for topical administration with enhanced skin retention has been formulated and evaluated. MATERIALS AND METHODS Chemicals and reagents Curcumin was purchased from Sigma Aldrich Corporation(St. Louis, USA), Sefsol 218 was obtained as a gift sample from Nikko Chemicals(Tokyo, Japan), Labrasol, Labrafac, Capryol 90 were obtained as gift samples from gattefosse (Saint-Priest, France). Tween 80 was purchased from Central Drug House, New Delhi, India. PEG was purchased from Spectrochem Pvt Ltd Mumbai, India. Water was obtained from the Milli-Q-water purification system (Millipore, MA). All other chemicals and reagents were of analytical grade and procured from Merck (Mumbai, India) and S.D. Fine Chem. (Mumbai, India). The Ethical Clarence has been taken from the Institutional Ethical cum Research board with vide letter no. SGR/2019-51- 20 Formulation Development and Optimization Solubility Studies Assessment of solubility of the drug was done to select the most appropriate oil, surfactant and co-surfactant for the nanoemulsion formulation containing curcumin. For determination of the solubility of curcumin the oils (Arachis Oil, Castor oil, Cinnamon oil, Clove oil, Corn oil, Olive oil, TPGS: Sefsol-218 Oil (1:1), Sunflower oil and Tea tree oil ), surfactants (Span 80, Tween 20, tween 60 and tween 80) and cosurfactants (Glycerol, Labrafac, Lauroglycol FCC, PEG 600 and Solutol HS 15) were taken in 5 ml stoppered vials and drug in excess quantities was put in the oils, surfactants and cosurfactants to produce supersaturated solutions and using a vortex mixer kept at 25±1°C in an incubator shaker mixed for 72h for the attainment of equilibrium, after which the samples were centrifuged for 15 min at 3000 rpm. The supernatant was collected after filtration through a 0.45µm membrane filter diluted with methanol if necessary and analysed by UV spectrophotometer at 420 nm. The experiment was done in triplicate. Miscibility studies were also performed to select the cosurfactants. For the conduct of the miscibility study, surfactant and co-surfactant were mixed in the ratio of 1:1 in 5ml vials with the help of a vortex mixer. These vials were then kept at 25±1°C in an incubator shaker for 72h for attaining equilibrium. After 72h the vials were kept overnight to ascertain the miscibility of the surfactants and co surfactants5. To confirm the miscibility, visual observations were done. The transparent or clear mixtures were considered for other studies. Construction of Pseudoternary phase diagrams for formulation selection For the formulation of nanoemulsions, an aqueous titration method was used and pseudo ternary phase diagrams were constructed with water, a mixture of surfactant and cosurfactant (Smix) as well as the oil phase. TheSmix was made in different ratios (1:1, 2:1, 3:1, 4:1, 5:1) by increasing the amount of surfactant added. Fourteen combinations of oil :Smix ratios (1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3.5, 1:3, 3:7, 1:2, 4:6, 7:3, 8:2, 9:1) were made. The aqueous phase was used for carrying out slow titration for all ratios of oil &Smix. The titrations were continued till a clear, transparent low viscosity nanoemulsion was obtained. The formulated nanoemulsions were kept undisturbed for 24 h to attain equilibrium and to eliminate metastable formulations.6 From the phase diagrams constructed, different combinations of placebo formulations were selected based on the presence of the minimum quantity of oil that could solubilise the drug dose and the minimum Smix quantity. For drug-containing nanoemulsions, the same procedure was followed except for the incorporation of the drug.   Thermodynamic stability studies                                          These stability studies are a very integral part of nanoemulsion formulation development as they help in choosing the nanoemulsions which are stable and also help in eliminating the metastable and unstable formulations. The following thermodynamic studies were done for the drug-loaded nanoemulsions.7 Centrifugation study In the centrifugation study, nanoemulsions were centrifuged at 25°C and 3000 rpm for 30 mins after which the presence of any physical instability, creaming or cracking them was Ssesses visually.  Heating cooling cycle             After passing the centrifugation study, the nanoemulsions were assessed for stability by performing the heating-cooling cycle. The temperature variation effect on nanoemulsion stability was checked by keeping the formulations between refrigerator temperature 4°C and 40°C for six cycles, with storage of not less than 48 hrs at each temperature. Freeze-thaw cycle             Further stability was assessed by freeze-thaw cycle in which the nanoemulsions were kept at -21°C and +25°C with storage for not less than 48 hrs at each temperature. Characterization of Nanoemulsion The optimized telmisartan loaded oral nanoemulsions were assessed for :  Viscosity measurement The nanoemulsion viscosity was evaluated by Brookfield Viscometer (Model DV-1 Prime) using spindle # 61 at 37± 0.5°C at 60 rpm. A two minutes wait time was used. The experiment was done in triplicate and the standard deviation ascertained. pH The pH of the nanoemulsions was checked using a digital pH meter. The experiment was done in triplicate and the standard deviation ascertained. Percentage transmittance For the percentage transmittance, a hundred times dilution was done for one ml of the formulation using the blank as distilled water and analyzed at 650nm by UV visible spectrophotometer (UV-1800, Shimadzu Corporation, Kyoto, Japan Shimadzu). The observations were taken in triplicate and the standard deviation was calculated.8 Refractive index Refractive indices of the nanoemulsions were evaluated using Abbe’s refractometer. The observations were taken in triplicate and the standard deviation was calculated. In vitro drug release and determination of release rate orders Franz diffusion cell, with treated dialysis membrane mounted between the donor and receptor compartment, was used for the study.35 ml of acetate buffer pH 5.5:PEG 600 (6:4) was used as the release media which was maintained at 37±1°C with continuous stirring at 75 rpm. Drug loaded nanoemulsion (1 ml) was placed over the dialysis membrane in the donor compartment. Samples (3ml) were taken at 0.5,1, 1.5,2,2.5 3,3.5, 4, 5, 6, 7 and 8 hrs and analyzed by using UV spectrophotometer (Shimadzu, 1800, Japan)9. The experiments were done in triplicate. To establish the kinetics of drug release of the formulation, data of in vitro drug release studies was plotted in different kinetic models: zero-order, First order, Higuchi model and Korsmeyer–Peppas model. The model which had the value of correlation coefficient near to 1 was confirmed to be the best fit model.10 In vitro skin permeation study and drug retention study of optimized nanoemulsion using excised rat skin Excised rat skin was used for the study after cleaning. The skin was held between the compartments of the Franz diffusion cell. Phosphate buffer pH 7.4 with methanol (6:4)(35ml), kept at 37 ± 1 °C with stirring at 75 rpm was used as the media. The nanoemulsion(1ml) was placed over the skin in the donor compartment. Samples (5 ml) were taken at time intervals of 1,1.5, 2,2.5, 3,3.5, 4, 5, 6, 7 and 8 hrs and analyzed at 428 nm using a UV spectrophotometer (Shimadzu, 1800, Japan). A graph was plotted between the cumulative amount of nanoemulsion permeated through the Wistar rat skin (Q, µg/cm2) as a function of time (hrs). From the slope and intercept of the straight line, drug flux (Jss, µg/cm2/h) was calculated. After dividing the flux with initial drug concentration (Co)  permeability coefficient (kp) was obtained.        Further, the formulation remaining on the skin was removed and the skin after the cleaning was weighed, cut into small pieces followed by sonication with methanol for 15 mins for extracting curcumin. The solution so obtained was centrifuged, filtered and the drug content (µg/cm2) was evaluated by using a UV spectrophotometer. Histopathological Study        To ascertain whether the curcumin loaded nanoemulsion was suitable for topical application histopathological study was performed. Freshly excised abdominal rat skin was taken and after washing with phosphate-buffered saline pH 7.4 mounted in a franz diffusion cell with the epidermis facing the upper side. 1ml of the nanoemulsion was placed over the skin in the donor compartment. The receptor compartment was filled with Phosphate buffer pH 7.4 with methanol (6:4) and left for 24 hours under constant magnetic stirring. After 24 hours, the rat skins were removed; blot dried and sent for histopathological evaluation using eosin and haematoxylin staining. The results were compared with those obtained for normal rat skin without the application of the formulation. Droplet size and zeta potential Malvern Zetasizer (Malvern, Worcestershire, UK) was used to determine the average particle size (z-average) and polydispersity index (PDI) of the developed nanoemulsion, after dilution and filtration using a 0.22µm membrane filter. The observations were taken in triplicate for each nanoemulsion evaluated. RESULTS Formulation Development and Optimization Solubility Studies The solubility of curcumin was evaluated in different oils and the results are given in Figure 1. Among the different oils evaluated, curcumin exhibited maximum solubility (8.01±0.026 mg/ml) in a combination of TPGS: Sefsol-218 Oil (1:1) and the combination was therefore chosen as the oily phase for the formulation of nanoemulsion. The solubility of curcumin in various surfactants was evaluated as per the given procedure. Curcumin exhibited the highest solubility in Tween 60. Based on the results of miscibility and solubility studies, Sefsol-18:TPGS (1:1), Tween 60 and Solutol HS 15were selected as oil, surfactant and cosurfactant respectively. Construction of Pseudoternary phase diagrams for formulation selection Placebo nanoemulsions were formulated using the chosen oil, surfactant and cosurfactants. Pseudo ternary phase diagrams were constructed for each Smix ratio prepared to identify the nanoemulsion region. When Smix 1:0 was used (Figure 4), a reduced nanoemulsion area was observed with macroemulsions comprising most of the region in the constructed pseudo ternary phase diagrams. As the cosurfactant amount was increased from 0 to 1 (Smix 1:1), the nanoemulsion region was observed to increase. An increase in the surfactant concentration from 1:1 to 4:1in Smix increased the nanoemulsion area. The maximum nanoemulsion area was obtained with Smix 4:1. Thermodynamic stability studies The placebo formulations selected from pseudo ternary phase diagrams were subjected to thermodynamic studies. In those placebo nanoemulsions which remained stable, the drug was added and then the drug-loaded formulations prepared were again subjected to thermodynamic stability studies. The formulations which exhibited turbidity and phase separation were discarded as they failed the tests for physical stability. Table 1 gives the compositions of stable drug-loaded nanoemulsion formulations which were taken up for further evaluation studies. Characterization of nanoemulsion formulations Viscosity The nanoemulsions exhibited viscosity in the range 17cp to 20cp. pH The nanoemulsions had a pH between 5 to 6, which was similar to the skinpH. Percentage Transmittance The percentage transmittances of all the developed formulations were found to be near 100% which confirmed the clarity and transparency of the optimized formulation.8 Refractive index The refractive index of nanoemulsions was close to that of sefsol-218:TPGS (1:1). Since no significant difference (pEnglishhttp://ijcrr.com/abstract.php?article_id=3631http://ijcrr.com/article_html.php?did=3631 Ahmad N, Ahmad R, Al-Qudaihi A, Alaseel SE, Fita IZ, Mohammed Saifuddin Khali MS and Pottoo FH.Preparation of a novel curcumin nanoemulsion by ultrasonication and its comparative effects in wound healing and the treatment of inflammation. RSC Adv 2019;9:20192-20206. Teichmann A, Heuschkel S, Jacobi U, Presse G, Neubert RH, Sterry W, et al. Comparison of stratum corneum penetration and localization of a lipophilic model drug applied in an o/w microemulsion and an amphiphilic cream. Eur J Pharm Biopharm 2007;67(3):699–706. Ahmad N, Alam MA, Ahmad FJ, Sarafroz M, Ansari K, Sharma S, Amir M.Ultrasonication techniques used for the preparation of novel eugenol–nanoemulsion in the treatment of wounds healings and anti-inflammatory. J Drug Del Sci Technol 2018;46:461–473. Anwer MK, Jamil S, Ibnouf EO, Shakeel F. Enhanced antibacterial effects of clove essential oil by nanoemulsion. J Oleo Sci 2014;63:347–354. Bali V, Ali M, Ali, J. Study of surfactant combinations and development of a novel nanoemulsion for minimising variations in bioavailability of ezetimibe. Colloids Surf B Biointerfaces 2010;76:410-420. Sharma S, Sahni JK, Ali J. Effect of high-pressure homogenization on formulation of TPGS loaded nanoemulsion of rutin – Pharmacodynamic and antioxidant studies. J Drug Del Sci Technol 2014;22(4):1-11. Mishra RK, Soni GC, Mishra RP. Nanoemulsion: A review article. World J Pharm Sci 2014;3(9):258-274. Wani RR, Patil MP, Chaudhari CA, Dhurjad PK, Shirsagar SJ. Microemulsion based gel: A novel approach in the delivery of hydrophobic drugs. Int J Pharm Res Scholars 2015;4(2):397-410. Kharwade RS, Mahajan NM. Formulation and Evaluation of Nanostructured Lipid Carriers Based Anti-Inflammatory Gel for Topical Drug Delivery System. Asian J Pharm Clin Res 2019;12(4):286–291. Bhagat KA, Bhura MRG, Shah SK. Formulation and evaluation of topical nanoemulgel of adapalene.World J Pharm Sci 2015;3(4):1013-1024. Dhawan B, Aggarwal G, Harikumar SL. Enhanced transdermal permeability of piroxicam through novel nanoemulgel formulation. Int J Pharm Invest 2015;4(2):65-76. Groves MJ, Mustafa RMA, Carless JE.  Phase studies of mixed phosphate surfactants, n-hexane and water. J Pharm Pharmacol 1974;26:616–623. Zhang QZ, Jiang XG, Jiang WM, Lu W, Su LN, Shi ZQ. Preparation of nimodipine-loaded microemulsion for intranasal delivery and evaluation on the targeting efficiency to the brain. Int J Pharm 2004;275:85-96. Baboota S, Alazki A, Kohli K, Ali J, Dixit N, Shakeel F. Development and evaluation of a microemulsion formulation for transdermal delivery of terbinafine. J Pharm Sci Technol 2007;61:276-285. Mate A, Ade P, Kharwade R, Pise S, More S. Formulation and evaluation of polyherbal gel for the management of acne. 2021;13(4):116-119. Wennerstrom H, Olsson U. Microemulsion as model systems. CR Chim 2009;12:4–17. Hussain A, Samad A, Singh SK, Ahsan MN, Faruk A, Ahmed FJ. Enhanced stability and permeation potential of nanoemulsion containing sefsol-218 oil for topical delivery of amphoterecin-B. Drug Dev Indust Pharm 2015;41(5):780-790. Ahmad J, Kohli K, Mir SR, Amin S. Formulation of Self –Nanoemulsifying Drug Delivery System for Telmisartan with improved dissolution and oral bioavailability. J Disper Sci Technol 2011;32:958-968.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareStudy to Compare the Efficacy of Orally Administered Melatonin and Clonidine for Attenuation of Hemodynamic Response During Laryngoscopy and Endotracheal Intubation in Gastrointestinal Surgeries English156161Sarmila Guha BanerjeeEnglish Pradipta SanyalEnglish Ujjwal BandyopadhyayEnglishIntroduction: The haemodynamic response to anaesthetic induction with endotracheal intubation may be harmful in patients with cardiovascular disease, increased intracranial pressure, or anomalies of cerebral vessels. Objective: A comparative study for attenuating reflex hypertension and tachycardia by using oral administration of Clonidine and Melatonin before induction of general anaesthesia. Methods: After obtaining the institutional ethical approval total of 76 patients scheduled for gastro-intestinal surgeries under general anaesthesia were divided into 2 groups, Group 1( 38 patients) received oral Melatonin 6 mg tab before surgery and Group 2 (38 patients) received oral Clonidine 100mcg before surgery. Heart rate, Systolic blood pressure(SBP), Diastolic blood pressure (DBP) and Mean arterial pressure (MAP) were recorded in two groups at different point of time i,e before premedication, immediately before induction, after tracheal intubation and every 3 min thereafter for 15 min. Incidence of perioperative and postoperative complications in 2 groups was also observed. Result: The results of observation were compiled, tabulated and statistically analyzed using the Chi-square test and unpaired t-test as and where applicable. Oral Melatonin was found better than oral Clonidine in lowering Systolic blood pressure(SBP), Diastolic blood pressure (DBP) and Mean arterial pressure (MAP) and heart rate changes associated with laryngoscopy and intubation. Occurrence of peri and postoperative complication in both the groups which were mild. Conclusion: From the observation of the present study, it can be concluded that oral Melatonin is a better agent than oral Clonidine in maintaining haemodynamic stability during gastrointestinal surgery under general anaesthesia. EnglishMelatonin, Clonidine, Haemodynamic stress response, Laryngoscopy, Intubation, PremedicationIntroduction                     In 1940, Reid and Bracefirst described the hemodynamic response to laryngoscopy and intubation. The mechanisms of the responses to laryngoscopy and orotracheal intubation are proposed to be by somatovisceral reflexes.1 Laryngoscopy and endotracheal intubation cause stimulation of proprioceptors at the base of the tongue, that cause reflex sympathetic activation of vagal and glossopharyngeal afferents. This activation cause hypertension, tachycardia and increased intracranial tension.2 TheseeffectscanbewelltoleratedbyASA (American Society of Anaesthesiology) grade1and 2 patients but can be detrimental for elderly patients, patients with cardiologicalandneurologicaldiseases.3 This haemodynamic surge is a problematic issue for gastrointestinal surgeries especially prolonged and major surgeries.3 A large array of pharmacotherapy has been tried to prevent laryngoscopic surge with varying degree of success such as Lignocaine, opioid,  vasodilator, Ca channel blocker, beta-blocker, Magnesium sulphate, Gabapentin, Pregabalin but none of the above-mentioned drugs has been proved fully efficacious to blunt haemodynamic stress response during laryngoscopy and endotracheal intubation, and all these agents have some limitations.4,5 Melatonin, (N_Acetyl_5_Methoxytryptamin) is an endogenous sleep-regulating hormone secreted by the pineal gland. Melatonin differs from other premedical because it exerts the sleep-promoting effect by amplifying day/night differences in alertness & sleep quality and displaying a modest sleep-inducing effect. The hypnotic, anti_nociceptive,  anxiolytic properties of Melatonin makes this neurohormone a useful medicine in anaesthesia and critical care.2 Melatonin 6 mg tablet if taken 120 minutes before operation then it can significantly decrease laryngoscopic surge.1,3 Apart from all the above mentioned benefits, melatonin has a short half-life so prolonged sedation is less likely, as well as melatonin does not have any abuse potential. Clonidine is a centrally acting alpha2 agonist. Clonidine causes stimulation of presynaptic alpha2 receptors, which decreases plasma renin and norepinephrine and thus blood pressure also decreases, the vagolytic effects of Clonidine causes a decrease in heart rate. Oral clonidine100 mcg tablet taken 120 minutes before operation can significantly reduce haemodynamic surge due to laryngoscopy and endotracheal intubation.3 MATERIALS AND METHODS Consenting patients of either sex, aged between 18 and 60 years, ASA physicalstatusIand2postedforelectivegastrointestinalsurgeries  under general anaesthesia with tracheal intubation were selected for the study. Single  laryngoscopic attempt and  laryngoscopic period Englishhttp://ijcrr.com/abstract.php?article_id=3632http://ijcrr.com/article_html.php?did=3632 1) Reid LC, Brace DE. Irritation of the respiratory tract and its reflex effect upon the heart. Surg Gynae Obstet 1940;70:157-62. 2) King BD, Hartis LC, Greifenstein FE, Elder JD, Dripps RD. Reflex circulatory response to direct laryngoscopy and tracheal intubation performed during general anaesthesia. Anaesthesiology 1951;12:556-66. 3) Boralessa H, Senior DF, Whitman JC. Cardiovascular response to intubation. Anaesthesia 1983;38:6237. 4) Pernerstorfer T, Krafft P, Fitzgerald RD, Krenn CG, Chiari A, Wagner O, et al. Stress response to tracheal intubation: direct laryngoscopy compared with blind oral intubation. Anaesthesia 1995;50(1):17-22. 5) Forbes AM, Dally FG. Acute hypertension during induction of anaesthesia and endotracheal intubation in normotensive man. Br J Anaesth 1970;42:618-24. 6) Fox Ej, Sklar gs, Hill CH, Vilanueva R, King BD. Complication related to the pressor response to endotracheal intubation. Anaesthesiology 1977;47:524-25. 7) Randel T, Hemodynamic response to intubation: what more do we have to know? Acta anaesthesiology Scand 2004;48:393-95. 8) Low JM, Harvey JT, Prys-Roberts C, Dagnino J. Studies of anaesthesia concerning hypertension. VII: Adrenergic responses to laryngoscopy. Br J Anaesth 1986;58(5):471-7. 9) Fassoulaki A, Kaniaris P. Does atropine premedication affect the cardiovascular response to laryngoscopy and intubation? Br J Anaesth 1982;54:1065-80. 10) Fassoulaki A, Kaniaris P. Intranasal administration of nitroglycerin attenuates the pressure response to laryngoscopy and intubation of the trachea. Br J Anaesth 1983;55:113. 11) Rani R, Nesargi SS. Oral Clonidine for Attenuating Hemodynamic Stress Response to Laryngoscopy and Intubation. Karnataka Anaesth J 2015; 1(2):50-54. 12) Raval D, Mehta M. Oral clonidine premedication for attenuation of haemodynamic response to laryngoscopy and intubation. Indian J Anaesth 2002;46:124-29. 13) Das M, Ray M, Mukherjee G.Clonidine attenuate haemodynamic response during laparoscopy. Indian J Anaesth 2007;51:205-210. 14) Hoseini VS, Yekta R, Marashi S, Marashi SM. Melatonin, Gabapentin and Clonidine in reducing anxiety and pain in cholecystectomy. Arch Anesth Crit Care 2015;15:120-125. 15) Gupta P, Jethava D, Choudhary R, Jethava DD. Role of melatonin in attenuation of haemodynamic responses to laryngoscopy and intubation. Indian J Anaesth 2016;60:712-18. 16) Naguib M, Gottumukkala V, Goldstein AP. Mini-Review: Melatonin and Anaesthesia, a clinical perspective. J Pineal Res 2007;42:12–21. 17) Taittonen MT, Kirvela OA, Santana R, Kanto JH. Effect of clonidine and dexmedetomidine premedication on perioperative oxygen consumption and haemodynamic state. Br J Anaesth 1997;78:400–406. 18) Ionescu D. Melatonin as premedication for laparoscopic cholecystectomy SAJAA 2008; 14:8-11. 19) Kurdi MS, Patel T. The role of melatonin in anaesthesia and critical care. Indian J Anaesth 2013;57:137-44. 20) Mihara T, Nakamura N, Ka K, Goto T. Melatonin for paediatric emergence reaction. Eur J Anaesth 2015;32:1–10. 21) Ahmed M, Hosam A, El-Din A, Kassaby M, Ismail A, Helmy A. Effects of Melatonin Premedication on the Hemodynamic Responses and Perfusion Index During Laryngoscopy and Endotracheal Intubation. Med J Cairo Univ 2014;81:859-867. 22) Cardinalii DP, Pagano ES, Bernasconi, Pablo AS. Disrupted chronobiology of sleep and cytoprotection in obesity;possible therapeutic values of melatonin. Act Nery Super Rediviva 2011;53(4):159-176.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcarePhytochemistry of Amaranthus viridis: GC-MS Analysis English162166Varalakshmi ThanikachalamEnglish Indira A. JayarajEnglishEnglishAmaranthus Viridis, Ayurveda, Inflammation, Antioxidant, Treatment, ChromatographyINTRODUCTION The world is endowed with a rich diversity of medicinal plants. About 80% of the world’s population uses herbs for medicinal purposes. Herbs have always been the principal form of medicine in the world. Some biologically active compounds isolated from herbs have been explored for the inhibition of the growth of pathogenic microbes because of their antimicrobial potential.1 The medicinal value and multiple biological properties of several plants are defined by their phytochemical constituents.2 Plants have provided a source of inspiration for novel drug compounds,3 as plant-derived medicines made large contributions to human health and well-being. Plant-derived medicines are widely used because they are relatively safer than synthetic alternatives, as they are easily available and cheaper.4 Many plant species have been evaluated for their antimicrobial activity in the past 20 years.5 They provide us with the key chemical structure for the development of new phytomedicine to be used for the treatment of disease. The active principles of many drugs found in plants are recognized as secondary metabolites.6 Amaranthus, commonly known as Green amaranth, is a multinational genus of herbs. In the last decade, amaranth is not only used in the common diet but also in the diet of people with celiac disease or allergies to typical cereals.7 Recently drug resistance to a human pathogenic organism has been reported worldwide. Medicinal plants are an expensive gift from nature as they are the sources of important therapeutic aids for alleviating human ailments. Gas Chromatography-Mass Spectrometry (GC-MS) is the best technique to identify the bioactive constituents of long-chain hydrocarbons, alcohols, acids, esters, alkaloids, steroids, amino acid and nitro compounds.8-11 Therefore, characterization of extracts of medicinal plants is necessary due to its numerous benefits to science and society. Nevertheless, Amaranthus has received notably less research attention as vegetables than grain amaranths. Hence, the present study was aimed at determining phytochemical constituents with the aid of the GC-MS technique and in vitro screening of pure extract of leaves from locally grown Amaranthus Viridis plants for their phytochemical study. The findings of this study provide important data on the bioactive substances of this underutilized vegetable, and thereby promoting their utilization in the food industry. MATERIALS AND METHODS Collection of samples Fresh plant leaves of Amaranthus Viridis was collected and authenticated by Prof.P.Jayaraman, Director, Plant Anatomy Research Centre, Chennai, Tamil Nadu, and India. A voucher specimen (Reg.No. PARC/2019/4054). The leaves were thoroughly washed through tap water and dried under shade for 3-5 days. The dried leaves were ground to a fine powder and stored in bags for further use. Preparation of extracts 50 grams of dried powder of Amaranthus Viridis leaves were packed in a separate round bottom flask for sample extraction using 500 ml water. The extraction was conducted with 20ml of water for 24 hours. At the end of the extraction, the crude extract was stored in refrigerator.12 Chemicals and reagents All chemicals and reagents were procured from certified suppliers and were of the highest analytical standard. Gas Chromatography-Mass Spectroscopy Gas chromatography-mass spectroscopy (GC-MS), a hyphenated sys­tem which is a very compatible technique and the most commonly used technique for the identification and quantification purpose was used. The unknown organic compounds in the complex mixture can be deter­mined by interpretation and also by matching the spectra with reference spectra. Preparation of extract The extract of the leaves was analyzed using Gas Chromatog­raphy Mass Spectroscopy for the identification of the phytochemical compounds present. A solvent blank analysis was first conducted using 1 μl of absolute ethanol.13 Then 1 μl of the reconstituted extract solution was employed for GC-MS analysis as previously described with modifications.14 Procedure Identification of bioactive compounds by GCMS The purified extract fractions were individually examined using GC SHIMADZU QP2010 system and gas chromatograph interfaced to a mass spectrometer equipped with Elite-1 fused silica capillary column. For GC-MS detection, an electron ionization energy system with ionization energy of 70eV was used. Helium was used as carrier gas at a constant flow rate of 1ml/min and an injection volume of 2μl was employed. The sample was run for 40 minutes with a solvent out time of 9.50 minutes. Mass spectra were taken with a scan-interval of 0.6 seconds. Interpretation of the mass spectrum was achieved by using the database for different bioactive compounds. GCMS analysis of bioactive compounds from the sample The leaf extract was subjected to Gas Chromatography and Mass Spectroscopy for the determination of bioactive volatile compounds. Identification of phytoconstituents Interpretation of the mass spectrum of GC-MS was done using the database of National Institute Standard and Technology (NIST) having more than 62,000 patterns.15, 16NIST library was used to calculate the mass spectrum. Quantitative determination was made by respective peak areas to TIC areas from the GC-MS. The principle name, molecular weight, retention time and peak area percentage of the test materials was ascertained. The column temperature was programmed from 75 - 260°C (rate = 6°C/min) with the lower and upper temperatures being held for 3 and 10 minutes respectively. Total GC running time was 43.2 min. The GC injector and MS transfer line temperatures were set at 280°C and 290°C respectively. All analysis was done in the split-less mode. Helium (99.9%) was used as a carrier gas (flow rate = 1.0 ml/min) and an injection volume of one μl was used for analysis. Major and essential compounds were identified by their retention times and mass fragmentation patterns. RESULTS Phenolic compounds can be defined as a large series of chemical constituents possessing at least one aromatic ring, bearing hydroxyl and other sub-constituents. GC- MS analysis is the most used method for the identification of plant phenolic compounds. Amaranthus Viridis leaf extracts are found to be a vital source of useful bioactive substances. These bioactive compounds are involved in various biological functions such as communication, infection, reproduction and self- defence. In the present study, we have identified bioactive compounds present in the extract fraction of leaf by GC-MS analysis and summarized in Table 1. The active principles with their retention time (RT), molecular weight (MW) and concentration (%) of the corresponding compounds were observed in the leaf extract. GC-MS chromatogram of the leaf extract of A. Viridis belonging to the family Amaranthaceae showed 40 peaks which indicate the presence of forty compounds. The spectra of the compounds were matched with Wiley 9.0 and the National Institute of Standards and Technology libraries. The compounds detected are presented in Table 1 DISCUSSION The plants contain large amounts of secondary metabolites that exert a wide range of biological activities on physiological systems. It was also reported that the activities of some plant constituents with compound nature of alkaloids, flavonoids, palmitic acid (hexadecanoic acid, ethyl ester and n-hexadecanoic acid, unsaturated fatty acid and linolenic (docosatetraenoic acid and octadecatrienoic acid) as antimicrobial, antioxidant, anti-inflammatory, hypocholesterolemic, cancer preventive, hepatoprotective, antiarthritic, antihistaminic, antieczemic, immunomodulatory and anticoronary.17 Secondary metabolites of the plants attract beneficially and repel harmful organisms, serve as phytoprotectants and respond to environmental changes. In general, the phytochemical contents (Table 1) were by the previous reports for some of the vegetables. These phytocompounds are responsible for various pharmacological actions of the leaves of the plant. Benzoic acid, 2-[(trimethylsilyl)oxy]-, trimethylsilyl ester (24.09%) is found as the major compound and forty nine minor compounds such as Nonamethyl, Phenyl-, Cyclopentasiloxane (6.10%), (+,-)-3.beta.-(acetyloxy)-3-ethynyl-1,2,3,4,4a.beta.,12a. (5.67%), [Bis (trimethylsilyl) methyl] diphenyl phosphine $$ Phosphine (5.59%), Benzoic acid, 2-[(trimethylsilyl) oxy]-, trimethylsilyl ester (5.32%), 1,2-Diphenyl tetramethyldisilane $$ Disilane,1,1,2,2-tetramethyl-1,2-diphenyl (4.98%), Benzophenone, 2-(trimethylsiloxy)- (CAS) Trimethylsilyl ether of O-hydroxybenzophenone (3.50%), 7-(P-Chlorophenyl)imino-6-(P-tolyl)-1,3-dimethyl-2,4 dioxo-1,2,3,4,6,7- hexahydro pyramid [4,5-d] pyramidine (3.14%), 4-.alpha.,20-dimethyl-3-.beta.-dimethyl-.. (2.84%), Silane, [1,3,5-benzenetriyltris (oxy)] tris [trimethyl- (CAS) Phloroglucinol tris MS (2.76%), 2-Isopropyl phenol- Trimethylsilyl- Ether (2.73%), 4H-1-Benzopyran-4-one, 2-(2,6-dimethoxyphenyl)- 5,6-dimethoxy- (CAS) Zapotin $$ Flavone (2.46%), (Z)-1-[(1&#39;,1&#39;-dimethylethyl)diphenylsilyl]-3-trimethylsilyloxyprop-1-ene (2.37%), Prosta-5,10,13-trien-1-oic acid, 15-[[(1,1-dimethylethyl) dimethyl ethylsily]oxy]-9-oxo- (2.19%), Silane, trimethyl (triphenylethenyl)- $$ (2.18%) and the remaining compounds peak area ranged from 1.84% to 0.16% The results from the current study indicate that methanol leaf extract of the Amaranthus Viridis tested by GC-MS analysis contained various types of compounds with potential pharmacological activity. The presence of various bioactive compounds justifies the use of Amaranthusviridisfor various ailments by traditional practitioners. From GC-MS data, identification of more compounds in their extract and it previously reported that these compounds have antibacterial, antifungal, antioxidant and anticancer activity but further researches should be made to isolate and purification of natural products in their extract. CONCLUSION The edible plant species Amaranthus viridis from the underutilized plant family had a rich amount of valuable ingredients that are beneficial for health. Further research work is required in more details about in vitro and in vivo investigations to establish which components of the extract are biologically active in terms of activity. The isolation of components from this readily available plant resource and its utilization as natural agents could be of high economic value. Hence, the identified plant components using GC-MS can be used as a tool for the identification of adulterants. The current pioneering study sug­gests that the extract is a potent therapeutic agent. It paves the way for the development of several treatment regimens based on this extract. Also, further research is necessary to identify and purify the active compounds responsible for therapeutic activity, as well as the unidentified compounds. ACKNOWLEDGEMENT We are thankful to the Department of Biochemistry, Kongunadu College, Coimbatore, Tamil Nadu, and India for providing facilities to carry out research work. The authors acknowledge the immense help received from the scholars whose articles are cited and included in references to this manuscript. The authors are also grateful to authors/ editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed Conflict of Interest: Authors declare no conflict of interest. Source of Funding: None Englishhttp://ijcrr.com/abstract.php?article_id=3633http://ijcrr.com/article_html.php?did=36331. Abukakar MA, Ukwuani AN, Shehu RA. Phytochemical screening and antibacterial activity of Tamarindus indica pulp extract. J Biochem 2008;11(2):134-138. 2. Fallah HSM, Alavian HR, Heydari MR, Abolmaali K. The Efficacy of Liv-52 on Liver Cirrhotic Patients: A Randomized, Double-Blind, Placebo-Controlled First Approach. J Phytomed 2005;12:619-624. 3. Velayutharaj A, Muthumani L, Senthilnathan R, Rajendran SM, Shivakumar R, Saraswathi R. Significance of Secreted Frizzled Related Protein 4 (SFRP4) in Type 2 Diabetic and non-Diabetic Subjects in the Rural Agricultural Population Who are Exposed to Pesticides for the Prediction of Diabetes Mellitus. Int J Curr Res Rev 2018;10(06):7-9. 4. Iwu MM, Duncan AR, Okunji CO. New Antimicrobials of Plant Origin.  Int J Janick Ed 1999;1:11. 5. Castello MC, Anita P, Naresh C, Madhuri S.Antimicrobial Activity of Crude Extracts from Plant Parts and Corresponding Calli of Bixaorellina. Indian J Exp Biotechnol 2002;40:1378-1381. 6. Ghani A. Introduction to Pharmacognosy. Ahmadu Bello University Press, Nigeria. 1990. 7.  Dobelis IN. Magic and Medicine of Plants. The Readers Digest Association Inc., 1993. 8. Subramanian S,  Ramakrishnan N. Chromatographic fingerprint analysis of Naringi crenulata by HPTLC technique. Asian Pac J Trop Biomed 2011;1:195-198. 9. Muthulakshmi A, Joshibhi-Margret R, Mohan VR. GC-MS analysis of bioactive components of Feronia elephant Correa (Rutaceae). J Appl Pharm Sci 2012;1:69-74. 10. Yamunadevi M, Wesely EG, Johnson M. Chromatographic fingerprint analysis of steroids in Aerva Lanasa L. by HPTLC technique. Asian Pac J Trop Biomed 2011;1:428-433. 11. Gopalakrishnan S, Vadivel E. GC-MS Analysis of some bioactive constituents of Mussaenda frondosa Linn. Int J Pharm Biosci 2011;2:313-320. 12. Paranthaman R, Praveen K, Kumaravel S. GC-MS Analysis of Phytochemicalsand Simultaneous Determination of Flavonoids in Amaranthus caudatus (Sirukeerai) by RP-HPLC. J Anal Bioanal Tec 2012;1:14. 13. Komansilan A,  Abadi A, Yanuwiadi B, Kaligis DA. Isolation and Identification ofBiolarvicide from Soursop (Annona muricata Linn) Seeds to Mosquito (Aedesaegypti) Larvae. Int J Engi Technol 2012;12(03):28–32. 14. Kumar PP, Kumaravel S, Lalitha C. Screening of antioxidant activity, total phenolics and GC-MS study of Vitex negundo. Afr J Biochem 2010;4:191-195. 15. Akubugwo IE, Obasi NA, Chinyere AE. Nutritional and chemical value of Amaranthus hybrid slaves from Afikpo Nigeria. Afr J Biotechnol 2007;6:2833-2839. 16. Akubugwo IE, Obasi AA, Chinyer GC, Ugbogu AE. Mineral and phytochemical contents in leaves of Amaranthus hybridus L and Solanum nigrumL. Subjected to different processing methods. Afr J Biochem Res 2008;8:40–44. 17. Rodrigues E, Tabach R, Galduróz JCF,  Negri G. Plants with possible anxiolytic and/or hypnotic effects indicated by three Brazilian cultures - Indians, Afro-Brazilians, and river-dwellers. Stud Nat Prod Chem 2008;35:549–595.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareFunctional Outcomes in Trans Femoral Amputees using Indigenously Developed Safe Gait Prosthetic Knee Joint English167171Veerendra K. ShandilyaEnglish Parth DevmurariEnglish Ashwinikumar V. ShandilyaEnglish Lata D. ParmarEnglishIntroduction: India has a large young population with limb loss, especially from low economic strata, trauma being the major cause, suddenly takes a toll on the finances of the amputees. A low-cost endo-skeletal prosthetic knee joint, named ‘Safe Gait’ has been designed and developed, with a mechanism of free knee joint with swing assist and a lock that can be operated at will and with ease by a patient at any point of time and terrain. Objective: The present study aims to assess the TFA using SGPKJ on the functional outcomes and compare to the patients wearing other imported prosthetic knee joint (OIPKJ) like polycentric knee joint, hydraulic knee joint, etc. It was hypothesized that SGPKJ would provide stability and mobility at a very low cost and thus could be an option for non-affording TFA. Methods: This was an ‘Open-label’ study. The participants who satisfied the inclusion criteria were divided into two groups based on their prosthetic options, they were compared on functional measures like single limb stance time in a sec, Time Up and Go Test, & Six Minutes’ Walk Test. Results: Show single-limb stance as 1.39 sec & 1.48 sec; Gait speed as 1.37 m/sec & 141m/sec in Safe Gait Prosthetic Knee Joint and Other Imported Prosthetic Knee Joint resp. There was no statistically significant difference between the two groups in other parameters viz. TUG & 6MWT. Conclusion: The participants with SGPKJ were found to be compatible with mobility and stability on various locomotor tests. English Functional outcomes, Six-minute walk test, Time up and go test, Transfemoral amputation, Prosthetic knee joints, Lower extremities amputationsIntroduction It is stated that 200-500 million major amputations each year are performed worldwide and 85% of these are lower extremities amputations (LEA).1 According to the sample survey of India 2011, approximately 2.2% of the population is living with a disability. That comes to around a 2.2million people living with disabilities. Limb loss is a life-changing event that can be overwhelming. Amputation (limb loss) is one of the commonly seen disabilities in our country. Trauma being the major cause, with young amputees in India, it has been observed that most of these youth belong to the lower strata of the society and face a hard time coping up with the mainstream. The prosthetic technology in recent years has grown by leap & bounds. The most advanced prosthesis incorporates advanced gadgets like microprocessors; these are available with exorbitant costs.2,3 Most endo-skeletal prosthetic knee joints available in India are imported and are expensive. The most common & economical prostheses – Jaipur foot Prostheses- are fitted & distributed in camps free of cost. These above-knee prostheses have an Exoskeletal system with a pair of Orthotic drop lock for all its prosthetic fittings, this results in a stiff knee gait.4 There are three major groups in which the transfemoral amputees (TFA) prosthesis have been classified, viz. Passive, damping & powered.5 Worldwide endo-skeletal Prosthetic knee joints supplied are either free/ functional or with a locking mechanism, amputees do not have the option to self-lock or unlock the prosthetic knee. In India, on uneven inclined terrain as the prosthetic knee axis moves anteriorly and will tend to buckle, for stability needs on varied terrain conditions, overcrowded/rush situations, health issues, etc. locking option must be available to the TFA. Further, the design has to be such that not only is the cost of the prosthesis very low, but also the recurring expenditure of maintenance becomes much lower than at present. An endo-skeletal prosthetic knee joint was thus designed & developed keeping in mind the above factors and has been named as ‘SAFE GAIT.’ Design considerations for Safe Gait Prosthetic Knee Joint Stainless Steel material was chosen for the manufacture of the knee joint because: 1. Titanium is costly and would take the cost too high. 2. Stainless steel would make the Knee Joint rust-proof (free of corrosion) and durable. 3. An investment casting has been done on the joint to make it cost-effective manufacturing. 4. Since there are no standard tests on knee joint testing available in the country, a prototype was manufactured with Stainless Steel 304 and a compression load test was done at ERDA (Energy Research Development Agency – an authentic autonomous body in the field of testing) Technical Features The Knee axis is behind (posterior) for stability Extension assist is done with an elastic cord which is easily interchangeable and tension can be adjusted for the swing phase. The lock mechanism works with a spring-loaded plunger, when the knob is shifted; it immediately locks the knee (in full extension) Load Limit - 120 KGs, UNB 12.9 Knee Flexion Range – 125 degree (approx.) Height (Knee centre – upper edge) - 22m Optional height (Knee-Centre with special coupling) -12mm Universal socket assembly features (which could be fitted to any prosthetic component manufacturer’s assembly system) - 30mm pylon tube fitting (Figure 1). The cost of the knee joint is only Rs. 2000/-this pilot study was planned to assess this safe gait prosthetic knee joint (SGPKJ) in trans femoral amputees (TFA) using simple functional tools. It was important to assess a single limb stance along with other functional tests like the ‘Timed Up and Go Test (TUG) and Six Minutes&#39; Walk test (6MWT). The Time Up Go test is used to assess individuals&#39; function in balance, transfer, walking, and turning around.6 Robert Gailey in 20027 is the first researcher to find 6MWT practical for measuring functional capabilities and endurance. 6MWT is also said to demonstrate good to excellent test-retest reliability as a measure of functional capacity in the population with lower limb amputees. The present pilot study aimed to assess the TFA using SGPKJ on the functional outcomes as mentioned and this was compared to the patients wearing other imported prosthetic knee joint (OIPKJ) like polycentric knee joint, hydraulic knee joint, etc.  It was hypothesized that SGPKJ would provide stability and mobility at a very low cost and thus could be an option for non-affording TFA. MATERIALS AND METHODS Study Design: Open-label study, Registered with CTRI Procedure After the institutional ethical clearance was obtained, the referred patients were approached. They were explained about the study and the prosthetic options available. All those patients agreeing to participate in the study were requested for written informed consent, they were provided with the written information sheet explaining the study. Following this, they were screened for inclusion & exclusion and Demographic data and education information with contact numbers were taken. Patients were divided into two groups based on: - On the economy (Price, fitting cost, annual maintenance cost) Function with the prosthesis Stability while weight-bearing with the prosthesis Quality of life with the prosthesis Inclusion criteria Above-knee amputees Age 16 years and above Having unilateral TFA Being able to read, write and understand & communicate Exclusion criteria Amputees with upper extremity amputations Trans-tibial /partial foot amputees Psychiatric disorders, hearing impaired and visually impaired persons with amputations. Other neuromuscular problems, acute illness or chronic illness, and renal dialysis. Mentally challenged persons with amputations Comparison GROUP A (1) - case (with ? safe gait prosthetic knee joint) GROUP B (2) - patients fitted with other imported prosthetic knee joints The simple outcome measures were used as follows: Single limb stance time in secs TUG 6MWT The above procedures were performed as described below: Single limb stance time was checked for a prosthetic limb. Eligible subjects were asked to stand with shoes on the prosthetic limb, with the normal limb raised allowed to touch the wall with the tip of a finger. For the test, standard procedure was followed i.e. each participant was asked to look at a point on the wall in front. The duration for which the participant was able to stand on his prosthetic limb with the other limb raised off the floor was timed and recorded. The test was terminated if the participant used his arms, moved his raised foot in an attempt to stabilize himself or shifted/rotated on his prosthetic leg/foot on the ground. The procedure was repeated 3 times, and the average of the 3 trials was recorded. In our study, we asked subjects to perform the test with the eyes open and ‘Gait belt’ in the parallel bar to ensure safety. The TUG test required the subjects to rise from a chair, walk 3 meters at a comfortable pace to a marked place on the floor, turn around at the mark, and walk back to the starting point and return to sitting in the chair. The test&#39;s score is the time he/she takes to complete the test. This test assesses time taken in four components i.e. raising from sitting, walking, turning, and sitting back. A six-minute walk test (6MWT) was performed as per guideline. At the outset, subjects were instructed to walk between starting and endpoints at a normal pace with their prosthesis in a marked 30 meters long corridor. Before the commencement of the test, the subjects were asked to sit comfortably for 10 minutes at least, following which vital parameters such as Pulse rate (PR), Respiratory rate (RR), and Rate of perceived exertion (RPE) were recorded. Vital parameters were taken before the test, post immediate, and 2 minutes after. The test was performed at a quiet and well-ventilated place so that commands at regular intervals could be audible to subjects. The test was carried out at room temperature and all necessary precautions like standby emergency services, test supervision by a senior therapist, and availability of a chair at a short distance were taken care of. Statistical analysis The data so collected were entered into Microsoft Excel for analysis. TUG, RPE, and 6MWT are assumed to be normal and are reported as mean (SD) whereas the age of patients and single-limb stance are assumed non-normal and hence are reported as median (IQR). An Independent t-test is used to compare TUG, RPE, and 6MWT between two types of knee joints whereas age and single-limb stance are compared using the Mann Whitney test. The significance level is set at 5% throughout the study. Results Total participants were 20 and all had similar comfortable ischial containment socket fittings with silicone socket having pin lock suspension and multi-axial foot with standard pylon; with 10 subjects fitted with SGPKJ all males & 10 subjects OIPKJ4- Polycentric knee joint, 3- four-bar linkage knee joint, 2- Stance control single-axis knee joint,1- Hydraulic knee joint. OIPKJ group had 8 males and 2 females. Single limb stance – 1.393 sec. & 1.488 sec in SGPKJ & OIPKJ resp. TUG – Average was 15.93 & 15.80 of SGPKJ and OIPKJ group       6MWT- Average RPE immediate posttest was 2.53 & 2.80 of SGPKJ and OIPKJ group SPO2- varied between 96 -97.55                                             DISTANCE WALKED- was 268.70 m and 251.90 m for SGPKJ and OIPKJ group resp. GAIT SPEED- 1.37m/s in the SGPKJ and 1.41m/s in the OIPKJ. Discussion Loss of limb is a huge trauma, physically, psychologically, and financially. It is known worldwide that lower limb amputations are common as compared to upper limbs. Transfemoral (above knee) amputations result in a greater loss of mobility and function due to the absence of knee and ankle-foot.8 Designers and innovators have focused their attention on developing prosthetic knee joints to provide near-normal functions. However, few designs have been successful.8 There is also not enough data on various functional / performance parameters despite several designs presented.9 In India, very few prosthetic knee designs have been developed. The Safe Gait Prosthetic Knee Joint designed and developed has been tested in the present study by comparing it with the Other Imported Prosthetic Knee Joints on various functional parameters using locomotor tests. Single-limb stance 30seconds it is 2 or stable.7 In this pilot study, the two groups of the same age (Table 1) were compared, almost all were males (Figure 2), there was no statistically significant difference in the single-limb stance on the prosthetic limb unsupported between the two groups (Table 2) the patients did well if allowed to stand with minimal support or with a walking aid. This shows that the SGPKJ was also providing similar stability as was provided by the imported knee joints. A successful knee joint is expected to provide stability in the stance phase for adequate step and stride in gait thus helping the amputee to walk faster.9 The stability of the prosthetic knee is provided by design and alignment and this is essential to prevent the knee from buckling especially on uneven terrain as is common in our country.9 Gait speed in the present study was also found to be 1.37m/s in the SGPKJ and 1.41m/s in the OIPKJ. According to the author Caroline 2016, ≤0.44m/s was indicative of non-community ambulator and states traumatic amputee as having gait speed of 1.3m/s.10 With regards to TUG, literature reports range from 7.2 to 102 seconds and maybe slower for geriatric people with TFA.10 In the present study there was no significant difference between the groups for TUG (Table3). The TUG ranged from 13.68 to 20.19. Although it is well above the age-matched normal. The TUG reported by authors 2016  for the prosthetic users is 12.5 (9-20.8).10 With regards to 6MWT also, the two groups were not found statistically different. The average 6MWD was 262.7m with SGPKJ and 254.5m with OIPKJ. This distance is much lesser compared to the age-matched normal. Caroline 201610 reported 6MWD in prosthetic users as 274 (156-371). The vital parameters i.e. pulse rate, respiratory rat (Figure 3) and RPE (Table 3) measured were also not significantly different between the two groups. According to the literature, 6MWT is reported to vary from 4 to 858m in cohorts with LLA.  The LLA’s with the inability to walk >200m nonstop are predicted to be inefficient community ambulator.10 To improve QOL, functional mobility, and develop functional independence following LLA, it is vital to undergo physiotherapy and rehabilitation as soon as possible.11 The above functional locomotor tests help identify the level of independence in the LLA and if found impaired could be targeted for improvement and modification by adopting various rehabilitation strategies like gait retraining, improve strength and balance, use of mobility aids, etc.10 Trauma is the major cause of young amputees in India however, India being the diabetes capital, diabetic foot ulcers are a high-risk factor apart from PVD for nontraumatic LEA. This indigenously developed SGPKJ would provide the option for millions in our country, as it is not only cost-effective, providing the ability to walk free at will but also have locking unlocking made easy for stability and mobility resulting in incompatibility on functional outcomes.12 Conclusion The most advanced gadgets like microprocessors cannot be competed with. However, SGPKJ indigenously made, found to be statistically compatible with OIPKJ available on the mobility and stability tests, provides hope of affordable option to millions in our country. Acknowledgement: Authors acknowledge the ‘Rehabs Clinic’ and ‘Sumandeep Vidyapeeth’. The authors also acknowledge the immense help received from the scholars whose articles are cited and included in references to this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Conflict of Interests: Authors declare NIL conflicts of interests. No financial support received. Englishhttp://ijcrr.com/abstract.php?article_id=3634http://ijcrr.com/article_html.php?did=3634 Creylman V, Knippels I, Janssen P, Biesbrouck E, Lechler K, Peeraer L. Assessment of transfemoral amputees using a passive microprocessor-controlled knee versus an active powered microprocessor-controlled knee for level walking. Biomed Engi 2016;15(3):142. Gailey RS, Roach KE, Applegate EB, Cho B, Cunniffe B, Licht S, et al. The amputee mobility predictor: an instrument to assess determinants of the lower-limb amputee&#39;s ability to ambulate. Arch Phys Med Rehab 2002;83(5):613-627. Uchytil J, Janda?ka D, Farana R, Zahradník D, Rosicky J, Janura M. Kinematics of gait using bionic and hydraulic knee joints in transfemoral amputees. Acta Gymnica 2017;47(3):130-137. Mahida H, Mahida D. Jaipur Foot: An Attempt to Replicate Jaipur Model in Surat City. NJCM India 2015;6:289-291. Fu H, Zhang X, Wang X, Yang R, Li J, Wang L, et al. A novel prosthetic knee joint with a parallel spring and damping mechanism. Int J Adv Robo Syst 2016;13(4):1729881416658174. Levin AZ. Functional outcome following amputation. Top Geriatr Rehabil 2004;20:253–261. Gailey RS, Roach KE, Applegate EB, Cho B, Cunniffe B, Licht S, et al. The amputee mobility predictor: an instrument to assess determinants of the lower-limb amputee&#39;s ability to ambulate. Arch Phys Med Rehab 2002;83(5):613-62 Wilson AB, BSME J. Recent advances in above-knee prosthetics. Artif Limbs 1968;12(2):1-27. Silver-Thorn MB, Glaister CL. Functional stability of transfemoral amputee gait using the 3R80 and Total Knee 2000 prosthetic knee units. JPO 2009;21(1):18-31 Roffman CE, Buchanan J, Allison GT. Locomotor performance during the rehabilitation of people with lower-limb amputation and prosthetic nonuse 12 months after discharge. Phys Ther 2016;96:985–994. Ülger Ö, Y?ld?r?m ?ahan T, Çelik SE. A systematic literature review of physiotherapy and rehabilitation approaches to lower-limb amputation. Physiother Theory Pract 2018;34:821–834. Nanjappa BA, Karthik P, Aroul TT, Smile SR. Risk factors for lower extremity amputation in patients with diabetic foot ulcers. Int J Curr Res Rev 2012;4(6):30-36.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareComparative Quality Analysis of Cefpodoxime Proxetil Branded Tablet Product with Available Indian Generic Products: Short-Term Accelerated Study English172184Darshana R. DumbhareEnglish Fahimuddin S. KaziEnglish Debarshi Kar MahapatraEnglish Ujwala N. MahajanEnglishEnglishCefpodoxime proxetil, Branded, Generic, Accelerated Stability Study, Impurity, DegradationINTRODUCTION The concept of quality assessment is an absolute system to create and follow the practices and strategies for providing the most consistent laboratory outcomes and to lessen the errors involved in the pre-analytical, analytical, and post-analytical stages.1 Among the two foremost elements, Quality Assessment is one of the universal quality management systems. Performance Improvement is one of the headways that have activities related to the activity of the other processes.2 Quality assessment service in context to growing organizational stages principally highlighting the data quality and the detection, particularly contained by the monetary sectors, that data correctness and veracity are critical to execution with lawmaking and regulatory approvals.3 Cefpodoxime is a safe, short course, effective, semi-synthetic, third-generation cephalosporin available for use as a prodrug known as Cefpodoxime proxetil.4 It is absorbed readily from the human gastrointestinal tract. It is an active drug given in various regimens for diverse infections caused by Gram-positive cocci such as streptococci, staphylococci, penicillinase-producing strains as well as Gram-negative bacteria such as Meningococci, Hemophilus, Gonococci, Klebsiella, Moraxella, E. coli, etc.5 It is highly recommended for treating sinusitis, otitis media, urinary tract infection, respiratory tract infections (upper and lower), skin infections, and soft tissue infections.6 The drug can be recommended as a parenteral cephalosporin with a maximum dose of 8–10 mg/kg/d.7 It is majorly excreted by the kidneys in an unchanged form. In renal compromised patients, dose adjustments are required.8 It attains sufficient stages beyond the minimum inhibitory concentration (MIC) in several body fluids, therefore, the doses need to be adjusted (single or double doses).9 A branded medication is an innovative creation that has been created by an innovator organization. When an innovator develops a new medication, the developed novel drug product must endure and pass recommended quality tests and evaluate to make sure its effectiveness in remedial situations to treat and safely administer for human use.10 As innovator organizations spend a substantial amount of resources to formulate a novel drug product, they are provided with the exclusive right to produce and market the medications for a given duration.11 When an organization is provided exclusive rights to produce and market, the medicament is credited to patent(s). For a given duration of the patent award to the original company, no other organizations have the right to manufacture the same drug.12 For this motive, a branded medicine is well known and that particular medication is generally very much trusted. A generic medicine may be defined as a drug product that corresponds to a meticulous reference listed drug product (RLD) or drug brand in the context of strength, type of dosage form, route of administration, performance characteristics, quality, and its application, which may be distinguished as a pharmaceutical equivalent, bioequivalent, and chemical equivalent to RLD.13 A generic medicine, is in reality, a replica of the innovative recognized drug product.14 Once the lawful exclusive rights for the original product have been exhausted over time, the organizations that innovated the product are devoid of the elite certified right to create and share out the medicine.15 Other organizations will produce the same adaptation of the identical medication with the equivalent excellence characteristics and can be put up for trade in the marketplace at a much reasonable cost.16 The present exploration involved investigating the quality attributes (Assay determination and Impurity testing), physicochemical test (Physical appearance of tablets, Packaging and labelling of tablets, Tablet diameter and thickness, Weight variation test, Friability Test, Hardness test, Disintegration test, and In vitro dissolution study), and accelerated stability study (1 month, 3 months, and 6 months under temperature 40°C ± 2°C and humidity 75% ± 5% RH) of a branded cefpodoxime proxetil product and five different generic cefpodoxime proxetil products available in the Indian market as per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines (Q2 and Q3) and the United States Pharmacopeia (USP) guidelines. A comparison between the branded product and the generic products has been made in terms of the quality attributes. MATERIALS AND METHODS Instruments The UV-Vis spectroscopic analysis was performed using the double-beam Shimadzu® Ultraviolet-Visible Spectrophotometer (Model: UV-1800, Japan) which was connected with a computer desktop system. The system has a spectral bandwidth of 1 nm with wavelength accuracy of ±0.3 nm and also comprises a pair of matched quartz cells having a 10 mm path length. All weighing of chemicals were carried out using Wensar® high precision electronic balance (Model: PGB100, USA). The sonication was done using the Transonic Digital S (Sonicator), USA. The hardness testing was achieved by using a Monsanto hardness tester (Model: Campbell, USA). Electrolab® disintegration tester USP (Model: ED2L) was employed for studying the disintegration of the tablets. Electrolab® dissolution tester (Model: TDT-08L) was utilized for studying the dissolution of the tablets. Electrolab® Roche Friabilator (Model: EF2) was used for studying the friability of the tablets. The HPLC study was carried out on a Waters® 2695 system with PDA detector 2996 on a reverse-phase Denali C18 column (150 mm × 4.6 mm dimension, 5 μm particle size). The system was equipped with EMPOWERS v.2 software comprising of a 20-μl loop manual rheodyne injector. Chemicals HPLC grade methanol, acetonitrile, glacial acetic acid, and water, as well as analytical grade glycine, sodium chloride, and ammonium acetate, were procured from Himedia Ltd., Mumbai, India. The cefpodoxime proxetil branded product and the five generic cefpodoxime proxetil products were purchased from various Pharmacies across the city limit of Nagpur, Maharashtra, India. Spectral analysis of cefpodoxime proxetil Preparation of stock solution Accurately weighed 25 mg of cefpodoxime proxetil was transferred in a 25 mL volumetric flask and further dissolved in methanol. The final volume of this stock solution (A) was made up to 25 mL with methanol to make 1000 µg/mL concentration. Preparation of dissolution medium pH 3.0 In a 1000 mL volumetric flask, 54.5 g of glycine and 42.6 g of sodium chloride were dissolved in 500 mL of water and 14.2 mL of hydrochloric acid was added with swirling. The content was allowed to cool, further diluted with water, and mixed well. 50 mL of the stock solution was transferred to a volumetric flask and diluted with 900 mL of water to obtain a solution. A pH of 3.0 ± 0.1 of the stock solution was adjusted with 10 N NaOH. Determination of λmax for dissolution From the above stock solution (A), 10 ml of the solution was taken and further diluted to 100 mL with dissolution medium pH 3.0 to make 100 µg/mL concentration (referred to as stock solution (B)). From the above solution, 25 mL content was taken and again diluted to 100 mL with dissolution medium pH 3.0 to make 25 µg/mL concentration (referred to as stock solution (C)). This solution was scanned in the range of 400 nm to 200 nm using a blank and the λmax was determined. Preparation of standard curve of cefpodoxime proxetil The dilutions were prepared from the stock solution (C) where the UV absorbance of 5 µg/mL, 10 µg/mL, 15 µg/mL, 20 µg/mL, and 25 µg/mL solutions were measured on UV-visible spectroscopy at 261 nm for the preparation of standard curve of cefpodoxime proxetil. Evaluation parameters for marketed branded and generic products The test methods given under the USP monograph for cefpodoxime proxetil tablets were employed for the evaluation of all products. Tablet description The color, shape, and size were examined by visual observation. Tablet Diameter and Thickness The diameter and thickness of tablets are important for determining the uniformity of tablet size. Thickness and diameter were measured using Screw Guage Micrometer, which permits accurate measurements and provides information on variation between tablets.17 Weight variation As per the USP, the weight variation test was performed by weighing 20 tablets individually, calculating the average weight, and comparing the individual tablet weight with that of the average. According to the specification outlined in USP, the test for the uniformity of weight for drug products; 130-324 mg is ±7.5% and >324 mg is ±5% of the average pass. Then, the tolerance limit for weight variation was estimated.18 Tablet hardness The tablets require a certain amount of strength and resistance to friability and also to withstand mechanical shocks during handling in manufacture, packaging, and shipping. It is the property of a tablet that is measured to assess its resistance to permanent deformation. The tablet hardness tester device (Monsanto tester) was used to test tablet hardness. The hardness of each branded tablet and generic tablets were measured in unit kg/cm2. Each sample was analyzed in a triplicate manner.19 Disintegration study Disintegration is the breakdown of the tablets into smaller particles or granules when it comes in contact with a solution. The time taken by a tablet to disintegrate was measured in a USP disintegration apparatus. For determining the disintegration time, one tablet was placed in each tube and the basket rack was positioned in 1 L simulated gastric fluid without enzymes at pH 1.2 under a temperature of 37°C ± 1°C. The frequency of cycles per minute was 28 to 32.20 Dissolution study The release rate of cefpodoxime proxetil film-coated tablets was determined by using USP dissolution testing apparatus-II (Paddle type). The dissolution test was performed using 900 mL (0.1 N HCl) volume at a temperature of 37°C ±0.5°C with 100 rpm stirring. An aliquot (10 mL) of the solution was collected from the dissolution vessel, filtered through a 0.45 µm Whatman filter paper, and diluted 2 mL of the filtered content with a dissolution medium at time intervals 10 min, 20 min, and 30 min. Each aliquot was replaced with a fresh dissolution medium to maintain the sink conditions. The absorbance of diluted solutions was measured at 261 nm.21 Accelerated stability studies The branded cefpodoxime proxetil product and generic cefpodoxime proxetil products were placed inside a polyvinyl chloride (PVC) container and aluminium foil was wrapped over it. The stability studies were performed under the accelerated conditions of temperature (40°C ± 2°C) and moisture (75% ± 5% RH) for the duration of 1 month, 3 months, and 6 months.22 Assay preparation Preparation of mobile solution A filtered and degassed mixture of 0.02 M ammonium acetate and acetonitrile was prepared in the ratio of 6:4. Preparation of diluent A filtered and degassed mixture of water and acetonitrile was prepared in the ratio of 6:4. Standard preparation 25 mg of cefpodoxime proxetil was weighed accurately and transferred in a 50 ml of volumetric flask. The content was dissolved in 5 mL of methanol, further diluted with the diluent to the desired volume and mixed. 5 mL of the above solution was transferred to 100 mL of the volumetric flask, further diluted with the diluent to the desired volume, mixed, and passed through a filter having 0.45-μm porosity. Assay procedure Finely powdered 20 tablets were accurately weighed and a portion of the powder equivalent to 50 mg of cefpodoxime proxetil was transferred to a 100 mL volumetric flask. The content was dissolved in 10 mL of methanol, further diluted with the diluent to the desired volume and mixed. 5 mL of the above solution was transferred to 100 mL of the volumetric flask, further diluted with the diluent to the desired volume, mixed, and passed through a filter having 0.45-μm porosity. Chromatographic system suitability The liquid chromatography system was equipped with a 235 nm detector having a 250 mm × 4.6 mm dimension packing column. The flow rate was 2 mL/min at ambient temperature (30°C). The standard and the test preparations were chromatographed and the peak responses were recorded. About 0.9 for S-epimer cefpodoxime proxetil and 1.0 for R-epimer cefpodoxime proxetil are the relative retention periods. The resolution, R, is not less than 2.5 between the S-epimer cefpodoxime proxetil and the R-epimer cefpodoxime proxetil. No more than 1.5 is the cefpodoxime proxetil R-epimer tailing factor and no more than 1.0 percent is the relative standard deviation measured for replication injections from the cefpodoxime proxetil S-epimer and cefpodoxime proxetil R-epimer regions. Not more than 1.0% for any peak at relative retention times is desirable. Not more than 0.5% of any other individual impurity and not more than 6.0% of total impurities is desired. Not more than 3.0% of any peak at a relative retention time and individual peaks having relative retention times not higher than 2.0 is desired. The percentage of each impurity in the portion of tablets was calculated by the formula: % purity = 2000 (CP/W) (rU/rS) C is the concentration (in mg per mL) of cefpodoxime proxetil in the standard preparation; P is the designated potency (in μg per mg) of cefpodoxim in USP cefpodoxime proxetil; W is the weight (in mg) of cefpodoxime proxetil taken to prepare the assay preparation; rU and rS are the peak response numbers obtained from the preparation of the assay and the standard preparation of cefpodoxime proxetil S-epimer and cefpodoxime proxetil R-epimer, respectively. Impurity profiling Preparation of Solution-A Solution A was prepared by taking filtered and degassed 0.02 M ammonium acetate. Preparation of Solution-B Solution A was prepared by taking filtered and degassed acetonitrile. Preparation of the mobile phase The mobile phase of the chromatographic system involved the utilization of variable mixtures of solution-A and solution-B. Preparation of diluent A filtered and degassed mixture of water and acetonitrile was prepared in the ratio of 2:1. System suitability solution A quantity of cefpodoxime proxetil was dissolved in the diluent to obtain a solution containing 10 µg/mL. A volume of methanol not exceeding 10% of the total volume in the final solution may be used to facilitate dissolution. Standard solution 50 mg of cefpodoxime proxetil was accurately weighed and transferred to a 50 ml of volumetric flask. The content was dissolved in 5 mL of methanol, using sonication if necessary, diluted with diluent to the desired volume, and mixed well. The solution was injected promptly and analyzed within 24 hours when stored at 8ºC temperature. Test solution 50 mg of cefpodoxime proxetil was accurately weighed and transferred to a 50 ml of volumetric flask. The content was dissolved in 10 mL of methanol, using sonication if necessary, diluted with diluent to the desired volume, and mixed well. The solution was injected promptly and analyzed within 24 hours when stored at 8ºC temperature. Chromatographic system The liquid chromatography system was equipped with a 261 nm detector having 250 mm × 4.6 mm dimension packing column (5 μm particle size). The flow rate was 2 mL/min. The standard and the test preparations were chromatographed and the peak responses were recorded according to the following gradient procedure: 0 min [equilibration (10 min), solution A (90%), solution B (10%)]; 0-10 min [linear gradient, solution A (90%→68%), solution B (10%→32%)]; 10-40 min [isocratic, solution A (68%), solution B (32%)]; 40-80 min [linear gradient, solution A (68%→50%), solution B (32%→50%)]; 80-85 min [isocratic, solution A (50%), solution B (50%)]; 85-90 [linear gradient, solution A (50%→25%), solution B (50%→75%)]; 90-95 [isocratic, solution A (25%), solution B (75%)]; 95-100 [linear gradient, solution A (25%→90%), solution B (75%→10%)]. The tailing factor and the relative standard deviation for replicate injection should not more than 2.0. Assay Procedure 20 μL of test solution was injected into the chromatography system and the chromatogram was recorded. All the measurements were done by the peak areas. The percentage of each impurity in the portion of cefpodoxime proxetil was obtained from the formula: % Impurity = 100 (ri / rs) Where ri is the peak area for each impurity and rs is the sum of the areas of all the peaks. RESULTS AND DISCUSSION Spectral analysis of cefpodoxime proxetil lmax determination The prominent wavelengths at 261.40 nm, 213.20 nm, and 208.80 nm were predominantly seen (Figure 1). An absorption maximum was found to be at 261 nm. Hence, 261 nm was selected as the lmax for further studies.   Prepared standard curve Cefpodoxime proxetil showed maximum absorption at wavelength 261 nm in glycine medium. The calibration curve was prepared by taking the UV absorption of solutions at 5-25 µg/mL concentration (Table 1). The details of calibration curve include y = 0.0294x + 0.0027 with R2 = 0.9994 (Figure 2). Evaluation parameters for marketed branded and generic products Tablet description The colour, shape, and size were examined by visual observation where the factory manufactured cefpodoxime proxetil products (1×10 tablets in blister packaging) had the same attributes as per pharmacopoeia recommendation and the manufacturer’s claim (Table 2). The visual appearance of the product is an important factor for patient compliance. The branded product was a white colour, uncoated, round shape tablet whereas the generic products were white color uncoated, caplet shape tablet (G1 product), pale yellow color, uncoated, round shape tablet (G2 product), White color, uncoated, round shape tablet (G4 product), and pale orange color, uncoated, round shape tablet (G3 and G5 products). After performing the accelerated stability studies, no changes were observed. The physical appearances and the physical stability of the products were found to be very uniform. Tablet Diameter and Thickness All the generics and the branded cefpodoxime proxetil tablets were found to be within the acceptable range of thickness (0.2-0.36 mm), length (11-14 mm), width (0.7 mm), and diameter (11 mm). The thickness and the diameter uniformity of the tablets are a necessary factor for the consumer requirements and also important for better packaging. No changes occurred in branded and generic products after the stability study (Table 3). Weight variation The average weight of 20 tablets was calculated and the individual tablet weight was compared to the average weight. The weight variation test is required to assure that the drug content in each unit dose is distributed in a narrow range around the label strength. If the drug substance forms the greater part of the oral solid dosage form, the weight variation reflects variation in the content of the active ingredient. The results indicate that five generics (G1-G5) products and one branded (B1) product possess acceptable uniformity of weight as per the USP limit. After the stability shelf period, there were no prominent changes observed (Table 4). Tablet hardness The hardness test showed the ability of tablets to withstand pressure or stress during handling, packaging, and transportation. The results indicated that the branded tablet product B1 had the highest strength; i.e. 1.96 kg/cm2 as compared to all the five generic products (G1-G5). The generic products G2 and G5 retained the hardness across the 6 months accelerated stability period while branded tablet product B1 as well as generics G1, G3, and G4 showed an increase in the hardness value after 6 months of the accelerated stability period. The reason for the augmentation of hardness value (breaking strength) may be due to the specific excipients used, moisture absorption, and temperature. Generic products G1, G2, G4, and G5 showed a hardness value of less than 2 kg/cm2, after accelerated stability studies which are not a good sign of compressibility and therefore the batches failed. Only branded tablet product B1 and generic product G3 showed a good sign for tablets dosage form (Table 5). Friability study 6 tablets of each product were used for the friability test. The branded product B1 and generic tablet products (G1-G5) demonstrated friability of not more than 1% in both initial study and for 6 months accelerated stability study. The friability values were observed to be decreased over time which may be due to retaining product hardness owing to moisture absorption and temperature. All these products passed the friability test (Table 6). Disintegration study The disintegration test is the most important step in the release of drugs from the immediate release dosage forms. The rate of disintegration is directly proportional to the rate of dissolution. The rate of disintegration is straightly influenced by the rate of influx of water into the tablets and also depends on the porosity of the tablets. The above results indicated that the branded product B1 and three generic products G2, G3, and G5 passed the disintegration test at initial levels but branded product B1 and two generic products G2 and G3 failed after the accelerated stability period according to the USP guidelines (Table 7). It was observed that the G1 generic product displayed out of limit at both initial and accelerated stability levels. Generic products G4 and G5 presented variable results after the 3rd month and 6th month of accelerated stability period which may be due to tablet to tablet variation. Dissolution study The dissolution test of the drug from a solid dosage form is important for drug bioavailability. In the dissolution study, 6 tablets of one branded product B1 and generic products (G1-G5) were tested for the % drug release, according to the method described for the cefpodoxime proxetil tablet in the USP monograph. It is stated in the monograph that the amount of cefpodoxime proxetil released within 30 minutes should not less than 70% of the stated amount. The in vitro dissolution rate is used to simulate the bioequivalency of different formulation of generics (cefpodoxime proxetil tablet) concerning the branded product. Initial dissolution study was performed and the results suggested that the branded product B1, generic products G1 and G3 passed the test while generic products G4 and G5 were at borderline; i.e.  ~71% while the generic batch G2 failed in the dissolution testing. The reason may be due to the altered physical characteristics and content non-uniformity. The 1st month accelerated stability samples were analyzed for the dissolution test. Branded product B1 passes with best results; i.e. 86.89% after 30 min. The generic products G1 and G3 were observed to release the drugs at 73.26% and 72.68%, respectively (Table 8). At the same time, generic product G5 passes the critical lower limit of drug release; i.e. 71.8%. However, generic products G2 (59.74%) and G4 (58.71%) failed to show desired dissolution results. A great effect of temperature and humidity may be the plausible reason. Generic product GS4 passed the initial study with a close limit but after 1-month stability, a decrease in the drug dissolution was perceived due to poor production batch. The 3rd month accelerated stability results presented that the branded product B1 passed the dissolution test while the generic products G1, G3, and G5 demonstrated the % drug release close to the limit; i.e. 70%. In contrast to it, two generic products G2 and G4 failed to meet the desired limit. According to Biopharmaceutics Classification System (BCS), cefpodoxime proxetil comes under Class-II (high permeability, low solubility) where it is expected that the dosage form should release >70% drug and as per specification (30 min). There was a decrease in the drug release for the branded product B1 and the generic products G1, G3, and G5, they closely met the required limit. The generic products G2 and G4 failed. The 6th month accelerated stability results concluded that the branded product B1, as well as generic products (GS1-GS5), failed the dissolution study. Only the branded product B1 complied with the dissolution test until 3rd-month stability while generic products G1, G3, and G5 passed at a boundary level till the 3rd month stability period. Generics G2 and G4 were regarded as failed products during the initial study to 6 months accelerated stability study (40°C ± 2°C and 75°C ± 5% RH). Assay of products All the five generics products and one branded product were assayed for the drug content according to the isocratic mode based HPLC method (0.02 M ammonium acetate and acetonitrile in the ratio of 6:4) outlined in the individual drug monographs of the USP (acceptance limit: 90-110%). The content of cefpodoxime proxetil equivalent to cefpodoxime was calculated from the peak areas of the chromatograms of test and reference standard solutions (Table 9). Englishhttp://ijcrr.com/abstract.php?article_id=3635http://ijcrr.com/article_html.php?did=3635 Brook RH, Williams KN, Avery AD. Quality assurance today and tomorrow: forecast for the future. Annal Inter Med 1976;85(6):809-17. Kessner DM. Quality assessment and assurance: early signs of cognitive dissonance. New Eng J Med 1978;298(7):381-6. Farris KB, Kirking DM. Assessing the quality of pharmaceutical care II. Application of concepts of quality assessment from medical care. Annal Pharmacother 1993;27(2):215-23. Frampton JE, Brogden RN, Langtry HD, Buckley MM. Cefpodoxime proxetil. Drugs. 1992;44(5):889-917. Chugh K, Agrawal S. Cefpodoxime: pharmacokinetics and therapeutic uses. Indian J Pediatr 2003;70(3):227-31. Todd WM. Cefpodoxime proxetil: a comprehensive review. Int J Antimicrob Agent 1994;4(1):37-62. Fulton B, Perry CM. Cefpodoxime proxetil. Paediatr Drugs 2001;3(2):137-58. Borin MT. A review of the pharmacokinetics of cefpodoxime proxetil. Drugs 1991;42(3):13-21. Chocas EC, Paap CM, Godley PJ. Cefpodoxime proxetil: a new, broad-spectrum, oral cephalosporin. Annal Pharmacother 1993;27(11):1369-77. Ahire K, Shukla MA, Gattani MA, Singh VI, Singh ME. A survey-based study in the current scenario of generic and branded medicines. Int J Pharm Pharm Sci 2013;5(3):705-11. Bera A, Mukherjee A. The importance of generic drugs in India. Int J Pharm Chem Biol Sci 2012;2(4):575-87. Dadhich A, Upadhyaya M. A review: exploring branded generic drugs by Indian pharmaceutical multinational companies as a new prospect. Pharmacophore 2011;2(6):271-5. Greene JA. Generic: The unbranding of modern medicine. JHU Press; 2014. Van der Merwe SE, Bredenkamp J. Originator and generic medicine: pricing and market share. Int J Pharm Healthc Mark 2013;7(2):104-19. Dylst P, Vulto A, Godman B, Simoens S. Generic medicines: solutions for a sustainable drug market? Appl Health Econ Health Policy 2013;11(5):437-43. Simoens S. International comparison of generic medicine prices. Curr Med Res Opin 2007;23(11):2647-54. Mahajan NM, Pardeshi A, Mahapatra DK, Darode A, Dumore NG. Hypromellose and Carbomer induce bioadhesion of Acyclovir tablet to vaginal mucosa. Indo Am J Pharm Res 2017;7(12):1108-18. Mahajan NM, Wadhwane P, Mahapatra DK. Rational designing of sustained-release matrix formulation of etodolac employing hypromellose, carbomer, eudragit and povidone. Int J Pharm Pharm Sci 2017;9(12):92-7. Patil MD, Mahapatra DK, Dangre PV. Formulation and in-vitro evaluation of once-daily sustained release matrix tablet of nifedipine using rate retardant polymers. Inventi Impact Pharm Tech 2016;4:190-6. Gangane PS, Kadam MM, Mahapatra DK, Mahajan NM, Mahajan UN. Design and Formulating Gliclazide Solid Dispersion Immediate Release Layer and Metformin Sustained Release Layer in Bilayer Tablet for the effective Postprandial Management of Diabetes Mellitus. Int J Pharm Sci Res 2018;9(9):3743-56. Gangane PS, Ghughuskar SH, Mahapatra DK, Mahajan NM. Evaluating the Role of Celosia argentea Powder and Fenugreek Seed Mucilage as Natural Super-Disintegrating Agents in Gliclazide Fast Disintegrating Tablets. Int J Curr Res Rev 2020;12(17):101-8. Godbole MD, Mahapatra DK, Khode PD. Fabrication and characterization of edible jelly formulation of stevioside: a nutraceutical or OTC aid for diabetic patients. Inventi Rapid: Nutraceut 2017;2:1-9.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareCapsicum Plant Leaves Disease Detection Using Convolution Neural Networks English185190Himanshu PantEnglish Manoj Chandra LohaniEnglish Janmejay PantEnglish Prachi PetshaliEnglishEnglishAccuracy, Capsicum, Computer vision, Classification, Convolutional neural networks, Leaf diseaseIndia is a rich country having a huge amount of natural and human resources.  Almost 70% of the Indian economy is based upon the agriculture and horticulture sectors.1 This agriculture sector contains numerous ingredients.  Plants are one of the significant factors among them. Fit and healthy plants provide health.2 However, plant diseases are bullying the maintenance of this natural resource. Production and economic losses increases due to plant diseases in the agriculture and forestry sector. Capsicum (Capsicum annum) is the most used food crop in the world. Bell pepper bacterial spot (a fungal disease in capsicum) has caused a momentous economic and commercial loss and just by eliminating 20% of this bacterial infection, the farmers may benefit from an extraordinary profit. Therefore, early detection and identification of capsicum bacterial diseases play the utmost important role to take timely measures for the quality of the plant. There are numerous ways to perceive plant pathologies. Some plants have no visible symptoms of diseases associated with them or diseases may appear only when too late to cure. So it is necessary to perform a classy analysis of the plant diseases in the laboratories by the experts using powerful microscopes or employing different electromagnetic spectrum that is not visible to humans.3 Early-stage identification and classification of crop plant disease is a major problem in agricultural practices. Farmers bear a great loss in the economy every year due to infection in the crops and plants cultivation.4 Therefore, fast, efficient, less expensive and accurate diagnosis is required to prevent bacterial infection in the capsicum. These accurate and automated methods may help to inhibit the loss of crops, improves the quality of the product and helps in the economic growth of the country as well.5 This research study focuses on disease detection and classification of capsicum plant (Bell pepper) based on the bacterial symptoms of the diseases that show unhealthy signs on the leaves of the plant. Capsicum plants are dumpy shrub in nature with woody trunks. These plants nurture with colourful fruits. Capsicum leaves disease detection model to perform various steps. The primary step is to obtain a feature vector and another important step is to classify the feature vectors of the given input data. Mostly, the identification of the disease is guessed first by the human’s visualization. Experts of the domain may be efficiently recognizing the disease present in the particular plant but in most cases, there is no domain professional present in the particular area to give feedback on the disease to the farmers after data analysis on the plant. Hence farmers are required a quick, cheap, accurate and automatic technique to detect the plant disease efficiently.6 Bacterial infection and fungus are the main reason for capsicum plant diseases. Numerous diseases may appear in the capsicum plant. This article considered the images of bacterially infected leaves along with the healthy leaves (HL) images. The Xanthomonas campestris (black rot) is the primary bacterial species of the capsicum plants. Therefore, digital Image-based plant disease identification and classification models have been developed in the literature for plants.7 Computer vision, artificial intelligence, machine learning and deep learning techniques are more popular research areas for object detection and classification from images, text and videos.8 Digital image processing technique minimized the inaccurate manual disease detection and improve the accuracy, feasibility and efficiency to predict the disease on a time from a plant.9 This paper leverages the identification and classification of the disease and healthy images from the capsicum plant using recent advancement in computer vision with the help of a convolutional neural network (CNN). The first aim of the current research was the collection of a sufficient capsicum image dataset from the field and then classifies the images into two categories (Bacterial spot images and healthy images). The overview of the proposed system architecture of capsicum plant disease identification and classification is shown in figure-1. Nowadays, researchers pay attention to convolutional neural network techniques due to their great performance in image classification. The advantage of the convolutional neural network is that it avoids extraction of complex hand-crafted features unlike traditional machine learning techniques and provides end-to-end learning.10 For image classification and accurate prediction, the CNN model provides a relationship between layers and spatial information of the image.11 Along this line, there are limited works on capsicum leaves disorders identification and classification using CNN. Some author inspected the capability of the deep CNN technique for the classification of numerous rice diseases. A total of 597 images has been considered and used the CNN model with three convolution layers, three stochastic pooling layers and a softmax layer at the end. The classification accuracy of 91.620% has been reported. After augmentation of the dataset, the model achieved 94.972%% accuracy. A total of 418 images belonging to the training dataset and 179 images belong to the test dataset out of 597. MATERIALS AND METHODS In this section, the authors performed the various steps and operations on the capsicum plant image dataset. To identify and classify the infected capsicum leaves from the huge dataset, numerous operations can be performed as shown in figure-2. These steps describe the complete architecture from image acquisition to image disease classification through which farmers can easily predict the healthy plant from the mixture of infected and bacterial image sets.  The methodology followed is discussed in detail. Dataset Descriptions In this article, a huge amount of the capsicum plant’s leaf images are required to identify and classify the diseases associated with it. The images are captured from the different agricultural fields and various gardens in Nainital, a district of Uttarakhand, India. These datasets are required for image disease classification research during the training, testing and validation phase. The images are acquired using numerous types of standard cameras, captured from both front and back end leaves. The Apple iPhone 8, the Samsung Galaxy M3 and Redmi note 5 pro cameras are used for image acquisition. It contains a collection of images taken in a different environment. A dataset containing 597 capsicums leaves of two image classes including bacterially infected leaves and healthy leaves. The sample images of both categories are shown in the figure-3. Data pre-processing The capsicum plant images are initially unlabelled and not in annotated form. The labelling process of the images is processed by their filenames with the word “HL” and “BS”. Where HL belongs to healthy capsicum leaves while BS represents bacterial spot capsicum leaves. The file naming convention is in the form of HL.1.jpg, Hl.2.jpg, BS.20.jpg, BS.23.jpg etc. Primarily RGB coloured images are taken as a sample. In the captured images dataset, some images are in landscape format, some are in portrait format, and the remaining images are in square format.12 To classify and identify the disease from the capsicum images standardized photo size are required. All the images must be reshaped before modelling so that the size and shape of all images would be the same.13 Keras image processing API is applied to achieve this standardization by uploading all images to the ImageDataGenerator class and reshapes them to 200×200 square photos. The labels of the images are also determined based on the filenames. After standardizations of all images into 200 x 200 sizes, the next objective is to pre-process images into standard directories using flow_from_directory() application programming interface (API). This API divides all data into separate train/ and test/ directories, and under each directory to have a subdirectory for each class, e.g. a train/HL/ and a train/BS/ subdirectories and the same procedure is applied for test images. Images are then organized under the subdirectories. EXPERIMENTAL SETUP AND PERFORMANCE ANALYSIS The experiment was designed to appraise the performance of the baseline convolution neural network model for the capsicum image dataset to classify and identify whether a specific image is infected or not.  This baseline CNN model is established to compare the model performance of other CNN models. The general architectural principles of the VGG models are used for the experiment.14This architecture involves assembling convolutional layers with small 3×3 filters followed by a max-pooling layer. A combination of convolutional layers and pooling layers form a block, and when the number of filters in each block is increased with the depth of the network, these blocks can be repeated. Padding is used on the convolutional layers to ensure the height and width shapes of the output feature maps matches the inputs. The authors applied this VGG architecture to the healthy capsicum plant image and bacterial spot image problem and then compare a developed model with this architecture using the first three blocks. In this designed architecture each layer is used the ReLU activation function and the he_uniform weight initialization. The designed baseline CNN model is fitted with stochastic gradient descent and start with a conservative learning rate of 0.001 and a momentum of 0.9.15The proposed model can be fit using train iterator and dataset validation is done by test iterator. The number of steps for the train and test iterators concerning one epoch must be specified for the model fitting and it can be calculated by dividing the total number of capsicum images (both HL and BS) in the train and test directories by the batch size of 64. Once the model is fitted, the performance and quality can be evaluated on the unseen testing dataset. The performance is measured in terms of accuracy.  The model accuracy of each block VGG model can be calculated by increasing the convolutional layer and pooling layer.16 One Block baseline Visual Geometry Group (VGG) model performance The one-block baseline Visual Geometry Group (VGG) model has only one convolutional layer with 32 filters followed by a max-pooling layer. The images are in the form of matrices and have to convert two-dimensional matrixes into dimensional form to create a neural network. For this purpose, the flattening technique is used.  The dense layer is used to create a fully connected neural network by which each input node is connected to all output nodes. The classification accuracy of one block VGG model to classify the capsicum image dataset to predict whether the given image is infected or not and the cross-entropy loss on the test and training dataset at the end of each epoch is shown in figure-4. The one-block VGG baseline model gives 84.358% prediction accuracy. Two Block baseline VGG model performance The working procedure of the two blocks VGG model is almost same but in two-block VGG model. It is the extend version of one block VGG model by adding a second block (convolutional layer and max pooling layer) with 64 filters (just double from the first convolutional layer filter) as shown in figure-5. After compiling, the model had achieved a small improvement in the performance of accuracy and prediction from 84.358% to 84.916%. Three-Block baseline VGG model performance The architecture of three-block VGG model extends the two block VGG model by adding a third block (additional convolutional layer and max pooling layer) with 128 filters (just double from second convolutional layer filter). Repeating the compilation process using these three layers, model achieved a great improvement in the performance of accuracy and prediction from 84.916% to 91.620%. The classification accuracy and cross-entropy loss with respect to epoch is shown in figure-6. Performance measurement after capsicum image data augmentation: Image data augmentation techniques are used to modify the training images in the dataset. It is an approach that allows us to increase the existing training dataset without collecting new. The augmentation process involves cropping the images, padding, and horizontal flipping of the images on the existing dataset to create a new one. Small changes in the input data of healthy capsicum images and bacterial spot capsicum images might generate a huge amount of new data by applying translation, rotation, small shifts and horizontal flips. The augmentation should be used only for training dataset using ImageDataGenerator in Keras. Images are augmented in the training dataset with small shifting in random horizontal and vertical shifts and used random horizontal flips which create a mirror image of a photo. When this baseline three-block VGG model is compiled with augmentation, the model performance is improved. It is observed that the model lift an excellent performance of about 10% from the baseline one block VGG performance which was 84.358%. Now after augmented model predicts perfectly whether a given image healthy or infected with 95.531% accuracy on the given dataset. The classification accuracy and cross-entropy loss concerning each epoch are shown in figure 7. To describe the performance of the developed classification VGG model with three layers after augmentation, the confusion matrix is used for which the true values are known on a set of test data. The performance of the classification algorithms in deep learning and machine learning model is summarized by the confusion matrix. This matrix allows the visualizing of the performance of the developed CNN model in the matrix form. It is also used for decision making for selecting the right observations and can help to reduce errors. This binary classification problem is used to categorized and identify the capsicum plant disease. The confusion matrix and the classification report for three block baseline VGG model after augmentation with 95.531% accuracy are shown in the table-1. CONCLUSION A convolutional neural network is performed to detect, identify and classify the capsicum plant disease in this research. Limited research has been done on capsicum plant disease classification and automation. In this paper different baseline, Visual Geometry Group (VGG) model with one two and three blocks of CNN was explored to classify the capsicum plant disease. Moreover, the designed model had applied augmentation on the training dataset to improve efficiency and accuracy. The proposed model can classify infected or healthy plants with a classification accuracy of 94.972%. The dataset is split into 80:20 ratios of training and testing respectively. This research article reconnoitred three different improvements to the baseline model. The performance of the different results can be summarized as given table-2. It is shown that the result may be improved when the three-block augmentation approaches further increases the number of training epochs. FUTURE SCOPE The performance of the proposed model can be further improved with a large dataset of capsicum plant images with both healthy and infected leaves. The CNN model is trained using the images captured from the natural environment by cameras. The purposed model has achieved 95.531% accuracy of classification and identification of the disease. This accuracy may be increased by applying the transfer learning model on pre-trained model like VGG 16, VGG 19, Alexnet, etc. In the future authors will apply computer vision techniques like image segmentation and object detection on plant leaves. Englishhttp://ijcrr.com/abstract.php?article_id=3636http://ijcrr.com/article_html.php?did=3636 India economic survey 2018: Farmers gain as agriculture mechanisation speeds up, but more R&D needed. The Financial Express. 29 January 2018. Retrieved 8 January 2019 Phadikar S, Sil J, Das AK. Rice diseases classification using feature selection and rule generation techniques. Comp Electr Agricult2013;90:76–85. Barbedo JGA. A review of the main challenges in automatic plant disease identification based on visible range images. Bio Syst Engi 2016;144:52-60. Al Bashis D. A Framework for Detection and Classification of Plant Leaf and Stem Diseases. IEEE International Conference on Signal and Image Processing (ICSIP), Chennai 2010, pp.113-118. Sannakki SS. Diagnosis and    Classification of    Grape    Leaf    Diseases using    Neural    Networks. IEEE proceedings of 4ICCCNT, 2013. Smith JS. An image-processing based algorithm to automatically identify plant disease visual symptoms. Biosyst Eng 2009;102:9–21. Liu K. Identification method of rice leaf blast using multilayer perception neural network. Transactions Chinese Soc Agricult Engi 2009;25(S2). Barbedo J. A review on the main challenges in automatic plant disease identification based on visible range images. Biosyst Eng 2016;144:52-60. Asefpour K. An artificial neural network approach to identify fungal diseases of cucumber (Cucumissativus L.) Plants using digital image processing. Arch Phytopathol Plant Protect 2013; 46(13):1580-1588. Lu. Identification of rice diseases using deep convolutional neural networks. Neurocomputing 2017;267:378–384. Arel. Deep Machine Learning - A New Frontier in Artificial Intelligence. IEEE Computational Intelligence Magazine. 2010;5(4):13-18. Dhakate M. Diagnosis of Pomegranate Plant Diseases using Neural Network. IEEE 5th National Conference on Computer Vision, Pattern Recognition, Image Processing and Graphics (NCVPRIPG), Patna2015. Nam Y. A representation and matching method for shape-based leaf image retrieval. J KIISE: Softw Appl 2005;32(11):1013-1021. Simonyan K. Very deep convolutional networks for large-scale image recognition. 2015. Available https://arxiv.org/cs/1409.1556 Wang L. Training deeper convolutional networks with deep supervision,” 20 Available https://arxiv.org/cs/1505.02496 Leaf recognition algorithm for plant classification using probabilistic neural network. Available http://flavia.sourceforge.net/

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareEffect of Clinical Pilates on Core Muscle Strength, Balance and Posture Control of a Recurrent Lacunar Stroke Patient – A Single Case Study English191195Surya VishnuramEnglish Kumaresan AbathsagayamEnglish Prathap SuganthirababuEnglishEnglish Pilates-based Exercises, Stroke, Balance, Posturehttp://ijcrr.com/abstract.php?article_id=3637http://ijcrr.com/article_html.php?did=3637

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareA Succinct Analysis for Deep Learning in Deep Vision and its Applications English196203Preethi NEnglish W. Jai SinghEnglishEnglish ConvNet, Stochastic Hopfield network, Generative graphical model, Social media analysis, Data processing, Deep Learning INTRODUCTION Deep vision is an integrative intelligent arena that oversees in what way deep tin be ended to build raised equal appreciation since deep pictures or chronicles. Since the viewpoint of building, it attempts to deep endeavours that the hominidgraphic structure tin do.1-3 Deep learning grants machine models of various taking care of layers to be told and address knowledge with various degrees of reflection addressing however the neural structure sees and grasps multimodal data, as needs are unquestionably getting included structures of colossal extension data. What perceived deep vision from the basic field of cutting edge picture getting ready around then remained a yearning toward isolate three-D structure after pictures by the area of accomplishing complete act thoughtful. Training during the 1970s moulded the initial basic aimed at countless the deep vision figuring that happen nowadays, as well as withdrawal of limits as of pictures, naming of appearances, non-polyhedral and polyhedral illustrating, depiction of articles as inter associates of humbler constructions, visual stream, and development approximation.4 The desire to form a structure that reproduces the human brain drove the elemental progression of neural frameworks. In 1943, McCulloch and Pitts5 endeavoured toward realizing however the neural structure might create uncommonly complicated models by victimization organized developed cells, known as neurons. The McCulloch and Pitts prototypical of a vegetative cell, known as an MCP prototypical, has created a vital duty toward the advance of pretending neural frameworks. Deep learning has drove fantastic strolls in briefing of computer vision complications, let&#39;s say, entity identification, movement following, activity acknowledgment, human skill estimation, and linguistics division.6-15 During this arrangement, we&#39;ll minimalistically evaluation the core progressions in deep learning models besides figuring for computer vision bids during the extraordinary circumstance, Three of the foremost vast sorts of deep learning prototypical with relevancy their significance in visual kind, that is, ConvNet, the "Boltzmann special" composed with Deep Belief Networks (DBNs) and Deep Boltzmann Machines (DBMs) and Stacked (Denoising) Autoencoders. This paper consists of surveys. In Section two, Deep Learning Approaches and Changes are audited. In section three, we tend to portray the uses of Applications of deep learning in deep vision. In Section four,  Deep learning challenges and directions are mentioned. At long last, Section five completes the paper with an outline of the results. DEEP LEARNING APPROACHES AND CHANGES ConvNet ConvNetvictimization the boner incline and accomplishing usually astounding ends up in a briefing of model affirmation tasks.16-18 ConvNet is spurred by the visual scheme sassembled, and expressly through its prototype planned in.19 The principle machine prototype dependent on procurable systems among neurons and logically created changes out of the copy are create inNeocognitron20, which delineates that once neurons through comparative limits are suitable on areas of the past pane by completely dissimilar zones, a method of change of location in vacillation is secured. A CNN incorporates 3 commonplace forms of neural layers, to be express, (a) ConvNet Phase, (b) combining layers, and (c) whole Merging layers. Variety of phase settle for substitute activity. (Figure1) shows a CNN building for a piece characteristic proof in copy mission. Each phase of a ConvNet changes info capacity to a yield capacity of somatic cell commencement, over the long-term agitative the last whole connected layers, achieving a mapping of the info to a 1D embody vector. CNN&#39;s are exceptionally productive in deep vision applications, to Illustrate, face affirmation, object revelation, driving idea to apply autonomy vehicles. (a) ConvNetPhase: Conv Net Phase, a Conv Net uses numerous bits to rotate the entire copy similarly to the midway component maps, delivering diverse component plots. As the advantages of the complication action, a couple of task 21,22have planned it as an additional for totally associated phase through the ultimate objective of achieving speedier learning events. (b) Combining Phase: Combining Phaseremainin charge of diminishing the three-dimensional estimations of the data capacity aimed at the following difficult phase. The combining phase doesn&#39;t impact the significance estimation of the capacity. This action achieved through this lateral indicated sub examining or down testing, by way of the diminishing of scope prompts a concurrent lack of evidence. In any case, mishap may be helpfully aimed at the framework because the decline in dimensions prompts fewer figures overhead used for the future phases of the framework, also besides that kills overflow. Typical combing and max combining are the maxima generally applied procedures. Distinct academic examination of max combining and ordinary combining shows is assumed, and exhibited the most extreme combing could provoke speedier intermixing, select dominating constant structures, and upgrade hypothesis.23,24 Different various assortments of the combining phase of the composition, inspired through changed motivation sand helping indisputable necessities. c) Completely Linked Layers. Following one or two complexity and combining phase, large phase intuition within the neural framework is achieved by strategies for wholly associated phase. Neurons in an exceedingly wholly associated phase take complete relationship with completely sanctionative within past phase, as per their tag recommends.  In the beginning, this point forward is non-commissioned with a structured growth followed by an inclination offset. wholly associated phase ineluctably change the 2nd article plots into a 1D embrace path. The result and path moreover may well remain addressed advancing to selected ranging orders used for game set up or might remain thought-about as per a path for additional method.25,26 ConvNet structure uses three strong attention: (a) area responsive range, (b) fixed burdens, and (c) Structural subsample. Taking into account close by responsive range, every component in a convolutional layer gets ideas of neighbouring components having a spot with the last layer. Thusly neurons are fit for removing simple visual features, for instance, boundaries or joints. The above-mentioned selections are formerly joined through the following complex phases acknowledge complex solicitation structures. Plus, straight forward part locators, that are helpful on a touch of a picture, are presumptively attending to be vital over the complete figure is ample through the chance of joint burdens. The joint burdens goal is a good deal of items toward own vague burdens. Decidedly, the elements of a convolutional layer sifted through sphere. Entireelementsset up supply a comparable game plan of burdens. Hence, every plane is in danger for building a selected part. The yields of the sphere are known as structural plots. Every convolutional layer involves one or two of planes,thus varied phase maps is created at every region. throughout the advancement on the part plot, these complete figures are checked through elements that are taken care of at staring at regions within the phase maps. This improvement is like a convolution action, trailed by an extra substance tendency period and sigmoid edge:   Here ???? states the  importance of the convolutional layer, the weight cross-section is denoted by W, and the inclination term is denoted by b. Completely associated neural frameworks, burden lattice is filled, that is, interfaces all commitment to the respective element through totally unlike burdens. For ConvNet, W is the burden system which is too little visible of the chance of tied burdens. Consequently, the kind of W has Here w is matrices taking comparative estimations with the units&#39; open fields. victimisation associate inadequate weight matrix diminishes the quantity of the framework&#39;s tunable parameters and thus grows its theory limit. increasing W with layer inputs takes once convolving the commitment with w, which might be seen as a trainable channel. If the commitment to ????−1convolutional layer is of estimation ????×???? and therefore the responsive field of units at a particular plane of convolutional layer ???? is of estimation ????×????, by then the created element guide is going to be a structure of estimations (????−????+1)×(????−????+1). Precisely, the phase of feature plot at(????,????) tract is going to be Here the scalar b. Using (2) and (3) consecutively for entirely (????,????) spots of knowledge, the half plot for the relating plane is made. The difficulties which will develop by preparing of CNNs needs to fix with the tremendous variety of strictures which has got acknowledged, that can incite the effort by overfitting. to the present finish, frameworks, maybe, random pooling, dropout, and information development are planned. additionally, CNNs are as typically as attainable assumed to pre-processing, which is, a technique that instates the framework by pre-processing parameters as hostile without aim set ones. Pretraining will enliven the educational technique and update the hypothesis limit of the framework. All things thought-about, CNNs were looked as if it would primarily trump commonplace AI methods is a very wide extent of Deep vision and model affirmation errands32, samples are given in Section3. Its splendid show got alongside the relative simplicity in preparing are the essential reasons that specify the unfathomable arrive of their predominance in the course of the most recent number of years. Generative graphical model and Stochastic Hopfield network with hidden units Deep Belief Networks and Deep Boltzmann Machines are Deep learning models that belong to the "Boltzmann family," as they utilize the Restricted Boltzmann Machine (RBM) as a knowledge module. The generative stochastic artificial neural network is also called the Restricted Boltzmann Machine (RBM). DBNs have an objectiveless relationship by two layers that structure an RBM and guiding relationship with the lesser layers. The generative graphical model has a directionless association among the full layers of the framework. An apt representation of DBNs and DBMS can be initiate in (Figure2). In going with subclasses, we will depict the major characteristics of DBNs and DBMs, in the wake of presenting their fundamental structure hinder, the RBM. Generative graphical model Deep Belief Networks (DBNs) are probabilistic generative models that provide a probability transport ended perceptible information and also names. That is shaped by loading RBMs and setting them up in a covetous way, that are planned in.27 A DBN from the start uses a helpful layer-by-layer energetic knowledge procedure to gift many frameworks, and, within the facet project, adjusts all plenty in conjunction with the right yields. DBNs are graphical models that add up the way to evacuate a big dynamic depiction of the readiness information. They model the be a part of t spread between watched vector x and also the ???? lined layers h???? as follows: where x =h0,????(h???? |h????+1)is Associate in Nursing unforeseen alternative aimed at the perceptible components at level ???? adjusted arranged the lined components of the RBM at equal????+1, and ????(h????−1|h????) remains that the taken for granted - lined joint unfold within the top-ranking RBM. The top two layers of a DBN structure a directionless outline n and the remainder of the layers structure a conviction facilitate by composed, top-down affiliations. In a DBM, all affiliations are directionless. The standard of m voracious layer-wise freelance coming up with is functional to DBNs by RBMs because the structure frustrates for every layer.28 A summary portrayal of the system is as follows: (1) Train the chief layer as an RBM which models the rough info x=h0 as its perceptible layer. (2) Practice that 1st layer to secure an outline of the info which may be used as data for the following layer. 2 typical game plans exist. This depiction is picked just like the mean authorization (h1=1|h0) or trial of (h1|h0). (3) Train the second layer as an RBM, which modified information (tests or mean commencement) as coming up with examples(for the conspicuous layer of that RBM). (4) Restate steps ((2)and(3)) for the right variety of layers, whenever multiplying upward either test or mean characteristics. (5) Fine-tune all the limits of this vital structure with relevancy a middle person for the DBN log-likelihood, or concerning a regulated coming up with live (consequent to adding further learning device to vary over the perceptive depiction into oversaw gauges, e.g., an instantaneous classifier). There are 2 essential inclinations within the above-depicted unsatiable learning methodology of the DBNs.29 First, it handles the trial of acceptable assurance of parameters, that once during a whereas will incite poor within reach optima, as desires are guaranteeing that the framework is befittingly conferred. Second, there&#39;s no essential for checked information since the system is freelance. Regardless, DBNs are in a like manner full of totally different deficiencies, parenthetically, the procedure value connected with putting in a DBN and also the manner that the strategies towards any improvement of the framework dependent on most outrageous chance preparing gauge are cloudy.28 to boot, a very important obstruction of DBNs is that they don&#39;t speak to the 2D structure of an information image, which can primarily impact the show and connection in deep vision and sight and sound examination problems. Regardless, a later assortment of the DBN, the CDBN is a generative graphical model, uses the three-dimensional info of neighbouring pixels by presenting convolutional RBMs, afterwards creating a change invariant generative model that with success scales with relevancy high dimensional photos.30 APPLICATIONS OF DEEP LEARNING IN DEEP VISION These days, employments of Deep learning join anyway are not obliged to NLP (e.g., sentence gathering, translation, etc.), visual data dealing with (e.g., Deep vision, blended media data assessment, etc.), talk and sound getting ready (e.g., overhaul, affirmation, etc.), relational association examination, and restorative administrations. This portion offers nuances to the unrelated techniques used for the respective application. Natural Language Processing NLP is a movement of estimations and frameworks that essentially based on demonstrating deep to fathom the human language. NLP endeavours fuse report portrayal, understanding, revise recognizing confirmation, content closeness, summary, and question answering. NLP progression is attempting a direct result of the multifaceted nature and unsure building of the human language. Furthermore, ordinary language is significantly setting express, where severe ramifications change subject to the kind of words, joke, and region distinction. Significant learning systems have starting late had the choice to show a couple of compelling undertakings in attaining high precision in NLP errands. Most NLP models follow a practically identical pre-planning step: (1) the information content is isolated into words by tokenization and a while later (2) these words are rehashed as vectors or n-grams. Addressing words in a low estimation is fundamental to make a precise perspective on similarities and differentiation among numerous words. The test shows up when there is essential to pick the length of words limited in each n-gram. This strategy is setting express and needs previous region data. A part of the outstandingly noteworthy systems in appreciating the most prominent NLP assignments are presented underneath. Sentiment Analysis This bit of NLP supervises looking at a book and organizing the propensity or evaluation of the author. Maximum datasets for end assessment are separate as either confident or undesirable, and reasonable enunciations are expelled by bias demand philosophies. The single commended model is the Standford Sentiment Treebank (SST)31, a dataset of film surveys set apart into five classes (going from negative to unfathomably positive). Near to the preface to SST, Socher et al.31 propose a Recursive Neural Tensor Network (RNTN) that usages word vectors and parses a tree to address an enunciation, getting the coordinated efforts among the parts with a tensor-based strategy work. This recursive methodology is perfect concerning sentence-level depiction since the emphasis reliably shows a tree-like structure. Kim32 improves the correctness for SST by next a substitute technique. Notwithstanding the way that CNN models were first made considering picture insistence and strategy, their execution in NLP has displayed to be a triumph, accomplishing brilliant outcomes. Kim grants a direct CNN model utilizing one convolution layer on masterminded word2vec vectors in a BoW structure. The representations were spared sensibly crucial with relatively few hyperparameters for change. By a mix of low tuning and pretrained task-express parameters, they understand how to accomplish high precision on two or three standards. Online life is a prominent wellspring of information while dissecting notions. Machine Translation Deep learning has accepted a noteworthy activity in the updates of customary customized translation systems. Cho et al.33 presented a new RNN-based encoding and unwinding configuration to set up the words in a Neural Machine Translation (NMT). The RNN Encoder-Decoder framework practices two RNNs: one plots a data progression into fixed-length vectors, however, the other RNN translates the vector into the goal pictures. The problem with the RNN Encoder-Decoder is the introduction fall as the data course of action of pictures extends. Bahdanau et al.29 address this question by introducing a dynamic-length vector and by together knowledge the alter and disentangle techniques. Their approach is to play out a matched mission to scan for syntactic structures that are commonly judicious for understanding. In any case, the starting late planned translation schemes are known to be computationally classy and incompetent in dealing with sentences covering unprecedented words. Paraphrase Identification Rework distinguishing proof is the path toward separating two sentences and foreseeing how equivalent they rely upon their essential covered semantics. A key part which is useful for a couple of NLP occupations, for instance, copyright encroachment acknowledgment, answers to questions, setting area, abstract, and region recognizing evidence. Socher et al.34 suggest the usage of spreading out Recursive Autoencoders (RAEs) to amount the resemblance of two sentences. Using syntactic trees to progress the part space, they measure both word-and articulation level comparable qualities. Notwithstanding the way that it is on a very basic level equivalent to RvNN, RAE is useful in independent request. Not in the slightest degree like RvNN, RAE registers a generation screw up in its place of a controlled score during the meeting of two vectors into a compositional vector. This article furthermore introduced a dynamic pooling layer that can consider and bunch two sentences of unlike sizes as either a translation or not. Visual Data Processing Deep learning techniques have developed the central bits of numerous front line intelligent media systems and Deep vision.35 Even more unequivocally, CNNs have exhibited basic results in different genuine endeavours, including picture taking care of, object acknowledgement, and video getting ready. This zone discusses more bits of knowledge concerning the most recent significant learning structures and estimations planned over the late years for visual data taking care of. Image Classification In 1998, LeCun et al. prevailing the essential type of LeNet-5.36 LeNet-5 is a standard CNN that joins two convolutional layers close by a subselection layer ultimately getting done through a complete relationship in the previous layer. Despite the way that, since the mid-2000s, LeNet-5 and other CNN strategies were hugely used in dissimilar issues, counting the division, area, and portrayal of pictures, they were nearly rejected by data mining and AI study get-togethers. Over a multi decade later, the CNN figuring has started its thriving in Deep vision systems. Exactly, AlexNet is seen as the first CNN prototype that significantly enhanced the image portrayal consequences on an amazingly colossal dataset (e.g., ImageNet). It was the victor of the ILSVRC 2012 and improved the best results from the prior years by for all intents and purposes 10% concerning the best five test bumble. To recover the effectiveness and the rapidity of setting up, a GPU execution of the CNN is used in this framework. Data increment and dropout methods are furthermore used to altogether lessen the overfitting issue. Object Detection and Semantic Segmentation Deep learning methodology accepts a noteworthy activity in the movement of article distinguishing proof starting late. Before that, the best article acknowledgement execution began from complex structures with a couple of low-level structures (e.g., SIFT, HOG, etc.) and huge level settings. In any case, with the methodology of new significant learning frameworks, object ID has in like manner showed up at another period of progress. These advances are driven by compelling methodologies, for instance, zone recommendation and section-based CNN (R-CNN).37 R-CNN defeats any obstruction among the article area and picture game plan by presenting section grounded thing repression strategies using deep frameworks. Besides, the move to learn and relating on a colossal dataset (e.g., ImageNet) is applied since the little thing acknowledgement datasets (e.g., PASCAL [46]) fuse lacking named data to set up a gigantic CNN sort out. In any case, in R-CNN, the readiness computational time and memory are over the top costly, particularly on novel ultra-significant frameworks (e.g., VGGNet). Video Processing Video examination has pulled in broad thought in the deep vision organize and is measured as a troublesome task since it consolidates mutually spatial and brief data. In an early slog, colossal degree YouTube accounts containing 487 game classes are used to set up a CNN model. The model fuses a multiresolution designing that employs the close-by development data in accounts and joins setting stream (for low-objectives picture illustrating) and fovea stream (for significant standards picture dealing with) modules to arrange chronicles. An occasion acknowledgement from game chronicles using significant learning is presented in.38 In that work, both spatial and brief data are determined using CNNs and feature mix through standardized Autoencoders. Starting late, another framework called Recurrent Convolution Networks (RCNs)39 was introduced for video dealing. It smears CNNs on video traces for pictorial comprehension and a short time later deals with the housings to RNNs for exploring transient information in accounts. Social Media Analysis Social Web Analysis The notoriety of various relational associations like Facebook and Twitter has engaged customers to part a ton of data with their photographs, considerations, and sentiments. Because of the way that significant knowledge has revealed hopeful execution on visual data and NLP, unmistakable significant learning methods have been grasped for relational association examination, including semantic evaluation, interface estimate, and crisis response.40-42 Semantic appraisal is a huge field in casual association assessment, which means helping machines with understanding the semantic significance of posts in relational associations. Though a collection of techniques have been planned to separate works in NLP, these strategies might disregard addressing a couple of standard difficulties in relational association assessment, for instance, spelling botches, abbreviated structures, unprecedented characters, and easy-going vernaculars.43 Twitter can be considered as the most routinely used wellspring of appraisal request for relational association examination. Generally speaking, feeling examination hopes to choose the mien of analysts. Consequently, SemEval has given a standard dataset reliant on Twitter and run the assessment game plan task since 2013.44 Another practically identical model is Amazon, which ongoing as an online book shop and is as of now the world&#39;s greatest online retailer. With an abundance of acquirement trades, a colossal proportion of evaluations are made by the clients, making the Amazon dataset a remarkable hotspot for tremendous extension estimation plan.44 Information Retrieval Deep adapting incredibly influences information recuperation. Deep Structured Semantic Modelling (DSSM) is planned for text recuperation and web search45, where the dormant semantic examination is driven by a DNN and the inquiries nearby the explore data are used to choose the eventual outcomes of the recuperation. The encoded requests and explore data are mapped into 30k-estimation by term hashing and a 128-estimation feature space is delivered by the multilayer nonlinear plans. The proposed DNN is set up to interface the offered inquiries to their semantic criticalness with the help of the explore data. Regardless, this proposed classical treats each term autonomously and disregards the relationship among the terms. DEEP LEARNING CHALLENGES AND DIRECTIONS With the extraordinary progression in profound learning and its assessment scenes existence at the centre of attention, profound learning has expanded excellent power in talk, language, and visual revelation structures. In any case, a couple of spaces are still faultless by DNNs owing to either their troublesome countryside or the nonattendance of data openness for the overall populace. This makes important possibilities and productive ground for compensating upcoming study streets. The most significant future AI issues won&#39;t have sufficient getting ready tests with names.46 Beside the zettabytes of starting at now open data, petabytes of data are incorporated every day. This exponential improvement is gathering data that can never be named by human aid. he current estimation is pleasing to coordinated learning, by and large by the immediately open imprints and the little sizes of current datasets. Regardless, with the brisk augmentations in the scope and unpredictability of information, independent learning will be the predominant course of action later on. Current profound learning models will in like manner need to conform to the rising issues, for instance, data sparsity, missing data, and disordered data to get the approached info over discernments as opposed to getting ready. Another achievement challenge looked by profound learning techniques is the decline of dimensionality without losing fundamental information required for request. In clinical applications like danger RNA sequencing examination, generally, the amount of tests in each imprint is far not the number of features. In current profound learning models, this causes outrageous overfitting issues and limits the fitting course of action of lacking cases. Current profound learning structures require broad proportions of computational advantages for the push toward the front line presentations. One system attempts to vanquish this test by using store preparing. Added different is to custom the slow systems that misuse medium and colossal datasets on separated getting ready. In rhythmic movement years, various researchers have moved fixation to develop equivalent and versatile profound learning frameworks. CONCLUSION Deep learning, another and fervently discussed issue in AI, can be described as a course of layers acting nonlinear getting ready to get comfortable with various degrees of data depictions. This article contemplates the top tier counts and methods in deep learning. A couple of disclosures of this article and likely future work are dense underneath: • Thoughdeep learning can recall a huge proportion of data and info, its feeble cognitive and perception of the data makes it a disclosure answer for certain applications. The interpretability of deep learning should be inspected later on. • Deep learning despite everything encounters issues in showing various erratic facts modalities at the equalperiod. Multimodal profound learning is extra notable heading in progressing deep learning research. • Disparate human personalities, deep adapting needs wide datasets (unmistakably named data) for coming down the machine and anticipating the inconspicuous information. This issue turns out to be all the more overwhelming when the existing datasets are pretty much nothing (e.g., social protection data) or when the data ought to be arranged ceaselessly. One-shot learning and zero-shot learning have been packed in the continuous hardly any years to help this issue. • In disdain of all the profound learning types of progress starting late, various applications are up &#39;til now flawless by profound learning or are first and foremost times of using the profound learning systems (e.g., disaster information the load up, cash, or clinical data assessment). Englishhttp://ijcrr.com/abstract.php?article_id=3638http://ijcrr.com/article_html.php?did=3638 Ballard DH, Brown CM. Deep Vision. Prentice-Hall, 1982 . Vandoni HT, Carlo E. Deep Vision : Evolution and Promise (PDF). CERN :21–25. Sonka M, Hlavac V, Boyle R. Image Processing, Analysis, and Machine Vision. Thomson, 2008. Szeliski R. Deep Vision: Algorithms and Applications. Springer Science & Business Media 2010:10–16. McCulloch WS, Pitts W. A logical calculus of the ideas immanent in nervousactivity. Bull Math Biol 1943;5(3):115–133. Ouyang W, Zeng X, Wang X. 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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareDifferent Interaction Analysis of Receptors and Ligands in Suppressing Diabetics English204208Balasankar KaravadiEnglish Premalatha JEnglish Vinothkumar CEnglish Rajasekar BEnglishIntroduction: Many recent studies are done on the efficiency of natural components for the treatment of diabetes, it has been found that many antioxidant components are present. Objective: The present study aims to study and identify the target sites which can be used as targets with appropriate ligands for diabetics. Methods: Structure-based drug design was performed and protein docking was done with known ligands. Analyzing the receptor-ligand interaction between the active site of the protein and the chemical molecules was performed. Results: All the compounds that satisfy the absorption, distribution, metabolism, excretion and toxicity (ADMET)properties are more favourable to bind with receptor. Discovery Studio tool was used to find the least energetic compound and based on the docking parameter against the receptor-ligand complex and which notified us of the acceptors and the donors in the analysis portion. from the study, we analysed that these components which contain bioactive compounds can be used in medical research. Nitenin showed a good dock score of 47.404 and can inhibit the overexpression of the CD62E gene. Conclusion: The finalized complex of Nitenincompound can be used for further medical studies in the preparation of drug molecule against diabetes. The docking study reveals that the antioxidant components that are present can be used in the treatment of diabetes. EnglishADMET, Diabetic, Ligand, Docking, DrugINTRODUCTION Diabetes is described by unusually significant levels of sugar (glucose) in the blood.1 At the point when the measure of glucose in the blood increments, e.g., after supper, it triggers the arrival of the hormone insulin from the pancreas. Insulin invigorates muscle and fat cells to expel glucose from the blood and animates the liver to use glucose, causing the glucose level to diminish to ordinary levels.2 In individuals with diabetes, glucose levels stay high. This might be because insulin isn&#39;t being created by any means, isn&#39;t made at adequate levels, or isn&#39;t as compelling as it ought to be. The most well-known types of diabetes are type 1 diabetes (5%), which is an immune system issue, and type 2 diabetes (95%), which is related to stoutness. Gestational diabetes is a type of diabetes that happens in pregnancy, and different types of diabetes are extremely uncommon and are brought about by a solitary quality transformation.3 For a long time, researchers have been scanning for intimations in our hereditary cosmetics that may clarify why a few people are bound to get diabetes than others.4 The hereditary scene of diabetes presents a portion of the qualities that have been proposed to assume a job in the improvement of diabetes.5      As the incendiary reaction advances, chemokines discharged by harmed tissue enter the veins and initiate the moving leukocytes, which are presently ready to firmly tie to the endothelial surface and start advancing into the tissue. P-selectin has a comparable capacity, however is communicated on the endothelial cell surface inside minutes as it is put away inside the cell as opposed to delivered on request.6 ADMET screening is accomplished for the characteristic segments that are recovered from the database for testing their harmfulness and to locate the best lead particle that fulfils the ADMET parameter conditions (Figure 1).7,8 The ADMET screening process is finished utilizing Disclosure STUDIO programming. The ligand particles for which the AMET must be done is downloaded from PUBMED then stacked in the revelation page and checked for ADMET results. Structure-based drug design is the method of performing docking using the known protein docking with known ligands. This has been the frequently used method of analyzing the receptor-ligand interaction between the active site of the protein and the chemical molecules.9 MATERIALS AND METHODS The template structure was obtained from protein data bank.10 The modeled structures were validated using SAVS, an online server.11 The CASTp server was used to analyze binding sites of the protein molecules. Further, on the basis of high throughput method lead molecules having more affinity with the target proteins were obtained from DrugPort database.12 Then the structurally similar compounds were obtained using PubChem database.3 Finally a datset was created for potential ligands inhibiting the target proteins using vegaZZ software. Accelrys Discovery Studio 2.0 was used to analyze specific protein-ligand docked complexes and finally toxicity of the ligand molecules were analysed using ADMET descriptors.4 RESULTS AND DISCUSSION Receptor Ligand Interaction: Ligand-Nitenin The docking score of this interaction is 47.404 and LigScore1_Dreiding and LigScore2_Dreiding values are 2.14 and 4.19,  -PMf value is 8.7 and the internal energy is 12.391 (Figure 2). Ligand- Fulvinervin A The docking score of this interaction is 44.994and LigScore1 and LigScore2 values are 1.34 and 4.29,  -PMF 9.49 value is and the internal energy is 364.27 (Figure 3). Ligand- Emoroidenone The docking score of this interaction is 39.125 and Lig1Score 1.15,Ligscore2 3.78  PMF value is 10.35 and the internal energy is -7.686 (Figure 4). From the results obtained it is analyzed that the antioxidants present in Tephrosia play a major role in the treatment of diabetes. It is concluded that they can use the CD62e receptor which increases estrogen production and bone mineral density. CONCLUSION In this study, we have noticed that the compounds present in herbal medicinal can be used as a diabetes medicine. The anti-diabetes properties related to the compound which use as a ligand to show the interaction between receptor CD62E structure tells that the compound can inhibit the overexpression of CD62E receptor by binding in the active site cavity. Discovery Studio tool was used to find the least energetic compound and based on the docking parameter against the receptor-ligand complex and which notified us of the acceptors and the donors in the analysis portion.  Finally, from the study, we can conclude that these components which contain bioactivity can be used in future medical research. Nitenin showed a good dock score of 47.404 and can inhibit the overexpression of the CD62E gene. The finalized complex of nitenin compound can be used for further medical studies in the preparation of drug molecule against diabetes. ACKNOWLEDGMENT: The authors acknowledge the scholars whose articles are cited and included in references to this manuscript. The authors are also thankful to authors/ editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. We acknowledge our institute management for their support. CONFLICT OF INTEREST: Nil Englishhttp://ijcrr.com/abstract.php?article_id=3639http://ijcrr.com/article_html.php?did=3639 Anderson DM, Maraskovsky E, Billingsley WL, Dougall WC, Tometsko ME, et al. Homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature 1997;390:175–179. Boyce BF, Xing L, Shakespeare W, Wang Y. Regulation of bone remodelling and emerging breakthrough drugs for osteoporosis and osteolytic bone metastases. Kidney Int Suppl 2003;12: 2–5. Simonet WS, Lacey DL, Dunstan CR, Kelley M, Chang MS, Luthy R, et al. Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell 1997;89:309–319. Ida Leida M, Ridwan M. Thaha, Andi SelviYusnitasari, Afsahyana Effect of Sap Palm (Borassus flabellifer) on Blood Glucose Level in Pre-Diabetic Patients. Int J Curr Res Rev 2020;12(24):96-100. Tolar J, Teitelbaum SL, Orchard PJ. Osteopetrosis. N Engl J Med 2004;351:2839–2849. Breuil V, Schmid-Antomarchi H, Schmid-Alliana A, Rezzonico R, Euller-Ziegler L, Rossi B. The receptor activator of nuclear factor (NF)-kappaB ligand (RANKL) is a new chemotactic factor for human monocytes. FASEB J 2003;17:1751–1753. Aswathy BK, Nisha H. Homology Modeling and Docking Studies to Identify the Targets in Pancreatic Cancer. Res J Pharm Tech 2017;10(7): 2032-2040. Aswathy BK, Martina V. Insilico Analysis of Inhibitors Related to Aggressive Behavior in Human Beings. Res J Pharm Tech 2018;11(4):1436-1441. Karavadi B, Suresh M. Homology modelling and molecular drug design approach in identifying drug targets of TIGR4 in Streptococcus pneumonia. Bio sci Biotechnol Res Asia 2014;11:517-522. Karavadi B, Suresh MX. In silicomodeling of capsular polysaccharidebiosynthesis protein and tyrosine kinase of G54 strain in Streptococcuspneumoniae and their ligand identification. Int J Pharm Pharm Sci 2014;6:547-550. Karavadi B, Suresh MX. Receptor identification and lead the molecular discovery of a phage-encoded protein in TCH8431/19A strain of Streptococcus pneumoniae: A computational approach. Int J Appl Pharm 2014;6:6-10. Karavadi B, Suresh XM. Homology modelling of polymerase and CPS biosynthesis proteins in CGSP14 strain of Streptococcus pneumonia and its ligand identification: An in silico approach. Asian J Pharm Clin Res 2014;7:162-165.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareThe Anticancer Activity of Artemisia Judaica Crude Extract in Human Hepatocellular Carcinoma HepG2 Cells by Induction of Apoptosis and Cell Cycle Arrest English209215Neima K. Al-SenosyEnglish Naglaa M. EbeedEnglish Lamiaa M. SalemEnglish Shenouda M. GirgisEnglish Ekram S. AhmadEnglishIntroduction: Cancer is one of the major causes of death around the world. Medicinal plants have been investigated across the world to exploit their potential anticancer activity on a large scale of applications. Objective: The present study is focused on the anticancer activity of the methanolic extract of the medicinal plant, Artemisia Judaica, on Human hepatocellular carcinoma (HepG2) and normal liver (THLE2) cell lines, as well as the mechanisms involved were also investigated. Methods: The collected aerial parts of the plant were extracted by maceration with methanol and the crude extract was collected and stored until use. Cytotoxicity and cell proliferation, cell cycle analysis and the expression level of apoptosis-related genes such as p21, CycB1, CDK1, p53, Bcl-2 and Bax in HepG2 cell lines were estimated using the MTT colourimetric, flow cytometry and quantitative real-time PCR (qRT-PCR) assays, respectively. Results: The results showed that A. Judaica extract (IC50 = 33.76μg/ml) displayed strong cytotoxicity and antiproliferative effect in HepG2 cancer cells. On the other hand, this extract exhibited no cytotoxic activity on the liver normal cell line (THLE2). Flow cytometric analysis of propidium iodide staining revealed that the treatment of HepG2 cells with A. Judaica led to an increase in G2/M phase cell cycle arrest. The qRT-PCR assay revealed that both cyclin B1 (cycB1) and cyclin-dependent kinase (CDK1) genes, as well Bcl-2 showed down-regulation expression levels in HepG2 treated with A. Judaica compared to the untreated cell line. Furthermore, the apoptotic mechanism activated by the plant extract resulted in up-regulation of p53, P21and Bax at mRNA level on HepG2 cell line. Conclusion: These results suggest that A. Judaica could be a promising candidate species as a natural source of anticancer molecules. EnglishArtemisia Judaica, Antiproliferation, Apoptosis, Anticancer, HepG2 cell lineINTRODUCTION Cancer is one of the major causes of death worldwide.  It is characterized by genetic alterations of normal cells which become malignant cells. Those are characterized by uncontrolled cell growth, immortality, invasiveness, and the ability to form distant metastasis. It was found that natural products may interfere with the carcinogenesis process by altering the behaviour of tumour cell and targeting cancer cells signalling pathways.1         Among different cancer types, liver cancer is the 4th most common cause of death from cancer worldwide, the incidence of human hepatocellular carcinoma (HCC) is increasing particularly in males in some countries. Hepatitis B and C viruses (HBV and HCV) and dietary aflatoxin intake remain the major causative factors of HCC, where different modes of cancer therapy for HCC have been tried such as surgery, chemotherapy, radiotherapy, as well, tremendous works have been done at the molecular level.2 The use of natural products and supplements of medicinal plants has very increased over the past three decades with more than 80% of people worldwide depend on them for some part of primary healthcare.3 The diet enriched with naturally occurring substances significantly reduces the risk for some cancers. Consequently, many drugs used for the treatment of cancer have been discovered from medicinal plants.4 Biologically active components of the medicinal plants target tumour cells by different mechanisms, resulting in angiogenesis, inhibition of carcinogenesis, cell cycle arrest, oxidative stress, autophagy or differentiation and apoptosis.5-9           Artemisia was found to possess sesquiterpene lactones and other derived phytochemicals as active components. Sesquiterpene lactones were used for their therapeutic and other properties.10 Recently, monoterpenes, sesquiterpenes, sesquiterpene lactones, flavonoids, coumarins, sterols, polyacetylenes have been isolated from Artemisia species.11 Some Artemisia species have shown that it possesses medicinal properties such as anti-bacterial and anti-cancer effects.12 Few species of Artemisia such as Artemisia Judaica, showed promising anticancer activities against the proliferation of cancer cell lines.13 The main effect of its essential oils is attributed to the main constitutes, the thujone, in this species,14  enriched with fraction has potential anticancer activities.15 A. Judaica (Arabic name, Shih Balady) is a perennial fragment shrub that is abundant in North Africa and Middle Eastern countries.16 As well in Saudi Arabia, Yemen and Egypt.17-19 This plant grows abundantly in different parts of the Arabian peninsula such as Saudi Arabia and Yemen,17,18 and has been used in traditional Egyptian medicine for the treatment of gastrointestinal disorders.19 Besides, the Artemisia species have been used in Iranian traditional medicine as an anti-infectious, anti-bacterial, gastric tonic, digestive and stomachic.20 Isolated compounds from A. Judaica have exhibited antiviral, antibacterial, antifungal, and cytoprotective effects,21-23 and for the treatment of hepatitis, cancer and menstrual-related disorders.24 The composition of A. Judaica includes artemisinic acid, sesquiterpene lactones, methyl wormwood, artemisinic alcohol, eucalyptol, Artemisia ketone, camphor, caryophyllene, piperitone and essential oil.25-26 Besides, diverse chemical components, such as flavonoids, coumarins, sterols, polyacetylenes, monoterpenes, polyphenols, sesquiterpenes and sesquiterpenes lactones, have been found in plants from the Artemisia genus.22,24            Sesquiterpene lactone is also a potent apoptotic inducer in cancer cells via multiple pathways. It is readily depleted intracellular glutathione (GSH), disrupts cellular redox balance, depletes intracellular thiols,27 triggers an intracellular reactive oxygen species (ROS),28 decreases anti-apoptotic Bcl-2 protein expression and induce apoptosis.29 Studies of the molecular mechanisms have shown that caspase and p53-independent activation, downregulation of Bcl-2, generation of ROS, reduction of mitochondrial membrane potential, increased amount of Bax protein, arrest of the cell cycle at G2/M-phase, suppression of the Notch 1, and inhibition of NF-κB are important mechanisms for the cytotoxic effects of Artemisia.30 As well, flow cytometric analysis for cell cycle revealed increment of G2/M phase cell cycle arrest after treatment of HepG2 cells with A. herba alba extract. The apoptotic mechanism was also activated by the crude extract of Artemisia included up-regulation of p53 and Bax and downregulation of Bcl-2 expression levels with no cytotoxic effect for normal cell lines exposed to the plant extract.31 Besides, Artemisia extract increased the number of cells in the G2/M phases, followed by caspase 3 upregulation, and apoptosis. Further, it is inhibited cancer cell proliferation and induced apoptosis.32 Therefore; the present study is designed to evaluate the anticancer activity of crude extract of A. Judaica, on human hepatocellular carcinoma (HepG2) cell line. The possible underlying antiproliferation mechanisms were investigated, by studying the effect of A. Judaica on cytotoxicity, cell cycle arrest and apoptosis-related genes in the human hepatocellular carcinoma (HepG2) cell line. MATERIALS AND METHODS Plant collection and extract preparation The aerial parts (leaves and stems) of A. Judaica were collected during the summer season (July 2020) from Wadi Gharandal, South Sina, Egypt. Taxonomical identification was confirmed by a botanist and voucher specimen, were deposited in the Herbarium of the National Research Center, Dokki, Cairo, Egypt. Herbs were washed and shade dried for a week and was milled to fine powder, then it was extracted with methanol at a ratio of 20 g dry powder in 200 ml of ethanol for 48 h using the maceration method. The liquid extract was filtered and concentrated under vacuum by soxhlet extraction, then stored in the dark at 4°C until use.                            Assessment of cytotoxic and anti-proliferous activities of A. Judaica Cell proliferation and viability of the cells were estimated using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) colourimetric assay. Human hepatocellular carcinoma (HepG2) and normal liver (THLE2) cell lines in exponential growth phase were seeded at a density of 1 x 104 cells per well (100 μl/well) onto a 96-well plate (Falcon, Franklin Likes, NJ, USA) in DMEM medium (GIBCO, Grand Island, New York, USA; Cat.no.A1049101). The cell density was adjusted by the trypan blue exclusion method. The whole compounds were cultured in different concentrations (ranged from 0 to 200 µg/mL) for 24 hours at 37 °C in a 5% CO2 with a 95% humidity incubator. Besides, different concentrations of cisplatin as a reference chemotherapeutic drug were added and the microplates were incubated for a further 48 hour in DMEM medium (200 µL). The medium was washed gently twice with ice-cold PBS and a volume of 200 μL MTT  [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, a yellow tetrazole, (Molecular probes, Eugene, Oregon, USA; Cat.no.V-13154)] was added to each well. The microplate was incubated at 37 °C for another 4 hours in a CO2 incubator. About 180 μL medium/MTT was removed and 100 μL of acidified isopropanol were added per well to solubilize the formazan produced. Finally, the microplate was incubated with shaking for 15 minutes. The absorbance of each well was measured at 630 nm using a microplate reader (ELX800,  Biokit, Spain). Assays were performed in triplicate on three independent experiments. Sigmoidal and dose-dependent curves were constructed to plot the results of the experiment. The concentration of the compounds inhibiting cancer cell growth by 50% of the control level (IC50) was calculated using this sigmoidal curve. Cell cycle analysis by flow cytometry To evaluate the effect of A. Judaica crude extract on the cancerous cell division of HepG2, the cell cycle analysis was carried out using the protocol of Applied Bio-system, USA. Cells were digested with warm Trypsin-EDTA + warm PBS-EDTA (0.25%) (500 µl + 500 µl) with incubation for 10 minutes at 37°C. The mixture was centrifuged at 450 rpm for 5 min, and then the supernatant was carefully removed. The mixture was washed twice in warm PBS and the cell pellet was re-suspended in 500 µl warm PBS, centrifuged and the supernatant was removed. A volume of 150 µl PBS + 350 µl ice-cold 70% ethanol was added and incubated at 4°C for 1 hour to fix the cells. To remove ethanol, the mixture was centrifuged at 350 rpm for 10 minutes and then the supernatant was carefully removed. The mixture was washed twice in warm PBS and the cells were re-suspended in 500 µl warm PBS, centrifuged and the supernatant was removed. The cells were re-suspended in 100 µl PBS and stored at 4° for up to 4 days. In the darkness, the cells were stained with 100 µl of propidium iodide (PI) solution + 50 µl RNase A solution (100 µg/ml) and incubated in darkness for 30-60 min. The stained cells were read in Attune flow cytometer (Applied Bio-system, USA). Determination of the expression levels of apoptosis-regulatory genes by quantitative real-time PCR RNA extraction was carried out utilizing an RNeasy Mini kit (Qiagen, Inc., Valencia, CA, USA) for HepG2 cell lines, or treated with IC50 of A. Judaica extract for 24 h. The procedure was performed according to the manufacturer’s instructions. The cDNA was synthesized from purified RNA with an RT2 First Strand Kit (Qiagen, Inc., Valencia, CA, USA) according to the manufacturer’s guidelines as the template for RT-qPCR. Corbett Rotor-Gene 6000 (Qiagen, Inc., Valencia, CA, USA) was used to perform quantitative real-time reverse transcriptase PCR (qRT-PCR). A final volume of 25 µl pre-mix was prepared to contain 12.5 µl of RT2 SYBR R Green ROXTM FAST master mix (Qiagen, Inc., Valencia, CA, USA), 1 µl of primers (RT2 qPCR Primer Assays, Qiagen, Inc., Valencia, CA, USA), 1 µl of cDNA, and 10.5 µl RNase-free water to make the final volume. The following primer pairs for target genes and the housekeeping β-actin gene was chosen from the Primer Bank website. The default PCR conditions were as follows: the PCR plate was run at 95?C for 10 min to activate the enzyme, 40 cycles of 15 s at 95?C (denaturation) followed by 30 s at 60?C (annealing and synthesis). Finally, the dissociation curve was constructed immediately after the PCR run to check and verify results. The housekeeping gene β-actin was used as a reference to calculate fold change in target gene expression. Statistical Analysis Data were expressed as means ± standard error (SEM). One way ANOVA using SPSS 18.0 software was used to detect the significant difference. Values were considered statistically significant when P≤0.05. Comparison of means was carried out with Tukey’s HSD test. RESULTS Cytotoxic activity of A. Judaica crude extract using MTT assay  The cytotoxic activity of A. Judaica crude extract was screened on human hepatocellular carcinoma cell line (HepG2) by MTT assay (Fig. 1). The results revealed that A. Judaica extract (IC50 = 33.76μg/ml), inhibited the proliferation and exhibited increasing in cytotoxic activity to HepG2 cell line in a dose-dependent manner, where, the percentage of cell viability declined up to 20% by A. Judaica dose increment, while the crude extract of A. Judaica didn’t induce growth inhibitory effect on normal liver cell line (THLE2) (IC50 = 1041μg/ml), where, a very low cytotoxic activity was noticed by A. Judaica against normal liver cell line. Effect of A. Judaica crude extract on cell cycle arrest using flow cytometry The DNA content in A.judaica treated HepG2 cells was assessed using the cell cycle phase distribution analysis. The cell cycle arrest was analyzed using flow cytometry (Fig. 2). According to the inhibition rate of liver cancer (HepG2) cell viability, the results revealed that the anti-proliferative effect of A. Judaica was associated with cell cycle arrest using flow cytometry-based cell cycle distribution (Table 1). Compared to the control group, the treatment with A.judaica crude extract at 33.76 μg/ml affected the cell cycle distribution on HepG2 cells, where, the G0/G1 phase showed decreased from 60% to 29% of cell cycle arrest, while the S-phase percentage increased slightly from 19% to 22% in the HepG2 and HepG2 treated with A.judaica crude extract, respectively. Interestingly, the percentage of HepG2 cells at the G2/M phase was increased (49%) compared to (21%) for the untreated cell line. These results suggested that A. Judaica inhibited the cellular proliferation of HepG2 cells via G2/M phase arrest of the cell cycle. The expression level of apoptosis-related genes modulated by A. Judaica  To evaluate the molecular mechanism of A. Judaica-induced apoptosis in HepG2 cells, the expression level of apoptosis-related genes such as p21, cyclin B1 (CycB1), cyclin-dependent kinase (CDK1), p53, Bcl-2 and Bax in HepG2 cells (Fig. 3) was estimated by quantitative real-time PCR (qRT-PCR). Results revealed that compared to the untreated HepG2 group, the expression level of p21, p53 and Bax was increased (upregulated) in A.judaica treated HepG2 cell line, whereas, the expression level of Bcl-2, CycB1 and CDK1genes was down-regulated (decreased) in HepG2 treated with A. Judaica compared to the untreated HepG2 cell line. DISCUSSION Medicinal herbs have shown their pharmacological activities as a new drug reaching the market.33,34 Different forms of cancer are constantly gaining resistance to current drugs, creating a need for the discovery of new drugs. A. Judaica extract showed remarkable cytotoxic activity against the tested liver cancer cell lines in a dose-dependent manner. This could be due to the presence of artemisinin and other sesquiterpene lactones, which have been found in A. Judaica.35,36 Artemisinin which is a sesquiterpene lactone demonstrated antitumor agent and anticancer activity when tested in vitro and in vivo.37,38 These results agree with previous studies reporting biologically active compounds from the medicinal plant A. Judaica possess sesquiterpene lactones, essential oils and other derived phytochemicals as active constituents which have shown medicinal properties such as anticancer activities (cytotoxicity and antiproliferation) for cancer cell lines, with no cytotoxic effect for normal cell lines exposed to the plant extract.31 The cell cycle arrest was analyzed using flow cytometry-based cell cycle distribution. The results indicated that A. Judaica extract inhibited the liver cancer (HepG2) cell cycle through triggering G2/M phase arrest involved transcriptional suppression, where the percentage of HepG2 cells at the G2/M phase was increased and reached about 50 % (49% ± 2.08) compared to 21% for the untreated cell line. That coincided with Lang et al.,32 who found that Artemisia extracts induced accumulation of multinucleated cancer cells within 24 h of treatment, increased the number of cells in the S and G2/M phases of the cell cycle, followed by loss of mitochondrial membrane potential, caspase 3 activation, and formation of an apoptotic cell population. These data indicated that A. Judaica exhibited an antiproliferative effect by a cell cycle blocking at the G2/M phase and apoptosis mediated cytotoxicity in carcinoma cells. However, Honda et al.,39 found that equipotent concentrations of A.judiaca extract highly arrested the cell cycle in the G0/G1 phase of cancer cell lines. The current study inspected the ability of A.judiaca crude extracts to induce apoptosis where it is known to be the most promising pathway for a cancer therapy strategy. Consequently, results showed that the extract had the highest cytotoxic activity, have ingredients or molecules involved in the activation mechanisms of one or more antiproliferative pathways. A. Judaica medicinal plant induces apoptotic process which is modulated by different tumour suppressor genes including p53. The P53 induces apoptotic cell death by direct or indirect change expression of the Bcl-2 family of proteins, Bcl-2 and Bax.40 The Bcl-2 gene is an anti-apoptotic gene that suppresses initiation steps of apoptosis via inhibition of the pro-apoptotic proteins.41 P53 may modulate susceptibility of cells to apoptosis by downregulation of BCL-2 and causing up-regulation of BAX 27 and that coincides with our findings. The cell cycle is mediated by the activation of a highly conserved protein kinase family, the cyclin-dependent kinases (CDKs).42 Cyclins can activate CDKs by forming complexes with Cdks, and these cyclin/CDK complexes are cell cycle regulators. Among them, the cyclin B1/CDK1 complex, in which B-type cyclins associate with CDK1, is considered as one of the primary regulators of transition from the G2 to M phase. This complex was originally discovered and defined as the maturation-promoting factor or M phase-promoting factor (MPF).43 In our study, the down-regulation of CycB1, Bcl-2 and CDK1 gene expression as responsible genes for Cyclins and cyclin-dependent kinase (the cell cycle regulators cyclin/CDK complexes) in HepG2 treated with A. Judaica compared to the untreated HepG2 cell line at G2/M phases of the cell cycle, confirm these findings. Sesquiterpene  lactone as active constituents of Artemisia was found to be a potent apoptotic inducer in cancer cells via multiple pathways especially in depletes intracellular GSH, disrupts cellular redox balance, depletes intracellular thiols, 27 triggers an intracellular reactive oxygen species that lead to mitochondrial dysfunction: loss of mitochondrial membrane potential, the onset of mitochondrial membrane transition, and release of mitochondrial pro-apoptotic proteins,28 decreases anti-apoptotic Bcl-2 protein expression and induce apoptosis.29 CONCLUSION Based on the findings of the study, the crude extract of A. Judaica was found to had anticancer properties specific for human tumour cells, without any toxicity on normal cells. These results indicated that A. Judaica has an antiproliferative effect on HepG2 cell lines.  Moreover, A. Judaica crude extract evaluated to identify the mechanisms behind the toxicity such as cell cycle blocking at the G2/M phase using flow cytometry. The real time-PCR to measure the mRNA levels of cyclin B1 and cyclin-dependent kinase (CDK1) genes (cell cycle regulator genes) and Bcl2, where those genes showed down-regulation expression levels in HepG2 treated with A. Judaica compared to the untreated cell line. Gene expression of ribonucleic acid (RNA) study confirms A. Judaica extract to induction of apoptosis via activation of p53, p21 and Bax and inactivated of Bcl2. Taken together, these data suggest that A. Judaica could be a promising candidate species as a source of anticancer molecules. Finally, natural products such as A.judaica are attractive sources for the development of new medicinal and therapeutic agents. Those with anticancer potential and apoptosis may be more selective and have weaker adverse effects on normal cells compared to chemotherapy used for cancer treatment. So, these research trends indicate that natural products will be among the most important sources of new drugs for cancer prevention and treatment in human being. Conflicts of Interest: The authors declare that they have no conflicts of interest related to this study. This work was financially supported by us. Acknowledgements: Authors acknowledge the immense help received from the scholars whose articles are cited and included in references to this manuscript. The authors are also grateful to authors/editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Source of funding: We expend from our account. Englishhttp://ijcrr.com/abstract.php?article_id=3640http://ijcrr.com/article_html.php?did=3640  Fabiani R. Antitumoral Properties of Natural Products: Editorial.  Molecules 2020;25:650.                               Tang ZY. Hepatocellular Carcinoma-Cause, Treatment and Metastasis: Review. 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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareStem Cell Research-Ethico-Legal Perspectives: Protection of Human Embryos English216222Lakshmi KolluEnglishStem Cell technology is a rapidly developing field that combines the efforts of Cell biologists, geneticists and medical practitioners, offers effective treatment for a variety of malignant and non-malignant diseases because their stimulation can be accelerated to develop into any type of cell or tissue or organ system. The Article studies the latest development in medical sciences is the use of Stem Cell therapy. There are serious ethical and religious questions involved in the use of embryo Cells as it could be considered as feticide because of the belief that life begins at conception. So the next best thing to go with the research of umbilical cord blood. Scientists believe that several incurable diseases can be treated and even cured by Stem Cell therapy. For this purpose, Stem cell banks have been set up in big cities like Chennai and Mumbai In these banks Stem Cells of newborn babies will be preserved for twenty years or more in liquid nitrogen. Rich Indians have accepted Stem Cell therapy as a health insurance cover for their progeny. Medical experts and Stem Cell banks claim that stored stem cells can cure Seventy life-threatening diseases including diabetics, leukaemia, blood cancer, and Alzheimer’s diseases. As Civilization progressed, the man had to fight against diseases caused by both external and internal agents. So he had to invent and prepare curative and preventive medicines by using intellectual mind performing through Researches. EnglishCord blood Cells, Ethical and moral and legal, Foetus, Human Embryonic Stem Cells.Introduction1-5 Research on human subjects involving cells and tissues derived from human embryos and foetuses must safeguard human rights, dignity, and fundamental freedom. This includes processes related to obtaining human tissues and cells for research, diagnosis, and therapy.  The fundamental tenets of beneficence, non-malfeasance, justice, and autonomy should be adhered to in all research involving human subjects. To achieve these objectives, all research involving the use of stem cells must be guided by the general principles laid down in the Ethical Guidelines for Biomedical Research on Human Participants published in 2006  by the Indian Council of Medical Research  (ICMR)  and specific principles related to stem cells as provided under these guidelines must be followed. Scientists began to investigate the possibility of Stem cell-based therapies to cure disease, which is often referred to as regenerative medicine. The major conflicting unethical issue identified with this research is the extraction of embryonic stem cells by embryo destruction.  The very embryo which can become a human being is destroyed at the onset of its potentiality of becoming one of us. Any research which stands to violate these principles is bound to suffer from moral and ethical controversies. The main goal of Human Embryonic Stem Cell Research is to identify the mechanisms that govern cell differentiation and to turn HESCs into specific cell types that can be used for treating debilitating and life-threatening diseases and injuries. The main goal of Human Embryonic Stem Cell Research is to identify the mechanisms that govern cell differentiation and to turn Human Embryonic Stem Cells into specific cell types that can be used for treating invigorating and life-threatening diseases and injuries. Sources and Technology Human Embryonic Stem Cells HESCs re-derived from the inner cell mass of the human blastocysts.  A blastocyst is formed five days after fertilization of the egg by the sperm. It has an outer shell that matures and if survives implantation becomes placental tissue and the inner cell mass becomes the tissues of the human body. The extraction of HESCs from inner cell mass for research purpose leads to the destruction of the embryo. The major source of human embryonic stem cell tissues are the spare or supernumerary embryos created during in-vitro fertilization as a part of infertility treatment. The other source is creating embryos with somatic cell nuclear transfer techniques (SCNT). The legislation of most countries including India allows the use of spare or supernumerary embryos either fresh or frozen created during in-vitro fertilization. Some countries with a more liberal view have allowed the creation of human embryos with SCNT as a source of embryonic tissues. The controversial issue in embryo research is concerned with which embryos are suitable and can be used for research. There is disagreement over whether it is appropriate to create embryos solely for research purposes, and what techniques should be used to create those embryos. Many people and government feel that an appropriate restriction on embryo research is to limit the use of embryos in research to those embryos that are surplus to infertility treatments. Legal Perspectives The extraction of embryonic tissue for research purpose involves the destruction of the embryo. Most of the arguments about the rightness and wrongness of embryo destruction are based on the moral status of the embryo. The moral wrongness associated with embryo destruction will not only make the research impermissible but also deny the potential benefits expected from this research.  Using human embryonic tissues for research poses a moral problem as it brings two highly valued but conflicting moral principles. The objective is to provide treatment to ease pain and suffering on one hand and the value of human life and dignity on the other. The extraction of stem cells from human embryos violates the second principle as it leads to the destruction of potential human life.  Both principles cannot coexist together, but which principle takes precedence is a rather contentious issue. the embryo should be considered from the moral or legal point of view as the main debatable issue associated with HESC research. The HFE Act,1990 permits research to be licenced on embryos until the formation of a primitive streak. That is taken to occur not less than fourteen days after fertilization. The HFE Act imposes several important restrictions on the use of embryos in research. These include the following: An embryo cannot be stored or used for research after the primitive streak, which is taken to be fourteen days from the mixing of gametes. The use or storage of an embryo requires a licence and licences can only be issued by the HFEA for certain purposes including Promoting advances in the treatment of infertility, miscarriages, contraception, or causes of congenital diseases. Human Fertilization and Embryology Act 2008 has added three new purposes to the original five sets out in the Act: Increasing knowledge about the development of embryos, serious disease, or enabling the application of any such knowledge in developing treatments for serious disease. It is not permitted to mix human and animal gametes or to place a human embryo in a non-human animal, except as permitted by the HFE Act 2008. The HFEA will only consider licences for research if a Research Ethics Committee has approved the research. Controversially the HFE Act permits research not only using embryos that are ‘spare’ following infertility treatment but also on embryos that are specifically created for research.    Most people who support research involving embryos, including the Warnock Committee whose report underpins the HFE Act, require that embryos be treated with respect. The House of Lords Committee on Stem Cell Research recently concluded that the HFE Act does demonstrate respect for the embryo used in research, for example, by only allowing embryo research to be performed where no alternative means are available, and only for one of the permitted purposes The HFE Act 2008 inserts a new section 4A into the 1990 Act: No person shall place in a woman- A human admixed embryo, Any embryo that is not human, Any gametes are other than human gametes. No person shall – Mix human gametes with animal gametes, Bring about the creation of a human admixed embryo, or Keep or use a human admixed embryo, except in pursuance of a licence. A licence cannot authorize keeping or using a human admixed embryo after the earliest of the following- The appearance of the primitive streak, or The end of the period of the two weeks beginning with the day on which the process of creating the human admixed embryo began, but not counting the time during which the human admixed embryo is stored. A licence cannot authorize placing a human admixed embryo in an animal. So the Act only allows the creation of admixed embryos up until the age of 14 years and does not allow such embryos to be placed in a woman. The creation of admixed embryo is only permitted if done under a licence. It’s very difficult to ascertain the moral status of the embryo as it varies. There are different opinions about this moral status. The leading persuasion deliberate that the embryo has the status of Persons, or  Potential persons,  or Divine creations,  or  Subjects of moral harm,  or the beginning of human life with intrinsic value, or organic material with no moral standing than other body parts. The development of human life or person is an evolving process starting from fertilization to the birth of a newborn. The early stages of development mostly compromise cellular differentiation whereas, in the end, the fetus assumes its full form both in physical and functional status. There is no clear-cut demarcation during this process of physical development as to when personhood is acquired. At one end of the horizon of purviews on this issue is the belief that the embryo, from the moment of conception, is created by God and is an individual who possesses their rights with the equal moral status as an adult human being. Those who hold this horizon, such as Catholic Bishop Richard Doerflinger, says that it is wrong to destroy embryos of any gestational age, for any purpose. This absolutist view is not shared by all those with religious beliefs. A substitute stance is that the embryo acquires a full personal identity, and the ethical rights that come with this status, step by step during the process of development occurring between conception and birth. It is so ethically acceptable, under these circumstances, to use embryos for research purpose. This read has been defended by some theologians of alternative faiths, together with Protestant, Christians, Jews, Muslims and Buddhists, and is additionally seconded by many folks who don’t have religious faith.  The embryo in its early stage is a cellular structure and don’t have the psychological, physiological, emotional and intellectual characteristics that we tend to attribute with individuality. Consequently, traces that if the human embryo does not fulfill the standard measure for personhood it does not possess any interests to be protected and thus may be instrumentally used for the benefit of other human persons.In the discussion about embryo research, the formation of the primitive streak is considered an important landmark point. The primitive streak, seen in the form of appearance of a surface thickening, is the first visible organization of the embryo which usually happens around fourteen days after fertilization. The primitive streak, seen in the form of appearance of a surface thickening, is the first visible organization of the embryo which usually happens around fourteen days after fertilization. The term ‘pre-embryo’ was introduced in 1985 to describe the early embryo up to this point.  One argument that was used to justify distinguishing the pre-embryo and the embryo proper was that the possibility of splitting the pre-embryo into two parts or twin parts. It appears, as per this argument, that the pre-embryo wasn’t a person, as personhood is commonly taken to imply indivisibility or individuality Others have argued that the concept of the pre-embryo is a rhetorical device invented to justify embryo research and that it creates an artificial division in what is, in reality, a continuous biological process of development. Some research workers argue that the formation of the central nervous system should be considered as the watershed for the definition of life since this implies that the possibility of sensation initially exists. Up to 14 days of the embryonic period, the blastocyst has no central nervous system and therefore, cannot be considered as sensate.  If we can remove organs from brain dead declared patients who are alive in some sense, then we can use two hundred-cell embryos as cell donors at the same moral status as brain dead individuals. It is argued that the early-stage embryo is not sufficiently personalized to possess the ethical and moral weightage of personhood. Global Legislation Governing Embryonic Stem Cell Research: Legislation governing human embryonic stem cell research is not uniform and varies from country to country. Most of them have allowed the use of spare or supernumerary embryos created during in-Vitro fertilization for this purpose but have prohibited the creation of human embryos specifically for research purposes. The use of spare or excess embryos is subjected to certain provisions like informed consent, donation of embryos without financial compensation and restrictions on the use of embryo not beyond fourteen days.1 Few countries have put prohibitions on buying and selling of gametes, fertilized eggs, embryos and foetal tissues. But some countries with a more liberal view have allowed the creation of human embryos for research purpose by somatic cell nuclear transfer technique. India has allowed the establishment of new HESC lines with spare, supernumerary embryowith prior approvalofthe Institutional Committee for Stem Cell Research and Therapyand Institutional Ethics Committee.2 Need for Definitive Legislation: Since the spare embryos created during infertility treatment are the most valuable source of embryos,India lacks in having definite legislation regulating artificial reproductive technologies (ART).  The existing guidelines directing stem cell research including embryonic Stem cell are prepared by the Indian Council for Medical Research. These recommending guidelines have two inherent defects.  One, these guidelines do not have any legal effect and second, it has no penal provisions for violating the rules/policies mentioned in these guidelines. The absence of effective legislation will raise serious objection regarding the rights of the donor of embryos, the number of spare embryos, quality of the embryos, preservation, and disposal of frozen embryos etc. The presence of definite enactment will help to regulate the activities of ART clinics b imposing strict accountability and responsibility through penal provisions.  Registration of ART clinics should be made mandatory and subjected to periodic supervision to ensure a high standard of norms, care, quality of treatment and facilities offered by them specifically because of use, disposal and preservation of embryos. The rights and autonomy of the donor couple and donor of gametes should be adequately protected.  Informed consent of the donor regarding the use and destruction of the spare embryo should be taken. The question of financial compensation given to them should be adequately addressed keeping into mind the relevant existing rules and regulations of the country. Legislation similarly on the lines of the Human Fertilization and Embryo Act, as prevalent in the United Kingdom, will help to lessen the problems associated with ART. Legal and Constitutional Status of the Embryo/Unborn Foetus: All humans are born free and equal in dignity and rights.3  The word born was used to exclude the foetus and embryo from granting human rights. An amendment was suggested and rejected that would have deleted the word “born”, as it was deliberated to protect the right to life from the moment of conception.4 Even the Convention on the Rights of the Child does not recognize the right to life until birth.  Thus, a foetus has no rights under UDHR. The main standard for the protection of human life in general international law is Article 6 of (CCPR).  Article 6 of the CCPR, in its first paragraph the  norm prescribes  that  every human  being  has  the  inherent  right  to  life. However, the phraseology of the norm doesn&#39;t outline the term “human being.5  The unborn foetus has the full potential to become a human being in the right environment. In the liberal interpretation of the above fundamental right, one can conclude that the unborn foetus, from conception to birth, has a right to life and it is immaterial whether the foetus is created in vitro or in vivo. Now, if this standard was applied to all forms of unborn life, not only would research with embryonic stem cells infringe upon art.6 CCPR, but the legality of abortion laws would also be highly debatable. The US Judiciary system has never ruled on the constitutional status of embryos outside of the body and most US states have no law on the matter. But the court has ruled that foetuses are not persons within the meaning of the 14th Amendment, and thus do not have constitutional rights as such. Presumably, that ruling would also extend to embryos as well.6 The person’s right shall be protected by law and, in general, from the moment of conception and not to be deprived of his life.7 But the Inter-American Commission on Human Rights, one of two adjudicatory bodies that interprets and monitors compliance with the American Convention, has clarified that this protection is not absolute.But the Inter-American Commission on Human Rights, one of  two  adjudicatory  bodies that interprets and monitors compliance with the American Convention,  has clarified  that  this protection is not absolute.8 Everyone’s right to life shall be secured under law as provided under the European Convention on Human Rights provides. In Paton v. United Kingdom, held that the Convention language “tends to support the perspective under Art 2 and acknowledged that recognition of an absolute right to life before birth would be averse to the object and purpose of the Convention. In Vo v. France, the European Court of Human Rights, which interprets and monitors compliance with the European Convention, affirmed that “the unborn child is not regarded as a ‘person’ directly protected by Article 2 of the convention and that if the unborn do have a ‘right’ to ‘life,’ it is implicitly limited by the mother’s rights and interests, including her rights to life, health, and privacy.” The above judgment brings forth another controversial issue of foetal rights versus maternal rights of autonomy. The liberalized abortion laws existing in different countries and so proposed by various organizations have determined the precedence of maternal rights over foetal rights.9 The fundamental right to life is guaranteed under Article 21of the Constitution of India that no person shall be deprived of his life or personal liberty except according to procedure established by law.  Even here the term “person” is not defined. The Indian Legal System provides for the protection of the rights of the foetus through provisions provided under the Indian Penal Code (IPC) which deals with miscarriage. Section 315 IPC deals with and Section 316 IPC deals with the penal provisions, the unborn child is protected from any act which prevents it from being born and provides punishment for causing its death which is considered equivalent to culpable homicide. In the above penal provisions, the unborn child is protected from any act which prevents it from being born and provides punishment for causing its death which is considered equivalent to culpable homicide. Section 416 of Code of Criminal Procedure (CrPC) 1973 provides for postponement of capital sentence of pregnant women and to commutes the sentence to life imprisonment in such circumstances. This provision is made to protect the life of the unborn foetus as it has nothing to do with the act committed by the pregnant woman. Here the legislation has considered the unborn foetus as a distinct and separate individual/entity with the right of protection against potential harm. This provision is made to protect the life of the unborn foetus as it has nothing to do with the act committed by the pregnant woman. Here the legislation has considered the unborn foetus as a distinct and separate individual/entity with the right of protection against potential harm. Here the statute has defined the unborn as a legal person by fiction and acquire benefit in receiving property as transferee. The unborn foetus is protected against potential harming in the same manner as the fundamental rights of non-Interference with personal life and bodily integrity guaranteed to a human person. If the embryo is granted the status of Personhood, then they too will have the right not to be harmed or killed with imposed obligations of not to do so. With the lack of clarity on the status of the embryo and deliberations put forth by constitutions of various countries and decision given by competent courts, it can be assumed that the foetuses are not the person and hence cannot enjoy fundamental constitutional rights meant for human beings or persons. Though the Indian Penal Code and Code of Criminal Procedure protect the foetus from potential harm the Indian Constitution is silent on this aspect of extending the fundamental rights to the unborn foetus in clear terms. The European Commission states that, despite the diversity of views on the moral status of the human embryo among its member states, one can find two conflicting tendencies emerging concerning the moral position of the embryo and the legal protection which should be afforded the embryo concerning scientific research. These two positions are: 1.  Human embryos have the equal moral status as human persons and consequently, are worthy of equal protection. 2.  Human embryos do not have the equal moral status as human persons and consequently have a relative worth of protection. The spare embryos which are the Outcome of infertility treatment are the essential source of embryonic tissue. These embryos can either be used for embryonic stem cell research or can be discarded as leftover material once the objective of infertility treatment is achieved. The transformation of discarded embryos into stem cells has been referred to by one scientist as the process of turning ‘garbage into gold.10 The child intending couples have to make the emotional, physical and financial investment to reap the benefits in terms of successful pregnancy through this beneficial outcome cannot be always guaranteed. That so considered ‘waste materials’ has economic value considering the initial substantive financial and emotional/physical cost incurred by these donors. Also, the potential commercial value associated with the result of embryonic stem cell research using such embryos might be tremendous. Pharmaceutical and Biotech companies will earn substantive commercial profit that may eventually flow from this work. This raises an important query about the right of the donor couple to seek or claim financial stake or compensation. Nevertheless, it is illegal under the Human Fertilization and Embryo Act (HFEA) of the United Kingdom (UK) for them to incur any financial reward for donating their embryos and they have no financial stake in any materials or procedures developed from their donation.11 Most commentators support a ban on the ‘sale of embryos.  For example, the European Group on Ethics in Science and New Technologies has stated that ‘embryos, as well as cadaveric tissues and foetal tissues, must not be bought or sold Measures should be taken to prevent such commercialization. It is illegal for gametes to be bought or sold. A rising number of biotech and pharmaceutical companies are gathering an array of ‘valuable’ bodily materials including DNA samples and umbilical cord blood (also used for stem cell research) from various corners of the globe for scientific and commercial exploitation. However, the issue of making payments to gamete donors or embryo donors remains ethically controversial as it may lead to “commodification of the body. In January 2003, representatives from several countries, including the US, met in London to consider initiating a Human Genome Project equivalent for stem cells. The idea would be to gather results of stem cell research efforts worldwide into a comprehensive stem-cell program with global reach, avoiding the legal quagmires and inefficiencies of each country moving forward independently. Roger Pederson, a senior stem-cell investigator at the UCSF Medical Center who moved to England the advantage of the more encouraging regulatory environment is championing the idea most vigorously. Bush&#39;s policy has stimulated considerable interest in the private sector to push for and support stem cell research. The foundations which have been established for Medical Research have given tens of millions of dollars to various laboratories, many in Europe. Several universities, teaching hospitals and biotechnology companies have also stepped up their involvement in the field, as have wealthy individuals. Late last year, an anonymous benefactor gave Stanford University twelve million dollars to build a stem-cell research centre, and Andrew s. Grove, the Intel chairman, gave the University of California-San Francisco five million dollars for such a centre. Exponents and opponents of embryonic stem cell research debate when human life begins, whether destroying embryos amounts to murder, and whether the moral good of deriving medical breakthroughs outweighs the ethical in injunctions. Therapeutic cloning is not acceptable to those who believe that a human being is created at the instant of fertilization. That belief is powerful and ultimately transcends scientific disagreement. The question is for our democracy on how tightly that spiritual belief should bind the hands of those who disagree with it. Under the Human Fertilisation and Embryo Act, import of embryos or gametes is regulated through the licensing arrangements for embryo research: a licensee who wishes to import embryos or gametes must apply for and receive authorization from the HFEA. Other tissue imports are regulated by the Human Tissue Act 2004, which relaxes the rules ordinarily applying to non-imported material. The rule that tissue may only be used under appropriate consent is specifically abrogated. Nonetheless, the Human Tissue Authority is preparing some guidance on good practice and scientists importing ES cell lines should seek the approval of the Steering Committee of the UK Stem Cell Bank. The regulatory system has thus provided a skeleton framework for monitoring the acceptability of using imported bodily material in stem cell research and therapy. Others believe that life begins with the emergence of personhood. That is the time when the emerging being first can sense its environment and respond to light, pain, pleasure, sound, and other external stimuli. This occurs many weeks into gestation in humans and far beyond the 5–6-day period when blastocysts would be harvested for stem cells. Indeed, there was a period that even the Roman Catholic church believed that life began at "quickening", the time at which a mother could sense the movement of an unborn child in the womb. Generally, this occurred around 40 days into gestation or 10 weeks after fertilization, just about the same time the organism ceases to be known as an embryo and is referred to as a fetus. This doctrine was modified in the late 19th century and presently it is thought that life commences now of conception for those who ascribe to the Roman Catholic faith. It is interesting to note that Sen. Orin Hatch (R-UT) has personally concluded that life does not begin now of fertilization because there is "zero chance" of becoming a human being until it is implanted.12 Since implantation happens after the blastocyst stage during which harvesting of stem cells takes place, Hatch is at peace with himself for condoning stem cell research for therapeutic purposes. The issues which have discussed so far are, for the most part, ones that relate either to embryo research in general or to Embryonic Stem cell research. However, some other issues arise in stem cell research. The important question is whether it is ethical or legal to use imported material where the consent process meets the rules of the country of the collection but not the standards of ethical sourcing that apply in the country of destination.12 CONCLUSION The Universal Declaration of Human Rights states that everyone has right to a standard of living adequate to the health and wellbeing of himself and his family. Art 25(2). Art 39(e) of the Indian Constitution enjoins the State to direct its policies to secure the health and strength of workers. Right to life, one of the fundamental rights, as enumerated under Indian Constitution, maintains the interrelationship between law and medicine. civilization progressed, the man had to fight against diseases caused by both external and internal agents. So he had to invent and prepare curative and preventive medicines by using intellectual mind performing through Researches. Medical jurisprudence deals with those relationships which are generally recognised as having legal consequences. Research on human subjects involving cells and tissues derived from human embryos and foetuses must safeguard human rights, dignity, and fundamental freedom. This includes processes related to obtaining human tissues and cells for research, diagnosis and therapy. The fundamental tenets of beneficence, non-malfeasance, Justice, and autonomy should be adhered to in all research involving human subjects. To achieve these objectives, all research involving the use of stem cells must be guided by the general principles laid down Niels Petersen.  The Legal Status of the Human Embryo in-vitro. in the Ethical Guidelines for Biomedical Research on Human Participants published in 2006 by the Indian Council of Medical Research (ICMR) and specific principles related to stem cells as provided under these guidelines must be followed. The Indian Medical Council for Research finalized a set of guidelines for Stem Cell Research and treatment. It is also planning to set up a National Stem Cell bank to help those who cannot afford the cost. Acknowledgement: The author is grateful to authors/editors/publishers of all those articles, journals, and books from which the literature for this article has been reviewed and discussed. No Conflict of Interest No Funding Englishhttp://ijcrr.com/abstract.php?article_id=3641http://ijcrr.com/article_html.php?did=3641 Sharma A. Cell Research in India: Emerging Scenario and Policy Concerns.  Asian Biotechnol DevRev2006;8(3):43-53 Lahiry S, Choudhury S, Sinha R, Chatterjee S. The National Guidelines for Stem Cell Research (2017): What academicians need to know?Perspect Clin Res 2019;10(4): 148–154. United Nations Universal Declaration on Human Rights, 1948. Article 1. http://www.ohchr.org/EN/UDHR/Documents/UDHR_Translations/eng.pdf CopelanR, ZampasC, Bruise E, DeVore J. Human Rights Begin at Birth:  International Law and the Claim of Fetal Rights. ReprodHealth Matters 2005;13(26):120-129. Petersen N. The Legal Status of the Human Embryo in vitro: General Human Rights Instruments. ZaöRV 2005;65:447-46. Robertson JA. Stem Cell Research: Ten Years of Controversy. J Law Med Ethics 2010;12:191-203. JoséS, Rica C. American convention on human rights. Adopted at the Inter-American Specialized Conference on Human Rights, 22 November 1969. Who’s Right to Life? Women’s Rights and Prenatal Protections Under Human Rights and Comparative Law.  http://reproductiverights.org/en/document/whose-right-to-lifewomens-rights-and-prenatal-protections-under-human-rights-andcomparative European Convention on Human Rights (Internet). http://www.echr.coe.int/Documents/ConventionENG.pdf. Visited on: 03-03-2016. See Article 2(1) Thompson C. Umbilical cords:  turning garbage into clinical gold. Science 1995;270(5243):1744-1745 Corrigan O, Liddell K, McMillan J, Stewart A, Wallace S. Ethical legal and social issues in stem cell research and therapy. A briefing paper from Cambridge Genetics Knowledge Park, 2nd ed.2006. Fiechte A. History of modern biotechnology.Springer-VerlagBerlin Heidelberg 2000, 153.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcareSocio-Economic Factors Influencing the Diabetic Patients Choice of Healthcare Services in Vellore, Tamil Nadu English4650P. Gokula KrishnanEnglish Savitha NEnglishIntroduction: Diabetes is a global health concern and also a major contributor to mortality in most developing countries. People usually choose the best available option to derive maximum satisfaction from healthcare services. Based on this the present study has been done to the factors influencing the choice of healthcare services of diabetic patients. Objective: The main aim of this study is to analyse the factors determining the diabetic patient’s choice of healthcare services. Methods: To analyse the socio-economic factors determining the diabetic patient’s choice of healthcare services, a binary logistic regression model was used. Results: Factors such as age, education, gender, monthly income and treatment cost were found to be the determinants. People’s choice of health care services was not identical and needs to be improved. At .000 p – valve treatment cost had a significant impact on healthcare choice. Conclusion: The study suggests that increasing formal education on patients with diabetes would help them in choosing the appropriate healthcare services and they would be able to afford healthcare charges on their own. EnglishDiabetes, Choice of Healthcare, Binary Logistic Regression Analysis, Health Economics, Diabetic Management, Health-care serviceIntroduction Diabetes is a global health concern and also a major contributor to mortality in most developing countries.1 In India, the second-most populous country on earth, 50 million people suffer from type 2 diabetes. Non-communicable diseases are the major cause of death in India, with diabetes topping this list. Lack of knowledge on the disease condition and also unavailability of healthcare services worsen the health outcomes and increase the health burden in many developing countries. Timely intervention and medication are required to prevent complications, but most patients are unable to access quality healthcare to manage diseases.2 Service quality is an important factor in the choice of health centres for patients. Increased awareness of diseases and their adverse effect on health has led to patients seeking better quality healthcare services.3 Availability of specialist healthcare services makes possible the long-term management of diseases. Previous studies have shown that people prefer hospitals service based on their income, socio-economic condition, and health status. Patients choose healthcare services for diabetes depending upon the quality of care provided to them. Both public and private sector hospitals provide healthcare services in India though their service quality varies widely. However, an assessment of how parents choose healthcare services for managing diabetes is necessary.4 Location-based studies on such choices are limited and only a few periodic surveys on patient choice were done in Vellore, a historic town in Tamil Nadu in India with a population of 4.8 lakh according to the 2011 census. Therefore the present paper focuses on the socio-economic factors that determine the choice of healthcare services for diabetic patients in Vellore. India boasts of vast public and private healthcare network. The government provides healthcare services free of cost through public hospitals in both urban and rural areas.5 Three-tier health infrastructure has been developed in rural areas where the size of the health centre depends upon the population. Mountainous areas are served by sub-centres, rural areas have primary health centres (PHC) and community health centres are present in both rural and urban areas. Tribal populations are provided by healthcare at no cost in sub-centres. PHCs operate in villages that come under the administration of village panchayat. PHCs are small clinical centres with limited infrastructure, providing basic medical aid to the rural people and focusing on creating awareness about health and healthcare promotions.6 Community health centres render services to people in rural and semi-urban regions. They have a comparatively bigger infrastructure than PHCs. Public healthcare in urban areas is provided through district hospitals and sub-division hospitals. The cost of care in privately run hospitals is high and also varies depending upon many factors.7 Materials and Methods Several studies have been done on factors influencing a diabetic patient’s decision to choose from available healthcare services.8 Most of the studies have been conducted in developed countries, and similar studies are very few in developing countries like India. To utilise healthcare services, people look at the desired goals of efficiency, equality, good quality and responsiveness to people’s needs. Here the emphasis is on how patients value their health. Socio-demographic factors such as age, education and gender influence the choice of patients in seeking healthcare services. In this study, the authors found male respondents to seek care in private hospitals more than in public hospitals. It was observed that females were less likely to tolerate the long waiting time in government hospitals. Older patients were seen to prefer public hospitals because the distance is a top priority, and disease pattern and being more demanding of hospital services than younger patients force them to come to public hospitals. Older patients have more experience of the available healthcare and knowledge of services rendered in hospitals.9 Mostly they tend to compare the services received between the past and the present before choosing health centres. Cowling et al. (2014) found sanitary conditions, employment area and gender to have a considerable impact on the patients’ choice of healthcare services. Their evaluation of services rendered and the resulting cure determines their specific choice of healthcare services.10 The work of Abdulaziz M et al. revealed that people having health insurance visit PHCs very less than people who did not have health insurance. Health insurance had a significant effect on hospital visits.11 In their study, age and gender did not have a significant effect on hospital choice, whereas education was found to have a significant influence. The objective of Oladigboluet et al. was to assess the socio-economic factors that influence people’s healthcare utilisation. They identified three important predictors such as social class, treatment cost and educational status to influence their utilisation of healthcare services.13 The odds of paying treatment fee are twice lower in the case of people from the lower social strata with informal education. Due to their difficulty in paying, poor people are largely deprived of quality healthcare services. According to Malik et al. people consider healthcare professionals’ efficiency, reputation and hospital effectiveness as important factors in their decision making.14 People were ready to pay the high cost and endure long waiting times for seeking care in high-quality hospitals. Islam M. state that healthcare plays an important role in keeping society in good health and helps improve public health. Social factors that determine the healthcare choices are birthplace, age, education, work and living environment.15 Most of these factors shape the quality, equality, quantity and equal distribution of resources in healthcare services. Source of Data Primary data was collected for the study through interviews. A meeting was arranged with respondents directly in their houses for a face-to-face interview to understand to which factors they pay attention to when choosing healthcare services. A total sample of 200 diabetic patients was selected for the study. Blocks with a high prevalence of diabetes within the Vellore municipal corporation limits were chosen for the study. A binary logistic regression model at 95.0 % CI was used for the analysis using SPSS software version 20. Hypotheses H01. There is no significant relationship between the choice of healthcare services and age, gender and monthly income of the respondents.16 H02. There is no significant relationship between the choice of healthcare services and the community of the respondents.17 General Linear Regression Model Logistic regression model was applied using the following equation: Y = β0 + β1 X1 + β2 X2 + β3 X3 + β4 X4 + β5 X5 + β6 X6 + + Ui                             ---------- (1) Here Choice of healthcare services as dependent variable (Y) and age, gender, monthly income of the respondents, insurance status, community as independent variables (X) as shown in Table 1. Binary Logistic Regression Model Logistic regression is one of the most significant models used for binary classification type variables.18 Logistic regression is mainly used for assigning discrete values like 0 and 1. In the present study, we assigned the value of 0 to a government hospital and 1 to a private hospital. The data should be linear. If it is a non-linear function, it is converted into a linear function by applying the following equation.22 ln(P1-P)= β0+ β1 X1 + β2 X2 + β3 X3 + β4 X4 +β5 X5+ Ui                    ---(2)     The term P/ 1 – P is called an odd ratio. Values β0,β1,β2,β3and β4are coefficients of the independent variables. Results and Analysis The results of the logistic regression analysis revealed that age, education, gender, marital status as well as a direct cost and indirect cost of care were significant determinants for patients in choosing healthcare services. Patients primarily consider cost and at times pay attention to quality as well. These factors had a strong effect on the choice of hospital for seeking diabetes treatment. Table 2 shows the demographic characteristics of the respondents of the study. 51% of the respondents were male and 49% were female. 30% of the respondents choose a government hospital and 70% choose a private hospital for the treatment. Table 1 shows the results of the binary logistic regression model. The variable ‘monthly income’ was found to be significant at .007. Variables having a significant value have an impact on choosing healthcare services. The unstandardized regression coefficient is expressed as the change in standard error.18 Discussion Table 1 shows the goodness of fit in the logistic regression model. B represents the values for the logistic regression equation for predicting the dependent variable from the independent variable22. They are identified as log-odds units. The prediction equation is Log (p/1 – p) = b0 + b1*× 1 + b2*×2 + b3*×3 + b4*×4 +ui                                                 - - - - (3) In Table 2 the least square regression coefficient value of 0.84 for education implies the predicted amount of the change in the dependent variable (choice of health care), for every variable, alters the value of independent variables. Therefore the logistic regression equation is written as log(p/1 – p) = 3.211 + 0.702 *Insurance + -.025*gender + -2.326*age (1) -1.372*age (2) – 1.673*age (3) + 1.534*education (3) + 19.8* community      - - - - (4) The reference group ‘Insurance’ is assigned a value of 0 and 1. So this coefficient represents the difference between Insurance 1 and Insurance 0. For every one-unit increase in monthly income, we expect a 0.000 increase in the log-odds of choice of health care, holding all other independent variables constant. A constant value is the expected value of the log-odds of choice of health care when all the predictor variables equal to zero. Hence we reject the first null hypothesis (H01) and the result predicts there is a relationship between the choice of healthcare services and age, gender and monthly income of the respondents. As there is no impact on choice of healthcare due to changes in the community, the second null hypothesis (H02) is accepted at a 95.0 % CI level. People from different community background choose their hospital not based on their community. Conclusion Patients choose healthcare services based on socio-economic factors such as age, education, gender, direct and indirect cost and insurance. The findings show that people’s choice of health care services was not identical and needs to be improved. Patients’ awareness of healthcare services has led to an improvement in their long-term management of diseases. This tends to change not only the quality of healthcare services but also the individual’s health behaviour. The study was performed considering only a limited number of socio-economic factors. Future studies taking into account more socio-economic variables are needed to validate our findings. A knowledge of factors that directly or indirectly influences the patient’s healthcare choice helps policymakers and managing authorities to understand the importance of their healthcare services from the provider’s perspective. The study suggests that increasing employment opportunity and formal education of patients with diabetes would help them in choosing the appropriate healthcare services and they would be able to afford healthcare charges on their own. Acknowledgement: The authors acknowledge the immense help received from the scholars whose articles are cited and included in references to this manuscript. The authors are much grateful to the authors/editors/publishers of all those articles and journals from where the literature for this article has been reviewed and discussed. Conflict of Interest: The authors declare there is no conflict of interest. Source of Funding: Nil Ethical Clearance: No experiment was done on humans or animals. Authors Contribution: Dr. N. Savitha and P. Gokulakrishnan contributed to the design and implementation of the research work. Englishhttp://ijcrr.com/abstract.php?article_id=3642http://ijcrr.com/article_html.php?did=3642 Al-Doghaither AH, Rahman A, Saeed BM, Magzoub AA. Factors influencing patient choice of hospitals in Riyadh, Saudi Arabia. R Soc Health J Health 2003;123(2):105-109. Al-Rubeaan K, Al-Manaa H, Khoja T, Al-Sharqawi A, Aburisheh K, Youssef A, et al.  Health care services provided to type 1 and type 2 diabetic patients in Saudi Arabia. Saudi Med J 2015;36(10):1216-1225. Alsubaie A, Almohaimede K, Aljadoa A, Jarallah O, Althnayan Y. Socioeconomic factors affecting patients′ utilization of primary care services at a tertiary teaching hospital in Riyadh, Saudi Arabia. J Family Community Med 2016;23(1):6-18. Chauhan V, Sharma A, Sagar M. Exploring patient choice in India: A study on hospital selection. Int J Healthc Manag 2019;1(11):89-103. Cowling K, Dandona, Dandona L. Social determinants of health in India: Progress and inequities across states.Int J Equity Health 2014;13(88):1-12. Hassan A, Mahmood K, Bukhsh HA. Healthcare System of Pakistan.Int J Adv Res. 2017; 1(4):171-73. Ewert B. Is patient choice the future of health care systems? Int J Health Policy Manag. 2013;1(3):227-228. Islam M. Social determinants of health and related inequalities: Confusion and implications. Front Public Health 2019;7:232-24 Khalid A, Al-Rubeaan, Hamad A. Al-Manaa, Tawfik A. Khoja, Ahmad H. Al-Sharqawi, Khaled H. Aburisheh, Amira M. Youssef, et al. Health care services provided to type 1 and type 2 diabetic patients in Saudi Arabia. Saudi Med J 2015;36(6):1216-1225. Hokshi MC, Patil B, Khanna, Neogi SB, Sharma J, Paul VK, et al. Health systems in India. J Perinatol 2016;36:9-12. Mohammad MA. Patient choice of a hospital: Implications for health policy and management. Int J Health Care Qual Assur 2014;27(2):152-164. Ul Haq NW, Taneja K, Adlakha N. Health System in India: Opportunities and Challenges for Enhancements. Int J Manag Bus 2013;9(12):74-82. Oladigbolu R, Oche M, Kaoje A, Gana G. Socio-economic factors influencing utilization of healthcare services in Sokoto, north-western Nigeri. Int J Trop Dis Health 2017;27(2):1- Rout SK, Sahu KS, Mahapatra S. Utilization of health care services in public and private healthcare in India: Causes and determinants. Int J Healthc Manag 2019;(5):1-8. Shumaila A, Iqbal J, Waris H, Ismail M, Naseer A. Health Care System in Pakistan: A Review. Res Pharm Sci 2016; 2(3):211-216. Siddique MK, Islam SM, Banik PC, Rawal LB. Diabetes knowledge and utilization of healthcare services among patients with type 2 diabetes mellitus in Dhaka, Bangladesh. BMC Health Serv Res 2017;17(1):552-558. Singh B, Singhi, R. Servqual. Impact on overall satisfaction and brand loyalty: An empirical study in Delhi-NCR hospitals. Int J Technol Manag 2018;17(1):49-60. Sperandei S. Understanding logistic regression analysis. Biochem Med 2014;5:12- Chatterjee S, Riewpaiboon A, Piyauthakit P, Riewpaiboon W, Boupaijit K, Panpuwong N, et al. Cost of diabetes and its complications in Thailand: A Complete Picture of Economic Burden. Health Soc Care Community 2011;19(3):289-298. Victoor A, Delnoij DM, Friele RD, Rademakers J. Determinants of patient choice of healthcare providers: A scoping review. BMC Health Serv Res 2012;12(1):25-38. Yadav A, Hui L, Ali M, Anis M. Analysis of healthcare data of Nepal hospital using multinomial logistic regression model. Int J Inf Technol 2016;11(2):2720-2730. 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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-52411307EnglishN2021April12HealthcarePhytochemical Composition and Antioxidant Activity of Two Varieties of Unripe Date Palm English106111M. GayathriEnglish S. ThilagavathiEnglishIntroduction: Date palm (Phoenix dactylifera) has been used for nutritional and medicinal purposes. They are used as medicine for the treatment of several diseases and also highly recommended for pregnant women before and after delivery. Objective: The present work aims to explore the phytochemicals present in unripened Dates (RD) and Yellow Dates (YD)and assess their antioxidant property. Methods: Different solvents such as aqueous, methanol, benzene, petroleum ether and chloroform were used to extract the phytochemicals present in it DPPH (2, 2-diphenyl-1-picryl-hydrazyl-hydrate), FRAP (ferric reducing antioxidant power), Chelation of Fe2+ and SOD (superoxide dismutase) were used to assess the antioxidant activity. Results: The screening of phytochemicals showed the presence of alkaloids, flavonoid, phenolic compounds, saponin, tannin, steroid and carbohydrate, however, the antioxidant activity revealed that the % inhibition of DPPH, FRAP, Chelation of Fe2+ and SODwere about 94.05 ± 2.73, 77.91 ± 2.96, 88.02 ± 2.50 and 82.50 ± 2.58 respectively for RD and 85.13 ± 2.74, 68.03 ± 2.66, 79.64 ± 4.0 and 72.30 ± 2.10 respectively for YD. Conclusion: Both YD and RD unripe date palm demonstrated high phytochemical content and are potent antioxidant activity. The analysis of qualitative and quantitative phytochemicals such as alkaloids, flavonoids, phenolic compounds, tannins, terpenoids and steroids revealed that the antioxidant activity of the YD and RD is due to these phytochemicals. EnglishPhoenix dactylifera, phytochemicals, antioxidant, flavonoids, phenolic compounds, alkaloidsINTRODUCTION             Phoneix dactyliferea commonly known as the date palm is an important plant in the scorched region of southwest Asia and North Africa.1 Dates are a good source of energy, vitamins and a group of elements like Phosphorus, Iron, Potassium and a significant amount of calcium.2 Besides its nutritional value date palm is also rich in phenolic compounds possessing antioxidant activity. The protection against oxidative stress in several diseases is mainly due to the presence of various antioxidants and vitamins present in the fruit. Antioxidants that can quench reactive free radicals can prevent the oxidation of other molecules and may therefore have health-promoting effects in the prevention of degenerative diseases.3 The interest in antioxidant has been increasing because of their high capacity in scavenging free radicals related to various diseases.4 Free radical oxidative stress has been implicated in the pathogenesis of a wide variety of clinical disorders such as cancer, diabetes, multiple sclerosis and arthritis.5 The human disease is believed to be due to the imbalance between oxidative, tissue damage and hence prevent diseases progression by antioxidant defence supplements.6 The in-vitro study reported that the aqueous extract of palm date fruit has antioxidative and anti-mutagenic properties.7 Flavonoids are known to possess anti-inflammatory, anti-oxidant, anti-allergic, hepatoprotective, anti-thrombotic, neuroprotective and anti-carcinogenic activities.8 However, studies deal with the fruit of date palm, not on the unripe or raw dates which were found to possess β – carotene the antioxidant vitamin. Here in this study, we determined the phytochemical content and their antioxidant property of two different varieties of unripe date palm. MATERIALS AND METHODS Sample collection             Two different varieties of PhoneixdactylifereanamelyBarhi (Yellow colour date, YD) and Khenaizi (Red colour Date, RD) were procured from Shariah date palm located in Dharmapuri district of Tamilnadu in India in June and July 2018. The selected dates were such that, they are uniform in size, free from physical damage and injuries by insects and fungal infection. The sample was washed, cut (uniform thickness) and dried using a cabinet drier at 65?C then it was ground into a fine powder and used for analysis. Preparation of extract Briefly, the samples were processed according to the procedure.9 The finely powdered samples of both varieties were extracted using a soxhlet extractor approximately 50gm of the sample was suspended in 100ml of 5 solvents aqueous, methanol, benzene, petroleum ether and chloroform at room temperature for 3 hours and the extract was collected. The extract was evaporated to dryness and stored at 4?C until use. Qualitative phytochemical screening The prepared date palm extract was screened for the presence of phytochemicals using the method.10 Quantitative phytochemical analysis             The Quantitative Phytochemical analysis was performed on testing different chemical group which are present in unripe date palm powder using the standard analytical procedure.11 Determination of antioxidant activities             The antioxidant activities of unripe date palm powder were measured using 4 different methods namely 1. DPPH(2, 2-diphenyl-1-picryl-hydrazyl-hydrate) free radical scavenging assay, 2. Ferrous ion chelating of activity, 3.Ferric reducing antioxidant power assay and 4.Superoxide dismutase activity method. DPPH free radical scavenging activity The antioxidant activities were measured in terms of hydrogen donating or radical scavenging ability using the stable radical DPPH.7 The analysis was carried out in a 96 well microtiter plate.  The analysis was done using 200 ml of DPPH solution to the 10 ml of both test solution and the standard solution and kept separately in the wells of the microtiter plate.  The final concentration of the test and standard solutions used were 250, 200, 150, 100, and 50 mg/ml. The plates were incubated at 37o C for 30 min and the absorbance of each solution was measured at 490 nm, using a 96 well microplate reader. The percentage inhibition of DPPH radical scavenging activity was calculated as follows. % Inhibition = [(Ab (blank) – As (sample) / Ab(blank)] * 100             Where Ab is the absorbents of blank As is the absorbents of sample. Measurement of ferrous ion chelating activity Iron-chelating abilities of the methanol extract of YD and RD and standards were estimated by the method.12 Four dilutions of dimethyl sulphano oxalate (DMSO) (20 mg/mL, 10 mg/mL, 5 mg/mL and 2.5 mg/mL) were prepared from the dried extracts. Briefly, 0.05 mL of each dilution was added to a 2.7 mL TRIS buffer (pH=7.4). Thereafter, 0.05 mL of 2 mM FeCl2 were added and vortexed for 15 sec. At 30 sec, the reaction was initiated by the addition of 5 mMferrozine (0.2 mL), the mixture was shaken vigorously at Vortex (VelpScientifica, UE) for 10 sec.  After 1 min beyond the addition of ferric chloride solution (FeCl2), the absorbance of the solution was measured spectrophotometrically at 562 nm. The ability of the sample extracts was calculated using the formula: chelating activity (%) = 100 x [(ACAS)/AC], where AC is the absorbance of the control, and AS is the absorbance of the sample. Ferric reducing antioxidant power assay Different concentrations of the methanol extract of RD and YD and its various fractions (10-50 μg/mL) was added to 2.5 mL of 0.2 M sodium phosphate buffer (pH 6.6) and 2.5 mL of 1% potassium ferricyanide [K3Fe(CN)6] solution. The reaction mixture was vortexed well and then incubated at 50°C for 20 min using a vortex shaker. At the end of the incubation, 2.5 mL of 10% trichloroacetic acid was added to the mixture and centrifuged at 3,000 rates per minute for 10 min. The supernatant (2.5 mL) was mixed with 2.5 mL of deionised water and 0.5 mL of 0.1% ferric chloride. The coloured solution was read at 700 nm against the blank concerning standard using Ultra violetSpectrophotometer. Here, ascorbic acid was used as a reference standard, the reducing power of the samples was compared with the reference standard. Superoxide dismutase activity The Superoxide radical scavenging activity was measured by the method of Fontana et. al. Based on this method, the activity is measured by the reduction of riboflavin/light/NBT (Nitro blue tetrazolium).  About 1 ml of reaction mixture contained Phosphate buffer, NADH, NBT and various Concentrations of sample solution. Mainly the method is based on the generation of superoxide radical by auto-oxidation of riboflavin in presence of light. The Nitro Blue Tetrazoliumwas reduced by Superoxide radical to a blue coloured formazan that can be measured at 560 nm. RESULTS AND DISCUSSION             YD and RD varieties of unripe date palm powder extracts were analyzed for their content of the phytochemical and antioxidant activity. Different Phytochemical have been found to possess a wide range of activities, which may help in protection against chronic diseases.13 Preliminary phytochemical of both varieties have been shown in Table 1. From the obtained results compared to all other extracts, the methanol extract showed the presence of more phytochemicals such as alkaloid, flavonoid, phenolic compounds, carbohydrates, saponin, steroid and tannin. RD contain a high concentration of phytochemicals than that of YD. Shah Alam Khan et al., (2016) shows the date variety such as fourth has the order of solvent according to the extraction of phenolic compounds was observed to be acetone ? methanol ? ethanol ? water while for khasab it was in the order of ethanol ? methanal ? acetone ? water.14 Alkaloid exhibit marked physiological effects when administrated to animals and hence their wide use in medicine for the development of drugs.15             The protective role of flavonoids was especially flavanols&flavonols from cardiovascular and cancer diseases have pointed.16 Tannin is reported to exhibit antiviral, antibacterial, antitumoral activities. It was also reported as some tannin was used as a diuretic.17 Saponin which is responsible for numerous pharmacological properties.18 Phenolics can manifest in type II diabetes by inhibiting the activities of α glycosidase and α amylase to increase blood glucose level.19 Steroids or Sterols of plants are called phytosterols and possess Myriad health benefits.20 The ratio and concentration of these constituents depend on the type of the fruit, stage of fruit picking, location and soil conditions. These also contribute to the organoleptic properties of the fruit.21             Table 2 shows that the phytochemical content of the RD seemed to be in a higher concentration than that of YD. The alkaloid content of RD was about 2.36 ± 0.13 and YD was 1.89 ± 0.97. The alkaloid produced analgesic anti-spasmodic and bactericidal effect.22 Flavonoids present in plants possess diverse health benefits which include antioxidant and radical scavenging activities, deduction of certain chronic disease, prevention of some cardiovascular disorders and certain types of a cancerous process.23 From the result the flavonoid content of RD was high as about 4.17 ± 0.19 and low in YD as 3.75 ± 0.03. The flavonoid and alkaloid content of the date palm fruit was about 34.29 and 5.20% respectively.24 Likewise the anthraquinone content of RD and YD was about 0.19 ± 1.16 and 0.14 ± 0.12 respectively. Triterpenoids and Saponin showed analgesic properties and central nervous system activities.25 The concentration of the value of saponin in both RD and YD were about 0.02 ± 0.08 and 0.01 ± 0.05 respectively, which is within the WHO permissible limit of 48.50 mg/100g as recommended 2003.26 The terpenoid content of RD was about 0.08 ± 1.07 and YD was about 0.05 ± 1.14. Similarly RD has a high content of Tannin at about 0.86 ± 0.15 than that of YD at about 0.49 ± 0.01. The tannin content of date palm fruit is about 7.51%.27 From the phytochemicals tannin is understood to react with protein to supply the standard tanning effect that’s important for the treatment of inflamed or ulcerated tissues.28 Invitro antioxidant activities of YD and RD date palm             Among the four antioxidant activity assay, the methanol extract of red dates shows more % inhibition than that of yellow dates. Several methods and modification have been used for the measurement of antioxidant activity. A single method is not enough to assay the whole antioxidant activity therefore application and combination of several tests recommended providing a better description. The scavenging activity of both samples was expressed as % inhibition and was compared with standard antioxidant as ascorbic acid. As a result, shows that the concentration of both samples increased the % inhibition as the concentration of sample increases. This means that the data indicating the hydrogen donating ability of the sample antioxidants such as phenolic compounds. This is similarly based on another study which relates the hydrogen donating ability using the DPPH method to the presence of phenolic and polyphenolic compounds.29 In the presence of hydrogen donors, DPPH is oxidized and stable free radical is formed from the scavengers. The methanol extract of RD showed higher % inhibition as 94.05 ± 2.73 for 100µg/1ml concentration, for the same concentration YD shows a lower effect as 85.13 ± 2.74. Date fruit has the second antioxidant activity among 28 fruits by using the FRAP test.30 FRAP assay of methanol extract of both samples were done. The value is compared to that of the standard antioxidant as vitamin C. RD had greater antioxidant activity as 77.91 ± 2.96 for 100µg/ml concentration, but the YD had only 68.03 ± 2.66 for the same concentration. Iron is essential for life because it is required for oxygen transport, respiration and the activity of many enzymes.31 Iron can generate free radicals from peroxides by Fenton reactions. So the production of these radicals can lead to lipid peroxidation, protein modification and DNA damage.32 Minimization of Fe2+ concentrations in the Fenton reaction affords protection against oxidative damage.27 Chelation of Fe 2+ ions activity of methanol extract of both samples were shown higher % inhibition compared to YD (79.64 ± 4.0), RD had higher value such as 88.02 ± 2.50 for 100µg/ ml concentration. Superoxide radical scavenging activity of methanol extract of RD had 82.50 ± 2.58 % inhibition, but the YD had only 72.30 ± 2.10 % inhibition for 100 µg/ml concentration. The antioxidant activity of RD and YD extract was estimated at different concentration in methanol extract and the result was graphically shown in figures 1-4. The graph shows the methanol extract of both the RD and YD varieties against the standard ascorbic acid. Among all the antioxidant activity assay the methanol extract of RD shows more effect than that of YD. CONCLUSION The detailed information from this study confirms that both YD and RD unripe date palm demonstrated high phytochemical content and are potent antioxidant activity. The analysis of qualitative and quantitative phytochemicals such as alkaloids, flavonoids, phenolic compounds, tannins, terpenoids and steroids revealed that the antioxidant activity of the YD and RD is due to these phytochemicals. The antioxidant activity was tested using various methods like DPPH, FRAP, Fe2+ and SOD proved that YD and RD are potent antioxidant and could be used as a supplement for the prevention of oxidative stress. Acknowledgement The author acknowledges the immense help received from the laboratory where the analysis had been done. The author is also thankful to the authors, editors, publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Source of funding: This article has been prepared without funding and as an independent study under the guidance of Dr. S. Thilagavathi Conflict of interest: The author expresses no conflict of interest Englishhttp://ijcrr.com/abstract.php?article_id=3643http://ijcrr.com/article_html.php?did=3643 Al-Farsi MA, Lee CY. Nutritional and functional properties of dates: a review. Crit Rev Food Sci Nutr 2008;48(10):877-887. Anwar-Shinwari M. Iron content of date fruits. J Colle Sci King Saud Uni 1987;18(1):5-13. Shahidi F, editor. Natural antioxidants: chemistry, health effects, and applications. The American Oil Chemists Society; 1997. Silva EM, Souza JN, Rogez H, Rees JF, Larondelle Y. 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