Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241923EnglishN2017November15HealthcareEvaluation of Salivary Total Protein in Insulin Dependent Diabetes Mellitus (IDDM) Patients
English0103Syed ShahbazEnglish Chandrika KattiEnglish Md. Munnawarulla KhanEnglish Kapil PawarEnglishAims: To evaluate salivary total protein in IDDM patients and to compare with healthy non-diabetic control group.
Methods and Material: The study consisted of 50 IDDM patients and 50 age matched healthy controls. All the subjects were
subjected to the salivary total protein estimations. Salivary total protein estimations were done by Biuret method, end point. All the estimations were performed using an auto analyser. Mean, standard deviation and students ‘t’ test were calculated. All these statistical analysis were performed by using SPSS 20 versionsoftware.
Results: The results showed elevated levels of salivarytotal protein in IDDM group compared to healthy controls.
Conclusion: There are definite changes in salivary composition with increased levels of salivary total protein in IDDM patients
compared to healthy controls.
EnglishIDDM, Saliva, Total proteinIntroduction
Diabetes mellitus (DM) is an endocrine disease characterized by a shortfall in the production of insulin with consequent alteration of the process of assimilation, metabolism and balance of blood glucose concentration.1 Although all forms of DM are characterized by hyperglycemia, the pathogenic mechanisms by which hyperglycemia arises differ widely. It is thus classified, as type IDDM (Insulin dependent diabetes mellitus) and NIDDM (Non-Insulin dependent diabetes mellitus).2IDDM is a multifactoral autoimmune disease for which susceptibility is determined by environmental and genetic factors. 3, 4It was estimated in 2010, that 76,000 children aged less than 15 years develop type I DM worldwide. The incidence rate of type I DM in India is 4.2/100,000 population per year. 1
Saliva has long been viewed as unique yet complex body fluids, like plasma or serum. Saliva is easy to collect by non-invasive methods and preservation is inexpensive. The diagnostic value of saliva lies in its components, flow and structure of glands. 3, 5Salivary total protein is a vital component of saliva, predominantly comprising proline-rich proteins.6, 7 Noticeably there is spurge of interest in non-invasive diagnostic testing. It has become apparent that saliva has many diagnostic uses especially in large scale screening.8With this consideration present study was done to evaluate salivary total protein in IDDM and compare with healthy non-diabetic controls.
Method and Materials:
The study was conducted on 50 patients with IDDM and 50 age-matched healthy non-diabetic individuals. The IDDM and control group consisted of 32 males and 18 females respectively. The patients included in the study were taken from Diabetic Care Centres. Patients with IDDM and individuals who volunteered to participate in the study were included and patients having systemic diseases other than diabetes mellitus were excluded from the study. The informed consent was obtained from both the groups, and ethical clearance was obtained from institutional ethical committee.
All the subjects included in the IDDM and control group were advised not to eat anything and until the samples were collected. The saliva samples from both study and control groups were collected at 8 am. The unstimulated whole saliva was collected by spitting method. The collected saliva was subjected to total protein estimation.The salivary total protein wasperformed by Biuret method, the peptide bonds of protein react with copper II ions in alkaline solution to form blue-violet complex, (biuret reaction). Each copper ion complexing with 5 or 6 peptide bonds, the colour formed is proportional to the protein concentration and is measured at 546nm (520-560nm).9, 10Allthe estimations were performed using reagent kits of Transasia Bio-Medical Ltd. and on an autoanalyser(ERBA, Mannheim, GERMANY).
Results
The study consisted of 50 diagnosed IDDM patients and 50 age-matched healthy controls. The IDDM and control group consisted of 32 males and 18 females, the age ranged from 4 to 16 years. Fasting saliva samples were collected with their consent and were evaluated for salivary total protein by using autoanalyser. The values obtained for salivary total protein were statistically analysed for mean and standard deviation (SD). Students ‘t’ test was employed to compare the values of IDDM and control group. All these statistical analysis were performed by using 20 Version SPSS software.
The mean salivary total protein level in control group was 131.42 mg/dl and SD 11.37 and the protein levels were ranged from 113 to 170 mg/dl.The mean salivary total protein level in IDDM group was 182.21 mg/dl and SD 9.02 and the levels were ranged from 162 to 192 mg/dl.The ‘t’ value for the comparison of salivary total protein in IDDM and control group was t= 24.74 and pEnglishhttp://ijcrr.com/abstract.php?article_id=2392http://ijcrr.com/article_html.php?did=2392
Gheena S., Chandrashekhar T, Pratibha Ramani. Salivary characteristics of diabetic children. Braz J Oral Sci 2011;10:93-7.
Ana Carolina U. Vasconcelos, Maria Sueli M. Soares, Paulo C. Almeida, Teresa C. Soares.Comparative study of the concentration of salivary and blood glucose in type 2 diabetic patients. Journal of oral science2010;52:293-8.
Roland Tisch and Hugh Macdevitt. Insulin dependent diabetes mellitus. Cell 1996;85:291-7.
Hedge A, Shenoy R, Prajwal D’Mello, Smitha A, Tintu A, Manjrekar P. Alternative markers of glycemic status in diabetes mellitus. Biomedical research 2010;21:252-6.
Maria-Sueli-Marques Soares, Mario-Marcio-Vasconcelos Batista-Filho, Marcele-Jardim Pimentel, Isabela-Albuquerque Passos, Eduardo Chimenos-Kustner. Determination of salivary glucose in healthy adults. Med Oral Patol Oral Cir Buccal2009;14:e510-3.
Krishna BA, Ashalatha G, Baghirath VP, Rajani Kanth AV, Malathi N. Saliva as a diagnostic biofluid-review. J OrofacSci 2010; 2:66-70.
Arati S. Panchbhai, Shirish S. Degwekar, Rahul R. Bhowle. Estimation of salivary glucose, salivary amylase, salivary total protein and salivary flowrate in diabetics in India. Journal of oral science2010;52:359-68.
Amer S, Yousuf M, Siddiqui PQR, Alam J. Salivary glucose concentrations in patients with diabetes mellitus- a minimally invasive technique for monitoring blood glucose levels. Pakistan journal of pharmaceutical sciences 2001;14:33-7.
Thomas L. Clinical laboratory diagnostics. 1st ed. Frankfurt: TH books Veriagsgessell chaft; 1998;131-7.
Tietz. N. W. Text book of clinical chemistry, W.B. Saunders, 1986;231-46.
Mandel ID. The diagnostic uses of saliva. J Oral Pathol Med 1990;19:119-25
American Diabetes Association. Diagnosis and classification of Diabetes Mellitus. Diabetes Care 2010;33:S62-9.
Herenia P. Lawrence. Salivary markers of systemic disease: noninvasive diagnosis of disease and monitoring general health. J Can Dent Assoc 2002;68:170-4.
Lopez ME, Colloca ME, Paez RG, Schallmach JN, Koss MA, Chervonagura A. Salivary characteristics of diabetic children. Braz Dent J 2003; 14:26-31.
Yavuzyilmaz E, Yumak O, Akdoqanli T, Yamalik N, Ozer N, Ersoy F, Yeniay I. The alterations of whole saliva constituents in patients with diabetes mellitus. Australian Dental Journal 1996;41:193-7.
Antonio D. Mata, Duarte Marques, Sara Rocha, Helena Francisco, Carolina Santos, Maria F. Mesquita, et al. Effects of diabetes mellitus on salivary secretion and its compostion in the human. Molecular and Cellular Biochemistry 2004;1-6.
Amita Nawalkar and Anilkumar Bhoweer. Alterations in whole saliva constituents in patients with diabetes mellitus and periodontal disease. Journal of Indian Academy of Oral Medicine and Radiology 2011;23:498-501.
Ben Aryeh H, Cohen M., Kanter Y., Szargel R., Laufer DZ. Salivary composition in diabetic patients. J Diabet Complications 1988;2:96-9.
Rosamund Harrison and William H. Bowen. Flow rate and organic constituents of whole saliva in insulin-dependent diabetic children and adolescents. Pediatric Dentistry 1987;4:287-91.
Maria A. Belazi, Galli-Tsinpoulou A, Drakoulakos D, Fleva A, Panayaiotis H. Papanayiotou. Salivary alterations in insulin-dependent diabetes mellitus. International Journal Of Pediatric Dentistry 1998;8:29-33.
Svante Tweetman, Tommy Nederfors, Birgitta Stahl Stefen Aronson. Two year longitudinal observations of salivary status and dental caries in children with insulin-dependent diabetes mellitus. Pediatr Dent 1992;14:184-88.
J. Tenovuo, O. Lehtonen, J. Vkari, H. Larjava, P. Vilja and P. Tuohimaa. Immunoglobulins and innate antimicrobial factors in whole saliva of patients with insulin-dependent diabetes mellitus. J Dent Res 1986;65:62-66.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241923EnglishN-0001November30HealthcarePrevalence of Anaemia in Child Bearing Women: A Challenge
English0408Shiv Lal SolankiEnglish Bhagraj CoudharyEnglish Bhagwan Ram VishnoiEnglishBackground: Anaemia is prevalent among population of all strata; whether rich or poor, educational profile high or low, it is prevalent everywhere. Iron deficiency anaemia is a serious public health problem having a significance impact on life of everybody. It affects the physical, mental and psycho-social development of the victim, causing generalized weakness, lethargy, lassitude, with effect on optimal work performance, amount to certain behavioural problems. Status of anaemia in child bearing age has areasonable impact on whole life span and has a reasonable impact on the outcome of pregnancy.
Objectives: 1. To assess the prevalence and associated factors of anaemia.
Materials and Methods: Across-sectional purposive study was conducted on 436 females of reproductive age group (15 to 49 years), residing in the field practice area of urban health training centre of department of community medicine, Geetanjali medical college and hospital, Udaipur, (Rajasthan). Study period was from May 2015 to April 2016.
Results: The prevalence of anaemia among the age group above 18 years was (73.8%), educated (72.4%), nuclear family (73.3%), married (87.1%), and low socio-economic status group V (79.9 %). Among anaemic subjects,(76%) were vegetarian, mean haemoglobin 10.11±1.03 gm %, and (3.2%) were severely anaemic. History of worm infestation having anaemia was (89.2%), and symptoms of easy fatigability among the anaemic were (84.8%)
Conclusion: The results of the study simplify the need to concentrate on anaemia prophylaxis measures, already in existence for masses and also for child bearing women. The component of health education including nutrition education also to be strengthened with awareness to the public.
EnglishHaemoglobin, Vegetarian, Worm infestation, HealthIntroduction
Anaemia is an important public health problem which affects masses, particularly child bearing women. It is estimated that (75%) of the anaemia is due to iron deficiency, followed by folate and Vitamin B121. The adolescent health was given priority by Government of India in RCH package since 1997.The anaemia is one of the major challenges to health sector for quality of day to day life. The prevalence of anaemia is disproportionally high in developing countries, because of lack of education, poverty, inadequate diet, poor health services, and early marriages. Report of WHO 2002 showed anaemia as one among the top 10 risks for infant mortality, maternal mortality, and preterm birth. The facts show that anaemia is one of the most prevalent diseases to combat, for increasing quality and life expectancy.Iron deficiency affects more people than any other condition. As many as (66-80%) of the world population may be iron deficient, over (30%) of the population are anaemic, mainly due to iron deficiency, frequently exacerbated by malaria and worm infestations2 in developing countries.According to world health organization, the global prevalence of anaemia is 24.8%, which means about 1.62 billion people world-wide are suffering with anaemia. The highest prevalence is in preschool age children (47.4%), while the lowest prevalence in men is (12.7%).Adolescent phase of life is the right path, due to the ever increasing evidence that control of anaemia in pregnancy is easy to achieve satisfactory if the iron status can be earned during adolescence. Anaemia occurs at all stages of the life cycle, but is more prevalent in pregnant women and young children. Anaemia is the most prevalent haematological disorder in women of reproductive age with increased rates of maternal, prenatal mortality, premature delivery, low birth weight, and other adverse outcomes. Moreover it has been shown to affect the cell-mediated immunity, mental development, learning capacity, resistance to infection and work performance.
Objective: 1. To assess the prevalence and associated factors of anaemia
Materials and methods
The present cross sectional study was conducted on females of reproductive age group residing in catchment area of urban field practice, ofdepartment ofcommunity medicine, Geetanjali Medical College and Hospital, Udaipur. In this community based study the population proportion 33% was used to arrive for the required sample size of 436 females. The 1st house was selected randomly by lottery system and thereafter every alternate house was picked up, till the required sample size was covered.The study instrument used was a preformat, pretested semi structured questionnaire. Haemoglobin estimation was done by electric impedance method. The purpose of the study was explained to the study subjects and the written consent was obtained. The study was conducted during May 2015 to April 2016.
Discussion
Table 1. The Socio-demographic characteristics:
a) Anaemia and age group: In our study the prevalence of anaemia above 18 years(late) was (73.8%), and below 18 (early) (71.2%), similar results were observed by Kaur S et al3, in late (64%) as compared to early (58%), study by Biradar S S et al4 in rural area of Belgaum showed higher prevalence in late ( 60%) as compared to early (38.9%)
b) Religion: The prevalence of anaemia in our study, among Hindus was slightly more (82.6%), compared to Muslims (80.8%), whereas Sachan B etal5 in their study, observed the prevalence (59.2%) in Hindu against (37.5%)Muslims.Similar results, Hindu(53.96%) and Muslims(47.92%), were seen by Bilkish N et al6 in their study on 416 females in rural area of Maharashtra.
c) Family composition: Our study shows the prevalence of anaemia was more (73.3%) in nuclear families, compared to joint families (69.9%). In contrast to our results Rawat CMS et al7 studied 504 females in rural area of Daurala PHC at Meerut, observed prevalence in joint family was higher (45.2%), than nuclear families (28.3%), Bilkish N et al6 also observed that the nuclear families were slightly less prevalent (51.79%) than joint families (52.08%)d) Education In present study the prevalence of anaemia was more among educated (72.4%) compared to illiterate (68.2%), this is supported by Bhanushali et al8 in their study among 387 females that among 104 anaemic females, the (55.8%) were literate. As education increases the prevalence of anaemia decreases, in contrast to our study Kaur S et al3observed, high prevalence among illiterate (66.7%), and Bilkish N et al6observed (71.32%) in illiterate and only (20.25%) in literate females. e) Marital status: Our study shows more prevalence of anaemia among married females (87.1%) compared to unmarried (70.9%). NFHS-3, Rajasthan9 also shows the prevalence of anaemia more in married (54.3%) compared to unmarried (49.9%).
f) Socio-economic status: It is observed that as socio-economic status increases prevalence of anaemia decreases, as in our study it is highest among class V (79.9%) and lowest among class III (67.9%). No female of class I was observed anaemic. Similar results were also observed by Kaur S et al3class V (73.4%), class II (54.3%), class I (41.7%). Rawat CMS et al7, revealed among class V (50%) and class I (27.3%). Biradar SS et al4 reported anaemia among females of class III was (4.1%) where as it was (43.1%) in class IV and (100%) in class V. The study by Bilkish N et al6 observed prevalence of anaemia among class IV and V (61.42%) and class I, II,III (56.62%).
Table 2 Mean haemoglobin level:
In our study the mean haemoglobin of those females who had anaemia was 10.11±1.03 gm% and that of non – anaemic females was 13.04±0.77 gm%. Similar results were observed by Biradar SS et al4 anaemic girls 10.9±1.04 gm%, and non-anaemic girls were 12.80±0.5gm%. Sen A et al10 studied the functional impact of anaemia in 350 young adolescent females from different schools in Vadodara and the mean haemoglobin level of total sample of females was 11.32g/dl; 10.67g/dl for anaemicand 12.68g/dl for non- anaemic.
Table 3 Severity of anaemia The present study reveals, (68.5%) females moderately anaemic, (28.3%) mild and (3.2%) severely anaemic. Study conducted by Siddharam S M et al11 had more prevalence of mild (54.9%), moderate (45.2%), and (4.92%) severe. Sharma et al12 observed similar results to our study, moderate (72%), mild (16.5%), and severe (11.5%), while the study ofToteja G S et al13observed moderate, mild, and severe (50.9%), (29.2%), (7.1%) prevalence of anaemia respectively.
Table 4 Anaemia and dietIn present study the prevalence of anaemia was more among vegetarian diet females (76%), compared to (63.3%) mixed diet. Bilkishet al6 showed prevalence of anaemia among vegetarian (55.08%) v/s (47.78%) among mixed diet, Kakkar R et al14 studied factors contributing to anaemia in 317 females in Bhopal, observed prevalence of anaemia was dependent on knowledge, literacy level, food habits, and non-vegetarian diet. Patel S et al15 in their study on 100 anaemic patients from Shri Krishna Hospital, Karamsad observed prevalence of anaemia among vegetarian was very high (84%), against non-vegetarian (16%).
Table 5 Anaemia and worm infestationIn this study the prevalence of anaemia was higher (89.2%) among females who gave a history of worm infestation as compared to those who did not give such history of worm infestation (69%). This difference was statistically highly significant. Our study is also supported by study of Goel S et al16 and Bilkish et al 6who observed that the females who had history of worm infestation were anaemic v/s non-anaemic(84.6%) v/s (43.8%) and (77.78%) v/s (50.75%) respectively. RamziM et al17 studied 363 school females in Kavar, Southern Iran, observed that only parasite infestation in the last three months had a 6.83 times more risk of anaemia than those without this history.
Table 6 Anaemia and presentation of symptoms In our study the prevalence of anaemia was higher (84.8%) among females who had symptoms of easy fatigability as compared to thosewho did not had such symptoms (60.8%), similarly with symptoms of reduced working capacity (82.4%) v/s (69.3%) with no such symptom, with symptom of palpitation (75.3%) v/s (71.3%) without palpitation. Goel S et al16 observed the signs and symptoms like headache, fatigue, dyspnoea, paraesthesia and attack ofsyncope, significantlymore prevalent in anaemic subjects.
Conclusion
The study was carried out to understand the current prevalence and associated factors of anaemia among females of child bearing age. To prevent anaemia, publicshould be made aware through conduction of health and nutrition education campaign, so that the current comprehensive health services may be utilised properly. Our study emphasised forvarious measures of social and economic development to be initiated for lowering the prevalence of anaemia to improve the quality of life of the females.
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 received and discussed.
Funding: No funding sources.
Conflict of interest: None declared
Ethical approval:This study was approved by the Institutional Ethics Committee.
Englishhttp://ijcrr.com/abstract.php?article_id=2393http://ijcrr.com/article_html.php?did=2393
Alem M, Enawgaw B, Gelaw A, Kena T, Seid M, Olkeba Y. Prevalence of anaemia and associate risk factors among pregnant women attending antenatal care in Azezo health centre Gondar town, Northwest Ethiopia. J Interdisipl Histopathol, 2013;1(3):137-144.
WHO (2004) Focusing on anaemia : Towards an integrated approach for effective anaemia control, Joint Statement by the World Health Organisation and the United Nations Children’s Fund, Geneva, WHO.
Kaur S, Deshmukh P R, Garg BS. Epidemiological correlates of nutritional anaemia in adolescent females of rural Wardha. Indian Journal of Community Medicine 2006;31(2):255-8.
Biradar S S, Biradar S P, Alatagi A C, Wantamutte A S, Malur P R. Prevalence of anaemia among Adolescent girls: A one year Cross-Sectional Study. Journal of Clinical and diagnostic Research 2012;6(3):372-7.
Sachan, Kishore B. Prevalence of anaemia in adolescent school going girls. Indian Journal of Community Medicine 1997;3(1):432-8.
Bilkish N Patavegar, MS Kamble, Sanjivani Langare Patil. Prevalence of anaemia and its epidemiliogical correlates among women of reproductive age in a rural setting. International Journal of Basic and Applied Medical Science 2014;4(2):155-9.
Rawat CMS, Garg SK, Singh JV, Bhatnagar M, Chopra H, Bajpai SK. Socio-demographic correlates of anaemia among adolescent girls in rural area of district Meerut (UP). Indian J. Comm. Med 2001;26:173-5
Bhanushali MM, Shirode AR, Joshi YM, Kadam VR. An intervention of iron deficiency anaemia in dietary behaviour among adolescent girls. International J of Pharmacy and Pharmaceutical Sciences 2011;3(1):40-2.
NFHS-3 India, 205-2006- National Family Health Survey-3. Anaemia among women and children by International Institute for population Sciences Mumbai.
Sen A, Kanani S J. Deleterious functional impact of anaemia on young adolescent school going girls. Indian journal of Paediatrics. 2006;43(2):220-6.
Siddharam SM, Venketesh GM and Thejeshwari HL (2011). A study of anaemia among adolescent girls in Rural area of Hassan district, Karnataka, South India. International Journal of Biological and Medical Research 2 92-4.
Sharma M K, Swami H M, Bhatia V, Verma A, Bhatia SPS, Kaur G. An epideological study of correlation of osteoarthritis in geriatric population of UT Chandigarh. Indian Journal of Community Medicine 2007;32(1):77-8.
Toteja G, Singh P. Micronutrient deficiency disorder in 16 districts in India. Report of an ICMR task for study. District Nutrition Project Part I.2001.
Kakkar R, Kakkar M, Kandpal SD. Depth of anaemia in adolescent school girls of Bhopal. Indian j of Community Health 2010;22:38-40.
Patel S, Shah M, Patel J, Kumar N. Iron deficiency anaemia in moderate to severely anaemic atients. Gujrat Medical Journal 2009;64(2) 15-8.
Goel S, Gupta BP. Low anaemia prevalence among adolescent of an urban hilly community. Indian Journal of Community Medicine.2007;32(1):67-8.
Ramzi M, Haghpanah S, Malekmakan N. Anaemia and iron deficiency anaemia in adolescent school going girls in Kavar Urban Area, Southern Iran. Iranian Red Crescent Medical Journal 2011;13(2):128-33.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241923EnglishN-0001November30HealthcareVitellogenin gene expression during the development of anautogenous malaria vector, Anopheles culicifacies A.
English0914Monika MiglaniEnglish S. K. GakharEnglishAim: Anautogenous mosquitoes require a blood meal before they can lay their first batch of eggs. Unluckily, this happens at the consequence of pathogen transmission and the spread of vector-borne diseases. The reproductive success of all oviparous species including insects depends on vitellogenin (Vg) biosynthesis and its accumulation in the developing oocytes. Therefore, during the present study, the temporal and spatial expression of the Vg gene has been studied in Anopheles culicifacies, which is the major vector in Indian subcontinent.
Methodology: The Dhera strain of An. culicifacies Species A was maintained in an insectary at appropriate rearing conditions. The RNA was extracted from the female fat bodies after blood feeding at different time intervals. The temporal and spatial expression patterns of the Vg gene was examined by Real-time PCR using SYBR Green dye.
Result: The An. culicifacies vitellogenin (AncuVg) showed significant and strong expression in fat body tissues only as compared to other tissues at specific time interval after blood feeding. Time and tissue specific expression of the gene showed induction of the gene at 3 h PBM and peak expression was observed after 24 h of blood feeding.
Discussion: The peak expression of Vg gene observed at 24h corresponds to the peak levels observed earlier for 20-E after 24h of blood meal and also corresponds with the timing of ookinete invasion and oocyst formation.
Conclusion: The molecular events of vitellogenesis as observed in case of An. culicifacies, may prove beneficial to develop novel control strategies to combat parasite transmission because the timing of Vg expression coincides with ookinetes invasion in general.
EnglishAnopheles, Fat body, Hematophagy, Mosquito, VitellogeninIntroduction
Hematophagy, or blood feeding behaviour is exhibited by most of the arthropod vectors of human pathogens both for reproduction and for transmission1. Hematophagous mosquitoes are broadly categorized into two groups with respect to egg development. Anautogenous mosquitoes require a blood meal before they can lay their first batch of eggs, whereas autogenous mosquitoes can use their protein reserves—stored in fat body tissue during larval development—to synthesize yolk proteins and develop one batch of eggs before taking a blood meal. One important distinction is that anautogenous mosquitoes must feed on vertebrate hosts more frequently than autogenous mosquitoes, thereby increasing their overall vectorial capacity. The understanding of the mechanisms underlying anautogeny is very crucial because this reproductive strategy is the driving force behind the transmission of disease to millions of people1-3.
These mosquitoes (Aedes and Anopheles species) also require a blood meal to synthesize vitellogenins during each gonotrophic cycle2,3. The developmental expression of Vgs has been documented in ananutogenous mosquitoes, Anopheles gambiae and Anopheles stephensi1,4-6 and in autogenous Culex tarsalis7. Similarly, the mechanisms underlying the molecular regulation of one of the three vitellogenin genes (VgA1) in anautogenous Aedes aegypti has also been characterized8-10. However, more such studies are required to confirm the pattern and to better understand the mechanism of vitellogenesis in other mosquito species.
An. culicifacies is considered to be the principal vector of malaria in India accounting for 60-70% of malaria cases in the country and exists as a species complex of five sib-ling species named A, B, C, D and E11-15. In the present study, the expression of the vitellogenin gene during different stages of development following blood meal has been studied. The present investigations complement our previous study where, the complete vitellogenin gene of An. culicifacies was cloned and characterized including its upstream regulatory region16.
Experimental Procedures
Mosquitoes
The Dhera strain of An. culicifacies Species A was maintained in an insectary at temperature of 28 ± 2°C, 75 ± 5% relative humidity and 14/10-h light-dark cycles17. Larvae were reared in enamel trays at a standard density of 300 larvae/450 ml of water and were fed on yeast extract and dog biscuits in the ratio of 2:3 (w/w). It passes through 4 instars for about 12-13 days. After pupation, the pupae were transferred to fresh bowl and were kept in cloth cages for emergence to adult mosquitoes.
Adult mosquitoes were maintained on 5% glucose and fed on rabbit blood for ovarian development. Mosquitoes were starved by denying access to sugar and water for 8 h before blood feeding and those that had fed to repletion were separated from the cohort and shifted to a new cage. After blood feeding 10-15 blood fed females were taken after regular time intervals to analyse gene expression profile. During the complete procedure, a bowl filled with water was placed in the cage to enable oviposition.
RNA isolation and cDNA synthesis
Total RNA was isolated from 5-7 day old sugar-fed males and non-blood fed and blood-fed females at several time points post blood meal (PBM) using TRI reagent (Sigma) according to the manufacturer’s protocol after homogenization with a motor-driven pellet pestle mixer. RNA samples also were extracted from immature stages and dissected tissues (i.e. fat body, midgut, salivary gland, ovaries). For each stage, tissues were dissected from 20 mosquitoes in a drop of DEPC treated water and were immediately processed for RNA isolation and stored at −80?C after treatment with amplification-grade RNase-free DNase I (Invitrogen) until used further for cDNA synthesis. All the plastic ware and glassware were treated with DEPC water before use.
RNA integrity was confirmed on a 1% agarose gel and was quantified using Nanodrop ND2000c spectrophotometer. For each stage 1 µg RNA was used for cDNA synthesis using the Superscript RT for PCR kit (Invitrogen) in 20µl volume with oligo(dT)20 primers at 37°C for 1hr followed by termination of the reaction at 70°C for 20 min. To minimize possible variations in reverse transcriptase efficiency, all cDNA synthesis were proceeded siumeltaneously.
Quantiative Real time PCR (qRT-PCR)
Real-time PCR was performed using SYBR Green dye on Real-Time PCR system (Applied Biosystem). In order to quantify relative gene expression, standard curves were generated, using 10-fold serial dilutions of cDNA pools containing high concentrations of the gene of interest. Each qPCR reaction was carried out in a total volume of 20 µL, using 2 µL cDNA template with SYBR Green chemistry and gene specific primers. The primer pair was derived from two different exons with an intervening intron in order to detect amplification products from genomic DNA that may have precipitated during the RNA preparation. Primers were designed using the primer premier 5.0 to yield amplification products ranging in size between 100 and 150 bp (Vg: forward, 5’-CCTACATGCGTTGTTGATGG-3’, reverse, 5’-TGACGACTATGCACTCCAGC-3’; beta-actin: forward, 5’-AGCGGGAAATCGTGCGTGAC-3’, reverse, 5’-CAATGGTGATGACCTGGCCAT-3’).
The amplification of the Vg gene and an endogenous control gene was performed simultaneously and the relative expression levels between Vg and endogenous control gene assessed. The relative abundance of the gene of interest in each RNA sample was estimated from the respective standard curves and the gene expression level was normalized against the β-actin expression level. Cycling conditions were kept constant for all assays. A 2-step qRT-PCR program described earlier18 was used in the amplification process. Melting curves were visually inspected to verify a single amplification product with no primer dimers.
Statistical Analysis
Statistical analysis was performed using Graph Pad Prism3.0 software19. All data were presented as the mean ± standard deviation. Differences between test samples and their respective controls were evaluated by unpaired Student’s t-test. The significant difference of expressions was shown at P < 0.05.
Results
The temporal and spatial expression patterns of the Vg gene were examined using qRT-PCR. The RNA was extracted from the female fat bodies after blood feeding at different time intervals. In addition, RNA was also extracted from midguts, ovaries and salivary glands. During the present study it was observed that the level of expression of the vitellogenin gene varied during different stages of development. For each gene (vitellogenin and beta-actin), a single peak on the melting curve was observed, indicating that target PCR products were amplified selectively and also the primer dimers and other products were not formed.
The expression was observed both in males and both sugar-fed and blood fed females excluding larvae and pupae samples (Fig. 1). The level of vitellogenin mRNA in these stages was negligible, indicating that the vitellogenin expression in male, and non-blood fed females is merely basal level transcription. It could also be ascribed to the presence of transcription initiation complex which could access the promoter region of the Vg genes. However, the Anopheles culicifacies vitellogenin (AncuVg) showed significant and strong expression in fat body tissues only as compared to other tissues viz. salivary glands, midgut and ovaries at specific time interval after blood feeding suggesting that the AncuVg is expressed only in fat bodies (Fig. 2).
The presence of the mRNA was detected in the fat body within 3 h of a blood meal. The expression increased several folds after blood feeding. The peak expression level was observed after 24h PBM then decreased gradually to almost negligible at 72 h PBM (Fig. 3). All the PCR products were also analysed by electrophoresis in ethidium bromide stained agarose gels (1%) (Figs. 4-6) showing expected results.
Discussion
Vg is synthesized abundantly in the female fat body in tissue-, sex- and stage-specific manner, secreted into haemolymph, and subsequently sequestered in the developing oocytes through receptor-mediated endocytosis20-22. During the present study, the AncuVg expression was found in males and non-blood-fed females at very insignificant levels as compared to that in blood fed females (Fig. 1), indicating that this gene may be expressing constitutively in all stages prior to blood feeding. However, the level of Vg RNA was too low to support the process of egg development. This may be because of the presence of certain rare synonymous codons that might have initiated the process of vitellogenesis after accumulating at the 5’ end in the absence of any external amino acid supply23. Also, as described earlier, the induction of Vg expression after a blood meal requires the Target of Rapamycin (TOR) pathway and ecdysone signaling24,25. Thus, once a female mosquito ingests blood, the vitellogenesis process may be triggered under the combined action of JH and 20-E along with the presence of enough amino acid residues.
The significant vitellogenin gene expression is thus restricted to adult female mosquitoes only. However, males of several species have been reported to express vitellogenin gene including Oncopeltus fasciatus26,Rhodnius prolixus27, and the sea urchin Strongylocentrotuspurpuratus28 but their physiological significance remains to be investigated. In addition, treatment with steroid hormones (estrogen for vertebrate and 20-HE for insects) also stimulates vitellogenin gene expression in males of the chicken, G. gallus29, the frog, X. laevis30, the fruit fly D. melanogaster31, the flesh fly Sarcophaga bullata32, An. gambie33, and Cherax quadricarinatus34. Thus, it is possible that an increase in tRNA levels especially those required to increase translation of rare synonymous codons has been stimulated by ecdysone along with the induction of Vg transcription after blood feeding.
Time and tissue specific expression of the gene showed induction of the gene at 3 h PBM and peak expression was observed after 24 h of blood feeding. These results of AncuVg gene are in accordance with the previous studies of Vg gene expression in fat bodies35-37. The temporal expression pattern of the Vg gene in An. culicifacies (Fig. 3) was found to be similar to that in An. gambiae6, VgA1 in Ae. aegypti9 and in Cx. tarsalis7. The only difference lies in the initiation of Vg expression by 3h after blood feeding in An. culicifacies and other autogenous mosquitoes as compared to at 12-24h after emergence in case of autogenous mosquitoes7
These results also are in accordance with the pattern shown during the hormonal regulation of anautogeny. The peak expression of Vg gene observed at 24h in the present study after blood meal corresponds to the peak levels observed earlier for 20-E after 24h of blood meal9 and also corresponds with the timing of ookinete invasion and oocyst formation as observed earlier in case of An. gambiae with infection of P. nigerensis6. This invasion was found to coincide with the time at which vitellogenesis is first affected. Similar results have also been observed with infection of P. berghei in An. gambiae38 and in An. stephensi4. A significant decrease in Vg mRNA level at 24h PBM was observed when the ookinetes were invading and transforming into oocysts6, indicating that the infection has a negative effect on vitellogenesis in the fat body, thus eventually causing a significant reduction in fecundity4. Thus, the peak observed at 24 h PBM in AncuVg gene indicate the possibility of similar hormonal regulation pathway in An. culicifacies and also correlating the similar mode of parasite infection as observed earlier in other mosquitoes.
Conclusion
The expression pattern of AncuVg shows that the gene is expressed significantly in response of blood meal. The results of present study indicates that the fat body is the only site of Vg gene expression in An. culicifacies female mosquitoes after blood feeding and that too showing its peak expression at 24h PBM coinciding with the time of ookinete invasion and oocyst formation as reported earlier. Thus, the molecular events of vitellogenesis as observed in case of An. culicifacies, may prove beneficial to develop novel control strategies to combat parasite transmission because of the similar timing of Vg expression and ookinetes invasion in general.
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.
Source of Finding
The author Monika Miglani highly acknowledges the financial assistance provided by Department of Biotechnology-Builder Programme, New Delhi (BT/PR4329/INF/22/144/2011).
Conflict of Interest
The authors declare they have no conflict of interest.
Englishhttp://ijcrr.com/abstract.php?article_id=2394http://ijcrr.com/article_html.php?did=2394
Dana AN, Hong YS, Kern MK, Hillenmeyer ME, Harker BW, Lobo NF, Hogan JR, Romans P, Collins FH, Gene expression patterns associated with blood-feeding in the malaria mosquito Anopheles gambiae. BMC Genomics, 6(2005) 5.
Hagedorn RH, Fallon AM and Laufer H, Vitellogenin synthesis by fat-body of mosquito Ae. aegypti - evidence for transcriptional control. Dev Biol, 31 (1974) 285.
Lu YH, Hagedorn HH, Egg development in the mosquito Anopheles albimanus. Int J Inver Rep Dev, 9 (1986) 79.
Ahmed AM, Maingon R, Romans P, Hurd H, Effects of malaria infection vitellogenesis in Anopheles gambiae during two gonotrophic cycles. Insect Mol Biol, 10 (2001) 347.
Nirmala X, Marinotti O, Sandoval J M, Phin S, Gakhar S, Jasinskiene N and James AA, Functional Characterization of the Promoter of the Vitellogenin Gene, AsVg1, of the Malaria Vector, Anopheles stephensi. Insect Biochem Mol Biol, 36(9) (2006) 694.
Chen XG, Marinotti O, Whitman L, Jasinskiene N, James AA, Romans P, The Anopheles gambiae vitellogenin gene (VGT2) promoter directs persistent accumulation of a reporter gene product in transgenic Anopheles stephensi following multiple blood meals. Am J Trop Med Hyg, 76(6) (2007) 1118.
Provost-Javier KN, Chen S, Rasgon JL, Vitellogenin gene expression in autogenous Culex tarsalis. Insect Mol Biol, 19(2010) 423.
Kokoza V, Ahmed A, Wimmer EA, Raikhel AS, Efficient transformation of the yellow fever mosquito Aedes aegypti using the piggyback transposable element vector pBac[3xP3-EGFP afm]. Insect Biochem Mol Biol, 31 (2001)1137.
Raikhel AS, Kokoza VA, Zhu J, Martin D, Wang SF, Li C, Molecular biology of mosquito vitellogenesis: from basic studies to genetic engineering of antipathogen immunity. Insect Biochem Mol Biol,32(2002) 1275.
Attardo GM, Hansen IA Raikhel AS, Nutritional regulation of vitellogenesis in mosquitoes: implications for anautogeny. Insect Biochem Mol Biol,35(2005) 661.
Green CA, Miles SJ, Chromosomal evidence for sibling species of the malaria vector Anopheles (Cellia) culicifacies Giles. J Trop Med Hyg, 83(1980) 75.
Subbarao SK, Vasantha K, Adak T, Sharma VP, Anopheles culicifacies complex: evidence for a new sibling species, species C. Ann Entomol Soc Am, 76(1983) 985.
Suguna SG, Tewari SC, Mani TR, Hiryan J, Reuben R, A cytogenetic description of a new species of the Anopheles culicifacies species complex. Genetica, 78(1989) 225.
Vasantha K, Subbarao SK, Sharma VP,Anopheles culicifacies complex: Population cytogenetic evidence for species D(Diptera: Culicidae). Ann Entomol Soc Am, 84(1991) 531.
Kar I, Subbarao SK, Eapen A, Ravindran J, Satyanarayana TS, Raghavendra K, Nanda N, Sharma VP, Evidence for a new malaria vector species, species E, within the Anophelesculicifacies complex (Diptera: Culicidae). J Med Entomol, 36(1999) 595.
Miglani M, Gakhar SK, Cloning and bioinformatical analysis of vitellogenin gene of the Indian malaria vector Anopheles culicifacies (Diptera: Culicidae). Acta Entomol Sin, 56(9) (2013) 1063.
Gakhar SK, Singh S, Shandilya H, Changes in soluble proteins during the development of malaria vector An. stephensi (Diptera: Insecta). Proc Natl Sci Acad USA, B63 (1997) 289.
Roelofs D, Overhein L, de Boer ME, Janssens TKS, van Straalen NM, Additive genetic variation of transcriptional regulation: metallothionein expression in the soil insect Orchesella cincta. Heredity, 96 (2006) 85.
Motulsky HJ, Analyzing Data with GraphPad Prism. GraphPad Software Inc., San Diego, CA www.graphpad.com, 1999.
Raikhel AS, Dhadialla TS, Accumulation of yolk proteins in insect oocytes. Annu Rev Entomol, 37 (1992) 217.
Sappington TW, Raikhel AS, Molecular characteristics of insect vitellogenins and vitellogenin receptors. Insect Biochem Mol Biol,28(1998) 277.
Snigirevskaya ES, Raikhel AS, Receptor-mediated endocytosis of yolk proteins in insect oocytes. In: Progress in Vitellogenesis, (A. S. Raikhel, ed.); series Reproductive Biology of Invertebrates (K.G. Adiyodi and R. G. Adiyodi, eds.) Vol. XII, Part B, pp. 199-228. Science Publishers, Inc., Enfield, USA- Plymouth, UK, 2005.
Isoe J, Hagedorn HH, Mosquito vitellogenin genes: Comparative sequence analysis, gene duplication, and the role of rare synonymous codon usage in regulating expression. J Insect Sci,7 (2007) 1.
Hansen IA, Attardo GM, Park JH, Peng Q, Raikhel AS, Target of rapamycin-mediated amino acid signaling in mosquito anautogeny. Proc Natl Sci Acad USA,101(2004) 10626.
Park JH, Attardo GM, Hansen IA, Raikhel AS, GATA factor translation is the final downstream step in the amino acid/target-of-rapamycin-mediated vitellogenin gene expression in the anautogenous mosquito Aedes aegypti. J Biol Chem,281(2006) 11167.
Kelly TJ, Telfer WH, Antigenic and electrophoredc variants of vitellogenin in Oncopeltus blood and their control by juvenile hormone. Dev Biol, 61 (1977) 58.
Chalaye D, Immunocheniical study of hemolymph and egg proteins of Rhodniusprolixus (Stal). Can J Zool, 57 (1979) 329.
Shyu AB, Raff RA, Blumenthal T, Expression of the vitellogenin gene in female and male sea urchin. Proc Natl Acad Sci USA, 83 (1986) 3865.
Deeley RG, Gordon JI, Bums AT, Mullinix KP, Binastein M, Goldberg RF,Primary activation of the vitellogenin gene in the rooster. J Biol Chem, 252 (1977) 8310.
Lanclos KD, Hamilton TH, Translation of hormone-induced messenger RNA in amphibian oocytes: I. Induction by estrogen of messenger RNA encoded for vitellogenic protein in the liver of the male African clawed toad (Xenopus laevis). Proc Natl Acad Sci U S A, 72 (1975) 3934.
Shirk PD, Minoo P, Posdethwait JH, 20-Hydroxyecdysone stimulates the accumulation of translatable yolk polypeptide gene transcript in adult male Drosophila melanogaster. Proc Natl Acad Sci U S A, 80 (1983) 186.
Huybrechts R, Deloof A, Induction of vitellogenin synthesis in male Sarcophaga bullata by ecdysterone. J Insect Physiol, 23 (1977) 1359.
Pondeville E, Maria A, Jacques J C, Bourgouin C, Villemant CD,Anopheles gambie males produce and transfer the vitellogenic steroid hormone 20- hydroxyecdysone to females during mating. Proc Natl Acad Sci USA,16 (105) (2008) 619.
Shechter A, Aflalo ED, Davis C, Sagi A, Expression of the reproductive female-specific vitellogenin gene in endocrinologically induced male and intersex Cherax quadricarinatus crayfish. Biol Reprod, 73 (2005) 72.
Cho KH, Cheon HM, Kokoza V, Raikhel AS, Regulatory region of the vitellogenin receptor gene sufficient for high-level, germ line cell-specific ovarian expression in transgenic Aedes aegypti mosquitoes. Insect Biochem Mol Biol, 36 (2006) 273.
Ye GY, Dong SZ, Song QS, Shi M, Chen XX, Hu C, Molecular cloning and developmental expression of the vitellogenin gene in the endoparasitoid, Pteromalus puparum.Insect Mol Biol, 17 (3) (2008) 227.
Horigane M, Shinoda T, Honda H, Taylor D, Characterization of a vitellogenin gene reveals two phase regulation of vitellogenesis by engorgement in the soft tick Ornithodorous moubata (Acari: Argasidae). Insect Mol Biol, 19 (2010) 501.
Richman AM. Dimopoulos G, Seeley D, Kafatos FC, Plasmodium activates the innate immune response of Anopheles gambiae mosquitoes. EMBO J, 16 (1997) 6114.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241923EnglishN-0001November30HealthcareEffect of Iron Overload on Gonadotrophins and Organ Sex Steroids in Pubertal Thalassemia
Patients
English1521Manali SinharayEnglish Surabhi MitraEnglish Anindya DasguptaEnglishBackground: Iron overload being one of the major adverse effect related to blood transfusion in β Thalassemia major,leads to various endocrinal disorders among which hypogonadism is noteworthy.Pituitary or gonadal iron deposition or both may lead to this.
Objectives: The main purpose of this study was to evaluate the effect of iron overload measured by serum ferritin on pituitary gonadotrophins (FSH, LH) and gonadal steroids (Estrogen in females,Testosterone in males) levels and the correlation between them in pubertal thalassemic subjects(cases).
Methods: Serum harvested from blood samples collected from 30 cases(15 males,15 females) and 30 controls were used to estimate Serum ferritin, LH, FSH, Estradiol in females and Testosterone in males.
Results: The serum LH (pEnglish? Thalassemia major, Gonadotrophins, Hypogonadism, Delayed pubertyIntroduction
β-thalassemia is the commonest single-gene disorder in the Indian population [1]. Ten percent of the total world thalassemics are born in India every year [2]. Certain communities in India, like Sindhis, Gujratis, Punjabis, and Bengalis, are more commonly affected with beta thalassemia, the incidence varying from 1 to 17% [3]
Iron overload which is a common complication of thalassaemic syndromes could lead per se to the development of organ damage and increased mortality [4]. In these patients, iron deposition in parenchymal tissues starts within 1 year of starting the regular transfusions [5]. Blood transfusions are important for survival of these patients, but chronic transfusions inevitably lead to iron overload as humans cannot remove excess iron actively. The cumulative effects of iron overload, if untreated, lead to significant morbidity and mortality [ 6]. The current management of thalassemia (TM) includes regular transfusion programs and chelation therapy. Current guidelines recommend a pretransfusion threshold not exceeding 9.5% g/dl, which seems to be associated with adequate marrow inhibition and a relatively low iron burden.[7] Research also indicates that iron overload can occur in patients with non-transfusion dependent thalassaemias(NTDT) [8]
Delayed puberty and hypogonadism are among the most common clinical consequences of iron overload. Iron deposition in the pituitary gonadotrophic cells leads to disruption of gonadotropin (LH and FSH) production. In the majority of well-chelated patients, the gonadal function is normal; however, gonadal iron deposition occasionally occurs. TM patients with a favorable genotype manifest less severe gonadal dysfunction, due to less iron loading. [9] The precise mechanism of iron overload-induced organ dysfunction leading to delayed puberty is not clear. It is still not known whether iron deposition in the pituitary gonadotrophic cells or gondal iron deposition or both causes the hypogonadism. Some studies[10] found significant difference in mean serum ferritin level between thalassemic patients with primary amenorrhea, irregular mense, hypogonadism and those without endocrinopathies. These findings yield the importance of iron overload in development of endocrine disorders among which hypogonadism is most frequent. In contrast, there are some othereports which have suggested no relation between the level of ferritin and some other endocrinopathies [11,12]. Based on the above mentioned information, we set out to study the effect of iron overload on pituitary gonadotrophins and the gonadal sex steroids in pubertal thalassemics and to conclude whether the former or later is most affected.
Aims and Objectives
1. To measure serum ferritin to detect iron overload pubertal thalassemic patients(cases) as well as in controls
2. To measure pituitary gonadotrophins-Luteinizing hormone (LH), Follicle Stimulating
Hormone (FSH) in the study subjects (cases and controls).
3. To measure gonadal steroids-Estrogen in female subjects, Testosterone in male subjects
5. To assess any correlation between serum ferritin, pituitary gonadotrophins and gonadal steroids in the case group.
Materials and Method
Study type: It was a hospital based observational study.
Study design: The study design was cross-sectional and non interventional.
Study population: The study population included pubertal thalassemic patients. The control
group were selected from the healthy relatives of the patients who will accompany them being age matched.
Selection criteria:
Cases were taken from the thalassemia unit of the institution as per the following criteria:
*INCLUSION CRITERIA
a) The age of the β-thalassemia major subjects were between the ages of 13to 17yrs to detect pubertal delay
b) They had already received multiple blood transfusions or iron therapy.
*EXCLUSION CRITERIA
a) The newly diagnosed patients were excluded.
b) The subject who were diabetic and had other inborn metabolic diseases.
c) Subject who had active infection and inflammation.
d) Subjects who were alcoholics and cigarette smokers.
Controls were selected from healthy individuals (without any metabolic diseases,
haematological diseases, chronic or active infection and inflammation).
Place of study: Selection of cases was done from the thalassemia unit of the institution.
Biochemical investigations and result analysis were performed in the dept. of Biochemistry,
Calcutta National Medical College, Kolkata.
Study duration: Two months after getting approval for the project as well as the instituitional ethical clearance certificate.
Sample size: Following inclusion and exclusion criteria thirty patients- fifteen male and fifteen female were selected by the method of convenience in each group of the cases and controls.
Ethical considerations: The study was conducted by strictly adhering to the guidelines from the Helsinki declaration, 1975 revised in 2000. Written and informed consents were obtained from participants as per protocol.
Data collection procedure: The valid written consents of the subjects were taken and proper ethical guidelines were followed. The patients were asked to come at a specific date. We collected demographic and anthropometric data and the history of menstruation, family history of diabetes, initiation and duration of blood transfusion, as well as chelation therapy. After the collection of the samples and carrying them to the Department of Biochemistry, the clotted blood was centrifuged at 1500 rpm for 5 min and the serum was harvested from which estimation of serum ferritin, Luteinizing hormone(LH), Follicle Stimulating hormone(FSH), Estradiol in female subjects, Testosterone in male subjects were done by ELISA.
Instruments required were:
1. ELISA Kit, ELISA Reader and Washer.
2. Micropipettes and microtips.
3. Syringes and cottons.
Principle of measurement:
Serum ferritin- Solid phase sandwitch assay method (CalBiotech Inc.)
Luteinizing hormone (LH), Follicle stimulating hormone(FSH)-Solid phase sandwitch ELISA (AccuDiag )
Estrogen, Testosterone-Solid phase competitive ELISA (DiaMetra-Italy)
In each analysis, standard and controls were tested along with the samples.
Confidentiality: Data would be kept strictly confidential and will be stored at least for three years.
Statistical analysis:
The data obtained from the above tests were analyzed for differences between the medians of the analytes studied between cases (pubertal thalassemics) and controls (neonates delivered by ND).
Kolmogorov and Smirnov method
The Kolmogorov and Smirnov method was used to check whether the data were normally distributed.
Mann–Whitney U-test
In order to study the significance in the differences between the two groups, the Mann–Whitney U-test was performed for all analytes, since the data did not pass the normality test.
Spearman’s correlation
Spearman’s correlations were done between all analytes. All of the tests were completed with a P Englishhttp://ijcrr.com/abstract.php?article_id=2395http://ijcrr.com/article_html.php?did=23951. Verma IC. The challenge of genetic disorders in India. In: Molecular geneticsand gene therapy- the new frontier,. Scientific Communications, Amsterdam 1994; pp11-20.
2. Bashyam MD, Bashyam L, Savithri GR et al. Molecular genetic analyses of beta thalassemia in South India reveal rare mutations in the beta globin gene. J Hum Genet 2004; 49: 408-413.
3. Gupta A, Hattori Y, Gupta UR et al. Molecular genetic testing of beta thalassemia patients of Indian origin and a novel 8bp deletion mutation at codons 36/37/38/39. Genet Test 2003 7(2):163-8.
4. Mariani R, Trombini P, Pozzi M, et al.– Iron metabolism in thalassemia and sickle cell
disease. Mediterr J Hematol Infect Dis. 2009; 1(1): e2009006
5.TaksandeA, Prabhu S, Venkatesh S–Cardiovascular aspect of beta thalassaemia. Cardiovasc Hematol AgentsMed Chem. 2012; 10:25-30
6. Cappellini MD – Exjade(R) (deferasirox, ICL670) in the treatment of chronic iron overload associated with blood transfusion. Ther Clin Risk Manag.2007; 3:291-9
7. De Sanctis V, Soliman AT, Elsedfy H, Skordis N, Kattamis C et al. Growth and endocrine disorders in thalassemia: The international network on endocrine complications in thalassemia (I-CET) position statement and guidelines. Indian J Endocrinol Metab. 2013 Jan-Feb; 17(1): 8–18.
8. Taher AT, Porter J, Viprakasit V, Kattamis A, Chuncharunee S, Sutcharitchan P. et al – Deferasirox reduces iron overload significantly in nontransfusion-dependent thalassemia: 1-year results from a prospective, randomized, double-blind, placebo-controlled study. Blood 2012; 120:970-7
9. Raiola G, Galati MC, De Sanctis V, Caruso Nicoletti M, Pintor C, De Simone M, et al. Growth and puberty in thalassemia major. J Pediatr Endocrinol Metab. 2003;16: 259–66.
10. AA Shamshirsaz et al.Metabolic and endocrinologic complications in beta-thalassemia major: a multicenter study in Tehran BMC Endocrine Disorders 2003, 3:4
11. Masala A, Meloni T, Gallisai D, Alagna S, Rovasio PP, Rassu S, Milia AF: Endocrine functioning in multitransfused prepubertal patients with homozygous beta thalassemia. J Clin Enocrinol Metab 1984, 58:667-70.
12. Zervas A, Katopodi A, Protonotariou A, Livadas S, Karagiorga M, Politis C, Tolis G: Assessment of thyroid function in two hundred patients with beta-thalassemia major. Thyoid 2002, 12:151-4.
13. S Sasima, O Boonsong, B Pongamorn Hypogonadism in thalassemia major patients. Journal of Clinical and Translational Endocrinology 5 (2016) ;42–45
14.Walter, P.B., Fung, E.B., Killilea, D.W., Jiang, Q., Hudes, M., Madden,J., Porter, J., Evans, P., Vichinsky, E. and Harmatz, P. Oxidative stress and inflammation in iron-overloaded patients with B-thalassaemia or sickle cell disease. British Journal of Haematology 2006;135, 254–264.
15.Chatterjee, R. and Katz, M. Reversible hypogonadotrophic hypogonadism in sexually infantile male thalassaemic patients with transfusional iron overload. Clinical Endocrinology 2000;53, 33–42.
16. Ellen B. Fung et al Increased prevalence of iron-overload associated endocrinopathy in thalassaemia versus sickle-cell disease., British Journal of Haematology 2006, 135, 574–582.
17.Chern JP, Lin KH, Tsai WY, Wang SC, Lu MY, Lin DT, et al. Hypogonadotropic hypogonadism and hematologic phenotype in patients with transfusion dependent beta-thalassemia. J Pediatr Hematol Oncol 2003;25(11):880–4.
18.Delvecchio M, Cavallo L. Growth and endocrine function in thalassemia major in childhood and adolescence. J Endocrinol Invest 2010;33(1):61–8.
19. Bronspiegel-Weintrob N, Olivieri NF, Tyler B, Andrews DF, Freedman MH, Holland FJ. Effect of age at the start of iron chelation therapy on gonadal function in beta-thalassemia major. N Engl J Med 1990; 323(11):713–19.
20. Jameson JL, Groot LJD. Endocrinology: adult and pediatric. 7th ed. Philadelphia: Elsevier; 2016.
21. Atkin SL, Hipkin LJ, Landolt AM, Jeffreys RV, Foy PM, White MC. Effect of cell density on hormonal secretion from human pituitary adenomas in vitro. Horm Res 1998; 49(5):203–9.
22. Canale V, Steinherz P, New M, Erlandson M. Endocrine function in thalassemia major. Ann N Y Acad Sci 1974;232(1):333–45.
23. U Rifki, K Erdal, A Murat, O Sehmus, A Murat, D Yaren, B Aydin. A rare presentation of transfusional hemochromatosis: hypogonadotropic hypogonadism. Case Reports in Endocrinology Vol. 2015, Article ID 493091, 4 .
24. Cappellini MD, Cohen A, Porter J, Taher A, Viprakasit V. Guidelines for the management of transfusion dependent thalassaemia. 3rd ed. Nicosia, Cyprus:
Thalassaemia International Federation; 2014.
25. Modell B., Khan, M., Darlison, M., Westwood, M.A., Ingram, D. andPennell, D.J. Improved survival of thalassaemia major in the UK and relation to T2* cardiovascular magnetic resonance. Journal of Cardiovascular Magnetic Resonance 2008;10, 42.
26. Styne DM, Grumbach MM. Physiology and disorder of puberty. In: Melmed S, Polonsky KS, Larsen PR, Kronenberg HM, editors. Williams textbook of endocrinology. 13th ed. Elservier; 2016. p. 1074–218.
27.De Sanctis V, Borsari G, Brachi S, Govoni M, Carandina G. Spermatogenesis in young adult patients with beta-thalassaemia major long-term treated with desferrioxamine Georgian Med News; 2008;156:74-7.
28.Zacharin M, Sabin MA, Nair VV, Dagabdhao P. Addition of recombinant follicle-stimulating hormone to human chorionic gonadotropin treatment in adolescents and young adults with hypogonadotropic hypogonadism promotes normal testicular growth and may promote early spermatogenesis. Fertil Steril. 2012; 98:836-42.
29.Soliman AT, Nasr I, Thabet A, Rizk MM, El Matary W. Human chorionic gonadotropin therapy in adolescent boys with constitutional delayed puberty vs those with beta-thalassemia major. Metabolism. 2005;54: 15-23.
30. Xu L, Freeman G, Cowling BJ, Schooling CM. Testosterone therapy and cardiovascular events among men: a systematic review and meta-analysis of placebo-controlled randomized trials. BMC Med 2013;11:108.
31. Smith JT, Acohido BV, Clifton DK, Steiner RA. KiSS-1 neurones are direct targets for leptin in the ob/ob mouse. J Neuroendocrinol 2006;18(4):298–303.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241923EnglishN-0001November30HealthcareA Case of Neurofibromatosis Type 1 Associated with Cervical Cord Ependymoma
English2224Ankur MittalEnglish Rattilal MeenaEnglish Neera SamarEnglish Satish KumarEnglish Ashish KhandelwalEnglishPatients with neurofibromatosis 1 (NF1) are predisposed to develop central nervous system tumours, due to the loss of neurofibromin, an inactivator of proto-oncogene RAS. We present a case of NF1 patient with a spinal cord ependymoma. She presents with 3 months history of increasing weakness in right Upper limb and lower limb associated with numbness with involvement of left half of the body since 15 days. Magnetic resonance imaging revealed heterogeneously enhancing solid cystic mass lesion in the intramedullary compartment of the cord extending from C2-C3 to D2-D3 levels with cord edema up to medulla and D11 level craniocaudally respectively, likely to represent ependymoma.
EnglishAutosomal dominant disorder, Ependymoma, Neurofibromatosis 1, NeurofibrominIntroduction
Neurofibromatosis 1 (NF1) is an autosomal dominant disorder caused by heterozygous mutations of the NF1 gene, which is located on chromosome 17q11.21, 2, 3. Mutations in NF1 result in loss of, an inactivator of proto-oncogene RAS, leading to increased proliferation and tumorigenesis, therefore, patients with NF1 are predisposed to develop innocent and malignant tumors4,5. In central nervous system, gliomas are the most common neoplasms in individuals with NF16, however, ependymoma with NF1 has rarely been reported. To date, only three cases have been reported in English literature7,8. Moreover, cervical spinal cord ependymoma, to the best of our knowledge, has never been reported occurring in NF1 patients previously. Recently, we experienced a case of cervical spinal cord ependymoma in a patient with NF1. In this report, we discuss the diagnosis, the clinical management, mechanisms of such a rare case with review of the literature.
Case report
A 21 Year old female patient was admitted to our department for the complaint of weakness of right half of the body since 3 months which was associated with numbness in right half of the body later on involving the left half of the body since 15 days. In her past medical history there were expeditiously increscent cutaneous neurofibromas respectively on her back, lower limb and upper limb which were present since childhood. In her family history, her mother and her maternal grandmother has the similar cutaneous neurofibromas for which they never consulted a doctor and also doesn’t have any neurological complaint in their lifetime. On physical examination, widespread café-au-lait spots, axillary and groin frecklings, cutaneous neurofibromas. No iris hamartomas had been found and mammary gland was normal. In neurological examination, her mental functions were normal, speech normal, all cranial nerve were normal, motor system examination reveals increased tone in both lower limbs, power was 4/5 in all four limbs, Deep tendon reflexes were absent in both upper limb and exaggerated in both lower limb. Planters were bilaterally extensor. Sensory system reveals diminished sensation for all the primary modalities of sensation. No neck rigidity. No cerebellar signs.
In RNT medical college all the investigations were performed. In which Haemoglobin- 13.5gm%; White blood count -8510/mcl; Platelets- 3.83 lacs/mcl; PBF-Normocytic normochromic; ESR- 29; Blood glucose- 103mg/dl; Urea-25mg/dl; creatinine-0.55mg/dl; liver function tests were normal; Lipid profile was normal; Serum electrolyte(Sodium=137; Potassium=4.6; Chloride=108; Calcium=8.7); Chest X-Ray Posterioanterior view and Electrocardiogram were normal; Ultrasonography abdomen does not show any significant abnormalities; Fundus examination was normal.
Magnetic resonant imaging study reveals heterogeneous solid-cystic mass lesion in the intramedullary compartment of the cord extending from C2-C3 to D2-D3 levels appears iso-intense on T1 weighted & heterogeneously hyper intense on T2 weighted sequence. There is heterogeneous enhancement on the post contrast study with non enhancing cystic component. There is cord edema craniocaudally up to medulla superiorly and D11 inferiorly appears isointense on T1 weighted and hyper intense on T2 weighted sequence represent ependymoma.
Magnetic resonant imaging brain doesn’t show any significant abnormality.
Discussion
Neurofibromatosis-1(NF1) occurs with approximately 1: 2000 to 1: 5000 in individuals3. The diagnosis of most NF1 patients is based on clinical manifestations. Diagnosis requires at least two major criteria - 2 or more neurofibromas or 1 plexiform neurofibroma, 6 or more café-au-lait patches, axillary or groin freckling, optic pathway glioma, lisch nodules in the Iris, a distinctive osseous lesion, a first degree relative with NF1. The clinical manifestations of our patient revealed a typical NF1.
Spinal cord ependymomas are the most common intramedullary tumours in adults which account for 60% in all spinal cord tumours, and cervical region are the most common localization they occur9,10,11. The clinical course of our patient is consistent with spinal cord ependymoma.
Gliomas are often associated with NF1, most with a low grade, mainly locate in the optic nerve12,13, and only 1% in the spinal cord12. The incidence of intramedullary gliomas in NF1 patients may be far more than their sporadic counterparts according to similar works14. Meanwhile, compared with NF215, 16,17), ependymomas were reported rarely to occur in NF1 patient so our patient is a rare case presented with manifestations mentioned above
The clinical courses of our patient and others revealed that there were no abnormality between ependymomas in NF1 patients and their sporadic counterparts. Concurrent NF1 glioma mechanism and NF1 genes may closely relate. This gene locates on the long arm of chromosome 17 in the area of 17q11.2, can encode and achieve the synthesis of. This protein as proto-oncogene RAS inhibitors, when NF1 gene function deficiency can result in tumour formation. In addition, NF1 germline mutation, can also lead to tumour form5,11, 18). However, specific mechanisms for NF1 complicated with glioma are still unclear, molecular mechanism of tumour gene and potential therapeutic targets for tumour may be becoming the trend and direction of research 4,18,19.
Conclusion
Ependymoma with NF1 is a rare situation, we first report a spinal cord ependymoma which occurred in a NF1 patient. Referencing other cases, we find that the clinical course of ependymomas in NF1 patients have no abnormality compared with their sporadic counterparts. We emphasize that a detailed history talking and physical examination to a NF1 patient are needed and a multidisciplinary cooperation is essential. Further elucidation on the molecular changes of NF1 that drive tumorigeness remains needed aiming to explore a potential therapeutic protocol.
Englishhttp://ijcrr.com/abstract.php?article_id=2396http://ijcrr.com/article_html.php?did=23961. Jett K, Friedman JM. Clinical and genetic aspects of neurofibromatosis 1. Genet Med. 2010;12:1–11.
2. Lakkis MM, Tennekoon GI. Neurofibromatosis type 1. I. General overview. J Neurosci Res.2000;62:755–763.
3. Rasmussen SA, Friedman JM. NF1 gene and neurofibromatosis 1. Am J Epidemiol. 2000;151:33–40.
4. Brems H, Beert E, de Ravel T, Legius E. Mechanisms in the pathogenesis of malignant tumours in neurofibromatosis type 1. Lancet Oncol. 2009;10:508–515.
5. Cnossen MH, van der Est MN, Breuning MH, van Asperen CJ, Breslau-Siderius EJ, van der Ploeg AT, et al. Deletions spanning the neurofibromatosis type 1 gene : implications for genotype-phenotype correlations in neurofibromatosis type 1? Hum Mutat. 1997;9:458–464.
6. Rodriguez FJ, Perry A, Gutmann DH, O'Neill BP, Leonard J, Bryant S, et al. Gliomas in neurofibromatosis type 1 : a clinicopathologic study of 100 patients. J Neuropathol Exp Neurol.2008;67:240–249.
7. Riffaud L, Vinchon M, Ragragui O, Delestret I, Ruchoux MM, Dhellemmes P. Hemispheric cerebral gliomas in children with NF1 : arguments for a long-term follow-up. Childs Nerv Syst. 2002;18:43–47.
8. Sharma AS, Emery ME, Metcalfe KM, Sabin HIS, Drake WMD. A case of thoracic cord ependymoma in neurofibromatosis type 1. Endocr Abstr. 2006;12:17.
9. Alkhani A, Blooshi M, Hassounah M. Outcome of surgery for intramedullary spinal ependymoma. Ann Saudi Med. 2008;28:109–113.
10. Eroes CA, Zausinger S, Kreth FW, Goldbrunner R, Tonn JC. Intramedullary low grade astrocytoma and ependymoma. Surgical results and predicting factors for clinical outcome. Acta Neurochir (Wien)2010;152:611–618.
11. Son DW, Song GS, Han IH, Choi BK. Primary extramedullary ependymoma of the cervical spine : case report and review of the literature. J Korean Neurosurg Soc. 2011;50:57–59.
12. Guillamo JS, Créange A, Kalifa C, Grill J, Rodriguez D, Doz F, et al. Prognostic factors of CNS tumours in Neurofibromatosis 1 (NF1) : a retrospective study of 104 patients. Brain. 2003;126(Pt 1):152–160
13. Rosenfeld A, Listernick R, Charrow J, Goldman S. Neurofibromatosis type 1 and high-grade tumours of the central nervous system. Childs Nerv Syst. 2010;26:663–667.
14. Rasmussen SA, Yang Q, Friedman JM. Mortality in neurofibromatosis 1 : an analysis using U.S. death certificates. Am J Hum Genet. 2001;68:1110–1118.
15. Aguilera DG, Mazewski C, Schniederjan MJ, Leong T, Boydston W, Macdonald TJ. Neurofibromatosis-2 and spinal cord ependymomas : report of two cases and review of the literature. Childs Nerv Syst.2011;27:757–764.
16. Lim BS, Park SQ, Chang UK, Kim MS. Spinal cord tanycytic ependymoma associated with neurofibromatosis type 2. J Clin Neurosci. 2010;17:922–924.
17. Ueki K, Sasaki T, Ishida T, Kirino T. Spinal tanycytic ependymoma associated with neurofibromatosis type 2--case report. Neurol Med Chir (Tokyo) 2001;41:513–516.
18. Graf N. Glioblastoma in children with NF1 : the need for basic research. Pediatr Blood Cancer.2010;54:870–871.
19. Dilworth JT, Kraniak JM, Wojtkowiak JW, Gibbs RA, Borch RF, Tainsky MA, et al. Molecular targets for emerging anti-tumour therapies for neurofibromatosis type 1. Biochem Pharmacol. 2006;72:1485–1492.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241923EnglishN2017December6HealthcareTinea Capitis Among Primary School Children: A Clinicomycological Study in a Rural Hospital in Central India
English2531Ruchita O. AttalEnglish Vijayshri DeotaleEnglish Akshay YadavEnglishAims and Objectives
1. To determine the clinical and mycological profile of tinea capitis Among Primary School students of Rural area
2. To correlate of KOH microscopy and culture for diagnosing Tinea capitis.
Methods and Results: A total of 323 Primary School students from six different Government rural schools were enrolled and screened for any infection on hair and scalp. Of these, 81 were clinically found to be of Tinea capitis. Woods lamp examination, 20% KOH microscopy and Fungal cultures were performed for all these cases. Non-inflammatory type of tinea capitis was predominant (95.06%) type and female students (55.5%) were affected more compared
to male(44%). In our study, the positivity by 20% KOH and Fungal culture was 40.74% and 11.11% respectively. The etiological agents were T. violacium, Microsporum canis and T. rubrum.
Conclusion: Tinea capitis was detected mostly in pupils with low level of educational and poor socio-economic status of their parents.
EnglishTinea capitis, Dermatophytes, KOH microscopyINTRODUCTION
Tinea capitis is a fungal infection of the scalp hair follicles and the surrounding skin caused by dermatophytes In fact, Tinea Capitis infection has been described as a “modern-day epidemic.” (Ginter-Hanselmayer G et al. 2011). It is the most commonly diagnosed dermatophytosis of childhood and is more frequently seen among prepubescent children. Mohrenschlager M et al.2005)
Dermatophytes genera include Trichophyton, Epidermophyton and Microspora and are characterized by their ability to invade the superficial layers of the epidermis, particularly the stratum corneum and the high keratin-concentration containing appendages, the hair and nails of the living host. The most common etiological agents include the members of the genera Microsporum and Trichophyton species. The infection may range from mild, almost subclinical, with slight erythema and a few patchy areas of scaling with dull gray hair stumps to a highly inflammatory reaction with folliculitis, kerion formation, and extensive areas of scarring and alopecia, sometimes accompanied by fever, malaise, and regional lymphadenopathy.
Tinea capitis is a common superficial fungal infection seen predominantly in children of school age in the developing countries. (Ahmed I et al.2006, Ayanbimpe GM et al.2008, Grover C et al 2010). WHO data has revealed that 7-33% of children of various age groups are affected (Mahe A et al. 2005). The epidemiology of Tinea capitis varies within different geographical areas throughout the world and in any given area, the species may change.
Hot humid tropical climates, low socioeconomic status, crowded living condition and poor hygiene contribute to an increased incidence of Tinea capitis. It is highly communicable and may reach epidemic proportions especially in overcrowded setups (Mahe et al. 2005, Ginter-Hanselmayer G et al. 2007).
Therefore, an increased level of surveillance in residential schools, hostels, orphanages are recommended. Very few research studies are available on prevalence and etiological agents of Tinea capitis in children from this part of India. Surveillance can be performed using Wood’s lamp for the detection of Tinea capitis based on the fact that some dermatophyte species produce characteristic fluorescence under UV light. The chemical responsible for the fluorescence is pteridine (Wolf FT et al 1957). Wood’s lamp is helpful in the diagnosis and treatment of an individual patient as well as for mass screening and control of epidemics in schools ( Halprin KM et al. 1967). Dermatophytes that cause fluorescence are generally members of the Microsporum genus. However, the absence of fluorescence does not necessarily rule out Tinea capitis as most Trichophyton species, with the exception of T. schoenleinii, are nonfluorescent.
On the basis of the type of hair invasion, dermatophytes are also classified as endothrix, ectothrix or favus. In endothrix infection the fungus grows completely within the hair shaft, the hyphae are converted to arthroconidia (spores) within the hair while the cuticle surface of the hair remains intact (Fuller et al. 2003). In ectothrix infection hair invasion develops in a manner similar to endothrix except that the hyphae destroy the hair cuticle and grow around the exterior of the hair shaft. Arthroconidia may develop both within and outside the hair shaft. Elongated hyphae, parallel to the long axis of the hair, persist within the hair. Favus is a rare type of Tinea Capitis characterized by typical honey-colored, cup-shaped, follicular crusts called scutula ( Brajesh et al. 2013).
AIMS AND OBJECTIVES
To determine clinical and mycological profile of Tinea capitis in children
To correlate KOH microscopy and culture in diagnosing Tinea capitis.
To determine the possible associated predisposing factors for fungal infections among them.
MATERIALS AND METHODOLOGY
Study design: A cross-sectional study was carried out in the Department of Microbiology of a Rural Hospital of central India, during the period of 1st June- 31st July 2015 after due approval from the Institutional Ethical Committee.
Setting: Study was carried out in the Microbiology department of a Rural Hospital of central India.
Study participants: All children aged between 6-10 years of age from the six different Government Primary schools, were screened for routine examination and investigations.
Sample size: All total 323 children belonging to classes’ I–V standard from six different Government Primary schools were included in this study.
Inclusion Criteria:
All Students of Classes I–V standard from six different Government Primary school and
With the consent of their parents/ respective teachers were included in this study.
Exclusion Criteria:
Students, not in the age group of 6–10 years
Students receiving anti-fungal treatment.
Not giving consent was excluded from the study.
Collection and Processing of Clinical Samples
Questionnaires were designed and adapted for collecting demographic and socio-economic aspects of the child. This included many factors like family size, family income, and history of antifungal treatment, parent’s educational level and parents’ employment. All areas of the scalp were thoroughly examined to assess the morphological types of Tinea capitis. Wood’s light was used for the diagnosis of Tinea capitis; however, the absence of fluorescence did not necessarily rule out Tinea capitis.
Collection of Hair plucking and scalp scrapping:
The specimens were collected and processed according to Weitzman and Summerbell. The lesions were thoroughly cleaned with 70% alcohol. The scalp scrapings were collected from the margins of the lesions with a sterile surgical blade (No.15). The affected dull and lusterless hairs were epilated with the help of a sterile forceps. All the specimens were collected in a clean white sterile paper. All samples were labeled appropriately and transported to the laboratory within 4 h after sample collection for direct microscopic examination and culture.
Direct Microscopy
Potassium hydroxide mounts
Direct microscopic examination of the scrapings and hairs was carried out by mounting with 1-2 drops of 20% KOH for 15-30 min. Slides were then microscopically evaluated for the presence of hyphae, arthrospore or conidia for either ectothrix or endothrix infection. Infection of the hair was described either as ectothrix (sheath of arthroconidia formed on the outside of the hair shaft) or as endothrix (arthroconidia formed within the hair shaft).
Mycology Culture
Each specimen was then inoculated on two separate Sabouraud's Dextrose Agar slopes containing chloramphenicol, one with and the other without cycloheximide (chloramphenicol-0.05 mg/mL, cycloheximide-0.5 mg/mL). The cultures were incubated at room temperature for 4-6 weeks and observed regularly for growth. The fungal isolates were then identified.
Identification of isolates
The cultural characteristics of the isolates were noted and identification was done on the basis of Duration of growth, surface morphology, pigment production on the reverse, the texture, whether fluffy, powdery, cottony or floccose, buff, whether the hyphae was radiating at the margins or whether their colony were folded. The microscopic characteristics of their hyphae were also noted. Microscopic examination in lacto phenol cotton blue preparation and slide culture was also done to confirm the isolate. Whenever necessary urease and hair penetration test were done. The clinical, microbiological and etiologic data was collected and correlated. Results so obtained were tabulated and statistical evaluation of the results was done.
Statistical analysis
The results obtained in this study are presented using descriptive statistics (frequency and percentage). We have used the odds ratio with 95% CI to find out the association of predisposing factors like gender, pets, parent’s education level with the proportion of mycoses among the pupils.
OBSERVATION and RESULTS
Of the 323 pupils so screened from six different rural schools, 81(25.1%) were found to be screened positive by Clinical examination and Wood’s lamp examination (Table 1). Amongst these 81 Tinea capitis cases, 27 were screened positive by Wood’s lamp examination and 54 by clinical examinations (Table 1).
The proportion of Tinea capitis among primary school children in relation to certain socio- dermographic variables is shown in Table 2. Amongst the 81 screened positive Tinea capitis cases, a slight female preponderance was seen, with females comprising 55.5% of the patients. Also it was noted that Tinea capitis cases were more (67.90%) among children from families with low educational level of parents i.e. primary school or less compared to 32.09% amongst those students with high educational level of their parents. Almost 50.61% of students gave a history of pets at home or prolonged contact with animals as most of the study population had farming business (Table 2). Bathing habits among children such as frequency of head bath, use of shampoos and use of hair oils was obtained from all the students. It was observed that the measures of personal hygiene were similar in both students with Tinea capitis and without Tinea capitis.
As per Table 2 odds for suffering with Tinea capitis (culture positive) are same in both males as well as females (Odds ratio=1) and children from families with low educational level of parents i.e. primary school or less were found to have slightly lesser odds (Odds ratio=0.94) for suffering with Tinea capitis (culture positive) as compared to those children who belong to families with high educational level of their parents i.e. secondary school. Children from families having pet were found to have slightly higher odds(Odds ratio=1.25) for suffering with Tinea capitis (culture positive) as compared to those children who belong to families having no pet.
However all these parameters we studied were not statistically significant as 95% Confidence interval is overlapping as well as including base value of 1.
Among the various clinical variants seen, non-inflammatory Tinea capitis i.e. Grey patch, Black dot and seborrhoeic types was more common (96.29%) than inflammatory TC (3.7%) Among the Non-inflammatory cases, seborrhoeic type was more common than the other types (Table 3).
Potassium hydroxide studies were carried out on hair samples from 81 clinically as well as Wood’s lamp positive (Figure 2) suspected cases of Tinea capitis. The findings are summarized in Figure 1.Fungal spores invading hair shafts could be seen in 40.74% (81) of the cases. An endothrix pattern of spore distribution (Figure 3) was more common (23.45%) than an ectothrix pattern (13.58%) only 3 cases (3.70%) showed spores in both the patterns simultaneously on initial examination. No fungal elements could be identified in 48 cases (59.25%). Furthermore, we compared the culture results with microscopic examination. In our study, we found 9.87% of Tinea capitis cases positive by both KOH and Culture, 30.86% by KOH only, 1.23% by Culture only while 58.02% cases were negative by both KOH and culture both (Table 4).
In our study, the total number of culture positive isolates were 9, out of which Trichophyton violacium (Figure 5)was the commonest isolate, [4(44.44%)], followed by Microsporum canis (Figure 4) [3(33.33%)]and Trichophyton rubrum (Figure 6) [2(22.22%)]. Most of the culture positivity was in seborrhoeic type of Tinea capitis i.e. 66.66%.
DISCUSSION
Tinea capitis is a common fungal infection in children of school age, particularly among those living in unhygienic conditions. In the present study the rate of tinea capitis was 25.07%as screened by clinical and wood’s lamp examination. The lower isolation may be due to geographical and climatic variations, as this year our area received lower rainfall and is less humid and drier; thus are less chances of acquiring fungal infections. Varying prevalence rates of tinea capitis ranging from 4.6% - 39.6% have been reported in studies from Nepal, Nigeria, Central Africa, India, Iran (Barbhuiya JN et al.2002, Rastegar Lari et al. 2005, Jha et al. 2006, Emele FE et al.2008, Hogewoning AA et al. 2011). These variations may be attributed to social, socio- economic and geographical variations; cosmetic factors play a role in the spread of infection (Higginis EM et al 2000).
Various conflicting views exist regarding the sexual predominance of Tinea capitis. Some authorities believe that Tinea capitis may be common in boys due to shorter hair, allowing easy access for circulating spores (Friedlander SF et al. 2003), while others believe that it may be more common in girls due to tight hair braiding (Chen BK et al.2001). In our study slightly higher proportion of female students (55.5% out of 81 screened positive cases) affected compared to male students (44.4%). This is in contrast with the studies carried out at Kenya ( Ayaya SO et al. 2001), Kolkata ( D Kundu et al. 2012) and Rajasthan (Kalla G et al. 1995) showing male preponderance.
The present study showed a relationship between level of parental education and the proportion of Tinea capitis in the population studied. The frequency of Tinea capitis was more in children whose parents had primary or no education (67.90%out of 81 screened positive cases) when compared with the frequency in children whose parents attained secondary and post secondary school education (32.09%). All the studied school children were from rural areas in and around Wardha district. Majority of the parents of infected children were involved in farming thus exposing them to environmental agents. The average family size was six members per family and the whole family resided in a single room. The normal level of hygiene was not maintained as the hair wash frequency was not more than twice a week in diagnosed cutaneous mycoses cases because of lack of awareness as well as limitation of water supply. The practice of sharing of combs allowed the spread of infection at a higher rate in Tinea capitis cases specifically. Majority children screened for infection of Tinea capitis belonged to overcrowded government schools. This points to the importance of health education in control of infections. Inadequate and poor infrastructures as was observed in all the six schools could be added as one of the major factors influencing transmission of infection amongst the pupils.
Most of the similar studies reveal that non-inflammatory types of Tinea capitis were more common (51%) than inflammatory variants (32%) ( Hussain I et al. 1994, Kalla G et al. 1995, Singal A et al. 2001, Jha BN et al. 2006). In our study, Seborrhoeic type was more common followed by Black dot and grey patch types., study by Bose et al (2011) and Singal A et al (2001) have reported seborrhoeic variant as the predominant type followed by black dot.
In the present study, examination of KOH-stained smears microscopically revealed different patterns of distribution of spores in the hair shaft. KOH positivity was seen in 40.74% of screened positive; out of which endothrix presentation (57.57%) was more common than ectothrix (33.3%) while 9.09% shows mixed pattern of spore arrangement. The fungal culture yielded positive result in 11.11% out of 81 screened positive cases. Isolation rates ranging from 24%-93% have been reported in previous studies from different geographical areas (Singal A et al. 2001, Jha BN et al. 2006, Al Samarai et al. 2007, Garg J et al 2009,Yazdanfar A et al. 2010, Bose et al. 2011, Azab MM et al. 2012). Culture specimens from all the cases were examined. No growth at the end of 6 weeks was recorded in 88.8% of the cases (72 cases). Of those showing growth of fungal elements, Trichophyton violaceum was the commonest isolate, [4(44.44%)] which has also been reported by various workers from India and other parts of the world as well (Jha BN et al. 2006, Jehangir M et al.1999) All the culture positive isolates were also shown positive results on KOH microscopy except one isolate of Trichophyton rubrum .
In the present study out of 323 students so screened, 25.07% were found to Tinea capitis by clinical and wood’s lamp examination. In this study a female preponderance was noted. Low socio-economic background and poor personal hygiene were significant pre- disposing factors of Tinea capitis among the studied students. The most common pattern of Tinea capitis noted was seborrhoeic type. Fungal culture results were positive among 11.11 %of children showing positive results on screening children. Among this Trichophyton violaceum(44.44%) was the commonest cause of Tinea capitis in our geographical area followed by Microsporum canis(33.33%) and Trichophyton rubrum (22.22%).
CONCLUSION
In our study, the sensitivity of direct microscopy considering culture as gold standard was found to be 88.89% and specificity was 65.28%. This emphasizes the need of performing both the tests.
Acknowledgement:
Source of Support: Nil.
Conflict of Interest: None declared.
Englishhttp://ijcrr.com/abstract.php?article_id=2397http://ijcrr.com/article_html.php?did=2397
Ahmed I, Ahmed Z, Sarwat N. Prevalence of tineacapitis and asymptomaticcarriage amongst school going children. Journal of Pakistan Association of Dermatologists. 2006; 16:215-219.
Al Samarai. Tinea Capitis among Iraqi Children: Public Health Implication Journal of Clinical and Diagnostic Research. 2007;1:476-482.
Ayanbimpe GM, Taghir H, Diya A, and Wapwera S. Tinea capitis among primary school children in some parts of central Nigeria. Mycoses.2008;51:336-340
Ayaya, S.O., Kamar, K.K. and Kakai, R. Aetiology of Tineacapitis. East Africa Medical journal; 2001. 78:531-5
Azab MM, Mahmoud NF, Abd Allah S, Alaa El Din, Hosny MS, Shehata AS and Mohamed RW. Dermatophytes isolated from clinical samples of children suffering from tinea capitis in Ismailia, Egypt. Australian Journal of Basic and Applied Sciences. 2012; 6:38-42
Barbhuiya JN, Das SK, Ghosh A, Dey SK, Lahiri A. Clinico-mycological study of superficial fungal infection in children in an urban clinic in Kolkata . Indian J Dermatol. 2002;47:221-23
Brajesh, K. J. and Mahadeva, S. M. Studies on invasive keratinophilic dermatophytes of human hair. Journal of Drug Delivery and Therapeutics; 20133(2): 70-74
Bose S, Kulkarni SG, Akhter I. The incidence of tinea capitis in a tertiary care rural hospital - a study. Journal of Clinical and Diagnostic Research. 2011;5:307-311
Chen BK, Friedlander SF. Tinea capitis update: a continuing conflict with an old adversary. Curr Opin Pediatr 2001;13:331-5
D. Kundu, L. Mandal, and G. Sen . Prevalence of Tinea capitis in school going children in Kolkata, West Bengal. J Nat SciBiol Med. 2012 Jul-Dec; 3(2): 152–155
Emele, F.E. and Oyeka, C.A. Tinea capitis among primary school children in Anambra state of Nigeria. Mycoses;200851:536–41
Friedlander SF, Rueda M, Chen BK, Caceros-Rios HW. Fungal, protozoal and helminthic infections. In: Schachner LA, Hansen RC, editors. Pediatric Dermatology. 3rd ed. Mosby; 2003: p 1093-140
Fuller, L., Child, F., Midgley, G. and Higgins, E. Diagnosis and management of scalp ringworm. British Medical Journal; 2003 326: 539-541.
Ginter-Hanselmayer G, Weger W, Ilkit M, Smolle J. Epidemiology of tineacapitis in Europe: current state and changing patterns. Mycoses. 2007;50:6-13
Ginter-Hanselmayer G, Seebacher C. Treatment of tineacapitis—a critical appraisal. J Dtsch Dermatol Ges.2011;9:109–114
Garg J, Tilak R, Garg A, Prakash P, Gulati AK, Nath G. Rapid detection of dermatophytes from skin and hair. BMC Research Notes 2009, 2:60
Grover C, Arora P, Manchanda V. Tineacapitis in the pediatric population: A study from north India. Indian J Dermatol Venereol Leprol. 2010;76:527- 532
Halprin KM. Diagnosis with Wood’s light. Tineacapitis and erythrasma. JAMA 1967;199:177
Higgnis EM, Faller LC, Smith CH. Guidelines for the management of Tinea capitis. Br J Dermatol 2000; 143: 53 – 58
Hogewoning AA, Adegnika AA, Bouwes Bavinck JN, Yazdanbakhsh M, Kremsner PG, van der Raaij-Helmer EMH, Staats CCG, Willemze R and Lavrijsen APM. Prevalence and causative fungal species of tinea capitis among schoolchildren in Gabon. Mycoses. 2011;54:354-359
Hussain I, Aman S, Haroon TS, Jahangir M, Nagi AH. Tinea capitis in Lahore, Pakistan. Int J Dermatol 1994;33:255-7
Jehangir M, Hussain I, Khurhid K, Haroon TS. A clinico etiologic Correlation in Tinea Capitis. International Journal of Dermatology 1999; 38: 275-278
Jha BN, Garg VK, Agrawal S, Khanal B, Agarwalla A. Tineacapitis in eastern Nepal. Int J Dermatol. 2006;45:100-102
Kalla G, Begra B, Solanki A, Goyal A, Batra A. Clinico-Mycological study of Tinea capitis in Desert district of Rajasthan. Indian J DermatolVenereolLeprol. 1995;61:342–5. [PubMed: 20953016]
Mahé A, and Hay RJ. In: Epidemiology and management of common skindiseases in children in developing countries, WHO bulletin. Ali Hussein. (ed). Geneva, Switzerland: World Health Organization Publications, 2005, page 22
Mohrenschlager M, Seidl HP, Ring J, et al. Pediatric tineacapitis: recognition and management. Am J Clin Dermatol. 2005;6(4):203–213
RastegarLari A, Akhlaghi L, Falahati M and Alaghehbandan R. Characteristics of dermatophytoses among children in an area south of Tehran, Iran. Mycoses. 2005;48:32-37.
Singal A, Rawat S, Bhattacharya S, Mohanty S, Baruah MC. Clinico- mycological profile of tinea capitis in North India and response to griseofulvin. J Dermatol 2001;28:22-6.
Wolf FT. Chemical nature of the fluorescent pigment produced in Microsporum-infected hair. Nature 1957;180:860-1.
Yazdanfar A. Tinea capitis in primary school children in Hamedan (West of Iran). International Journal of Medicine and Medical Sciences. 2010;2:29-033
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241923EnglishN-0001November30TechnologyOntology Based Optimization with Adaptive Network Based Fuzzy Inference System Based Health Maintenance Systemv
English3240R. JayaEnglish C. S. PillaiEnglish R. Jagadeesh KannanEnglishIn late years, economic growth and globalization have driven significant modernization of Ayurvedic Medicine in India. According to Ayurvedic Medicine, human body is divided into three constitutions/tridoshas based on the hypothesis of Prakriti. The categories of Prakriti are written based on the philosophy of tridoshas namely Vata, Pitta and Kapha. Ayurvedic clinicians’ records/documents the consultation of patient along with patient’s Prakriti in a paper. This is neither exchanged nor made accessible to other clinicians’. Due to rapid advancement in electronic media, the necessity of digital Ayurvedic patient management systems could help in diagnosing the patient before the consultation of clinicians. Henceforth, an electronic health record system is proposed that uses ontology oriented procedure to compute the physical constitutions of a human body. Specifically, a rule based system is developed and evaluated using Particle Swarm Optimization - Adaptive Network based Fuzzy Inference system (PSO-ANFIS). The ontological inputs to the fuzzy interface includes Doshas, food and lifestyle recommendations, obtained from clinical and textual data. Automated system created using ontology oriented PSO-ANFIS interface supports food, herbs, diet and life style recommendation to the patients’. The proposed model is evaluated with different patient and the sample is tested with the clinicians. The results proved that the proposed method fits with better results than the conventional ones.
EnglishParticle swarm optimization, Adaptive network based fuzzy inference system, ANFIS-PSO Markup LanguageINTRODUCTION
The core aim of data mining is to discover the meaningful knowledge from the data collection. The data mining theories are often represented in the form of models. Each model has its own process that builds the entire system. A reliable model holds the essence as specific formulation that may discover the knowledge from junk data. The model is designed in such a way that it acts as an interface between the human and machine world. Thus providing a model, a better understanding about the outer world. The certain models are designed strictly for production and diagnostic purpose [1] in the field of medicine. Data mining provides technical and practical solutions that provides better analysis of medical data. With such solutions, the construction of prediction models could be an easier task for diagnostic purpose [2].
Globalization in India has improve the evidence based research and the ingenuities required to regulate the procedure for Ayurvedic medicines. Ayurvedic not only considers the disease examination but also the patients and their diet, food, herbs and life style recommendation. The prominent method for examining the patient in Ayurvedic medicine is Prakriti. Prakriti represents the unique constitution, personality with the globe, represented in the form of three dimensions namely: Dhosas, Kapha and Vata. If the Prakriti of an individual are in correct proportion then the patient is considered healthy or vice versa. Clinicians assess the patients through Prakriti profile for customizing the treatment and its prior diagnosis. However, the Prakriti of a patient is recorded in a paper after the consultation. This is not usually exchanged between the clinicians [3].
The increased use of electronic devices further improved the interoperability of the health records in electronic format. This provided the use of electronic health system for prior diagnosis before consultation with the clinicians. The electronic records that posses the standards of Prakriti constituents and other associated datasets modernizes the Ayurvedic treatments. Clinicians analyzes the person health through their experience and skill, however, this is not available with all clinicians. Hence, an intelligence system for interpretation and analysis of health related issues is made precise with the available datasets. This combination of datasets could help in diagnosing the disease that is exact or precise in nature [4].
The aim of the research is to improve the degree of precision in diagnosing the ailments through the concept of fuzzy systems. The fuzzy rule base associated with logical systems represents the fuzziness of the input parameter. Further improvement in analyzing the ailments could be done through the concept of ontology. Ontology represents the knowledge in form of a tree with concept and its linked instances. To handle the vagueness and uncertainty of ontological data, fuzzy logic is combined with input dataset. However, to improve the matching pattern with more precise results, Particle Swarm Optimization (PSO) is used.
Since each individual is born with specific constitution, Ayurveda proposed three major Prakriti. The dominance of each constitution provides a particular Dosha to each individual according to which the body behaves. The present research works involves discovery of suitable Prakriti method that fits well in diagnosing the patient ailments. The solution to such ailments is provided through Prakriti constitutional medicine. Hence, an automated system is defined with strict rules that involves predefined ontological datasets in fuzzy rule base. The proposed method involves Adaptive Neuro Fuzzy Interference System (ANFIS) to diagnose the diseases with defined rule pattern. To improve the training pattern of the ANFIS, Particle Swarm Optimization (PSO) is used. The Diagnosis is carried out after performing the rules based on the inputs given at the user end. The outputs purely depends on the rule selection from the predefined ontological datasets. This system helps well as a diagnostic tool in finding out the ailments of the patients and treating them with one of the constitutional medicine.
The other parts of the paper includes: section 2 deals with the related works, section 3 deals with the collection of data and the procedure of using ANFIS and PSO algorithm to handle the datasets and to improve the training pattern. Section 4 and 5 describes the evaluation of the proposed method and finally, section 6 concludes the entire paper with future direction.
RELATED WORKS
Ranjit et al. [5] proposed fuzzy expert system on the physical constituents from human body namely Vatt, Pitt and Kapha [15]. The entire operations is carried out to assist the Ayurvedic diagnosis and treatment. Here fuzzy based expert system is used to calculate the three constituents. Begic et al. [6] proposed genetic algorithm based ANFIS expert system to predict the dermatological disease in human beings. Here, nine features are taken as inputs to the classifier for diagnosing the disease in real time. Farooque et al. [7] compared the performance of data mining algorithms like naïve bayes, knn classifier, logistic regression and decision tree for Ayurvedic Prakruti Temperament. However, Yim and Joo [8] used fuzzy logic designer to diagnose 8 constitutions in human body. This method is a Korean way of diagnosing the patients. The technique evaluated assist the physicians in determining the disease. The 8 constitution of human body is considered in the present research with more detailed parameters from [12]. Chattopadhyay [9] Mamdani’s Fuzzy logic controller (FLC) on a Feed Forward Multilayer Neural Net is used to diagnose the depression in human beings. Appaih et al. [10] used ANFIS for diagnosing the malaria disease. So, this proves how the ANFIS system is efficient in medical applications. However, to improve the effectiveness and computational capability can be improved with the addition of other meta-heuristic approaches. Karaboga and Kaya [11] proposed Ant colony optimization (ACO) algorithm to improve the training phase of ANFIS. Similar procedure is implemented in our proposed system but with PSO algorithm to improve the training phase using swarming procedure at every iterations. A ontology based automated clinical systems [13, 16, 17, 18, 22, 25, 26, 27, 29,30,31,32,33,34] has proved better in health care system and the association of fuzzy further enhanced the results [14, 20, 21, 23, 24, 28]. Hence the use of ontology is inbuilt in the proposed system with fuzzy logic for Prakriti ailments. The method in [19] uses PSO to build the ontological structure, however, the present system involves the use of PSO in improving the ANFIS training pattern.
PROPOSED METHODOLOGY
A three categories of questions is prepared for determining the Prakriti in Ayurvedic medicine. These categories include anatomical, psychological and physiological questions of each individuals or patients. Suitable questionnaires are prepared and the scores related to each questions is calculated. Sample questionnaire is attached under appendix A.The patients are allowed to fill the questionnaire that includes parameters like season details and patient details. The Prakriti concept is unique from other medical concepts. Hence, Prakriti is determined with its associated parameters using proposed ontological datasets, easily. The entire process is carried out in PSO – ANFIS interface that helps in improved accuracy of the test datasets.
3.1 ONTOLOGY – HEALTH MAINTENANCE FRAMEWORK
The health maintenance system is proposed with ontology rule based system that address the diagnostic problems using ontological representation. This includes the core principles of Ayurvedic medicine, Prakriti Doshas, Prakriti categories, herbs, diet, diseases, symptoms and diagnosis (Figure 1).
The figure represents the Prakriti system with input output interface that is controlled by a rule base system that involves ANFIS – PSO logic (Figure 3). The input parameters involves: user profile input, season input and determined anatomical, psychological and physiological Questions. These input ontology parameters is prepared through questionnaires, since it is not available in web resources, however, certain literatures related to Prakriti records the related resources. Depending on the Prakriti determination using ANFIS-PSO Markup Language (APML), the output parameters includes attributes like Food, Diet, Herbs and Lifestyle recommendation to the patients.
Database consist of set of different data related to the recommendation of user that is collected from various resources like questionnaires and from clinicians. A user interface is created via APML and delivers the Doshas from database that consist of herbs and usage, probable disease, health problems, food and diet style recommendation and life style. The output is delivered through the Prakriti determination through ANFIS-PSO system. The entire process is started upon the input from the user input interface. Here to improve the Prakriti determination in proposed method, the ontology is integrated with fuzzy logic for providing best recommendations to the user/patient. Fuzzy ontology generates ontology through fuzzy logic, fuzzy sets and variables. This predicts the uncertain things in an effective way and the knowledge extraction occurs based on the membership value of the variables.
The proposed system works in the following manner:
User provides information about age, height, weight and sex.
User enters his/her Prakriti type through recommended questionnaires.
User/patient enters the current season.
Fuzzy ontology is applied over the collected inputs from an individual.
PSO is applied over the results of fuzzy trained patterns to improve accuracy.
A fuzzy profile for the user is created.
For the recommendation purpose, fuzzy ontology and the fuzzy profile ontology is created finally.
Then the recommendations like: Food, Diet, Herbs and Lifestyle recommendation is suggested to the patients.
3.2 FUZZY ONTOLOGY
The Fuzzy ontology is similar to the procedure as mentioned in [20]. The fuzzy ontology system provides a semantic relationship between the associations of the same domain. A flexible fuzzy query searches the database contents and compares it with the domain ontology that represents the queried attributes in the domain. The ordering of resulting dataset is done through the degree of similarity calculated from the query subject and the database content in the ontology.
Input process divides the query and each subquery is sent to the corresponding process module according to the query attributes domain. Output processing combines the answers obtained from the different modules. Each row accomplishment degree in the resulting dataset should be higher than a user-given threshold [20].
3.3 Particle swarm optimization (PSO)
Particle Swarm Algorithm (PSO) solves the problem formulation iteratively by improving the solution w.r.t the given ontology. The optimization in PSO is carried out through n particles/individuals of candidate solution. The particles are moved around in the search space with specified mathematical formulation over its position and velocity. Here, each particle is assigned with random position in N-dimensional space. The position and velocity of the particle is denoted as xi and vi over solution space.
The movement of each particle is guided by its local best position to attain the best position in the search space. The position of previous best local or personal solution is denoted as Pbesti. The best positions are updated frequently to all the particles and move the swarm towards best solution in the search space. Similarly, the best global position is denoted as Gbesti. This is considered as the best particle among all particles in the search space. Here, each particle in search space is considered as a vector V that moves in the range of [-Vmax, Vmax]. The movement of particle in search space is controlled by the factor V that avoids excessive roaming. The velocity of the particle is updated using:
Vi(t+1) = ωVi(t) + Car1[Xi(t) - pi(t)] + Cbr2[Xi(t) – gd(t)] (1)
At each iteration the particle moves towards new position, hence, the range of particle should not exceed [min X, max X]. The position of the particle is thus updated using:
Xi(t+1) = Xi(t) + Vi(t+1) (2)
The process gets repeated until the stopping criterion is defined or reached and
ω = (ωmax – (ωmax–ωmin)/Imax)I
where,
i = 1,2,..., M
Total number of particles
d = 1, 2,..., D
Total dimension of particles the solution space
r1and r2 Î [0,1]
Random numbers distributed uniformly in the search space
Ca and Cb
Learning factors (value is taken as 2.0)
Ca represent individual cognition component that represents the particle’s ability in search space.
Cb represents social communication component that gets influenced from the social environment.
ω
Inertial component that limits the particle’s velocity in search space. The value of inertial component is 0.4
ωmax
Final inertial weights
ωmin
Initial Inertial weights
Imax
Total Iterations
I
Current Iteration
The algorithm for PSO in ontology evaluation is shown in following table.
Algorithm 1: Particle swarm optimization (PSO)
Step 1:
Step 2:
Step 3:
Step 4:
Step 5:
Step 6:
Step 7:
INITIALIZE the position and velocity, randomly to all the particles in the search space.
EVALUATE the xi and vi of each particle. The local (Pbesti) and global (Gbesti) position is evaluated
UPDATE the xi and vi of each particle using Eq.(1) and Eq.(2).
CALCULATE the xi and vi of each particle.
UPDATE Pbesti
For each particle
IF value of new position is better than that Pbesti,
THEN replace its Pbesti by new position.
UPDATE Gbesti
For each particle
IF value of new position is better than that Gbesti,
THEN replace its Gbesti by new position.
REPEAT the process until the maximum iterations
IF solution converges
THEN the value of Gbesti is considered as output
IF ELSE GOTO Step 3.
END
3.4 ADAPTIVE NEURO-FUZZY INFERENCE SYSTEM (ANFIS)
The ANFIS network is used to classify the Prakriti diagnosis with predefined set of rules. The ANFIS system possess fuzzy model with rules that formulates the input output datasets. The second model is the neural network model, which is integrated with Fuzzy model that improves the learning capability of nonlinear functions.
The ANFIS system consists of two inputs and a single output interface with if-then rule type. The rules are specified as follows:
If u is A and v is B then q = f (u,v) (3)
where,
Depending on the input entropy results, the ANFIS predicts accurately the class of disease to which it belongs. As a result, if the value is greater than 0 then the patients are marked as diseased and vice versa.
3.5 ANFIS-PSO CLASSIFIER
The algorithm of ANFIS-PSO classifier is described stepwise. Here, the recombination of the predicted signals is carried out to find the suitable category of Prakriti.
Step 1: Form a matrix with Prakriti data set in columns matrix. The column array possess patients profile collected from real time datasets. In this first step, the matrix will contain seven columns (represents seven Prakriti constitutional types) corresponding to collected datasets of Prakriti determination.
Step 2: Select total columns in previous array represents the values of real input data.
Step 3: Train the ANFIS system using the input data that uses a combination of the least-squares and back propagation gradient descent method. Training allows the proposed method that adjusts its parameters as submitted inputs or outputs. The acquired knowledge through the process of training is then tested through a new dataset or the testing set. The ANFIS network generalize for its accurate output over the testing data. It is undesirable in order to over-train the proposed system, since it represents that it works only on training dataset and does not generalizes on training dataset. A large training datasets is thus avoided that over-trains the system during the process of learning. To obtaining better accuracy, PSO trains the membership function parameters of fuzzy system
Step 4: A vector of N-dimensions is created that equals the total membership functions. The vector contains membership function parameters that is optimized by PSO. The mean squared error is used for defining the fitness function.
Step 5: The parameters of PSO is defined in Table.1. The Parameters are randomly initialized at first stage and gets updated using PSO. During each iteration, parameters are being updated and once the entire parameters gets updated, the initial parameter update is considered further and so on. The updated parameters are grouped in the form of vectors that gets updated during each iteration.
The PSO algorithm for optimizing the parameters/fuzzy membership functions is shown below:
Initialization of the population position and speed in swarm. For each particle, there obtains a random initialization of particle position and velocity. The size of the vector is same as the problem size.
Assess ability of each particle, Pbest. If the particle value is better than its present value, particle’s current position or Pbest is reset and individual value is updated. Location of best particles is made reset, if the best of all particles’ value is better than overall current Gbest value.
Measure each particle’s fitness, Pbest and store the best fitness value of each particle Gbest.
Modify the speed through the positions of Pbest and Gbest.
Update the particles.
End the process if the conditions are verified. When the current iteration reaches the maximum, then the iterations are stopped and the best solution is collected.
Sixth step: Extraction of the ANFIS output using the PSO parameters.
Seventh step: The final output provides the results of the predicted values of this approach.
IV.RESULTS
The proposed method is implemented over Jena framework that supports RDF, OWL and rule inference engine. Additionally, fuzzy logic with PSO training pattern is coded and applied over J7ena framework.
The evaluation of ontology depends on quality and correctness. Standard metrics like recall, precision and F-score is used for performance measurement. Let S – size of ground truth value, D – Number of correct values that is extracted by proposed system and N denotes the number of total values returned by the proposed system. The evaluation metrics is shown in table 2.
The results of the automated system is checked with a clinician to evaluate its correctness. Here, if the value of the prediction system is found to be 1, then the patient is said to have a specific disease and vice versa. Depending on the disease, the Prakriti type is found and it is divided into following class:
The set of test samples are tested after the system is trained by the PSO-ANFIS procedure. Initially, the epoch or the iterations of PSO algorithm is set as 200 and the total test samples for inspection is taken as 10. It is seen that the proposed PSO-ANFIS method has 9 matched samples which is higher than normal ANFIS [12] method with 7 samples. The total epoch is increased to 300 with other set of 10 samples. It is found that the proposed method has all the 10 samples matched and ANFIS has got 8 samples matched. Likewise, the test samples are increased for 15 over 200 and 300 iterations. From the test results, it is found that the proposed method attains 14 and 15 matched samples and the ANFIS method has12 and 13 matched samples respectively for 200 and 300 iterations.
The test success of the proposed method with the existing ANFIS is tested to see the errors present during the training and testing process. Similar inspected and epoch pattern is carried out to find the training, testing and check error. It is found from the results that the error rate of training, testing and check error for the proposed PSO-ANFIS system produces less error than the ANFIS system. It could also be interpreted that the use of PSO algorithm over the training samples further improved the test performance than the existing ANFIS.
DISCUSSIONS
It is found from the table 5 that the results obtained has better improvements in all performance parameters than the conventional ANFIS system. This proves that PSO classifier helped in training the samples of the fuzzy ontology output. Specifically, the variations of parameters like recall and F-measure has a considerable impact than the existing system. Further investigations on performance through FMeasure value as a benchmarking point has carried forward that enables the system to perform well over the defects in existing system.. Finally, a drift in performance is found that shows that the accuracy of PSO-ontology-ANFIS is further improved than the conventional ANFIS type.
CONCLUSIONS
In the present research, comparisons are carried ou
Englishhttp://ijcrr.com/abstract.php?article_id=2398http://ijcrr.com/article_html.php?did=2398
Tang PN, Steinbach M, Kumar V. Introduction to Data Mining. Pearson Addison: Wesley Bostan; 2005.
Bellazzi R, Zupan B. Predictive data mining in clinical medicine: Current issues and guidelines. International Journal of medical informatics. 2008; 77(2):81–97.
Ranjit K, Vishu M, Prateek A, Kumar SS, Amandeep K. Fuzzy Expert System for Identifying the Physical Constituents of a Human Body. Indian Journal of Science and Technology. 2016 Jul 27;9(28).
Shital, V., and Sambare, S. S. (2015). Study of Diet Recommendation System based on Fuzzy Logic and Ontology. International Journal of Computer Applications, 132(12), 20-24.
Ranjit K, Vishu M, Prateek A, Kumar SS, Amandeep K. Fuzzy Expert System for Identifying the Physical Constituents of a Human Body. Indian Journal of Science and Technology. 2016 Jul 27;9(28).
Begic FL, Avdagic K, Omanovic S. GA-ANFIS Expert System Prototype for Prediction of Dermatological Diseases. Studies in health technology and informatics. 2014 Dec;210:622-6.
Farooque MM, Aref M, Khan MI, Mohammed S. Data Mining Application in Classification Scheme of Human Subjects According to Ayurvedic Prakruti–Temperament. Indian Journal of Science and Technology. 2016 Apr 18;9(13).
Yim J, Joo J. Implementation of an Inference System to Classify Persons into Eight Constitutions. Indian Journal of Science and Technology. 2015 Oct 14;8(26).
Chattopadhyay S. A neuro-fuzzy approach for the diagnosis of depression. Applied Computing and Informatics. 2014 Feb 4.
Appiah R, Panford JK, Riverson K. Implementation of Adaptive Neuro Fuzzy Inference System for Malaria Diagnosis (Case Study: Kwesimintsim Polyclinic). International Journal of Computer Applications. 2015 Jan 1;115(7).
Karaboga D, Kaya E. An adaptive and hybrid artificial bee colony algorithm (aABC) for ANFIS training. Applied Soft Computing. 2016 Dec 31;49:423-36
http://ecmed.org/board/content.asp?bsNo=18andlng=en
Kuziemsky, C. E., and Lau, F. (2010). A four stage approach for ontology-based health information system design. Artificial Intelligence in Medicine, 50(3), 133-148.
El-Sappagh, S., Elmogy, M., and Riad, A. M. (2015). A fuzzy-ontology-oriented case-based reasoning framework for semantic diabetes diagnosis. Artificial intelligence in medicine, 65(3), 179-208.
Stranieri, A., Butler-Henderson, K., Sahama, T., Perera, P. K., Da Silva, J. L., Pelonio, D., Manjunath, SS., and Raghavachar, D. (2016). A visual grid to digitally record an Ayurvedic Prakriti assessment; a first step toward integrated electronic health records. Journal of Traditional and Complementary Medicine.
Riaño, D., Real, F., López-Vallverdú, J. A., Campana, F., Ercolani, S., Mecocci, P., Annicchiarico, R., and Caltagirone, C. (2012). An ontology-based personalization of health-care knowledge to support clinical decisions for chronically ill patients. Journal of biomedical informatics, 45(3), 429-446.
Saalmann, P., Zuccolotto, M., da Silva, T. R., Wagner, C., Giacomolli, A., Hellingrath, B., and Pereira, C. E. (2016). Application Potentials for an Ontology-based Integration of Intelligent Maintenance Systems and Spare Parts Supply Chain Planning. Procedia CIRP, 41, 270-275.
Grandi, F. (2016). Dynamic class hierarchy management for multi-version ontology-based personalization. Journal of Computer and System Sciences, 82(1), 69-90.
Pancerz, K., and Lewicki, A. (2014). Encoding symbolic features in simple decision systems over ontological graphs for PSO and neural network based classifiers. Neurocomputing, 144, 338-345.
Martínez-Cruz, C., Noguera, J. M., and Vila, M. A. (2016). Flexible queries on relational databases using fuzzy logic and ontologies. Information Sciences, 366, 150-164.
Karthikeyan, N. K. (2016). Fuzzy service conceptual ontology system for cloud service recommendation. Computers and Electrical Engineering.
Viale, P., Bora, J. J., Benegui, M., and Basualdo, M. (2016). Human endocrine system modeling based on ontologies. Knowledge-Based Systems, 111, 113-132.
Liu, J., Zheng, B. J., Luo, L. M., Zhou, J. S., Zhang, Y., and Yu, Z. T. (2016). Ontology Representation and Mapping of Common Fuzzy Knowledge. Neurocomputing.
Ali, F., Kwak, K. S., and Kim, Y. G. (2016). Opinion mining based on fuzzy domain ontology and Support Vector Machine: a proposal to automate online review classification. Applied Soft Computing.
Medina-Oliva, G., Voisin, A., Monnin, M., and Leger, J. B. (2014). Predictive diagnosis based on a fleet-wide ontology approach. Knowledge-Based Systems, 68, 40-57.
Koukias, A., and Kiritsis, D. (2015). Rule-based Mechanism to Optimize Asset Management Using a Technical Documentation Ontology. IFAC-PapersOnLine, 48(3), 1001-1006.
Arch-int, N., and Arch-int, S. (2013). Semantic ontology mapping for interoperability of learning resource systems using a rule-based reasoning approach. Expert Systems with Applications, 40(18), 7428-7443.
Ochs, C., He, Z., Zheng, L., Geller, J., Perl, Y., Hripcsak, G., and Musen, M. A. (2016). Utilizing a structural meta-ontology for family-based quality assurance of the BioPortal ontologies. Journal of biomedical informatics, 61, 63-76.
Ying, H. (1998). Sufficient conditions on uniform approximation of multivariate functions by general Takagi-Sugeno fuzzy systems with linear rule consequent. IEEE Transactions on Systems, Man, and Cybernetics-Part A: Systems and Humans, 28(4), 515-520.
Jaya R et al,Ontology based Decision Support System for Identifying the Physical Constituents of a Human Body, International Journal of Control Theory and Applications, Volume 9 • Number 51 • December 2016
Jaya R et al, A Narrative Review of Research on Traditional Indian Medicine and Role of Prakriti in Lifestyle Recommendation, International Journal of Control Theory and Applications, Volume 9 • Number 51 • December 2016
Jaya R,et al,Ontology Based Virtual Assistant for Diet Recommendation, IJCSMC, Vol. 6, Issue. 4, april 2017
Jaya R et al, Implementation Of Clinical Decision Support System For The Ayurveda Medicine, , IJCSMC ,Vol. 5, Issue. 9, September 2016, pg.29-33
Jaya R et al ,Ayurveda Medicine Roles in Healthcare Medicine, and Ayurveda Towards Ayurinformatics, IJCSMC, Volume 4 Issue 12, ISSN 2320-088X