Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareALTERED LIPID PARAMETERS AND THEIR RELATIONSHIP WITH THYROID STIMULATING HORMONE IN SUBCLINICAL HYPOTHYROIDISM English0105Bhavesh SadariyaEnglish Suman JainEnglish Sonal SoganiEnglishBackground: Subclinical hypothyroidism (SCH) is a common thyroid disease affecting large population of the world. Diversit persists regarding the level of lipid parameters in patients with SCH. Aim and Objectives: Aim of the present study was to estimate the level of lipid parameters in patients with subclinical hypothyroidism of Udaipur population and to see their relationship with TSH values. Materials and Methods: The present study was conducted on 75 patients of subclinical hypothyroidism and 100 euthyroid. Fasting venous blood was collected from each participant and analyzed for thyroid profile (T3, T4 and TSH) and lipid parameters (total cholesterol-TC, triglyceride-TG and high density lipoprotein - HDL). Low density lipoprotein (LDL) and very low density lipoprotein (VLDL) were calculated by using the Friedewald’s formula. Results: Mean values of total cholesterol, triglyceride, LDL, VLDL, TC/HDL ratio and LDL/HDL ratio were significantly higher in SCH patients as compared to euthyroid. Mean of Serum HDL was also higher in SCH patients than euthyroid, but was not statistically significant. All lipid parameters were positively correlated with TSH values in SCH patients. Mean values of TC, TG, HDL, LDL and VLDL were higher in female patients as compared to male patients, but were not statistically significant. Conclusion: Subclinical hypothyroidism is associated with dyslipidemia having raise in all lipid parameters, which may increases the risk of atherosclerosis and cardiovascular disease development. EnglishHypothyroidism, Cardiovascular disease, Euthyroid, Subclinical, DyslipidemiaINTRODUCTION Worldwide, thyroid gland disorders are most abundant public health problems.1 From total national burden of endocrine disorders, magnitude of thyroid gland disorders constitute approximately 42 millions in India.2 Thyroid disorders results from either hyper or hypo secretion of tri-iodothyronine (T3) and thyroxine (T4) from thyroid gland. Subclinical hypothyroidism (SCH) is defined as an elevated serum thyroid stimulating hormone (TSH) associated with normal serum concentration of thyroid hormone (total or free T4 and T3).3 Patients of SCH are asymptomatic or have minimal symptoms. SCH is more common in elderly people with twice in women than men.4 World-wide prevalence of SCH is about 7.5% to 8.5% in women and 2.8% to 4.4% in men.5 An epidemiological study in eight cities of India shows that prevalence of SCH is 8.73% in females and 7.17% in males.6 It is well known that Thyroid hormones and TSH affects synthesis, fate and mobilization of lipids.7 Several previous studies have shown elevated levels of total cholesterol, triglyceride and LDL-C in SCH patients 8, 9, while some study does not support this findings.10 This shows diversity regarding level of lipid parameters in subclinical hypothyroidism. So, our study was aimed to estimate the levels of lipid parameters in patients with subclinical hypothyroidism of our population and to see their relationship with TSH values. MATERIALS AND METHODS The present study was conducted at Biochemistry laboratory, Pacific institute of Medical Sciences, Udaipur. 75 patients of subclinical hypothyroidism and 100 age and sex matched healthy controls were included in the study. Inclusion criteria was patients having serum TSH > 5.0 mIU/L with normal serum T3 and serum T4 level. Patients having diabetes mellitus, renal impairment, alcohol addiction, taking treatment for thyroid dysfunction, psychotic disorders or lipid abnormalities, female patients taking oral contraceptive pills and body mass index > 30 kg/m2 were excluded from the study. Present study was approved by human ethical committee of Pacific Institute of Medical Sciences, Udaipur. History and informed consent was taken from each participants of the study. Fasting blood samples were collected from the SCH patients and from healthy controls and after centrifuging were analyzed for fasting blood glucose, serum creatinine, lipid profile (Total cholesterol, Triglyceride, High density lipoprotein) and thyroid profile (serum TSH, serum T3 and serum T4). Thyroid profile was estimated by using Cobas e 411 analyzer. Rest of the parameters were analyzed by using Cobas C 111 analyzer. Quality controls were done before analyzing all the parameters. Methods of estimating parameters: Total cholesterol: CHOD-POD enzymatic method, Triglyceride: GPO-POD enzymatic method, HDL: Homogeneous enzymatic method, Thyroid profile (TSH, T3 and T4): Electrochemiluminescence immunoassay (ECLIA) method. LDL and VLDL were calculated using the Friedewald’s formula. STATISTICAL METHODS Stastical analysis was performed using Graph pad Prism 5.0. Data were presented as mean ± SD (standard deviation). Unpaired t test was applied to compare the results between euthyroid and SCH patients and of male and female results of patients and controls. One way analysis of variance (ANNOVA) was applied to the result data of different group of patients. Correlation of TSH with lipid parameters was done by Pearson correlation coefficient. Results of the study were discussed at 95% confidence interval; Interpretation of the test results was done according to p value (p < 0.05 – significant, p < 0.001 – highly significant and p ≥ 0.05 – not significant). RESULTS Present study includes 100 healthy controls and 75 subclinical hypothyroidism patients. Control subjects include 59% females and 41% males, while SCH patients comprise 65.3% females and 36.7% males. Mean age of SCH patients was 49.21 ± 12.33 years while that of euthyroid was 47.13 ± 13.98 years (p = 0.3095). TSH level was significantly higher in SCH patients (7.449 ± 2.57 μIU/ml) as compared to Euthyroid subjects (2.581 ± 0.95 μIU/ml, p < 0.0001). Level of serum T4 was decrease in SCH patients (7.139 ± 1.49 μg/ dl) as compared to controls (8.104 ± 1.32μg/dl, p < 0.0001), but was within normal reference limits. SCH patients shows decrease in level of T3 (1.108 ± 0.19 ng/ml) as compared to controls (1.218 ± 0.24 ng/ml, p=0.0016), but was within normal reference range. Mean value of serum Total Cholesterol, Triglyceride, LDL and VLDL were significantly higher in SCH patients as compared to control subjects (p < 0.0001). Mean of TC/HDL and LDL/HDL ratio shows significant difference, while mean value of HDL does not show significant difference between SCH patients and control subjects (Table -1). SCH patients were randomly divided into three age groups: Group-1 (Age 20-42 years), Group-2 (Age 43-56 years) and Group-3 (Age 57-80 years). Values of lipid parameters among these groups were compared by one way analysis of variance (Table-2). All lipid parameters were correlated with TSH values in SCH patients by using Pearson correlation coefficient. Among all lipid parameters, Total cholesterol (r=0.189) show highest positive correlation with TSH values (Table-3). We have also compared the results of lipid parameters and thyroid hormone between male and female of SCH and control subjects. Male and female subjects of euthyroid show significant difference in values of total cholesterol (p=0.0161), HDL (p< 0.0001), TC/HDL (0.0060) and LDL/ HDL (0.0196). Mean value of lipid parameters were higher in female as compared to males of SCH patients, but were not statistically significant (Figure-1). DISCUSSION Subclinical hypothyroidism is a laboratory diagnostic condition, which is detected by Serum TSH measurement. Hallowell et al. showed that SCH usually occurs in 40-60 years of age range.11 In our study mean age of the SCH patients was 49.21 years. In present study SCH patients comprises 65.3% women and 36.7% men, which shows that SCH is more common in women that in men. Thyroid hormone plays significant role in cholesterol synthesis and uptake, lipoprotein metabolism and lipolysis by inducing the expressions of different enzymes.12-16 Alteration in level of lipid parameters in SCH patients is unclear. The present study shows that levels of all lipid parameters were significantly increased in SCH patients as compared to the euthyroid subjects, except increase in HDL was not statistically significant. Similar findings were observed by various studies: Adriana Santi et al. concluded that the levels of TC, LDLC and TC/HDL ratio were significantly increase in the subjects with subclinical hypothyroidism than euthyroid.17 In a study conducted by Laway BA et al. found that levels of TC, TG and VLDL were significantly higher in SCH patients as compared to euthyroid subjects.8 Study conducted by Michalopoulou18 showed increase in average level of serum HDL in subclinical hypothyroidism. There is an evidence of increased risk for atherosclerosis and myocardial infarction in elderly female SCH patients.19 The possible explanation of our results are: Decreased thyroid function results in reduction in cell surface receptor for LDL20 and decrease in their activity (abbas et al., 2008) leads to decreased LDL and IDL catabolism; these increases TC and LDL concentration. Decreased activity of hepatic lipase results in increase level of TG rich lipoproteins; these causes high VLDL and TG concentration.21 Tan et al. (1998) showed altered metabolism of HDL in thyroid dysfunction. Our study shows that TSH is positively correlated with values of all the lipid parameters estimated in SCH patients, though it is not good correlation. A Regmi et al.22 showed that values of TC, HDL and LDL, while Shashi A et al.23 found that values of TC, TG and LDL were positively correlated with TSH values in Subclinical hypothyroidism. A study conducted by Ali M. Nouh et al. in a Murzok, Libya Population found that TSH was positively correlated with lipid profile values in subjects with thyroid dysfunction.24 CONCLUSION From the present study, it is concluded that subclinical hypothyroidism shows secondary dyslipidemia which is associated with increase in levels of total cholesterol, triglyceride, LDL, VLDL, TC/HDL and LDL/HDL ratio. Although good cholesterol (HDL) level increase in SCH, it is not significant. These increases risk of atherosclerosis and cardiovascular disease development. So, all dyslipidemic patients, particularly women with 40-60 years of age should be tested for thyroid dysfunction and accordingly be treated. This may help to reduce mortality and morbidity due to atherosclerosis and cardiovascular diseases. ACKNOWLEDGEMENT Authors are thankful to all participants of the study and to the technical staff of biochemistry laboratory of Pacific Institute of Medical Sciences, Udaipur for their help and support during the study. 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. Englishhttp://ijcrr.com/abstract.php?article_id=439http://ijcrr.com/article_html.php?did=4391. Heuck CC, Kallner A, Kanagasabapathy AS and Riesen W. Diagnosis and monitoring of the disease of the thyroid. WHO Document (WHO/DIL/00.4) 2000; 8-9. 2. N. Kochupillai. Clinical endocrinology in India. Current Science. vol.79, no.8, 25 Oct 2000. 3. Mcdermott MT and Ridgway EC. Subclinical hypothyroidism is mild thyroid failure and should be treated. The journal of clinical endocrinology and metabolism 86(10):4585–4590. 4. Tunbridge WMG, Evered DC, Hall R, Appleton D, Brewis M, Clark F, et al. The spectrum of thyroid disease in a community: the Wickham survey. Clin Endocrinol (Oxf). 1977 Dec; 7(6):481-93. 5. Canaris GJ, Manowitz NR, Mayor G and Ridgway EC. The Colorado Thyroid Disease Prevalence Study. Arch Intern Med. 2000; 160:526-534. 6. Unnikrishnan AG, Kalra S, Sahay RK, Bantwal G and John M. Prevalence of hypothyroidism in adults: An epidemiological study in eight cities of India. Indian J Endocrinol Metab. 2013 Jul-Aug; 17(4): 647–652. 7. Pucci E, Chiovato L and Pinchera A. Thyroid and lipid metabolism. INT J OBESITY. 2000; 24:S109–12. 8. Laway BA, War FA, Shah S, Misgar RA and Kotwal SK. Alteration of Lipid Parameters in Patients with Subclinical Hypothyroidism. Int J Endocrinol Metab. 2014 July; 12(3): e17496. 9. Pirich C, Mullner M and Sinzinger H. Prevalence and relevance of thyroid dysfunction in 1922 cholesterol screening participants. J Clin Epidemiol. 2000 Jun; 53(6):623-9. 10. Hueston WJ and Pearson WS. Subclinical Hypothyroidism and the Risk of Hypercholesterolemia. ANNALS OF FAMILY MEDICINE VOL. 2, NO. 4, JULY/AUGUST 2004. 11. Hollowell JG, Staehing NW, Flanders WD, Hannon WH, Gunter EW, Spencer CA et al. Serum TSH, T4, and thyroid antibodies in the United States Population (1988 to 1994): National Health and Nutrition Examination Survey (NHANES III). J Clin Endocrinol Metabol 2002; 87: 489-99. 12. Simonet WS and Ness GC. Transcriptional and posttranscriptional regulation of rat hepatic 3-hydroxy-3-methylglutaryl-coenzyme A reductase by thyroid hormones. J Biol Chem. 1988; 263: 12448-53. 13. Shin D-J and Osborne TF. Thyroid Hormone Regulation and Cholesterol Metabolism Are Connected through Sterol Regulatory Element-binding Protein-2 (SREBP-2). J Biol Chem. 2003; 278: 34114-8. 14. Saffari B, Ong JM and Kern PA. Regulation of adipose tissue lipoprotein lipase gene expression by thyroid hormone in rats. J Lipid Res. 1992; 33: 241-9. 15. Berti JA, Amaral ME, Boschero AC, Nunes VS, Harada LM, Castilho LN, et al. Thyroid hormone increases plasma cholesteryl ester transfer protein activity and plasma high-density lipoprotein removal rate in transgenic mice. Metabolism. 2001; 50: 530-6. 16. Gagnon A, Antunes TT, Ly T, Pongsuwan P, Gavin C, Lochnan HA, et al. Thyroid-stimulating hormone stimulates lipolysis in adipocytes in culture and raises serum free fatty acid levels in vivo. Metabolism. 2010; 59: 547-53. 17. Santi A, Duarte MMMF, de Menezes CC and Loro VL. Association of Lipids with Oxidative Stress Biomarkers in Subclinical Hypothyroidism. Int J Endocrinol. 2012; 2012: 7p. 18. Michalopoulou G, Alevizaki M, Piperingos G, Mitsibounas D, Mantzos E, Adampoulos P, et al. High serum cholesterol levels in persons with ‘high normal’ TSH levels: should one extendthe definition of subclinical hypothyroidism? Eur J Endocrinol 1998; 138:141-145. 19. Hak AE, Pols HAP, Visser TJ, Drexhage HA, Hoffman A and Witteman JCM. Subclinical hypothyroidism is an independent risk factor for atherosclerosis and myocardial infarction in elderly women: the Rotterdam Study. Annals of Internal Medicine 2000; 132: 270–278. 20. Thompson GR, Soutar AK, Spengel FA, Jadhav A, Gavigan SJP and Myant NB. Defects of receptor mediated low density lipoprotein catabolism in homozygous familial hypercholesterolemia and hypothyroidism. Proc Natl Acad Sci U S A. 1981 Apr; 78(4):2591-5. 21. Brenta G, Berg G, Arias P, Zago V, Schnitman M, Muzzio ML et al. Lipoprotein alterations, hepatic lipase activity, and insulin sensitivity in subclinical hypothyroidism: response to L-T(4) treatment. Thyroid. 2007 May; 17(5):453-60. 22. A Regmi, B Shah, BR Rai and A Pandeya. Serum lipid profile in patients with thyroid disorders in central Nepal. Nepal Med Coll J 2010; 12(4): 253-256. 23. Shashi A and Sharma N. Lipid profile abnormalities in hypothyroidism. I.J.S.N., vol. 3(2) 2012: 354-360. 24. Nouh AM, Eshnaf IAM and Basher MA. Prevalence of Thyroid Dysfunction and Its Effect on Serum Lipid Profiles in a Murzok, Libya Population. Thyroid Science 3(10):CLS1-6, 2008.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareA DETAILED STUDY OF ABSENCE OF ANAEMIA IN THE EARLY STAGE OF HOOKWORM INFECTION DIAGNOSED BY DOING UPPER GASTRO-INTESTINAL ENDOSCOPY IN OUR INSTITUTE English0610Govindarajalu GanesanEnglishObjective: Anaemia occurs very commonly in hookworm infection. Many studies have also shown the presence of anaemia in hookworm infection. But so far detailed study was not done to know about the absence of anaemia in the early stage of hookworm infection. Hence a detailed study was done to know about the absence of anaemia in the early stage of hookworm infection diagnosed by doing upper gastro-intestinal endoscopy in our institute. Methods: A study of 1100 patients who had undergone upper gastro-intestinal endoscopy for a period of four and half years from May 2009 to October 2013 was carried out in our institute. In all these 1100 patients, the first and second part of duodenum were carefully examined to find out the presence of hookworms. In all the patients who were found to have hookworms in duodenum, investigations were done to know about the presence or absence of anaemia and eosinophilia except in the very few patients who were lost for follow up. The results were found as given below. Results: Out of these 1100 patients, as many as 18 patients were found to have hookworms in duodenum while doing upper gastro-intestinal endoscopy. But 4 patients were lost for follow up. The remaining 14 patients were taken into consideration for our study. Out of these14 patients, 9 patients had anaemia but 5patients did not have anaemia. Absence of anaemia indicates minimal loss of blood due to very low burden of hookworms which can occur only in the early stage of hookworm infection. Hence absence of anaemia is suggestive of early stage of hookworm infection. But majority of the patients who did not have anaemia due to early stage of hookworm infection (4 out of 5patients) had significant eosinophilia. Conclusion: Hence, in the early stage of hookworm infection eosinophilia alone can be present without the presence of anaemia. Hence eosinophilia is an extremely important indicator of early stage of hookworm infection which is not associated with anaemia. Hence upper gastro-intestinal endoscopy should be done to confirm the presence of hookworms in all patients with eosinophilia even when there no anaemia in tropical countries. EnglishAbsence of anaemia, Early stage of hookworm infection, Presence of eosinophilia, Upper gastro-intestinal endoscopyhttp://ijcrr.com/abstract.php?article_id=440http://ijcrr.com/article_html.php?did=440

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareA STUDY OF HEART RATE, BLOOD PRESSURE AND HEART RATE VARIABILITY AT REST, IN NORMOTENSIVE AND HYPERTENSIVE ADULT MALE SUBJECTS English1114Satish S. PatilEnglish GnanajyothiEnglishBackground: Essential hypertension is associated with cardiovascular autonomic imbalance. Heart Rate Variability (HRV) is a non-invasive tool to monitor the functioning of cardiovascular autonomic nervous system (ANS). Objectives: To compare Heart Rate, Blood Pressure and Heart Rate Variability in normotensive and hypertensive adult males. Material and Methods: The study was conducted on 30 normotensive and 30 hypertensive adult males matched for baseline anthropometric characteristics. ECG was recorded and using HRV software, analysis of time domain measures Mean RRI (RR Interval); RMSSD (square root of the mean of the sum of the squares of differences between adjacent NN intervals) and frequency domain measures, LF (Low Frequency); HF (High Frequency); and LF/ HF (Low Frequency/High Frequency) ratio were analysed. Results: The Heart Rate Variability parameters were significantly reduced in hypertensive subjects as compared to normotensives. Conclusion: Reduced Heart Rate Variability indicates impaired regulation of the cardiovascular autonomic function in the hypertensives. EnglishHypertension, Autonomic nervous system, Heart rate variabilityINTRODUCTION The heart has rich innervations from the sympathetic and parasympathetic divisions of the autonomic nervous system (ANS). Although the heart is capable of intrinsic regulation of cardiac rhythm, electrical conduction and contractility, these functions are largely under the control of the ANS. The heart rate and its fluctuations reflect changes in cardiac autonomic control. This neural link creates the basis of assessment of cardiac autonomic regulation through measurement of heart rate variability (HRV) 1 . Heart rate variability conventionally describes the beat-to beat fluctuations in the heart rate 2 . HRV is primarily due to the changing modulation of parasympathetic and sympathetic control of the heart and may therefore be considered as an estimate of autonomic heart rate control 3 . Measurements of HRV might assess progressive alterations in the sympathovagal balance observed in essential hypertension 4 . Hypertension is the most common human cardiovascular disease, characterized by systolic blood pressure (SBP) of > 140 mmHg and/ or diastolic blood pressure (DBP) of > 90 mmHg 5 . Worldwide it is estimated to cause 7.1 million premature deaths each year 6 . Hypertension doubles the risk of cardiovascular diseases, including coronary heart disease, congestive heart failure, ischemic and hemorrhagic stroke, renal failure, and peripheral arterial disease 7 .The pathogenesis of essential hypertension is not clearly understood but is believed to be due to renal, neurogenic, vascular and genetic factors; in reality it has a multifactorial aetiology 8 . The present work represents a study of the autonomic modulation of the heart, using the analysis of the variability in heart rate of normotensive and hypertensive subjects. MATERIALS AND METHODS In this study 30 normotensive and 30 hypertensive male subjects, in the age group of 30-60 years, were included. The study protocol was approved by the Institutional Ethical Committee. Inclusion Criteria: In the normotensive group, subjects with normal blood pressure, normal electrocardiogram(ECG ) and in good health as evaluated by general physical examination without any known respiratory, cardiovascular illness, or any disorder which can interfere the autonomic responses were included. In the hypertensive group known hypertensive on treatment with normal ECG were included. Exclusion Criteria: Subjects with diabetes mellitus, symptomatic coronary disease, congestive cardiac failure, arrhythmias, any systemic illness and those with h/o tobacco and alcohol consumption were excluded. Study Design: The study protocol was explained to the subjects and consent was obtained. During the first visit the anthropometric data was obtained. Each subject is given specific dates to visit autonomic laboratory. A day before the test subjects were advised to have their dinner before 9:00 pm and to refrain from any kind of stress. Also instructed not to have coffee, tea and cola 12 hours before the tests and to have light breakfast two hours before the tests. In the laboratory the subject is asked to relax in supine position for 30 minutes and then the tests were performed using ECG V: 52 [HRV analysis software], manufactured by NIVIQURE Meditech pvt Ltd. Bengaluru. ECG V: 52 is a Computerized Data Acquisition System used in conjunction with PC/Laptop. Resting Heart Rate: The subject was made to lie down in supine position. ECG leads were connected using electrodes from the subject to the ECG V: 52. The resting heart rate was recorded on a computerized ECG from lead II, at a speed of 30 mm/sec. Blood Pressure: Blood pressure was measured with digital electronic blood pressure monitor in supine position after a period of rest for 5 minutes. Lowest of 3 reading at intervals of 2 minutes was considered. Heart Rate Variability Analysis: Recording was standardized and instructions followed as per the guidelines of Task Force of The European Society of Cardiology and The North American Society of Pacing and Electrophysiology 9 . A chest lead ECG was recorded using ECG V: 52 for 5 minutes in supine rest with eyes closed, which is simultaneously analyzed by the software. Beat-to-beat variations in instantaneous heart rate were derived offline using a rate-detector algorithm. Briefly, a 5-min ECG was acquired at a sampling rate of 1000 Hz during supine rest with the subjects breathing normally at 12–18 per minute.RR intervals were plotted using the ECG V: 52 software. An RR series was extracted using a rate-detector algorithm after exclusion of artefacts and ectopic. A stationary 256 second RR series was chosen for analysis. In the time domain, the standard deviation of normal-to-normal RR intervals (SDNN) was taken as an index of overall HRV. The RR series was resampled at 4 Hz, the mean and trend removed, a Hann window applied and the 1024 data point series transformed by fast Fourier transformation. Low frequency (LF) and high frequency (HF) spectral powers were determined by integrating the power spectrum between 0.04 and 0.15 Hz and 0.15 and 0.4 Hz respectively. Spectral powers are expressed in absolute units of milliseconds squared. Statistical Analysis: All data is expressed as Mean ± SD. Student‘t’ test used to compare the data of normotensive and hypertensive subjects. Mann-Whitney test used to analyze HRV. p value < 0.05 considered statistically significant and p value < 0.01 as highly significant. Results: Subjects of both the groups were matched for age and BMI (Table-1). The resting heart rate (pEnglishhttp://ijcrr.com/abstract.php?article_id=441http://ijcrr.com/article_html.php?did=4411. Juha EK Hartikanianen, Kari UO Tahvanainen and Tom A Kuusela. Short term measurement of Heart Rate Variability. Malik (Ed), Clinical guide to cardiac autonomic tests. 1998; 149-176. 2. Akselrod S, Gordon D, Ubel FA, Shannon DC, Berger AC, Cohen RJ. Power spectrum analysis of heart rate fluctuation; a quantative probe of beat to beat. Cardiovascular Control Science. 1981;213:220-222. 3. Kleiger RE, Miller JP, Bigger JT, Jr, and Moss AJ. Decreased heart rate variability and its association with increased mortality after acute myocardial infarction. Am J Cardiol. 1987; 59:256- 262. 4. M. Malik and A.J. Camm. Heart rate variability and Clinical Cardiology. Br Heart J. 1994; 71:3-6. 5. Chobanian AV et al. Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure. National Heart, Lung and Blood Institute; National High BloodPressure Education Program Coordinating Committee. Seventh Report of the JointNational Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure. Hypertension. 2003; 42:1206-1252. 6. World Health Organisation (2003) and International Society of Hypertension statement on management of hypertension. World Health Organisation and International Society of Hypertension writing, group. Journal of Hypertension. November 2003; 21(11):1983-1992. 7. Harrison’s Principles of Internal Medicine;17th edition; Part 9 - Disorders of the CVS; Section 5- Vascular Disease; Chapter 241- Hypertensive vascular Disease. 8. Davidson’s Principals and Practice of medicine, 20 th edition, Hypertension, part 2,609. 9. Heart rate variability: Standards of measurement, physiological interpretation, and clinical use: Task Force of The European Society of Cardiology and The North American Society of Pacing and Electrophysiology. European Heart Journal. 1996; 17: 354- 381. 10. Asuman H. Kaftan and Osman Kaftan. QT Intervals and Heart Rate Variability in Hypertensive Patients. Jpn Heart J. March 2000; 41:173-182. 11. Gianfranco Piccirillo et al. Factors influencing heart rate variability power spectral analysis during controlled breathing in patients with chronic heart failure or hypertension and in healthy normotensive subjects. Clinical Science (2004) 107, 183–190. 12. Jagmeet P. Singh, Martin G. Larson, Hisako Tsuji, Jane C. Evans, Christopher J., O’Donnell and Daniel Levy. Reduced Heart Rate Variability and New-Onset Hypertension: Insights into Pathogenesis of Hypertension: The Framingham Heart Study. Hypertension. 1998; 32: 293-297. 13. R Virtanen, A Jula, T Kuusela, H Helenius and L-M VoipioPulkki. Reduced heart rate variability in hypertension: associations with lifestyle factors and plasma renin activity. Journal of Human Hypertension.2003; 17: 171–179. 14. Berntson GC, Cacioppo JT, Quigley KS. Cardiac psychophysiology and autonomic space in humans: empirical perspectives and conceptual implications. Psychol Bull. 1993; 114: 296-322. 15. Levy MN, Zieske H. autonomic control of cardiac pacemaker activity and atriventricular transmission. J Appl Physiol. 1969; 27: 465-470. 16. Giuseppe Schillaci, Matteo Pirro and Elmo Mannarino. Assessing cardiovascular risk: Should we discard diastolic blood pressure? Circulation. 2009; 119: 210-212. 17. Mohamed Faisal Lufti. Heart rate variability. Sudan JMS. 2001; 6(1): 43-50. 18. Rehnuma Tabassum, Noorzahan Begum, Sultana Ferdousi, Shelina Begum, Taskina Ali. Heart Rate Variability in Patients with Essential Hypertension. J Bangladesh Soc Physiol. 2010 June; 5(1): 1-7. 19. Harald M. Stauss, Physiologic mechanisms of heart rate variability. Rev Bras Hipertens. 2007;14 (1): 8-15. 20. Rehnuma Tabassum, Noorzahan Begum, Sultana Ferdousi, Shelina Begum, Taskina Ali. Power Spectral Analysis of Heart Rate Variability in Hypertensive Male. J Bangladesh Soc Physiol. 2011 June; 6(1): 32-38.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareSTUDY ON REPRODUCTIVE HEALTH AWARENESS AMONG ADOLESCENT GIRLS IN URBAN AND RURAL FIELD PRACTICE AREAS OF OSMANIA MEDICAL COLLEGE English1518G. Sudha RaniEnglish B. Babu RaoEnglishIntroduction: Reproductive health is a state of complete physical, mental and social wellbeing and not merely absence of disease or infirmity in all matters relating to reproductive system and its functions and process. RCH phase II programme implementation plan, the Ministry of Health and Family Welfare approved a technical strategy on adolescent reproductive and sexual health. The strategy focuses on reorganising the public health system in order to meet the service needs of adolescents. Objectives: To assess the awareness level of adolescent girls regarding various reproductive health issues and to impart health education about menstrual hygiene among the adolescent girls. Materials and Methods: A total of 760 adolescent girls (380 in each area) were interviewed. Menstrual history and history of other bleeding conditions are taken and interviewed about awareness on reproductive health issues. Results: In the present study only 37.4% in rural area and 48.7% in urban area had prior knowledge on menstruation (before attaining it), and 61.3% of the rural subjects and 59.5% of the urban subjects did not know from which organ the menstrual blood was coming during menstruation. In those who knew about menstruation before attaining, mother was the main source of information in both rural (12.9) and urban (18.2) area. Majority of the rural respondents used cloth as absorbent during menstruation. Majority of the urban respondents used sanitary pads as absorbent. Majority girls whose mothers were illiterate or having low educational status used cloth or both as absorbent during menstruation in both rural and urban area (statistically significant in both areas). Majority of girls followed restrictions during menstruation. Most of the rural girls (42.9%) knew about the leucorrhoea as a white discharge when compared to urban girls (35.0%). Significant percentage of study subjects found to know that infection was one of the causes of leucorrhoea in urban and rural area with 42.1% and 30.1% respectively. In this study 47.7% in rural area and 54.5% in urban area knew correct legal age of marriage in girls. Conclusion: There is poor knowledge regarding reproductive health issues in adolescent girls in both rural and urban areas. Hence there is urgent need to intervene in early adolescent period by imparting knowledge on reproductive health. EnglishAdolescent girls, Menstruation, Leucorrhoea, Legal age of marriageINTRODUCTION Adolescence, the second decade of life, is a period in which an individual undergoes major physical and psychological changes. The word ‘Adolescent’ has been derived from Latin word ‘Adolescere’ which means ‘to grow to maturity’. Adolescent is considered to be, no longer a child, and not yet an adult. WHO defines Adolescence as 10-19 years old, ‘Youth’ as15- 24 years old and ‘Young People’ as 10-24 years old. The adolescence has been divided into two phases: ‘early’ (10-14 years) and ‘late’ (15-19 years)1 . Adolescents in India represent over 1/5th (22.3%) of total population. Reproductive health is a state of complete physical, mental and social wellbeing and not merely absence of disease or infirmity in all matters relating to reproductive system and its functions and process2. Reproductive health, therefore, implies that people are able to have a responsible, satisfying and safe sex life and that they have the capability to reproduce and the freedom to decide if, when and how often to do so. Implicit in this are the right of men and women to be informed of and to have access to safe, effective, affordable and acceptable methods of fertility regulation of their choice, and the right of access to appropriate health care services that will enable women to go safely through pregnancy and child birth and provide couples with the best chance of having a healthy infant According to the WHO, “Reproductive and sexual ill-health accounts for 20% of the global burden of ill-health for women, and 14% for men. Hospital based retrospective studies in India shows that primary amenorrhoea, menstrual disorders, leucorrhoea, and genital infections are most frequent complaints of adolescents3 . Menstrual hygiene is another important issue that every girl and woman has to deal with in her life. There is lack of information on the process of menstruation, the physical and psychological changes associated with puberty and proper requirements for managing menstruation. The taboos surrounding this issue in the society prevents girls and women from articulating their needs and the problems of poor menstrual hygiene have been ignored or misunderstood. Good menstrual hygiene is crucial for the health, education, and dignity of girls and women as it is an important risk factor for RTI. This is an important sanitation issue which has long been in the closet and there was a long standing need to openly discuss it. The National Population Policy 2000 has recognised adolescents as an underserved, vulnerable group that need to be served especially by providing reproductive information and services .The health needs of adolescents have neither been recognised, researched nor addressed adequately, in particularly their reproductive health needs which are often misunderstood ,unrecognised or under estimated4 . So in RCH phase II programme implementation plan, the Ministry of Health and Family Welfare approved a technical strategy on adolescent reproductive and sexual health. The strategy focuses on reorganising the public health system in order to meet the service needs of adolescents. Hence there is urgent need to intervene in early adolescent period by imparting knowledge on reproductive health. Girl child, the women of tomorrow is a nation’s asset. Give her an opportunity to develop as her development is the development of nation. To conclude, awareness regarding reproductive health during adolescents will go a long way in improving health of future mothers and building an effective and sustainable nation. OBJECTIVES 1. To assess the awareness level of adolescent girls regarding various reproductive health issues. 2. To impart health education about menstrual hygiene among the adolescent girls. MATERIALS AND METHODS Study design: Community based cross sectional study, Study setting:-OMC field area harazpenta [urban] patencheru [rural], study population:- adolescents girls[11-19yrs] of age in harazpenta and patencheru. Sample size is 360 adolescents in urban area and 360 adolescents in rural area. Study period is one year [Jan 2012-Dec 2012]1month. Questanaire method data analysis:-by using epinfo 3.5.1, ms excel statistical test:-chi-square, percent. RESULT Majority of adolescent girls from rural area were in 14-16 yrs age group where as in urban area they were in 12-14 yrs age group and most of them were students. About 79.2% of urban and 77.6% of rural the study subjects were belongs to Hindu religion and most of the respondents belong to upper lower (42%) and lower middle (28.7%) in rural and urban areas respectively. The mean age of menarche was 12.45 and 12.46 in rural and urban areas respectively. In the present study only 37.4% in rural area and 48.7% in urban area had prior knowledge on menstruation (before attaining it), and 61.3% of the rural subjects and 59.5% of the urban subjects did not know from which organ the menstrual blood was coming during menstruation. This shows that girls in both rural and urban area had poor knowledge on menstruation. In those who knew about menstruation before attaining, mother was the main source of information in both rural (12.9) and urban (18.2) area. Majority of the rural respondents used cloth as absorbent during menstruation. Majority of the urban respondents used sanitary pads as absorbent. Majority girls whose mothers were illiterate or having low educational status used cloth or both as absorbent during menstruation in both rural and urban area (statistically significant in both areas). Majority of girls followed restrictions during menstruation. Most of the rural girls (42.9%) knew about the leucorrhoea as a white discharge when compared to urban girls (35.0%). Significant percentage of study subjects found to know that infection was one of the causes of leucorrhoea in urban and rural area with 42.1% and 30.1% respectively. Some other misconceptions found in them were eating heat items, raw rice and pain abdomen. In this study 47.7% in rural area and 54.5% in urban area knew correct legal age of marriage in girls. DISCUSSION A study done by Parvathi Nair5 showed that nearly half (45.7%) of the girls who had attained menarche and 29% of pre-pubertal subjects said that they had prior knowledge about menstruation, Mothers (41%) were the most common source of information about menstruation, followed by elder sisters (22.4%), friends (21%), relatives (6.7%), television (4.4%), books (3.3%), and doctors (1.1%) which were similar findings in the present study. Drakshayani Devi K, Venkata Ramaiah P6 conducted study on menstrual hygiene and found that, about 50% knew that hormones were responsible for menstruation. Most students (51) knew that menstrual bleeding originated from the uterus. Other sites mentioned were abdomen, intestines, and kidneys. Singh MM, Devi R, Gupta SS 7 showed in their study that the major sources of information were television (73.1%), radio (37.1%) and parents (36.1) A study by Parvathi Nair5 , majority (74.8%) of the girls used homemade sanitary pads, nearly 24% used ready-made sanitary pads, while 1.5% used cotton wool. Drakshayani Devi K, Venkata Ramaiah P 6 stated that all but one used old cloth during menstruation, 25 reused the cloth, 16 disposed of the used cloth through Dhoby, 13 put it into a canal. Saritha agarwal, Alfia fathima, C.M. Singh8 in their study on knowledge and attitude of adolescent girls towards reproductive health and related problem stated that only 10% of girls know about leucorrhoea. Sharma , Shipra Nagar and Goldy Chopra9 that the percentage of girls knowing about the ideal child bearing age was (43.7%) perceived the age of 26 to 30 years as ideal child-bearing age followed by 31.2% for 18 to 25 years. CONCLUSION In the present study only 37.4% in rural area and 48.7% in urban area had prior knowledge on menstruation (before attaining it), and 61.3% of the rural subjects and 59.5% of the urban subjects did not know from which organ the menstrual blood was coming during menstruation. This shows that girls in both rural and urban area had poor knowledge on menstruation. • Those who knew about menstruation before attaining, mother was the main source of information in both rural (12.9) and urban (18.2) area. • The mean age of menarche was 12.45 and 12.46 in rural and urban areas respectively. • In the present study majority of the rural respondents used cloth as absorbent during menstruation. Majority of the urban respondents used sanitary pads as absorbent. Majority girls whose mothers were illiterate or having low educational status used cloth or both as absorbent during menstruation in both rural and urban area. • Majority of girls followed restrictions during menstruation. • There is poor knowledge regarding reproductive health issues in adolescent girls in both rural and urban areas. Hence there is urgent need to intervene in early adolescent period by imparting knowledge on reproductive health. ACKNOWLEDGMENT • 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 the authors/ editors/publishers of all those articles, journals, and books from where the literature for the article has been reviewed and discussed. Englishhttp://ijcrr.com/abstract.php?article_id=442http://ijcrr.com/article_html.php?did=4421. Text book of public health and community medicine. 1st ed. Pune: Department of community medicine armed forces medical college, in collaboration with World Health Organisation; 2009. p. 856. 2. Progress45(1998). Available online at www.who.int/reproductive-health/hrp/progress/45/prog45.pdf. Surffed on 22nd July 2010. 3. Patki S, Pandit SN, Niyogi GM, Patker VD. Amenorrhea and genital anomalies in adolescents. J Obs Gyn Ind 1993;43:234- 240. 4. National population policy 2000, dept of H and FW, ministry of H and FW government of India 2000. 5. Parvathi Nair. Awareness and practices of menstruation and pubertal changes amongst unmarried female adolescents in a rural area of East Delhi. Indian J Community Med [serial online] 2007 [cited 2008 Nov 7];32:156-7. Available from: http://www. ijcm.org.in/text.asp?2007/32/2/156/35668. 6. Drubashayani Devi K, Venkata-Ramaiah P. A study on menstrual hygiene among rural adolescent girls. Ind J Med Sci 1994;48:139-43. 7. Singh MM, Devi R, Gupta SS. Awareness and health seeking behaviour of rural adolescent school girls on menstrual and reproductive health problems in Delhi. Indian J Med Sci 1999 Oct;53(10):439-43. 8. Saritha agarwal, Alfia fathima, Singh CM. Knowledge and attitude of adolescent girls towards reproductive health and related problem. Indian J Prev Soc Med 2007;38(12):36-41. 9. Shubhangna Sharma, Shipra Nagar, Goldy Chopra. Health Awareness of Rural Adolescent Girls: An Intervention Study in Himachal Pradesh. J Soc Sci 2009;21(2): 99-104.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareEFFECT OF SETTING ON THE PROPERTIES OF PROTEINS FROM THREADFIN BREAM (NEMIPTERUS JAPONICUS) FISH MINCE English1924B. U. SupreethaEnglish M. V. ChandraEnglish B. A. ShamasundarEnglishIn the present investigation effect of setting on the properties of proteins and gel forming ability from threadfin bream (Nemipterus japonicus) fish mince has been assessed. The term setting refers to obtaining a soft elastic gel upon incubating the fish mince mixed with sodium chloride (NaCl) at less than 40 °C for a known duration. When the set meat is subjected to heat processing at 90 °C for 45 min, yields a gel with higher strength. The setting was carried out at a temperature at 35 °C for different durations. The gel strength of the gel obtained from set meat at 35 °C for 45 min was found to be 643.8 while obtained from unset meat was 264.5 g.cm. The setting of fish mince was accompanied by decrease in protein solubility, free-sulfhydryl content, Ca2+-ATPase enzyme activity and viscosity. EnglishThreadfin bream mince, Setting and gel forming abilityINTRODUCTION Gelation is a process and gel is an end product. In order to improve the gel forming ability of proteins from fish species a process known as setting is employed in many of the fish processing and surimi industry. The term surimi refers to separated fish mince subjected for water washing, partially dehydrated and mixed with cryoprotectants, frozen and frozen stored (Yin et al., 2014). Surimi is used to prepare different analogue products, like crab leg sticks, lobster/shrimp moulded products, kamaboko and related products. The characteristic features of these products are its texture which is viscoelastic in nature and are contributed by myosin, the major myofibrillar protein fractions (Xiang et al., 2011). The setting is a process wherein, the fish mince is mixed with sodium chloride and incubated at a temperature below 40 ?C for certain duration resulting in the formation of a translucent, soft elastic gel (Benjakul et al., 2004; Liu et al., 2007). This process is called ‘setting’ and the meat is referred to as set meat. Cross-linking of myosin heavy chain is attributed to transglutaminase enzyme. The enzyme catalyzes the cross-linking reaction with the formation of isopeptide, ε-γ-glutamyl-lysine (EGL) linkages which form soft elastic gel (Binsi and Shamasundar, 2012). When the set meat is subjected to heating at 90 ?C for 45 min duration, yields a strong thermally induced visco elastic gel (Roussel and Cheflel, 1990) as compared to unset meat. Such inter and intra molecular covalent cross-linking of myosin molecules result in higher gel elasticity. The setting ability is species specific and the presence of endogenous TGase enzyme will determine the extent of process (Benjakul et al., 2004). Apart from the cross-linking of myosin heavy chain (MHC) during setting process may initiate certain changes in the properties of proteins like solubility, free-sulfhydyl content and viscosity (Niwa et al., 1991). As water washing is an essential step in the surimi preparation where many soluble protein fractions including TGase are removed. Such water washed mince (surimi) may have a little setting ability. It is common to add TGase enzyme from different source to initiate the setting process when surimi is used as base material for the preparation of products (Nowsad et al., 1997). It will be of interest to study the gel forming ability of fish mince in relation to setting process without washing the mince. It has been reported that unwashed mince will have TGase enzyme and can be made use for setting and enhancing the gel forming ability (Lanier, 2000). In the present investigation the effect of setting on properties of proteins from threadfin bream (Nemipterus japonicus) fish mince have been studied. MATERIALS AND METHODS Materials Fresh threadfin breams (Nemipterus japonicus) were procured from the local fish market, Mangalore, India. The fishes were washed with chilled potable water (3-4 ?C) and iced in the ratio of 1:1 (fish: ice) and transported to laboratory. The fishes were dressed to remove head and entrails and washed with chilled water. The semi dressed fishes were used for further analyses. Methodology Setting experiment Meat was separated from the threadfin bream fishes manually avoiding scales and bones and used for the study. The separated meat was macerated in a pre-cooled pestle and mortar where temperature was maintained at 4 ?C using ice bath. Sodium chloride at 2.5 % (w/w) was mixed with fish mince and macerated for 5 min to get a homogenous sol. The sol was subjected for setting at 35 ?C for 15, 30 and 45 min setting duration in an incubator which was thermostatically controlled (Orbitek, Scigenics biotech Company, Chennai, India). Gel preparation About 100 g of set meat (as mentioned above) was stuffed into krehalon casing (copolymer of vinyledene chloride and vinyl chloride) of 50 mm x 250 mm (dia x length) using a hand stuffer. The stuffed casings were sealed with aluminium clips using a ringer machine (Seinco, TX/ 8037, Barcelona, Spain). The heat processing was carried out at a temperature of 90 ± 2 ?C for 45 min to get thermally induced gels. The gels were cooled at room temperature and stored in a refrigerated condition at a temperature of 6 ± 2 ?C overnight prior to measuring the gel strength. The gels from meat without setting were prepared and served as control. ANALYSIS Protein solubility The solubility of total proteins in the unset (control) and set threadfin bream fish mince was assessed using high ionic strength buffer (phosphate buffer 50mM, pH 7.5 containing 1.0 M sodium chloride) as solvent. This solvent here afterwards will be referred to as extraction buffer (EB).The ratio of fish mince to extraction buffer used was 1: 10 (mince: buffer) and homogenized using laboratory homogenizer (Ultra-Turrax T125, Janke and Kunkel, Staufen, Germany) at 9000 rpm for 2 min. The slurry was centrifuged at 12000 x g at 4 ?C for 30 min using high speed refrigerated centrifuge (Sorvall Legend XTR centrifuge, Thermo Fisher Scientific, New Hampshere, USA). The protein concentration was determined according to the method as described by Lowry et al. (1951). The solubility of the protein was expressed as mg/ ml. Free sulfhydryl content Free sulfhydryl content was determined according to the method as described by Ellman (1959). The absorbance was measured at 412 nm was using double beam spectrophotometer (Spectro UV- VIS, Labomed, Inc., Los Angeles, CA, USA). The free sulfhdryl content was expressed as mM/g of meat. Calcium activated ATPase activity Calcium activated ATPase activity of mince was determined according to the method as described by Noguchi and Matsumoto (1970) and expressed as µmol of Pi/g meat/min at 27 ?C. The inorganic phosphorus released was estimated according to the process as described by Tausky and Shorr (1952). Sodium do-doecyl sulphate Poly acryl amide gel Electrophoresis (SDS - PAGE) Dialysis: In order to remove the sodium chloride in the set meat, dialysis was carried out. About 5.0 g of set mince meat was placed in a dialysis tubing (Sigma, Seamless dialysis tubing), having a width of 23 mm and dia of 15 mm. Dialysis was carried out at 4 ± 2 ?C using deionised water. The duration of dialysis was 6-8 h with frequent change of water for complete removal of sodium chloride. The meat free from sodium chloride was used for SDS-PAGE analysis. The method followed was as described by Laemmlli et al. (1970). The molecular weight of the protein bands obtained in the sample was approximated by measuring the relative mobility of the standard protein molecular weight markers (high molecular weight markers from Sigma, St. Louis, MO, USA). Viscosity Viscosity was determined using rotational viscometer (Brookfield Viscometer DV - II + Pro, Brookfield engineering labs, inc., Middleboro, U.S.A) at a constant temperature of 25 ± 1 ?C. The total proteins were extracted using extraction buffer as described previously. The supernatant obtained was diluted using extraction buffer to get a protein concentration of 2mg/ml. A known quantity of protein solution (7ml) was used for viscosity measurement. The sample was taken in sample holder and spherical spindle (SC - 18) was immersed in sample holder and attached to viscometer. The sample was equilibrated to the desired temperature (25 ±1 ?C) using circulatory constant temperature water bath (CB 2000 V, Cyber lab Corporation, Mumbai, India). The rotation of the spindle was carried out at 100 rpm and corresponding viscosity was recorded and expressed in mPa.s. Gel strength measurements Gel strength of the gel obtained from unset (control) and set mince was measured by using Texture profile analyser (TA - XT plus Stable Micro System, Surrey, England) with 50 kg of load cell. Prior to analysis, the refrigerated stored samples were equilibrated to ambient temperature for 30 min. A spherical probe with 5 mm dia was used for penetration at a constant test speed of 1.1 mm / sec with 10 g of trigger force and target distance of 15 mm. The parameters measured were breaking force (g) and breaking distance [deformation (cm)]. The gel strength was calculated by multiplying breaking force x deformation and expressed as g.cm. Statistical analysis To know the effect of setting on the physicochemical properties of proteins from threadfin bream (Nemipterus japonicus) fish mince and the gel strength, the statistical analysis was carried out. Analysis of variance (ANOVA) was tested using SPSS (version 21) software package (IBM SPSS). The test of significance was analysed using Tukey’s test at a level of 0.05. RESULTS AND DISCUSSION Protein solubility: The protein solubility of unset and set meat in EB as a function of setting duration at 35 ?C is given in Fig. 1A. The protein solubility decreased with increase in setting duration. The setting process initiates the formation of ε-(γ-glutamyl) lysine (EGL) isopeptide, which is a covalent bond and perhaps would have contributed for reduction in solubility. Decrease in solubility of proteins as a function of setting duration has been reported for bigeye snapper, threadfin bream, baracuda and bigeye croaker (Benjakul et al., 2003). Free sulfhydryl content: The free-sulfhydryl content of fish mince as a function of setting is given in Fig. 1B. A concomitant decrease in free-SH content is an indication of the formation of disulfide (-S-S-) bonds. The intermolecular disulfide bonding during setting is a result of oxidation of sulfhydryl groups in the presence of oxidants or metal ions (Ellman, 1959). From the results it is evident that the reduction in the solubility of protein during setting has been contributed by cross-linking of MHC through isopeptide and disulfide bonds. Calcium activated ATPase activity: Any conformational changes in proteins initiated by aggregation will be reflected in the Ca2+-ATPase enzyme activity of myosin molecule. The Ca2+- ATPase enzyme activity of the fish mince as a function of setting duration is given Fig. 1C. The reduction in Ca2+- ATPase enzyme activity was found to be significant (P < 0.05) with setting duration and it could be attributed to changes in the conformational status of myosin molecules as affected by setting process. The cross-linking of MHC is primarily an aggregation process involving globular head (Samejima et al., 1981). The ATPase enzyme activity is related to 95 KDa of globular head and it is reasonable to expect that cross-linking has altered the conformation of this particular site resulting in reduced Ca2+-ATPase enzyme activity. Viscosity: The viscosity of total protein from unset (control) and set at different durations were done at the protein concentration of 2 mg/ml. The results are given Fig. 1D. The viscosity of unset threadfin bream was 2.98 mPa.s. Nearly 2.6 fold decrease in viscosity values was recorded at the end of 45 min of setting. Viscosity is an intrinsic property of protein molecules chiefly dependent on the length and diameter ratio of the molecule. The probable reason for decrease in viscosity as a function of setting could be due to the nature of protein could get into solvent during extraction. It is well known that aggregated protein especially with formation of iso-peptide and disulfide bonds it may not be possible to extract in EB. The initial high value of viscosity in the unset meat includes all the fractions of the protein whereas in the set meat it may be protein fractions not involve in setting. Hence, the decrease in viscosity values is due to dissociation of protein which is not involved in setting. Sodium do-doecyl sulphate Poly acryl amide gel Electrophoresis (SDS - PAGE): The SDS-PAGE pattern of total proteins from unset (control) and set mince at 35 ?C for 30 and 45 min setting duration is given in Fig. 2. The SDS-Page pattern of unset meat indicated multiple bands in the molecular weight range of 205-25 KDa (lane B). The 205 KDa indicates myosin heavy chain (MHC). The pattern of set meat clearly indicated a decrease intensity of MHC and a band on the top of the gel which represents cross-linked MHC (lane C and D). The decrease in intensity of MHC band is an indication of aggregation and hence will not be observed in the pattern. The low-molecular weight fractions in the range of 66- 24 KDa appear to have not been affected by setting process. Gel strength measurements: The gel strength of thermally induced gel from unset (control) and set mince is given in Table 1. The gel from unset (control) mince recorded a value of 264 g.cm, while the gel from mince set at 35 ?C different durations the values were in the range of 492-643 g.cm. The results suggest that the gel strength values of gel were proportional to setting duration. This can be attributed to polymerisation of MHC by the action of endogenous TGase. CONCLUSION The gel forming ability of unwashed threadfin bream (Nemipterus japonicus) fish mince could be enhanced by setting process at 35 ? C for 45 min. The setting process initiated aggregation reaction as indicated by reduction in free-sulfhydryl content and solubility. The aggregation / cross-linking of MHC are further confirmed by SDS-page pattern. ACKNOWLEDGEMENT The financial assistance from European Union, Brussels [Grant No: 289282-Secure fish] for carrying out this work is gratefully acknowledged. Data are mean of triplicates and value in parenthesis are standard deviation, n=3. Different letters denotes the significant differences among treatments (P < 0.05). Figure 1 A: Protein solubility of threadfin bream (Nemipterus japonicus) mince set at 35 °C for different durations in high ionic buffer B: Free- sulfhydryl content of threadfin bream (Nemipterus japonicus) mince set at 35 °C for different durations C: Ca2+-ATPase enzyme activity of threadfin bream (Nemipterus japonicus) mince set at 35 °C for different durations D: Viscosity of threadfin bream (Nemipterus japonicus) mince set at 35 °C for different durations at 2mg/ml protein concentration\ Figure 2: SDS-PAGE pattern of threadfin bream (Nemipterus japonicus) mince set at 35 °C temperature for 30 and 45 min setting durations Lane A: Molecular weight markers Lane B: Threadfin bream mince unset (control) Lane C: Threadfin bream set at 35 °C 30 min Lane D: Threadfin bream set at 35 °C 45 min Englishhttp://ijcrr.com/abstract.php?article_id=443http://ijcrr.com/article_html.php?did=4431. Benjakul S., Visessanguan W. and Pecharat, S. 2004. Suwari gel properties as affected by transglutaminase activator and inhibitors. Food Sci. Nutr., 22:27-107. 2. Binsi P. K. and Shamasundar B. A. 2012. Purification and characterisation of transglutaminase from four fish species: Effect of added transglutaminase on the viscoelastic behaviour of fish mince. Food Chem., 132 (4):1922-1929. 3. Ellman, G.L. 1959. Tissues sulfhydryl groups. Arch. Biochem. Biophys., 82: 70-77. 4. Laemmlli U.K., 1970. Cleavage of structural protein during assembly of the head bacteriophage T4. Nature., 227: 680-685. 5. Lanier T.C. 2000. Surimi gelation chemistry. In: J.W. (Ed) surimi and surimi seafood. Marcel Dekker , New York., pp 99-145. 6. Liu R., Zhao S., Xiong S., Xie B. and Liu H. 2007. Studies on fish and pork paste gelation by dynamic rheology and circular dichroism. J. Food Sci., 72: 399-403. 7. Lowry O.H., Rosebrough, N.J., Farr, A.L. and Randall R.J. 1951. Protein measurement with folin phenol reagent. J. Boil. Chem., 193(1): 256-275. 8. Niwa E., Nowsad, A.A and Kanoh, S. Changes in viscoelastic properties of salted fish flesh sol during its low temperature setting. Nippon suisan Gakk., 57 (12): 2333-2336. 9. Noguchi S. and Matsumoto, J. J. 1970. Studies on the control of the denaturation of the fish muscle proteins during the frozen storage. I. Preventive effect of Na–glutamate. Bull. Jap. Soc. Sci. Fish., 36: 1078-1087. 10. Nowsad A.K., K Anoh S and Niwa E.1997. Phscico-chemical protperites of a Suwari gel from Alaska Pollack surimi with iodoacetic acid added. Bull.Fac. Bioresources, Mie Univ.,13: 25- 31. 11. Roussel H. and Cheflel J.C. 1990. Mechanism of gelation of sardine protein influence of thermal processing and various additives on the texture and protein solubility of kamaboko gel. Int. J. Food Sci. Technol., 25: 260-280. 12. Samejima K., Ishioroshi, M., and Yasui T. 1981. Relative roles of the head and tail portions of the molecule in heat-induced gelation of myosin. J. Food Sci., 46: 1412-1418. 13. Tausky H. H. and Shorr E. 1952. A micro colorimetric method for determination of inorganic phosphorous. J. Biol. Chem., 202: 675-685. 14. Xiang, D.S and Holley R.A. 2011. Factors influencing gel formation by myofibrillar proteins in muscle foods. Comprehensive Rev in Food Sci and Food Safety., 10: 33-51. 15. Yin T. and Park J.W . 2014. Effects of nano-scaled fish bone on the gelation properties of Alaska Pollock surimi. Food Chem., 150: 463-468.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareKNOWLEDGE, ATTITUDE AND PRACTICES OF PEOPLE TOWARDS MALARIA IN TRIBAL COMMUNITIES OF JAWHAR, MAHARASHTRA, INDIA English2530Suhas KadamEnglish Ambadas AdhavEnglish Balu MoteEnglish Shrikant KalaskarEnglish Thirumugam M.English Ravindra KurbudeEnglish Sushil PatilEnglishBackground: Malaria continues to be one of the major public health concerns that India is facing since decades. Early identification and management of malaria prevents its complications and subsequent mortality due to it. Several studies across the globe showed that, misconception of malaria still exists and practices of controlling malaria have been unsatisfactory indicating further need of exploring the knowledge, attitude and practices of malaria among the people of high Annual Parasite Incidence (API) states, district and blocks. Aim: The aim of the study is to assess the levels of knowledge, attitude and practices for prevention of malaria in tribal of Jawhar, Maharashtra. Methodology: A cross sectional community based KAP sample survey was carried out with the help of structured questionnaire. Data was collected by face to face interview by using a structured questionnaire from 140 participants by Systematic Random Sampling. Results: The results show, 24 participants (17.1 %) have a low knowledge level and 94 (67.1%) obtained an average level and 22 (15.7 %) have high level of knowledge on components like symptoms, causes and transmission of malaria. 118 participants (84.3 %) have the average level score for attitude component. 88 (62.9 %) of the participants have an average level, while only 14 (10.0 %) found to have low level score on practices component while 38 (27.1 %) found high level practices score. Knowledge on etiology and symptoms of malaria is found to be optimum level, while knowledge regarding the complications and preventive methods were found low among the participants. Health care system is the major source of knowledge for the tribal community. Practices of sanitation and correct health seeking behavior were found to be optimum but the sampled tribal community were found to be not co-relating its importance of health as a whole. Conclusion: Health education program directed towards community should be emphasized on a timely basis to improve the knowledge, attitude, and practice regarding malaria and its prevention in Tribal Communities of Jawhar. Capacity building among the government health workers on preventive part of malaria and complication management for malaria is the need of the hour time, as they are the major contact point for tribals. EnglishKnowledge, Attitude, Practices, Malaria, Tribal, Jawhar, Thane, Maharashtra, IndiaINTRODUCTION India achieved growth in terms of economic development but on the contrary India is facing a challenge of ‘triple burden’ of diseases with 21.0% of world global burden of diseases. On one hand India is facing challenge of maternal and child health, communicable diseases (sometimes termed as “Diseases of Poverty”) while on the other hand Diabetes, Heart Diseases and Mental Health Diseases (sometimes termed as the “Diseases of Life Styles”). (1) The incidence of India accounted 58.0 % of cases in the South East Asia Region of World Health Organization (WHO). (2) The Lancet on 20 November 2010 reported 205000 malaria deaths per year in India before the age 70 years with cumulative probability of death 1.8%. The report also added that 90.0 % of deaths were occurred in rural area. In India the high risk fatality associated with the malaria were noted from Orissa, followed by Chhattisgarh, Jharkhand and Assam. (3)Rural and more specifically tribal parts of all the state is more affected as compared to the urban population. Annual Parasite Index (API) for the Maharashtra for year 2011 was 36 per 1000 person. Deaths associated with malaria were recorded as 200 (2010) and 118 (2011) deaths in Maharashtra.(National Vector Borne Disease Control Programme)According to the health status report of Government of Maharashtra 2010, the proportion of dangerous Plasmodium falciparum was 25.7% across the state. It was and is significantly higher in the tribal districts of Gadchiroli, Gondia and across the Vidarbha and North Maharashtra followed by the tribal of Thane (Western Maharashtra). (5) Knowledge is the important component in health seeking behavior. Ignorance, low awareness and not prioritizing health are some of the issues in early identification and prompt treatment of diseases. Studies focusing on knowledge, attitude and practices (KAP) showed that direct interaction with community plays an important role in circumventing malaria problem.(6,7)Researchers have included the direct or indirect measures of people’s knowledge of malaria in equations describing the risk of infection and suggest that health education would offer promise of influencing individuals to adopt better attitude and practices towards preventive measures.(8) A community profiling was done as a part of data collection among the medical practitioners of the Jawhar Block in which they reported malaria as a very prevalent illness in this community. This is complemented by the findings from the National Vector Borne Disease Control Programme (NVBDCP). To achieve the targets of reducing malaria prevalence and preventing malaria epidemic, it is imperative to have active community participation which in turn depends on people’s knowledge and attitude towards the disease. The major public health problems are the contaminated water supply which leads to the unhealthy surroundings paving was for vector breeding and other communicable diseases in these rural and tribal regions. In fact human behavior as a major contributing factor has been largely neglected in research all over the globe on vector borne diseases in part because of the long-standing separation of the behavioral disorder from the physical and biomedical disciplines. So this study was conceptualized to explore the levels of knowledge, attitude and practices towards malaria in tribal communities of Jawhar, Maharashtra, India. Methodology: A cross sectional community based KAP sample survey was carried out with the help of structured questionnaire. Multi stage sampling was used in this study. Sakur PHC was selected out of four PHCs of Jawhar block of Thane by Simple Random Sampling. Six villages under the Sakur PHC, Jawhar were then further selected by Simple Random Sampling. 20.0 % households from every village were selected by using Systematic Random Sampling to get desired sample of nearly 140 households. Every 5th household in the village was interviewed by administration of structured questionnaire. In case of unavailability of households, the next household was included in the study for data collection. A total of 140 households (20.0 %) from these six villages were considered as samples in this study. Structured questionnaire was designed, pre-tested and standardized before the start of data collection. Ethical Clearance: The permission from the institutional ethical committee was taken before starting the study. The purpose of the study was very carefully explained to study participants and informed consent was obtained. Confidentiality about the identity of the participant was maintained throughout the study. One individual per household (head of household) was interviewed face to face and responses were recorded in a structured close ended questionnaire. In case if head of the household was not present then other available member of the household of age > 15 years were interviewed. Gender equity during interview was considered throughout the data collection. Both sexes were given preferences for interview without any discrimination. Data was analyzed with the help of SPSS software version 20 package and Microsoft Office Excel. Descriptive statistics and cross tabs were carried out. RESULTS Findings from study are presented under the sections, sociodemographic profile, knowledge component, attitude component and practice component regarding malaria prevention. The table 1 indicates the socio-demographic characteristics of the participants. It was found that more than 50% of respondents were involved in farming and 27 % agricultural labour/ practices. Majority (68%) of the participants were in the age group of 26 to 45 years of age. Almost all (98%) of the population were found to be in with the education below primary level, of which illiterate were nearly 60%. Majority of the respondents were living in Semi-Pacca houses (56%) and Kaccha house (31%) with around 73 % of the families reported of having family size of 5 or more members. It was found that 63 % of respondent were male in this study. Knowledge The majority sample correctly understood that malaria is transmitted by mosquitoes. Table 2 shows that 95.7 % of the population knows the correct cause of the malaria. As per the preventive perspective, community knowledge of malaria symptoms, such as fever accompanied by shivering, fever on alternating days, headaches, vomiting and rigors is very important for malarial control. Knowledge of the individual about the different symptoms described above is different for the different symptoms. Identification response was found 100% for fever and 83 % for feeling cold which are the basic symptoms for malaria. Among other symptoms only headache has been identified by the majority of the respondents and for the rest of the symptoms it was a low response. The knowledge level of individuals towards symptoms of malaria is low to average. Most of them could not identify other symptoms which are very important in the identification of fever related to malaria. Attitude Table 3 indicates that of 30 % individuals perceived that malaria is a severe disease and person can die due to its serious consequences.73 % of the respondents reported to always seek care when they get these symptoms and remaining 27 % of the respondents reported to seek care only sometime, during the symptoms. About 95 percent of the participants reported removal of stagnant as a preventive measure for malaria.   Practices: The percentage of population following important preventive practices to prevent malaria include, mosquito nets (39 %), indoor residual spraying (8 %), allowing the insecticidal fogging inside the house, cleaning the surroundings (62 %) as shown in table 3. In some practices, the percentage is very low, which is a cause of concern and should be addressed KAP Scale scores: To assess the final level of knowledge, attitude, and practices we have developed a scale based on how many symptoms, attitude and practices the individual have and set the minimum (low) level of score, average level and maximum (high) level of score. (Table 4) This scale has been developed by adopting the KAP study of malaria in Myanmar. (9) The changes have been made according to the demand of the community setting. In the developed scale mean is about the scale only and not describing the mean of the score obtained. Mean is used only to divide the scale equally for the low, average, and high values. So according to these scale ratings the results for the total score for knowledge, attitude and practices have obtained. One correct answer of respondent will get one mark. Score of each category again divided into three categories such as low, Average and high score viz. Knowledge and Practices score were low (1-3), Average (4-6) and high (7-9), while score for attitude was low (1-4) average (5-8) and high (9- 12), as the total questions for knowledge (9), attitude (12) and practices (9) were asked during the interviews of the participants. In the present study 67 % were found to be having an average knowledge on malaria 17.1 % have been found to have low knowledge level. Attitude was found at average level in 84 % of respondents and only 10 % reported low attitude levels. In the practices component, 63 % of the respondents were found to be of average level, while only 10 % found a low level score. DISCUSSION Health education is an important component in creating awareness and disseminating knowledge to the community. Slogans, IEC materials were available in Government of Maharashtra for increasing awareness of malaria health education in the community. The awareness regarding fever, cold, body ache and headache was found to be of optimal level from this study in tribal community of Jawhar indicating reach of Information Education and Communication (IEC) by the Government programme for prevention of communicable diseases under National Vector-Born Disease Control Programme (NVBDCP). Source of information of malaria mainly found as Government health care system and Doctors, this clearly depicts that Government is the major player of health care provision in the tribal areas. Tribals in Jawhar were found to be living in hilly and remote area where availability of infrastructures such as TV, radio etc. is less. Hence the tribal community is dependent mainly on Government healthcare system for the information, knowledge on disease transmission, prevention etc. Backwardness of tribal community as compared to other communities of rural and urban area is a known fact due to limited connectivity with external world like Television, Radio etc. Research regarding sanitation practices showed knowledge or awareness of sanitation helps indeed it is the good health seeking practices which are associated with the “odds” of communicable diseases. (10)Tribal people are aware or have the average knowledge of symptoms of malaria while they are not having high knowledge of prevention of malaria. This is an area which requires improvement as most of the respondents were found to be below the primary level of education. Further knowledge sharing on malaria through various health-education methods will help not only the tribal community to have strong healthy quality of life at the same time mortality due to malaria can be prevented. Though the respondents reported to be following good health seeking practices such as keeping surrounding clean, sanitation; indeed they fail to co-relate importance of these practices with their health status. So further upgrading their knowledge in area of sanitation, health hygiene and importance of behavioral change is needed. As knowledge and practices of the tribal people is a proxy indicator for the available social support or work done by the community health workers (CHWs) in area of health education, there is need of further capacity building among the health workers regarding the complications and treatment part of communicable diseases. (11) The respondent (more than 90%) had satisfactory knowledge regarding the aetiolgy, symptoms and treatment of malaria, majority of respondent (97%) understood that malaria is transmitted by mosquitoes. Studies that were conducted in other part of India in Rajasthan showed the same result and optimum knowledge of malaria in the community.(12)This could due to reach of IEC activities by Government or various organizations; also it could due to acquired knowledge from peers. Knowledge, attitude and practice study in India revels that there is discrepancy among the knowledge of Malaria associated with geography (hilly, tribal, urban, rural setting) and there is difference in different religions, because of their different health seeking behavior associated with their own cultural practices. (13) When we compared tribal community of Jawhar with other backward communities of tribal of India and Sub-Saharan countries then level of knowledge of Jawhar community is at average level.(14–17) CONCLUSION The knowledge level in the community was found to be average. Our findings suggest a need for a health education program aimed at the local community to increase knowledge level to high. Both attitude and practices were found to be average. Specific practices and attitudes changing trainings like use of mosquito nets, IRS spraying, standing water management through group discussion will help impart knowledge to curtail the spread of disease. An aware-ness programme about the common breeding sites (standing water) of Anopheles mosquitoes and ways to manage the complications is the need of an hour. This will improve attitude of the community to maintain their environment. It is recommended to conduct campaigns (IEC programs) for bringing in positive attitude towards Indoor insecticidal fogging in the community. Eventually, tribal of Jawhar needs health education on management of complications, prompt treatment and preventive part for control this communicable disease. Capacity building of the Community Health Workers (CHWs) is very much needed, as there were the main primary sources of knowledge and information dissemination at health care facilities. ACKNOWLEDGMENTS Authors duly thank School of Health System Studies, Tata Institute of Social Sciences, and Mumbai, India for providing the opportunity to conduct research and for permitting us to publish the contents of the research as an article in IJCRR and are also grateful to BAIF, Jawhar, and Maharashtra, India for field coordination. 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 Funding: Nil Conflict of interest: All author(s) declare(s) that there is no conflict of interest regarding the publication of this paper. Englishhttp://ijcrr.com/abstract.php?article_id=444http://ijcrr.com/article_html.php?did=4441. Deaton A. The Great Escape: Health, Wealth, and the Origins of Inequality. 2013. 376 p. 2. World Health Organization. World Malaria Report. 2014. 228 p. 3. World Health Organization. Country Profiles. 2014. 4. National Vector Borne Disease Control Programme. Malaria in India [Internet]. Available from: http://www.malariasite.com/ malaria-india/ 5. National Vector Borne Disease Control Programme. Annual Report [Internet]. 2012. Available from: http://www.nvbdcp.gov.in/ Doc/Annual-report-NVBDCP-2012.pdf 6. Ahorlu CK, Dunyo SK, Afari E a, Koram K a, Nkrumah FK. Malaria-related beliefs and behaviour in southern Ghana: implications for treatment, prevention and control. Trop Med Int Health. 1997;2(5):488–99. 7. Singh N, Singh MP, Saxena A, Sharma VP, Kalra NL. Knowledge, attitude, beliefs and practices (KABP) study related to malaria and intervention strategies in ethnic tribals of Mandla (Madhya Pradesh). Current Science. 1998. 8. Tyagi P, Roy A, Malhotra MS. Knowledge, awareness and practices towards malaria in communities of rural, semi-rural and bordering areas of east Delhi (India). J Vector Borne Dis. 2005;42(1):30–5. 9. Hla-Shein, Than-Tun-Sein, Soe-Soe, Tin-Aung, Ne-Win K-S-A. The level of knowledge, attitude and practice in relation to Malaria in O0-Do village, Myanmar. Southeast Asian J Trop Med Public Heal. 1998;29(3). 10. Sharma DC. India’s BJP Government and health: 1 year on. Lancet [Internet]. Elsevier Ltd; 2015;385(9982):2031–2. Available from: http://linkinghub.elsevier.com/retrieve/pii/ S0140673615609771 11. Perry HB, Zulliger R, Rogers MM. Community health workers in low-, middle-, and high-income countries: an overview of their history, recent evolution, and current effectiveness. Annu Rev Public Health [Internet]. 2014;35:399–421. Available from: http://www.ncbi.nlm.nih.gov/pubmed/24387091 12. Yadav SP, Yadav S, Kuma P. Knowledge , treatment-seeking behaviour and socio-economic impact of malaria in the desert of Rajasthan, India. South African J Infect Dis [Internet]. 2013;28(1):41–7. Available from: http://www.sajei.co.za/index. php/SAJEI/article/view/399 13. Sharma AK, Bhasin S, Chaturvedi S. Predictors of knowledge about malaria in India. J Vector Borne Dis. 2007;44(3):189–97. 14. Forero D a, Chaparro PE, Vallejo AF, Benavides Y, Gutiérrez JB, Arévalo-Herrera M, et al. Knowledge, attitudes and practices of malaria in Colombia. Malar J [Internet]. 2014;13(1):165. Available from: http://www.malariajournal.com/content/13/1/165 15. Yadav SP, Tyagi BK, Ramnath T. Knowledge, attitude and practice towards malaria in rural communities of the epidemic-prone Thar Desert, Northwestern India. J Commun Dis. 1999;31(2):127–36. 16. Paulander J, Olsson H, Lemma H, Getachew A, San Sebastian M. Knowledge, attitudes and practice about malaria in rural Tigray, Ethiopia. Glob Health Action. 2009;2(1):1–7. 17. Aderaw Z, Gedefaw M. Knowledge, Attitude, and Practice of the Community towards Malaria Prevention and Control Options in Anti-Malaria Association Intervention Zones of Amahara National Regional State, Ethiopia. Glob J Med Res Dis. 2013;13(5).

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareEVALUATION OF PARTIALLY PURIFIED SUBABUL PROTEASE INHIBITORS AS BIO INSECTICIDAL TOOL WITH POTENTIAL FOR THE CONTROL OF SPODOPTERA LITURA English3137Arti VasudevEnglish Satwinder K. SohalEnglishThe present study investigated the growth inhibitory potential of partially purified protease inhibitors from Subabul (Leucaena leucocephala) on Common cutworm Spodoptera litura (Fabricius) which has attained major pest status in India. Second instar larvae given treated diet (25, 50 100, 200, 400 and 800 μg/ml) showed a decrease in larval period with increase in concentration when compared with control whereas total developmental period increased at lower concentrations but decreased at 400 and 800μg/ml in comparison to control. The percentage pupation and emergence of females was inhibited and longevity of adults was reduced with increase in concentration. Percentage of male emergence increased significantly at highest concentration in comparison to control. No egg laying was observed at 400 and 800μg/ml concentrations. A significant decline in percent hatching upto 200μg/ml was noticed. The relative growth rate, consumption rate and efficiency of conversion of ingested and digested food was significantly reduced. The present findings clearly confirmed the potential of the inhibitor for pest control. EnglishLeucaena leucocephala, Spodoptera litura, Growth and development, Bioinsecticide, Protease inhibitorsINTRODUCTION All organisms when attacked by their enemies use effective counter measures to defend themselves. Plants have also evolved mechanisms to cope with predatory insects and pathogens. Exploiting these mechanisms will undeniably contribute towards enhancing insect pest resistance. Plants produce different types of proteins to protect themselves from their enemies especially phytophagous insects. There are numerous examples of these proteins like serine and thiol protease inhibitors, amylase inhibitors, lectins and enzymes (Hilder and Boulter 1999). These proteins interrupt pest’s essential amino-acid metabolism by inhibition of protein digestion (Hilder et al., 1992). Many insects particularly those belonging to the order Lepidoptera depend on serine proteases like trypsin, chymotrypsin, and elastase as primary protein digestive enzymes. Much research has focused on exploiting this plant defense mechanism for crop protection. Studies using artificial diets incorporated with plant-derived PIs targeting a particular class of proteinase(s) in the insect gut have established that these proteins retard growth and development in Lepidopteran pests (Boulter, 1993; Pandey et al., 2014). Transforming plant genomes with these proteinaceous PIs provides an ecofriendly and safe approach to pest control (Reckel et al., 1997). However, because of the variability of the insect proteinases and the restricted range of action of the proteinase inhibitors (Ortego et al., 1996), the expression of a particular PI in plants may not yield the desired result and therefore might not be an excellent candidate for biotechnology. Thus there is a continuous search for new inhibitors that are competent to combat pest adaptation via antimetabolic activity. Subabul which belongs to family Leguminoceae, is an ornamental tree used as fodder and firewood. It is reported to exhibit inhibitory activities for plasmin, human plasma kallikrein, trypsin, chymotrypsin and factor XIIa (Oliva et al., 2000). Previous study on STI showed significant reduction in growth and larval development of Helicoverpa armigera by Bhavani et al., 2007. We examined the effects of Subabul protease inhibitors on the growth and development of S. litura larvae. Furthermore, we investigated the effects of this inhibitor on food consumption, absorption and utilization, as well as its effects on the midgut and fecal proteolytic activity of larvae fed on an artificial diet, corroborating novel data on adaptation/resistance of insects to proteinase inhibitors. S. litura is a polyphagous noctuid commonly known as the cluster cater- pillar, tobacco cutworm and tropical armyworm. The larvae have a wide range of host plants of over 40 mostly dicotyledonous plant families, resulting in 69% reduction in yield. It is a polyphagous pest of many forage crops and vegetables in China, Japan and has recently attained the status of a moajor pest on agricultural crops in India (Gokulkrishnan et al., 2012). In India, S. litura is found more in the fields of ground nut, tomato, chilli, bhendi and cotton (Elumalai et al., 2010). MATERIALS AND METHODS Partial purification of PIs Mature Subabul seeds were collected from university campus and defatted using acetone (1:10) w/v for 2h then air dried for 15mins. Defatted seed powder (1:20) w/v was stirred for 1h at room temperature with extraction buffer i.e. 0.1M sodium phosphate buffer, pH7.5. This slurry was then centrifuged at 12,000rpm for 30mins at 40 C. Total soluble protein was incubated at 700 C for 10mins followed by centrifugation at 12,000rpm for 30mins at 40 C. Precipitates were dissolved in minimum amount of extraction buffer and were dialyzed against same extraction buffer using a membrane (cutoff Mr 12000-14000 Da) for 24h at 40 C. The dialyzed sample was treated as partially purified PI which was then subjected to further analysis of protein content as well as for trypsin inhibitory activity. Trypsin inhibition assay Protienase inhibitory activity was determined according to Paulino da Silva et al., 2001 using BApNA (N-α-benzoylDL-arginine p-nitroanilide) as substrate. Protein estimation Protein estimation was done both in defatted and partially purified preparations by the method of Lowry et al., (1951) using bovine serum albumin (BSA) as the standard, for preparing various test concentrations (25, 50, 100, 200, 400, 800µg/ml) for bioassay and nutritional assay studies. Insect culture Egg masses and larvae of S. litura were collected from the cauliflower fields around Amritsar (Punjab), India and subsequent generations were reared in the laboratory at 25±20 C temperature, 65±5% relative humidity (RH) and 12:12 (D: L) photoperiod. The rearing was carried out in glass jars (15cm×10cm) on castor leaves. Rearing on artificial diet was done as reported by Koul et al., (1997). Bioassay studies All experiments were conducted in B.O.D. incubator maintained at conditions mentioned earlier. Bioassay experiments were conducted as described by Vasudev and Sohal (2013). The various concentrations of partially purified PIs used were 25, 50, 100, 200, 400 and 800µg/ml. There were 6 replications with 5 larvae (n=210) in each replication for each concentration. Nutritional analysis Nutritional indices of S. litura were determined by following the procedure of Koul et al., (2005). Dry weights of larvae, diet left and fecal matter were taken by incubating the larvae at the end of experiments at 60°C for 72h inside an incubator. Nutritional indices thus obtained were calculated as proposed by Waldbauer (1968) using dry weights. Statistical analysis All the bioassays and nutritional assays were performed in six replicates and the values were represented as mean ± SE. The data were subjected to analysis of variance (ANOVA). Statistical differences were determined by Tukey’s post hoc test. RESULTS Proteinase inhibitors partially purified from Subabul exhibited 14.86 trypsin inhibitory units (TIU) per mg protein with 92.43% inhibitory activity against bovine trypsin (Table1). Bioassay studies The effect of STI was noticed on developmental physiology of S. litura larvae when they were weighed in their final larval stage after feeding on diet supplemented with different PI concentrations. Larval weight decreased in a dose dependent manner where they weighed 363.4mg less at 800µg/ml in comparison to control (Table 2). Also, an increase was observed in larval period with increase in concentration of STI supplemented diet fed to the second instar larvae. At low concentrations of 25 and 50µg/ml no drastic increase was noticed, whereas at 800µg/ml larval period prolonged significantly by 2.68days in comparison to control larvae (Table 2). Prepupal mortality was noticed at 400 and 800µg/ ml (Fig. 4a). Rearing of larvae on diet amended with STI adversely affected %pupation as only 36.67% larvae pupated at the highest concentration in comparison to 86.67% pupation observed in the larvae reared on control diet (Table 2). Pupal duration showed no particular trend as it increased at lower concentrations (25-100µg/ml) but with increase in PI amount in diet (200-800µg/ml), it decreased significantly when compared with control (Table 2). Pupal weight decreased effectively at all the concentrations when compared with control (Table 2). The pupa formed at 400 and 800µg/ ml concentrations were small in size (Fig. 4b). The adverse effect of STI in its partially purified form was also observed on male and female emergence along with their performance. While the female emergence decreased to 34.52% of the control, the percentage of males emerged from treated larvae was 1.97 times more than control at highest concentration (Table 3). Longevity of adults declined significantly by 4.11days at 800µg/ml when compared with control group. The number of eggs laid by adult females emerged from treated larvae was 593.30/female at 200µg/ml in comparison to control where 939.20eggs/female were laid. At high concentrations, no egg laying was seen by emerged females. Also the eggs laid by emerged females from treated larvae were not much viable as only 33.33% eggs hatched at 200µg/ml (Table 3). Aberrations in adults were more pronounced at 400 and 800µg/ml concentrations where adult half emerged from pupa or adults with deformity in wings were more often seen (Fig. 4c andd). Nutritional indices Nutritional analysis indicates considerable effect of partially purified protease inhibitors from Subabul on food utilization by S. litura. As is apparent from Table 4, there was noteworthy decline in relative growth and consumption rate of S. litura larvae as well as efficiency of conversion of ingested and digested food after ingestion of PI treated diet. Amended diet resulted in 47-80% reduction in RGR over the control. With the increasing concentration of STI, the RGR reduced significantly in comparison to control. Similarly, significant reduction in food consumption rate was recorded where it showed negative correlation with concentration. The concentrations of 400 and 800µg/ml caused 67.38 and 82.36% reduction in RCR over control. Adverse effects of partially purified PI were also recorded on ECI and ECD parameters. Both decreased significantly as concentration of PI increased in diet. Approximate digestibility (AD) increased in a dose response manner in all the treatments when compared with control (Fig. 5A). Metabolic cost to metabolize the ingested PI increased as the amount of PI in diet increased and was maximum (91.20%) at 800µg/ml in comparison to control (43.97%) (Fig.5B). DISCUSSION Partially purified Subabul PIs affected negatively growth and development of S. litura. Similar findings have been reported by Pompermayer et al., 2001 where soybean inhibitor in its partially purified form significantly affected the growth as well as development of Diaterea saccharalis larvae when given in artificial diet. Also trypsin inhibitor partially purified from Theobroma cacao seeds at 0.25% increased the larval and pupal period resulting in morphological abnormalities in adults of D. saccharalis and Anticarsia gemmatalis whereas in Heliothis virescens it caused a significant decline in pupal weight (Paulillo et al., 2012). The larval stage in Lepidoptera is an actively feeding stage whereby they accumulate efficiently all the nutrients present in the food. Any disturbance in essential nutrient accumulation efficiency will directly affect the larval weight and size which clearly accounts for the negative impact of PI on later life stages of S. liura as has been observed in the present study. Since achieving a critical weight is essential for pupation, the delayed larval period observed in the present work could be due to less consumption of diet by the larvae of S. litura indicating the antifeedant nature of the extract. The decrease in the weight of the pupae formed from larvae of S. litura fed on STI diet seems to have adversely affected the reproductive capacity of the emerged females. Achieving high pupal mass is significant as there exists a strong association between adult body weight and its reproductive potential (Tammaru et al. 1996a). Negative effect on fecundity and fertility has previously been documented on S. litura with partially purified PIs from B. oleracea (Vasudev and Sohal, 2013). Any interference in the protein assimilation at larval stage subsequently affects the egg laying capacity of female moths and morphological deformities in adults. Less fecundity at higher concentrations, less number of female adults at all treatments, absence of egg hatching at high concentrations in present study could be accounted for by the less bioavailability of protein or amino acids in food or inability on part of the insect to assimilate the digested food. Similar impact of PIs on nutritional indices of lepidopteran larvae has been reported by several researchers (Da Silva et al., 2012; Mittal et al., 2014; Singh et al., 2014). Nutritional indices and its analysis can form the basis to understand the behavioral and physiological aspects of insect-plant interactions (Lazarevic and Peric-Mataruga, 2003). Our data showed that S. litura larvae fed on STI diet had a low relative consumption as well as growth rate which indicated that less food was utilized by the larvae. This index shows that the feeding rate is directly connected to larval weight (Srinivasan and Uthamasamy, 2005). Our findings also demonstrated a dose dependent decrease in mean larval weight which correlates with the decrease in RCR. Consecutively the larvae took a longer time to pupate and were smaller in size, weighing much less than control which was evident from the low RGR, ECI and ECD. As a result the fecundity and longevity of the adult moths was also severely affected. ECI which is generally a measure of an insect’s capacity to utilize the food ingested for growth and development (Koul et al., 2004) was highest for control and lowest for high-est treatment. This indicated a sign of poor availability of proteins in diet supplemented with PI. Change in ECD also points towards the overall increase or decrease of the proportion of digested food metabolized for energy (Koul et al., 2004). In the current study, the larvae fed on the PI amended diet had reduced value of ECD which suggests that these larvae were actually not as competent in turning digested food into biomass. Approximate digestibility indicates ability of an insect to absorb food through the stomach wall. Increase in AD is an indicative of attempts made by insect to make up for inferior nutritive value of the food and to accomplish the desired growth rate. CONCLUSION In conclusion, partially purified Subabul proteinase inhibitors showed strong anti metabolic and growth inhibitory activity against major pest S. litura. Dietary utilization experiments clearly revealed the growth deterrent impact of the PI; thereby signifying its possible importance to control insect pest populations. These results indicate that future finding of PI from non host plant can be of great importance in environment safe pest management programmes. ACKNOWLEDGEMENT The authors acknowledge the Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar for identifying the seeds. The authors would also like to thank University Grants Commission for providing University with Potential for Excellence research fellowship under the PhD programme of Guru Nanak Dev University, Amritsar. Also authors acknowledge the immense help received from the scholars whose articles are cited in the manuscript. The authors are grateful to author/ editors/ publishers of all those articles, journal and books from where the literature for this manuscript has been reviewed and discussed. Source of funding Authors acknowledge the funds received from University Grants Commission, Delhi under University with potential for excellence (UPE). Conflict of Interest Authors declare that there is conflict of interest.     Englishhttp://ijcrr.com/abstract.php?article_id=445http://ijcrr.com/article_html.php?did=4451. 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Certain essential oil against the field pest army worm, Spodoptera litura (Lepidopetra: noctuidae). International Journal of Recent Scientific Research, 2: 56-62. 11. Gokulkrishnan, J., Krishnappa, K., Elumalai, K. (2012). Effect of plant oil formulations against armyworm, Spodoptera litura (Fab.), Cotton bollworm, Helicoverpa armigera (Hub.) and fruit borer, Earias vitella (Fab.) (Lepidopetra:Noctuidae). International Journal of Current Life Sciences, 2: 1-4. 12. Hilder, V.A., Boulter, D. (1999). Genetic engineering of crop plants for insect resistance a critical review. Crop Protection, 18, 177-191. 13. Hilder, V.A., Gatehouse, A.M.R., Boulter, D. (1992). Transgenic plants conferring insect tolerance: protease inhibitor approach. In: Kung, S., Wu, R., editors. Transgenic plants. New York (NY): Academic Press, pp. 310-338. 14. Koul, O., Shankar, J.S., Mehta, N., Taneja, S.C., Tripathi, A.K., Dhar, K.L. (1997). Bioefficacy of crude extracts of Aglaia species (Meliaceae) and some active fractions against lepidopteran larvae. Journal of Applied Entomology, 121: 245–248. 15. Koul, O., Singh, G., Singh, R., Multani, J. (2005). Bioefficacy and mode of action of aglaroxin A from Aglaia elaeagnoidea (syn. A. roxburghiana) against Helicoverpa armigera and Spodoptera litura, Entomologia Experimentalis et Applicata, 114: 197–204. 16. Koul, O., Singh, G., Singh, R., Singh, J., Daniewski, W., Berlozecki, S. (2004). Bioefficacy and mode-of-action of some limonoids of salannin group from Azadirachta indica A. Juss and their role in a multi component system against lepidopteran larvae. Journal of Biosciences, 29: 409–416. 17. Lazarevic, J., Peric-Mataruga, V. (2003). Nutritive stress effects on growth and digestive physiology of Lymantria dispar larvae. Yugoslav Medical Biochemistry, 22: 53–59. 18. Lowry, O.H., Rosenbrough, N.J., Farr, A.L., Randall, R.J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193: 265–275. 19. Macedo, M.L.R., Freire, M.D.G.M., Cabrini, E.C., Toyama, M.H., Novello, J.C., Marangoni, S. (2003). A trypsin inhibitor from Peltophorum dubium seeds active against pest protease and its effect on the survival of Anagasta kuehniella (Lepidoptera:Pyralidae). Biochimica Biophysica Acta, 1621: 170-182. 20. Macedo, M.L.R., Mello, G.C., Freire, M.G.M., Novello, J.C., Marangoni, S., Matos, D.G.G. (2002). Effect of a trypsin inhibitor from Dimorphandra mollis seeds on the development of Callosobruchus maculatus. Plant Physiology and Biochemistry, 40: 891-898. 21. Mittal, A., Kansal, R., Kalia, V., Tripathi, M., Gupta, V.K. (2014). A kidney bean trypsin inhibitor with an insecticidal potential against Helicoverpa armigera and Spodoptera litura, Acta Physiologiae Plantarum, 36: 525-539. 22. Nandeesha, P., Prasad, D.T. (2001). Characterization of serine proteinase inhibitor from subabul (Leucaena leucocephala, L.) sedds. Journal of Plant Biochemistry and Biotechnology, 10: 75- 78. 23. Oliva, M.L., Souza-pinto, J.C., Batista, I.F., araujo, M.s., Silveria, V.F., Aureswald, E.A., mantele, R., Eckerskorn, C., Sampaio, M.U., Sampaio, C.A. (2000). Leucaena leucocephala serine proteinase inhibitor: primary structure and action on blood coagulation kinin release and rat paw edema. Biochimica et Biophysica Acta, 1477: 64-74. 24. Ortego, F., Novillo, C., Castafiera, P. (1996). Characterization and distribution of digestive proteases of the stalk com borer Sesamia nonagrioides Lef. (Lepidoptera: Noctuidae). Archives of Insect Biochemistry and Physiology, 33: 163–180. 25. Pandey, P. K., Singh, D., Singh, S., Khan, M.Y., Jamal, F. (2014). A nonhost peptidase inhibitor of 14 kDa from Butea monosperma (Lam.) Taub. seeds affects negatively the growth and developmental physiology of Helicoverpa armigera. Biochemistry Research International, http://dx.doi.org/10.1155/2014/361821 26. Paulillo, L.C.M.S., Sebbenn, A.M., Derbyshire, M.T.V.C., GoesNeto, A., Brotto, M.A.P., Figueira, A. (2012). Evaluation of in vitro and in vivo effects of semipurified proteinase inhibitors from Theobroma seeds on midgut protease activity of lepidopteran pest insects. Archives of Insect Biochemistry and Physiology, 81: 34-52. 27. Paulino da Silva, L., Leite, J.R.S.A., Bloch, C. Jr., Maria de Freitas, S. (2001). Stability of a black eyed pea trypsin/chymotrypsin inhibitor (BTCI). Protein and Peptide Letters, 8: 33–38. 28. Pompermayer, P., Lopes, A.R., Terra, W.R., Parra, J.R.P., Falco, M.C., Silva-Filho, M.C. (2001). Effects of soybean proteinase inhibitor on development, survival and reproductive potential of the sugarcane borer, Diatrea sachharalis, Entomologia Experimentalis et Applicata, 99: 79-85. 29. Reckel, G.R.K., Kramer, K.J., Baker, J.E., Kanost, M.R., Fabrick, J.A., Behnke, G. A., 1997. Proteinase inhibitors and resistance of transgenic plants to insects, in: N. Carozzi, M. Koziel (Eds.), Advances in Insect Control. The Role of Transgenic Plants. Taylor and Francis, London, pp. 157– 183. 30. Singh, D., Jamal, F., Pandey, P.K. (2014). Kinetic assessment and effect on developmental physiology of a trypsin inhibitor from Eugenia jambolana (Jambul) seeds on Helicoverpa armigera (HÜBNER), Archives of Insect Biochemistry and Physiology, 85: 94-113. 31. Srinivasan, R., Uthamasamy, S. (2005). Studies to elucidate antibiosis resistance in selected tomato accessions against fruitworm, Helicoverpa armigera Hubner Resistant Pest Management Newsletter, 14: 24–26. 32. Tammaru, T., Kaitaniemi, P., Ruohomäki, K. (1996). Realized fecundity in Epirrita autumnata (Lepidoptera: Geometridae): relation to body size and consequences to population dynamics. Oikos, 77: 407–416. 33. Vasudev, A., Sohal, S.K. (2013). Bioinsecticidal potential of partially purified proteinase inhibitors from Brassica oleracea (L.) against Spodoptera litura (Fab.) (Lepidoptera: Noctuidae), Efflatounia,. 13: 1-7. 34. Waldbauer, G.P. (1968). The consumption and utilization of food by insects, Advances in Insect Physiology, 5: 229–288.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareMODIFIED SCHIRMER TEST SCREENING FOR DRY EYE DISEASE IN RURAL POPULATION English3840Virendra Singh LodhaEnglishObjective: The present study was undertaken to screen rural patients for dry eye disease by a modified Schirmer 1 test without anaesthesia with a cut off of 10 mm wetting of Schirmer test strip. Methods: Schirmer 1 test without anaesthesia was performed on 140 patients (65 males and 75 females) over 40 years of age from rural background after obtaining informed consent and after applying some exclusion criteria. The test was aborted as soon wetting of 10 mm of Schirmer test strip occurred and time noted. The patients who did not achieve 10 mm of wetting on Schirmer test over 5 minute’s time were labelled Schirmer test positive. Results: The incidence of positive Schirmer1 test (without anaesthesia) was 16.4% in the studied group of 140 patients with a higher incidence in females over 50 years of age. Out of 140 patients screened, 63% patients achieved a wetting of 10 mm of Schirmer test strip in 2 or less than 2 minutes. Conclusion: Modified Schirmer 1 test (without anaesthesia) with a cut off limit of 10 mm can be a better option for dry eye disease screening in large volume based ophthalmic practice and eye camps as it reduces test time and causes less discomfort to the patient, and the procedure does not require any equipment other than readily available Schirmer test strips. EnglishSchirmer test, Dry eye diseaseINTRODUCTION Dry eye disease is a common ocular disorder affecting a significant percentage of population, estimated to be 5 - >30.1, 2 International Dry Eye Workshop (2007) defined dry eye as a multifactorial disease of tears and ocular surface that results in discomfort, visual disturbance and tear film instability with potential damage to the ocular surface and is accompanied by increased osmolarity of tear film and inflammation of ocular surface.3 The prevalence of dry eye disease is likely to increase in future due to increase in life expectancy, environmental pollution, increased use of medications, increased incidence of diabetes mellitus and life style changes. Dry eye disease results from either decreased tear production or increased evaporation of tears. Various methods used to diagnose dry eye disease include Schirmer test, Phenol Red Thread test, tear film break up time (TBUT),tear meniscus height, epithelial staining with rose Bengal and lissamine green, tear osmolarity, impression cytology and symptom questionnaires like ocular surface disease index (OSDI), dry eye questionnaire (DEQ) etc. Different tests have been shown to have different specificity and sensitivity. Tear osmolarity determination is the most reliable test to diagnose dry eye but is expensive to perform and time consuming. Only Schirmer test is one such test which is simple to perform and does not require slit lamp or other equipment and can be performed easily by a trained ophthalmic technician. Schirmer introduced the test in 1903(4) and since then it has been modified by many investigators of which Schirmer 1 test is the most popular of the variants and is carried out without anaesthesia, it measures both reflex and basal tear secretions.(5) In India rural people seek eye check up mostly in eye camps where they turn out in large numbers. The eye care professional has to examine large number of patients hence it becomes difficult to perform tests for dry eye disease which require more time, equipment and expensive. Hence the present study was undertaken to find out the prevalence of positive Schirmer 1 test in rural population, attending free outpatient services of the eye department of Pacific Medical College and Hospital, Udaipur. MATERIALS AND METHODS This study was conducted after obtaining approval from institutional ethics committee. After obtaining informed consent, a total of 140 patients from nearby villages, aged 40 years and above visiting eye department of Pacific Medical College and Hospital, Udaipur from September 2014 to December 2014 were included in the study. Exclusion criteria: Patients with history of any systemic disease, who were on any systemic medications (Allopathic, Ayurvedic, Homeopathic or Unani), history of eye surgery, on topical ocular medications, eye lid deformities, trichiasis, trachoma or other active ocular infection or inflammation, glaucoma, corneal opacities or degenerations and who were unwilling to undergo Schirmer test were excluded from this study. After performing routine ophthalmic examination which included visual acuity measurement, refraction, slit lamp examination and undilated fundus examination, Schirmer 1 test without anaesthesia was performed after explaining the procedure to the patient. Intra ocular pressure was recorded after performing Schirmer test and the patients with high intraocular pressure (>21) were not included in the study. Commercially available Schirmer test strips were used for the test. The test strip was inserted at the junction of medial two third and lateral one third of the lower conjunctival fornix of both eyes and the patient was instructed to keep eyes open, look straight and blink normally. The test strip was removed from the eye as soon as a wetting of 10 mm occurred and time noted. The patients whose wetting did not cross 10 mm mark at completion of 5 minutes were labelled as Schirmer 1 test positive. RESULTS Schirmer 1 test without anaesthesia was performed in 140 patients, 65 male and 75 female, table 1 shows age group wise distribution with maximum patients (67.85%) belonging to age group 40 – 50 years of age of which 11% males and 14% females were Schirmer 1 test positive. The incidence of positive Schirmer 1 test was 20% in the groups over 50 years of age in males. In female patients the incidence was 26.6% in the age group of 51 – 60 years and 30% in the age group of over 60 years with an incidence of 28% in the all females over 50 years of age. Table 2 shows that out of 117 Schirmer test negative patients, 88 (75%) required less than 2 or 2 minutes to achieve 10 mm wetting of the test strip (63% of total patients screened) and 29 patients (25%) required more than 2 minutes for 10 mm wetting to occur. It shows that only 37% patients including Schirmer test positive patients require the test to be continued for more than 2 minutes. None of the patients in this study complained of any ocular discomfort due to test procedure DISCUSSION Since its introduction in1903 by Schirmer6 , many variations of this test have been used clinically; especially test with closed or open eyes, with or without anaesthesia and type of paper and its strip size. Presently Schirmer 1 test without anaesthesia is the most popular test for measuring aqueous tear production with commercially available Schirmer test strips. To minimise ocular discomfort and increasing the co nvenience in performing the test in less time Nelson proposed to do it for 1- minute time with a cut off value of 6mm.7 Bawazeer and Hodge demonstrated that 1-minute test with anaesthesia correlated highly with the 5-minute test with anaesthesia.8 In the present study Schirmer 1 test without anaesthesia was performed to find out the incidence of positive Schirmer test with a 10 mm cut off (wetting of less than 10 mm in five minutes) and the incidence of negative Schirmer test to study the incidence of dry eye disease in rural population. The test was aborted as soon as 10 mm wetting of test strip occurred and time noted as 2 minutes or more. It was found that in the age group of 40-50 years the test was positive in 11% of males and 14% females with combined incidence of 12.6% in this group. In the age group of 51-60 years 20% males and 26.6% females were found to have positive Schirmer test (dry eye) with a combined incidence of 24% in this age group. In the age group of over 60 years 20% males and 30% females were found to be positive on Schirmer Test. The overall incidence of dry eye in the present study was found to be 16.4%, with 13.8% in males and 18.6% in females wherein only Schirmer 1 test without anaesthesia and with a cut off value of 10 mm was used for the diagnosis in rural population. In studies done in northern and eastern India prevalence of dry eye disease in hospital based population is reported to be between 18.4% and 40.8% with different diagnostic criteria being used.9,10,11 The lower incidence in the present study may be due to the fact that only Schirmer test was used as a screening tool. In the present study Schirmer test was aborted before 5 minutes if a wetting of 10 mm occurred earlier. Out of 117 patients who were negative on Schirmer test (wetting of 10 mm) 88 patients (75%) required less than 2 minutes or less and 29 patients (25%) required more than 2 minutes and from a total of 140 patients tested 63% required less than or 2 minutes of the test time if it was aborted as soon as 10 mm wetting of the test strip occurred. Similar observation has been made in an earlier study and authors concluded that 2- Minute Schirmer 1 test with anaesthesia could be used instead of the standard 5-minute rest.12 However in Schirmer 1 test with anaesthesia reflex tear production cannot be ruled out hence the present study was done with Schirmer 1 test without anaesthesia. CONCLUSION From the results of this study it can be concluded that a modified Schirmer 1 test (without anaesthesia) with a cut off limit of 10 mm wetting of the test strip, reduces the test time significantly which is more convenient for both, the eye care professional and the patient, and hence can be a good tool for dry eye disease screening in large volume based ophthalmic practice or eye camps. ACKNOWLEDGEMENTS 1. The author is thankful to the management of Pacific Medical College and Hospital, Udaipur, India for allowing me to conduct this study. 2. Author acknowledges the immense help received from the scholars whose articles are cited and included in references of this manuscript. The author is 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. Englishhttp://ijcrr.com/abstract.php?article_id=446http://ijcrr.com/article_html.php?did=4461. Smith JA, Albenz J, Beglet C et al. The epidemiology of dry eye disease: Report of the epidemiology subcommittee of the International Dry Eye Disease Workshop (2007), Ocul Surf. 2007, 5: 93–107. 2. Lemp MA, Advances in understanding and managing dry eye disease, Am J Ophthalmol. 2008 Sept; 146(3): 350–356. 3. International Dry eye Disease Workshop (DEWS). The definition and Classification and classification of dry eye disease: report of the Definition and Classification Subcommittee of the International Dry Eye Workshop Ocul Surf. 2007, 5: 75–92. 4. Savini G, Prabhawasat P, Kozima T, Grueterich M, Espana E, Goto e. The challange of dry eye diagnosis. Clin Ophthalmol. 2008; 2(1):31–35. 5. Smith J, Nichols KK, Baldwin EK. Current patterns in the use of diagnostic tests in dry eye evaluation. Cornea. 2008; 27(6):656– 662. 6. Schirmer O. Studien zur physiologie und pathologie der tranenabsonderung Und tranenabfuhr. Graefes Arch Clin Exp Ophthalmol. 1903; 56:197–291. 7. Nelson PS. A shorter Schirmer tear test. Optom Mon. 1982; 73:568–9. 8. Bawazeer AM, Hodge WG. One-minute Shirmer test with anaeasthesia. Cornea. 2003; 22 (4): 285–287. 9. Sahai A, Malik P. Dry Eye:Prevalence and attributable risk factors in a Hospital based population. Ind J ophthal 2005; 53: 87– 91. 10. Gupta N, Prasad I, Jain R, D’Souza P. Estimating the prevalence of dry eye Indian patients attending tertiary ophthalmology clinic. Ann Trop Med Parasitol 2010; 104: 247–255. 11. Basak SK, Pal PP, Basak S, Bandhyopadhyay A, Choudhary S, Sar S. Prevalence of Dry Eye Diseases in hospital-based population in West Bengal, Eastern India. J Indian Med Assoc. 2012; 110: 789–794. 12. Suphakasem S, Lekskul M, Rangsin R. Assessment of different wetting time and paper strip size of Schirmer test in dry eye patients. J Med Assoc Thai 2012, 95 Suppl 5: S107–110.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcarePHYSIOCHEMICAL PROPERTIES OF SOME PAEDIATRIC FORMULATIONS OF ARTEMETHER -LUMEFANTRINE PRESCRIBED FOR UNCOMPLICATED PLASMODIUM FALCIPARUM MALARIA. English4145Awofisayo Sunday O.English Okhamafe Augustine O.English Arhewoh Mathew I.EnglishPhysicochemical properties bothering on the quality of powder for paediatric suspension (PPS) of artemether-lumefantrine (AL) were evaluated. The moisture content, viscosity, total solid and chemical contents were determined. The assay was analyzed simultaneously for artemether and lumefantrine using high pressure liquid chromatography (chromosil C18 column/ UV detection at 216 nm). Acetonitrile: 25 mM potassium dihydrogen phosphate (70: 30%, v/v) and nevirapine served as mobile phase and internal standard, respectively. Statistical analysis was done using students t-test to compare the parameters for the products at CI, 95%. The artemether and lumefantrine contents varied from 40.3-112.54% and 71.9 – 91.4%, respectively. The range of values (mean) of moisture content, viscosity, pH and total solid were 2.9-6.9 (4.68)% , 99.1-193.8(124.7) mPa.s, 3.5-7.8 (4.7) and 93.1- 97.1 (95.3) %, respectively. The results showed statistical different outcomes (P < 0.05). PPS products sampled vary widely in their physicochemical properties. EnglishArtemether-lumefantrine, Paediatric formulations, Physicochemical properties, Uncomplicated malariaINTRODUCTION The incidence of malaria worldwide is estimated to be within the range of 300 – 500 million clinical cases each year with about 90% of these occurring in Africa (WHO, 2013). Malaria is estimated to kill between 1.1 and 2.7 million people worldwide each year, about one million of whom are African children under the age of five (Cesar, 2009). As Plasmodium falciparum causes millions of clinical episodes and infant deaths yearly in Africa, it is therefore of vital importance that antimalarial drugs used for treatments are genuine and of high quality (Amin and Kokwaro, 2007; Awofisayo et al., 2010). The high prevalence of substandard antimalarials in the African retail sector is of great importance in view of their frequent use for fever/malaria treatment (Bapner et al., 1996; WHO, 1999; 2006; Helin-Tanninen, 2001). The past decade has seen increased interest in specific population-targeted and individualized-drug development. Several legislative initiatives in the US (e.g., The Best Pharmaceuticals for Children Act) and Europe (e.g., Paediatric Investigation Plans as indicated in Paediatric Regulation EC 1901/2006), supported by the International Conference of Harmonization (ICH) and World Health Organization, were recently taken to stimulate and improve pharmaceutical care for infants, children and adolescents (EMEA, 2006; Zajieek, 2009; Vandercruyssen et al., 2004). There is need for availability of paediatric formulations of artemether-lumefantrine (AL) that permit accurate dosing and enhance patients’ compliance (USP, 2014). In spite of increased artemether use in treating malaria in endemic areas, the report of therapeutic failure is rising and scientific literature is still limited regarding analytical methods aimed at quantitation of the drug in pharmaceutical products. The United States Pharmacopeia, 2014 (Pingala and Mangaokar, 2013; USP, 2014) contains monographs of pure artemether and the parenteral form. Lumefantrine, the co-formulated. drug in fixed dose artemisinin – based combination therapy (ACT) has been analyzed by a chromatographic method involving the principle of tandem instrumental application with liquid chromatography – mass spectrophotometry (LC-MS) (Khalil et al., 2011). Quality indices of powder for paediatric suspension (PPS) formulations such as viscosity, pH and moisture content that may influence the physical and chemical stability of drug products, such as artemether and lumefantrine, will require systematic evaluation. The multisource nature of drug production may, however, be responsible for any observable physicochemical differences. The aim of this present work was to evaluate the physicochemical factors influencing the quality of PPS of fixed dose AL products sold in Nigeria. MATERIALS AND METHODS Chemicals AL PPS generic products coded (ALA - ALF), sourced from registered pharmacy outlets in Uyo, southeastern Nigeria; nevirapine was used as internal standard (IS), while acetonitrile and potassium dihydrogen phosphate, as mobile phase, were all products of Sigma Aldrich, Germany. Artemether and lumefantrine reference powders were obtained from Qimdis, France. All the reagents employed were of analytical grade.  Physical Observation The samples were visually examined to assess characteristics such as odour, colour and texture of powder. Viscosity Determination To reconstitute the powder products of AL, 20mL of distilled water was added, shaken well and made up to 60 ml mark with water. The viscosity of the reconstituted PPS products was evaluated using a viscometer (Mettler Toledo, Germany). Twenty milliliters of the suspension was placed between the cone and the basal plate at standard temperature condition, 32o C and rotation at 5 rpm for 5 min. The water content of the PPS products was determined using 1 g of powder for the analysis in a moisture analyzer and heating up to 105o C. Measurement was performed in triplicates for each drug product. pH Determination The pH of the reconstituted products was measured (n = 3) by dipping the probe of the device directly into the reconstituted products using a pH/mV meter (Mettler Toledo, Germany) at temperature of 25o C. Total Solid The reconstituted products were shaken up. After flocculation, 20 mL samples were taken with a pipette from the same depth and added to a porcelain dish of known weight, W1. This was evaporated to dryness by placing the dish with its content first on a water bath and subsequently in an oven (Galenkemp No. 335, England). The samples were intermittently cooled in a dessicator and weighed until a constant weight, W2, was obtained. The difference in weights (W2- W1) was calculated and the total solid percent determined from equation 1. Chemical content determination The internal standard was prepared by dissolving 20 mg of nevirapine in 10 mL of deionized water and made up to mark in a 1 L volumetric flask resulting in 20 mg/ml solution. Approximately 250 mg and 1500 mg of artemether and lumefantrine reference powders, respectively, were transferred into a 100-mL volumetric flask and made up to mark. The solutions were sonicated and then diluted to 1 L in a volumetric flask to produce 0.25 and 1.5 mg/mL stock solutions, respectively. The stock solutions produced were further diluted with acetonitrile: acetic acid (99: 2, v/v) to obtain the working solutions. The working solutions were spiked with IS solution to give uniform amount of the IS in the working solution. One sample was taken from one bottle and three bottles were sampled. High pressure liquid chromatographic (HPLC) system was used to assay the samples (HPLC Peak 7000 system with analytical chromosil column C18, 250 x 46 mm, Rheodyne manual samples injectors, Germany). Data Analysis All statistical analyses were performed using SPSS version 13.0. The values for each parameter evaluated were compared among brands and with the reference product coded ALA using one sample t-test and statistical difference was taken at PEnglishhttp://ijcrr.com/abstract.php?article_id=447http://ijcrr.com/article_html.php?did=4471. Amin A.A., Kokwaro G.O. 2007. Antimalarial drug quality in Africa. J Clin PharmTherap, 32: 429–440. 2. Awofisayo S.O., Willie E., Umoh E. 2010. Quality control evaluation of multi-source artemether- lumefantrine tablets prescribed for uncomplicated multi-drug resistant malaria. Ind J Nov Drug Del, 4: 153-157. 3. Bapner J.S., Tripathi C.D., Tekur U.1996. Drug utilization pattern in third world. Pharmaco-economics, 9:286-294. 4. Cesar I.C, Pianetti G.A. 2009. Quantitation of artemether in pharmaceutical raw material and injections by high performance liquid chromatography. Braz J Pharm Sci., 45: 738-742. 5. European Medicine Evaluation Agency (EMEA). 2006. EMEA/ CHMP/PEG/19481/2005. Reflection paper formulation of choice for the paediatric population; EMEA, London. 6. Food and Drug Administration (FDA). 2007. Food and Drug Administration Amendments Act, FDA, Silver Spring. 7. Helin-Tanninen M., Naaranlahti T., Kontra K., Wallenius K. 2007. Enteral suspension of nifedipine for neonates. Part 1. Formulation of nifedipine suspension for hospital use. J Clin Pharm Therap., 26: 49–57. 8. Khalil I.F., Abildrup U., Alifrangis L.H. 2011. Measurement of lumefantrine and its metabolite in plasma by high performance liquid chromatography with ultraviolet detection. J Pharm Biomed Anal., 54:168–172. 9. Lavoie F., Cartilier L., Thibert R. 2002. New methods characterizing avalanche behaviour to determine powder flow. Pharm Res.,19: 887-893. 10. Pingala S.G., Mangaonkar K.V. 2013. Quantification of lumefantrine in human plasma using LC-MS/MS and its application to bioequivalence study. J Pharm., 13:1-8. 11. Sridhar B., Rao K.H., Srinivas T.V. 2010. A validated reverse phase HPLC method for the simultaneous estimation of artemether and lumefantrine in pharmaceutical dosage forms. Int J Adv Pharm Sci., 1: 95-99. 12. United States Pharmacopoeia. 2014. The National Formulary, USP 38 NF13; United States Pharmacopeia Convention, Inc.: Rockville, MD, USA. 13. Vandercruyssen K., D’Hont M., Vergote V. 2004. LC-UV/MS Quality analytics of paediatrics artemether formulations. J Pharm Anal., 4 (I): 37 – 52. 14. World Health Organization. 2006. Counterfeit Medicines. Facts sheet no 275. 2006. Revised February 2006. 15. World Health Organization. 1999. Counterfeit Drugs; Guidelines for the development of measures to combat counterfeit drugs. Geneva: WHO, pp 1 – 60. 16. World Health Organization. 2013. World Malaria Report; Geneva, Switzerland. 17. Zajieek A. 2009. The national institutes of health and the best pharmaceuticals for children act. Paed Drugs., (II): 45 – 47.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareA COMPARATIVE STUDY OF DENTAL ARCH FORM IN CASES WITH THUMB SUCKING, TONGUE THRUSTING AND NORMAL ARCH FORM USING EUCLIDEAN DISTANCE MATRIX ANALYSIS English4656D. Ram MundadaEnglish R.H. KambleEnglish Anuradha RajkuwarEnglish Sunita ShrivastavaEnglish Narendra SharmaEnglish Rizwan GilaniEnglishTo compare dental arch form between the cases with thumb sucking, tongue thrusting and normal arch form using Euclidean distance matrix analysis” Materials and Method: The sample consist of 30 patients ranging in age from 13 to 17 years were divided into three groups - Group 1- Subjects with thumb sucking habit, Group 2- Subjects with tongue thrusting habit and Group 3- Subjects without any history of habit. Study model impression was made and analysis of recorded data was carried out using a WinEDMA to compare the arch shape, size, inter-canine and inter-molar width. The statistical significance of the form difference was tested by using a “bootstrap” procedure. Result: There was a significant arch-shape difference in the maxillary arches between thumb sucking subjects and subjects with no habits. Whereas significant arch-shape differences were found in the mandibular arch of tongue thrusting individual and those with no any habits. On comparing thumb sucking and tongue thrusting subjects, there was a significant arch-shape difference in the maxillary and mandibular arches. The arch size of tongue thrusting subjects was larger as compared to thumb sucking subjects. Conclusion: Expansion of narrow maxillary arch width in anterior region should be considered to harmonize with normal mandibular arch form in thumb-sucking subjects, whereas expansion of maxillary arch width I posterior region should be considered to harmonize with wider mandibular arch form in tongue thrusting subjects. EnglishArch form, Thumb sucking, Tongue thrusting, EDMAINTRODUCTION Nature of equilibrium of the natural dentition is of primary importance to an orthodontist, whose concern is to achieve ideal and stable dental arches. Orthodontists are also concerned with patients oral habits since these may lead to abnormal growth and development of craniofacial structures.19 Arch form is a unique expression of individual development and probably no universal design will ever be able to account for many small but significant variations in the arch shape of the individual. It is commonly believed that the dental arch form is initially shaped by the configuration of the supporting bone and following the eruption of the teeth, by the circum-oral musculature and intraoral functional forces.17 Dental arch form consists of dental units arranged in unique positions along a compound curve, which represents a steady state of equilibrium delimited by the counterbalancing force fields of the tongue and the circum-oral tissues. The primary determinants of arch form morphology are the muscle forces of the resting state in contraction to the intermittent forces of muscle in functioning states. Considering the circum-oral structure as an elastic envelope, the lips and the cheeks exert counterbalancing inward tension against the teeth.19 Abnormal pressure habits changes the alveolar bone morphology and regulates the teeth in that bone. However these changes are taking place in living bone, one cannot revoke abnormal pressure habits as an etiology factor in developing malocclusion.12, 13 Studies evaluating the effects of digit sucking found that the shape of maxillary arch was modified in thumb- and digitsuckers by elongation of the anterior segment. This acts to produce spacing, labial inclination, and protrusion of the maxillary incisors. The creation of an excessive overjet in turn fosters abnormal lip and tongue muscle activity. The thumb sucking is normal in the first two or three years of life but may cause permanent damage if continued beyond this time. Maxillary and mandibular inter-canine and intermolar width, and anterior and posterior transverse inter-arch discrepancies in prolonged sucking habits showed association with the narrow maxillary inter -canine and inter- molar width, increased posterior transverse discrepancies and increase prevalence of posterior cross bite. Various methods have been used to compare the dental-arch forms in different studies but it was noted that Euclidean Distance Matrix Analysis (EDMA) has been extensively used in craniofacial morphological studies, as it provides a good measurement of form differences, separates the contributions of size and shape. It was found that prolonged thumb-sucking leads to a reduction in maxillary arch width particularly in canine region but no method provides good information about the major variations in the arch forms.15,16 Therefore, the present was conducted to evaluate and compare dental arch form in cases with thumb-sucking, tongue thrusting and normal arch form using “Euclidean Distance Matrix Analysis (EDMA)” MATERIALS AND METHOD Sample selection Thirty patients, ranging in age from 13 to 17 years, were selected from the outdoor patients Department of Orthodontics, Sharad Pawar Dental College and students of Datta Meghe Institute of Medical Sciences (DU), Sawangi (M), and Wardha. The selected patients were divided into three groups comprising of 10 patients each as Group 1- Subjects with thumb sucking habit, Group 2- Subjects with tongue thrusting habit and Group 3- Subjects without any history of habit Study model impressions were made replicating all minute details, poured in dental stone and proper bases were formed. The data was processed at CAD-CAM centre, Department of Mechanical Engineering, VNIT (Regional Engineering College), Nagpur. YM-2115, three dimensional co-ordinate measuring machine was used for identification of landmarks on the study cast(Fig. A, Plate No. 1).11,21 It runs in the range of 150 mm X 200 mm X 100 mm and the accuracy of 3 orthogonal axes was 0.01 mm. The frictionless air bearing and touch trigger probe (0.5 mm) was used to identify the measuring point (i.e. anatomic point of each tooth) and to record the corresponding X,Y and Z coordinates to a computer data file. The analysis of recorded data was carried out using a WinEDMA version 1.0.1 beta as given by Cole TM.22 Method Fourteen landmarks (midpoints of incisal edges, canine cusps and buccal cusps of premolars and first molars) were selected to represent the dental-arch form for maxillary and mandibular arch of each subject.1 The mesial contact point of the maxillary central incisors to the mesio-buccal cusp of the maxillary first molars was selected as the maxillary standard plane (Fig.1). All measuring points were marked by the YM-2115, three dimensional co-ordinate measuring machine (Fig. B, Plate No. 1). The corresponding x, y and z coordinates were recorded in a computer data file (Plate No.2, 3, 4, 5). Fourteen landmarks representing both arches were projected to the corresponding standard plane. Then, a 2-dimensional EDMA was prepared to compare the arch forms between the 3 groups. In this study, the EDMA for arch form comparison was calculated as described by Lele and Richtsmeier1 and Ferrario et al6-8 using WinEDMA computer programme22. The procedure was as follows: All possible linear distances between pairs of landmarks were computed from the coordinates of the corresponding standard plane in each subject. This produced 10 maxillary matrices and 10 mandibular matrices of 91 distances (14x [14-1]÷2) in each group. Form matrices were then averaged for each arch in each group, thus obtaining 6 mean form matrices of maxillary and mandibular arches. Statistical analysis: The 91 ratio were then sorted from lowest to highest and the statistics, T= maximum ratio/minimum ration and M= medium ratio, were calculated. T represented the total range of arch-shape differences between the groups and M was a measure of general size difference by the form-difference matrices. The statistical significance of the form difference (i.e. Ho = similarity of forms, Ha = difference between forms) was tested by using a “bootstrap” procedure. The level of significance was set at 5%. The null hypothesis was rejected if the observed T statistics were in an extreme tail of the distribution- equal to or less than 5% (P =< 0.05). RESULTS There was a significant arch-shape difference in the maxillary arches between Group I and Group III subjects [(P1 Englishhttp://ijcrr.com/abstract.php?article_id=448http://ijcrr.com/article_html.php?did=4481. Lele S, Richtsmeier JT. On comparing biological shapes: detection of influential landmarks. Am J Phys Anthropol 1992; 87:49- 65 2. Lele S. Some comments on coordinate- free and scale-invariant methods in morphometrics. Am J Phys Anthropol 1991; 85:407- 17. 3. Lele S, Richtsmeier JT. Euclidean Distance Matrix Analysis: Confidence Intervals for form and growth differences. Am J Phys Anthropol 1995; 98:73-86. 4. Hens SM. Growth and sexual dimorphism in orangutan crania: a three-dimensional approach. Am J Phys Anthropol 2003; 121:19-29. 5. Singh GD, Hodge MR. Bimaxillary morphometry of patients with Class II Division 1 malocclusion treated with Twin Block appliance. Angle Orthod 2002; 72:402-9. 6. Ferrari VF, S forza C, Miani A Jr, Serrao G. Dental arch symmetry in young healthy human subjects evaluated by Euclidean distance matrix analysis. Arch Oral Biol 1993;38:189-94. 7. Ferrari VF, S forza C, Miani A Jr, Tartaglia G. Maxillary verus mandibular arch form differences in human permanent dentition assessed by Euclidean distance matrix analysis. Arch Oral Biol 1994;39:135-9. 8. Ferrari VF, S forza C, Miani A, Tartaglia G. Human dental arch shape evaluated by Euclidean-distance matrix analysis. Am J Phys Anthropol 1993;90:445-53  9. Bell A, Ayoub AF. Assessment of the accuracy of a three dimensional imaging system for archiving dental study models. J Orthod 2003;30:219-23. 10. Nie Q, Lin J. A comparison of dental arch forms between Class II division 1 and normal occlusion assessed by Euclidean distance matrix analysis. Am J Orthod Dentofac Orthop 2006;129:528- 35. 11. Braun S, Hnat WP, Fender DE, Legan HL. The form of the human dental arch. Angle Orthod 1998;68(1):29-36. 12. Takayuki K, Takashi O. Diagnosis and Management of Oral Dysfunction. World J Orthod 2000;1:125-133. 13. Klein E. Pressure habits, etiological factors in malocclusion. Am J Orthod 1952;38:569-587. 14. Aznar T, Galan A.F, Marin I., Dominguez A. Dental arch diameters and relationships to oral habits. Angle Orthod 2006;76:441- 445. 15. Larsson E. Sucking, chewing and feeding habits and the development of crossbite: a longitudinal study of girls from birth to 3 years of age. Angle Orthod. 2001;71(2):116-119. 16. Ogaard B, Larsson E, Lindsten R. The effect of sucking habits, cohort, sex, inter-canine arch widths, and breast or bottle feeding on posterior crossbite in Norwegian and Swedish 3-year-old children. Am J Orthod Dentofac Orthop 1994;106:161-166. 17. Strang RH. The fallacy of denture expansion as a treatment procedure. Angle orthod 1949;49:12-7. 18. Ruttle AT, William Quigley, Crouch JT and George E. Ewan. A serial study of the effects of digit-sucking. J Dent Res 1953;32(6):739-748. 19. Brader AC. Dental arch form related with intraoral forces: PR=C. Am J Orthod 1972;61(6):541-561. 20. Paola Cozza, Tiziano Baccetti, Lorenzo Franchi, Manuela Mucedero, Antonella Polimeni. Transverse features of subjects with sucking habits and facial hyperdivergency in the mixed dentition. Am J Orthod Dentofac Orthop 2007;132:226-9. 21. Nie Q, Lin J. Comparison of intermaxillary tooth size discrepancies among different malocclusion groups. Am J Orthod Dentofac Orthop 1999;116:539-44. 22. Cole TM (2002). WinEDMA: Software for Eulidean distance matrix analysis. Version 1.0.1 beta. Kansas city: University of Missouri- Kansas City School of Medicine. 23. Weinstein S. Minimal force in tooth movement. Am J Orthod 1967;53:881-903. 24. Straub WJ. Malfunction of the tongue Part I. The abnormal swallowing habit: It’s cause, effects and results in relation to orthodontic treatment and stretch therapy. Am J Orthod 1960;46(6):404-24. 25. Tulley WJ. A critical appraisal of tongue thrusting. Am J Orthod 1969;55(6):640-50. 26. Cleall JF. Deglutition: A study of form and function. Am J Orthod 1965;51(8):566-594. 27. Hanson ML, Cohen MS. Effects of form and function on swallowing and the developing dentition. Am J Orthod 1973;64(1):63-82. 28. Subtelny J.D, Subtelny J.D. Oral Habits- Studies in Form, Function and Therapy. Am J Orthod Dentofac Orthop 1973;43(4):347- 383

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareCRVO: OPTIC NERVE DIAMETER ALONG WITH VARIABLE EJECTION FRACTION A RISK FACTOR? English5760Sanjoy ChowdhuryEnglish Hitesh PatelEnglish Pratik BhosaleEnglish Nilanjan ChowdhuryEnglishBackground: Retinal vein occlusion is second commonest retinal vasculopathy. Narrow optic nerve diameter is a risk factor. A variation in ventricular ejection fraction can alter pressure inside any blood vessel. Purpose: To study ejection fraction in all cases of Retinal Vein Occlusion Methods: Prospective nonrandomised cohort study since 1997 to 2012.Standard protocol was followed in all cases of RVO.A/B Scan and echocardiography was done to assess optic nerve diameter and ejection fraction respectively. Equal number of matched controls was registered. Results: 82 cases of RVO, mean age 48.5, male preponderance in below 40 years. Variable ejection between preserved (pEF>50%) and reduced (rEFEnglishRetinal vein occlusion, Optic nerve diameter, Ejection fractionINTRODUCTION Obstructions in the retinal venous flow by thrombus formation, external compression or disease of wall of the veins lead to Retinal vein occlusion. This is the second commonest retinal vasculopathy exceeded only by diabetic retinopathy. [1] Engorgement and dilation of retinal vein with secondary intraretinalhaemorrhages along with intraretinal edema, retinal ischemia, retinal exudates andmacular edema are the main features of the disease.(Figure 1,1a) .Conclusive pathogenesis of thisdisorder with visually distressing consequences is still a matter of research. Histopathological studieshave shown thrombus formation at or near lamina cribrosa. [2]Presumablyeyes with shorter lamina cribrosa and narrow scleral canal are vulnerableto thrombus formation [3]. This anatomical risk factor combined with alteredcardiovascular haemodynamic can be the reason behind CRVO. A variationin ventricular ejection fraction can alter pressure inside any blood vessel [4].The aim of the study is to evaluate optic nerve diameter and cardiac ejectionfraction in CRVO. MATERIALS AND METHODS All the cases with central retinal vein occlusion attending Bokaro GeneralHospital from1997 to 2012were included in this study. Methods comprised of detailed history including that of any systemic disease in each case along with proper demographic data. Routine ophthalmic examination was performed and documented properly. Ophthalmic ultrasound (A/B Scan) was the tool to assess optic nerve diameter and cardiac ejection fraction was measured by echocardiography during this study. Equal numberof age and sex matched controls that came for refraction was registered.Optic nerve diameter was measured with A and B scans at the level oflamina cribrosa. Method ology is elaborated in Figure 2, 3, 4. Axial length of each eye was measured by biometry as shown in figure 4a. Echo cardiacparameters were recorded by echocardiography specialist whowas unaware of the case. Ejection fraction was measured on each follow up. Difference between Lowest Efr and highest Efr was calculated and statisticallyanalysed by univariate regression. Other covariates were also recorded e.g. Blood pressure was measured once, in a seated position, after at least 5 minutesrest. Hypertension was defined as known treated hypertension confirmed bycurrent use of antihypertensive medications and/or a systolic blood pressure(BP) of 140 mmHg and/or diastolic blood pressure of 90 mmHg. Diabeteswas defined as a self-reported history of diabetes confirmed by current antidiabetic therapy and/or fasting blood glucose of 7 mmol/L. Body mass index(BMI) was defined as weight/height2 in kg/ m2. A BMI between 25 and 30 kg/m2 was classified as overweight, and a BMI greater than 30 kg/m2 as obese.Fasting blood samples were obtained for the measurement of serum creatinineand plasma glucose. Plasma triglycerides and total cholesterol levels weremeasured by routine enzymatic methods. Renal function was assessed fromestimates of glomerular filtration rate using Modification of Diet in RenalDisease (MDRD) formula, based on plasma creatinine. Data management and analysis The data forms were checked for accuracy and completeness in the field before data entry.Continuous variables were presented as mean ± SD.Differences in continuous variables among the cases and controls were compared with independent samples t-test.Differences in categorical variables between cohorts and controls were compared with chi-square test p value 50%) and reduced (rEF50% at least 4 follow ups over aperiod of 3 months in 52 cases of CRVO which was significant (pEnglishhttp://ijcrr.com/abstract.php?article_id=449http://ijcrr.com/article_html.php?did=4491. Klein R, Moss SE, Meuer SM, Klein BE. The 15-year cumulative incidence of retinal vein occlusion: the Beaver Dam Eye Study. Arch Ophthalmol 2008; 126:513–8. 2. Green WR, Chan CC, Hutchins GM, Terry JM. Central retinal vein occlusion: aprospective histopathology study of 29 eyes in 28 cases. Retina 1981; 1:27–55. 3. Sanjoy Chowdhury. Optic nerve diameter in central retinal vein occlusion: A riskfactor? AIOC 2000 Proceedings (Col. Rangachari award 2000). 4. J W Kiel. The ocular circulation. Edited by DE Granger and J Granger.Collouquiumseries in integrated systems physiology: from molecule to function. P47. Morganand Claypool Life sciences 2011. 5. Hayreh SS. Classification of central retinal vein occlusion. Ophthalmology 1983; 90:458–74. Abbreviations used: RVO: Retinal Vein Occlusion pEF: preserved ejection fraction rEF: reduced ejection fraction CRVO: Central Retinal Vein Occlusion HFNEF: Heart failure with normal ejection fraction HFpEF: Heart Failure with preserved ejection fraction OND: Optic Nerve Diameter

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241718EnglishN2015September20HealthcareTYPE 2 DIABETES: A REVIEW OF CURRENT TRENDS English6166Ashwini N. S.EnglishDiabetes mellitus type 2 (DM) is a chronic metabolic disorder in which the frequency has become worldwide. Because of this, there is always an epidemic in some countries of the world, with the number of people affected is expected that within the next decade to double due to the increasing aging of the population, thus increasing the existing burden of health care providers, particularly in the less developed countries. This review is based on a search Medline, Cochrane Database of Systemic journals and references to the literature list at the bottom.Subject heading and key wordsused include type 2 diabetes, the prevalence, the current diagnosis and therapy in progress. Onlyarticles in English were included. Early detection and diagnosis is still for the World Health Organization (WHO) and the American Diabetes Association (ADA) criteria include clinical and laboratory parameters. No cure has been found yet for the disease;Treatment details, however, include lifestyle changes, treatment of obesity, oral hypoglycemic agents and insulin sensitizers such as metformin, a biguanidethat reduces insulin resistance, is still the firstline drug specifically for overweight patients.Other effective drugs includesnonsulfonylureasecretagogues, thiazolidinediones, inhibitors of alpha-glucosidase and insulin.Recent research in the pathophysiology of type 2 diabetes has led to the introduction of new drugs such as glucagon-like peptide 1 analogoues: Dipeptidyl peptidase-IV inhibitors of the sodium-glucose cotransporter 2 and 11s-1-hydroxysteroid dehydrogenase, glucokinase activators of insulin release and pancreatic-G-protein-coupled fatty-acid-receptor agonists, glucagon receptor antagonists, inhibitors of the hepatic metabolism of glucose production and fast release bromocriptine. Inhaled insulin has been approved in 2006, but was withdrawn from the market due to low patronage. EnglishType 2 diabetes, Diagnosis, Administration, New medicinesINTRODUCTION Diabetes mellitus (DM) is perhaps one of the oldest diseases known to mankind. It was first mention in Egyptian manuscript about 3000 years ago1 . In 1936 the difference between type 1 and type 2 DM madesignificantly2 .Type 2 diabetes is described (as non-insulin dependent DM earlier) as a component of the metabolic syndrome in 19883 . Type 2 diabetes,the most common form of DM characterized by hyperglycemia, insulin resistance and insulin deficiency4 .Type 2 DM results from the interaction of genetic, environmental and behavioral risk factors5, 6. People with type 2diabetes are more susceptible to various forms of short- and long-term complications that often lead to premature death. This trend of increasing morbidity and mortality is seen in patients with diabetes type 2 because of truism that type DM, insidious onset and late recognition, particularly in poor developing countries such resources Africa7 . Epidemiology It is estimated that 366 million people had DM in 2011; 2030 will be 552 million increased 8.The number of people with type 2 diabetes is increasing in all countries with 80% of people with DM living in low and middle income countries.DM caused 4.6 million deaths in 20118 . It is estimated that 439 million people suffer from type 2 diabetes since year 20309 . The incidence of type 2 diabetes ranges from one geographic area to another, due to lifestyleand environmental risk factors10. The literature has shown that there is little available data on type 2 diabetes prevalence in Africa as a whole.Study data trends based tip of Africa to show a dramatic increase in the prevalence of rural and urban areas, and both gender equally11. The majority of the weight in Africa appears to be type 2 DM, to be less than 10% of cases ofDMis type 1 DM11.A 2011 Centre for Disease Control and Prevention (CDC) report estimates that DM affects about 25.8 million people in the United States (7.8% of the population) in 2010, with 90% to 95% of which is type 2 -DM12.It is expected that  the prevalence of diabetes in patients with type 2 diabetes, it is important that adults will increase in the next two decades, and much of the increase will be in developing countries, where the majority of patients are aged between 45 and 64 years13.It is estimated that the state equal to or even greater than the previous one in the developing countries, which will be completed double taxation arising from the current trend of moving from non-communicable diseases contacts Lifestyle, Genetics, and Medical Conditions Type 2 diabetes is caused primarily by lifestyle factors and genetics15. A number of lifestyle factors are known to develop type 2 diabetes. These are physical inactivity, lack of exercise, cigarettesmoking and generous consumption of alcohol16. Obesity has been found to contribute about 55% of cases of type 2 DM17.The increased rate of childhood obesity between the 1960s and 2000s is believed to have led to an increase of type 2 diabetes in children and adolescents18. Environmental toxins may contribute to the recent increase in the rate of type 2 diabetes.Found a weak positive correlation between the concentration in the urine of bisphenol A, a component of some plastics, and the incidence of type 2 DM19.There is a strong hereditary genetic connection in type 2 DM with relatives (especially first degree) with type 2 diabetes increases the risk of type 2 diabetes significantly. Agreement between monozygotic twins is close to 100%, and about 25% of People with the disease have a family history of DM20.Recently discovered genes associated significantly with type 2 diabetes include TCF7L2, PPARG, FTO, KCNJ11, NOTCH2, WFS1, CDKAL1, IGF2BP2, SLC30A8 and JAZF1 HHEX. KCNJ11 (potassium internal channel correction, subfamily J, member 11), encodes the islet ATPsensitive potassium channel Kir6.2 and TCF7L2 (transcription factor 7-like 2) regulates proglucagon gene expression and thus production glucagonlike peptide-121. Furthermore, obesity (which is an independent risk factor for type 2 diabetes) is strongly inherited22.Monogenic forms as Maturityonset diabetes of the young (MODY), constitutes up to 5% of cases23. There are many diseases that can potentially cause or exacerbate type 2 diabetes, including obesity, hypertension, high cholesterol (combined hyperlipidemia), often referred to as conditions of metabolic syndrome (also known as syndrome X, Reaven’s syndrome known) 24. Other causes include acromegaly Cushing’s syndrome, thyrotoxicosis, pheochromocytoma, chronic pancreatitis, cancer and drugs25. Additional factors found that the risk for type 2 diabetes with increasing age, diet rich in fat 26 and a less active lifestyle27. Pathophysiology Type 2 diabetes is an insulin sensitivity due to insulin resistance, reduced insulin production, and pancreatic beta cells can failure28, 29. This results in a reduction in glucose transport in the liver, muscle and fat cells. There is an increase in the distribution of fat in hyperglycemia. The inclusion of the modified alpha-cell function has been detected recently in the pathophysiology of type 2 DM30. As a result of dysfunction, glucagon and hepatic glucose levels which are increased during fasting, is not removed with a meal. Since insufficient insulin levels and increased insulin resistance that results in hyperglycemia. The incretins are important mediators of the intestine of the release of insulin, and in the case of GLP-1, suppress glucagon. Although the activity of GIP is impaired in individuals with type 2 diabetes remain intact GLP-1 insulinotropic action and hence the GLP-1 is a potentially useful therapeutic option30.However, since the GIP; GLP-1 is rapidly inactivated by DPP-IV in vivo. Two therapeutic approaches have been developed to solve this problem: GLP-1 analogues with increased half-life and DPPIV inhibitors that prevent the breakdown of endogenous GLP-1 and GIP30.The two classes of agents have shown promise improved, with the ability to not only normalize postprandial glucose and fasting, but also for the function and beta cell mass. Investigations are under way, the role of mitochondrial dysfunction in the development of insulin resistance and etiology of type 2 DM31. Also very important is adipose tissue, as endocrine organ hypothesis(various Adipokytokines secretion, ieleptin, TNF-alpha,resistin and adiponectin resistance Insulin involved and possibly a dysfunction of beta cells) 30. The majority of people with type 2 diabetes are overweight, with central visceral obesity. Therefore, fat plays an important role in the pathogenesis of type 2 diabetes, although the prevailing theory used to explain this connection, the gate / visceral case gives a key role in non-esterified high fatty acid concentrations, two new theories resulting trumpets fat storage syndrome (triglyceride accumulation in muscle, liver and pancreatic cells). These two assumptions are the framework for the study of the interaction of insulin resistance and beta cell dysfunction in type 2 diabetes and between us obesogenicenvironment and the risk of DM in the next decade30. Screening and Diagnosis Tests for screening and diagnosis of DM are readily available. The recommended test is the same as for the diagnosis of prediabetes or DM32. Although approximately 25% of patients already type 2 diabetes have microvascular complications at diagnosis, suggesting a diagnosis, so it is a positive effect equivalent to that had the disease for more than five years old, when diagnosis33. It is still in the American Diabetes Association (ADA) guidelines of 1997 or the World Health Organization (WHO) national criteria diabetic group in 2006, which is for a single elevated glucose reading levels with the primary symptoms (polyuria, polydipsia, and polyphagia weight loss) Otherwise set values twice or fasting plasma glucose (FPG) ≥7.0 mmol / L (126 mg / dL) or Oral glucose tolerance test (OGTT), two hours after the oral dose of plasma glucose ≥11.1 mmol/L (200 mg/dL) 32 recommen-dations from the 1997 ADA for the diagnosis of DM emphasis fasting glucose, whereas WHO concentrates on OGTT32. The glycosylated hemoglobin (HbA1c), and fructosamine, is always useful to determine the control of blood glucose with time. But doctors often practiced using other measures than those recommended. In July 2009, the International Expert Committee (IEC)recommended additional diagnostic criteria for HbA1c result ≥6.5% for DM. The Commission proposed that the term prediabetes can be eliminated, but defines the range of HbA1c ≥6.0% and detection Englishhttp://ijcrr.com/abstract.php?article_id=450http://ijcrr.com/article_html.php?did=4501. Ahmed AM. History of diabetes mellitus. Saudi Med J 2002 Apr; 23(4): 373- 378. 2. Diabetes mellitus history- from ancient to modern times. http://science.jrank.org/pages/2044/Diabetes-Mellitus.html. 3. Patlak M. New weapons to combat an ancient disease: treating diabetes. FASEB J 2002 Dec; 16(14): 1853. 4. Maitra A, Abbas AK. Endocrine system. In: Kumar V, Fausto N, Abbas AK (eds). Robbins and Cotran Pathologic basis of disease (7th ed) 2005. Philadelphia, Saunders; 1156-1226. 5. Chen L, Magliano DJ, Zimmet PZ. The worldwide epidemiology of type 2 diabetes mellitus: present and future perspectives. Nature reviews endocrinology. www.nature.com/uidfinder. 6. Genetic basis of type 1 and type2 diabetes, obesity, and their complications. Advances and emerging opportunities in diabetes research: a Strategic Planning report of the DMICC. www2. niddk.nih.gov/NR. 7. Azevedo M, Alla S. Diabetes in sub-saharan Africa: kenya, mali, mozambique, Nigeria, South Africa and zambia. Int J Diabetes DevCtries 2008 Oct; 28(4): 101-108. 8. Global burden of diabetes. International Diabetes federation. Diabetic atlas fifth edition 2011, Brussels.http://www.idf.org/ diabetesatlas. 9. Chamnan P, Simmons RK, Forouhi NG, Luben R. KhawKy, Wareham NJ et al. Incidence of type 2 diabetes using proposed HbA1c diagnostic criteria in the EPIC-Norflok cohort: Implication for preventive strategies. http://care.diabetesjournal.org. 10. Zimmet P, Alberti KG, Shaw J. Global and societal implications of the diabetes epidemic. Nature 2001 Dec; 414(6865):782-787. 11. Mbanya JC. The burden of type 2 diabetes mellitus in the African diaspora.www.medscape.com/view article/560718_2. 12. Department of Health and Human Services. Centres for Disease Control and Prevention, 2011. National diabetes fact sheet: national estimates and general information on diabetes and prediabetes in the United States, 2011. http://www.cdc.gov/diabetes/ pubs/pdf/ndfs_2011.pdf. 13. Wild S, Roglic G, Green A, Sicree R, King H. Global prevalence of diabetes: estimate for the year 2000 and projections for 2030. Diabetes Care 2004; 127(5): 1047-1053 . 14. Yach D, Hawkes C, Gould CL, Hofman KJ. The global burden of chronic diseases: overcoming impediments to prevention and control. JAMA 2004 Jun; 291(21): 2616-2622. 15. Ripsin CM, Kang H, Urban RJ. Management of blood glucose in type 2 diabetes mellitus. Am Fam Physician 2009 Jan; 79(1): 29-36. 16. Hu FB, Manson JE, Stampfer MJ, Colditz G, Liu S, Solomon CG, et al. Diet, lifestyle, and the risk of type 2 diabetes mellitus in women. N Engl J Med 2001 Sep; 345(11): 790-797. 17. Prevalence of overweight and obesity among adults with diagnosed Diabetes United States, 1988-1994 and 1999-2000”Centers for Disease Control and Prevention (CDC) (November 2004) MMWR. Morbidity and Mortality Weekly Report; 53(45): 1066-1068. 18. Barlow SE and the Expert committee. Expert committee recommendations regarding the prevention, assessment and treatment of childhood and adolescent overweight and obesity: Summary report. Paediatrics 2007; 120: S164-S192 . 19. Lang IA, Galloway TS, Scarlett A, Henley WE, Depledge M, Wallace RB, et al. Association of urinary bisphenol A concentration with medical disorders and laboratory abnormalities in adults. JAMA 2008 Sep; 300(11): 1303-1310. 20. Rother KI. Diabetes treatment–bridging the divide. N Engl J Med 2007 Apr; 356(15): 1499-1501. 21. McCarthy MI. Genomics, type 2 diabetes, and obesity. N Engl J Med 2010 Dec; 363(24): 2339-2350. 22. Walley AJ, Blakemore AI, Froguel P. Genetics of obesity and the prediction of risk for health. Hum Mol Genet 2006 Oct; 15(Spec No 2): R124-R130. 23. Camastra S, Bonora E, Del Prato S, Rett K, Weck M, Ferrannini E; EGIR (European Group for the Study of Insulin Resistance). Effect of obesity and insulin resistance on resting and glucoseinduced thermogenesis in man.Int J ObesRelatMetabDisord 1999 Dec; 23(12):1307-1313. 24. Alberti KG, Zimmet P, Shaw J; IDF Epidemiology Task Force Consensus Group. The metabolic syndrome–a new worldwide definition. Lancet 2005 Sep; 366(9491):1059-1062. 25. Powers AC. Diabetes mellitus. In: Fauci AS, Braunwauld E, Kasper DL, Hauser SL, Longo DL, Jameson JL, Loscalzo J (eds). Harrison’s Principles of Internal Medicine.17th ed, New York, McGraw-Hill; 2008: 2275-2304. 26. Jack L Jr, Boseman L, Vinicor F. Aging Americans and diabetes. A public health and clinical response. Geriatrics 2004 Apr;59(4):14-17. 27. Lovejoy JC. The influence of dietary fat on insulin resistance. CurrDiab Rep 2002 Oct; 2(5):435-440 28. Kahn CR. 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