International journal of Clinical Biochemistry and Research

Page No. 1-5

Authors: PM Abraham Sam Rajan

Introduction: Clinical biochemistry laboratory uses many techniques for measuring the concentration of specific biochemical substances. All these techniques are subject to interference from variety of sources. The aim of our study is to see the changes in concentration of serum bilirubin on exposure to special blue light of 440nm for six hours and to study the usefulness of photolysis for the elimination of bilirubin interference in the estimation of creatinine, glucose, and alkaline phosphatase. Method: We have taken 218 samples and the samples were analyzed for the estimation of total bilirubin, direct bilirubin, creatinine, glucose and alkaline phosphatase before and after photolysis. Results & Conclusion: There was significant decrease in the concentration of total and direct bilirubin after photolysis. There was a significant negative interference of creatinine and glucose and significant positive interference of alkaline phosphatase with an increasing concentration of bilirubin which was also eliminated by photolysis.

Page No. 11-14

Authors: Asmabi M, Netravathi Basappa Sajjan, Tousif Devale, Shivaleela M Birasar, Shireen Quadri

Background: The leading cause of chronic renal disease and end stage renal disease is diabetic nephropathy, so early recognition is very important. The important biochemical test for diagnosis of renal function is estimation of glomerular filtration rate (GFR). The accurate methods of GFR estimation are Inulin clearance and 24h urine collection, which are expensive and time consuming and require experts to perform. Hence some formulas have been developed for the GFR measurement. Objective: To find out the prevalence of renal dysfuntion based on the levels of serum creatinne and estimated GFR, calcuted using Cockcroft-Gault (CG) and Modification of Diet in Renal Disease (MDRD) equations in type 2 diabetic patients. Materials and Methods: A case control study was carried out on 50 type 2 diabetic patients and 50 healthy controls. Renal function was analyzed through serum creatinine and GFR estimated using the CG and MDRD equations. Results: Among the 50 diabetic patients, 34% show high level of serum creatinine i.e. > 1.4 mg/dl, while the low eGFR that is < 90ml/mim is seen in 58% of diabetic patients by MDRD equation and 64% of patients by CG equation. The mean serum Creatinine level is significantly high p value (<0.001) in diabetic patients when compared with controls. The eGFR is significantly low p value (< 0.001) in diabetic patients when compared with controls. Conclusion: The prevalence of renal dysfunction based on serum creatinine level is low when compared with eGFR by MDRD and CG equation, indicating the significance of these equations in estimating GFR. Hence for the better assessment of renal function these equations can be used as compliment to the levels of serum creatinine alone.

Page No. 19-21

Authors: Deepa Singh

Studies have shown that Vitamin D is necessary for normal insulin secretion. Vitamin D reduces insulin resistance through regulation of the insulin receptor gene and its effects on calcium and phosphorus metabolism. Therefore, deficiency of vitamin D results in decreased insulin secretion and increased insulin resistance, both of which are characteristics of type 2 Diabetes mellitus. The predisposition of Vitamin D deficiency to Glucose intolerance via altered insulin secretion may be done either through a direct action via Vitamin D receptor (VDR) activation or indirectly via calcemic hormones and also via inflammation. The study is aimed to measure and correlate Vitamin D status with serum level of fasting and postprandial Glucose, Total Cholesterol and Triacylglycerol level in individuals of Udaipur. The study was carried out on total 300 vegetarian subjects, out of which 190 were females and 110 were males with an average age of 54.25±8.94 years. Subjects were divided into two categories depending upon their serum total Vitamin D value. Category A included subjects with total Vitamin D value below 10 ng/ml and category B included subjects with total Vitamin D value between 30-70 ng/ml. The subjects with total Vitamin D value between 10-30 ng/ml were excluded from the study. Results clearly indicated that Category A with Vitamin D value below 10 ng/ml had significantly higher serum fasting, post prandial Glucose, Total Cholesterol and Triacylglycerol level as compared to subjects with Vitamin D value between 30-70 ng/ml (category B) with significance (p value <0.05). Vitamin D deficiency can lead to dysregulation of Glucose and lipid metabolism. Supplementation of Vitamin D in Vitamin D deficient individuals may reduce the risk of Diabetes mellitus and dyslipidaemia by improving insulin secretion and Glucose tolerance.

Page No. 25-29

Authors: Dhiraj J. Trivedi, Vishwaraj PK

Aim: To evaluate erythrocyte surface electric charge in β- thalassemia by using Alcian Blue dye binding. Objective: Estimate the binding of AB dye to Sialic acid on the surface of thalassemia and normal, healthy erythrocytes. Compare negative charge due to Sialic acid on the surface of erythrocyte of thalassemia with healthy control subjects. Result: The study was performed on 60 thalassemia subjects (39 male, 21 female, mean age 8.3years) and 30 age and sex matched normal control subjects (17 male, 13 female, mean age 6.8years). Hematological parameters analyzed on beta thalassemia subjects’ shows significant difference among control and Thalassemia group. Levels of HbA2 showed a marginal difference (p=0.041). Cationic dye, Alcian Blue, in PBS (pH7.4) was used as a reagent to demonstrate binding of the dye to Sialic acid. Results of dye bound to thalassemic erythrocytes was significantly less (p<0.05) as compared to binding on erythrocytes of the control group. Conclusion: We believe, our data support the perception that there is decreased negative charge on the erythrocyte membrane in thalassemia reflected by cationic Alcian Blue dye binding.

Page No. 33-38

Authors: Chakrabarti C, Jatin D. Patel, Hitesh Mewada

Aim: Evaluation of different alternatives for Blood Gas Analysis of paediatric Cardiac surgical patients. Objectives: Arterial blood gas (ABG) analysis is very important tool for assessment of the clinical condition of patients in various critical conditions; however, arterial puncture may associate with many complications, particularly in paediatric patients. This study is conducted to determine whether venous blood gas (VBG) values can replace arterial blood gas values in paediatric age group patients undergoing cardiac surgery for congenital heart disease. To study correlation between arterial blood gas and venous blood gas we have examined arterial as well as venous blood pH, PCo2, PO2, HCO3- , Base excess, Ct O2, SO2 and Ct CO2. Method: The study was conducted during June 2008 to December 2010 at an academic tertiary care hospital in Ahmedabad where DM and MCH super specialist are available as a consultant. Totally, 50 paediatric patients suffering from congenital heart disease included in this study and arterial as well as venous blood sample were collected for blood gas analysis. Result: There was no significant difference observed between arterial and venous blood respectively in pH (7.37 +/-0.04, 7.33+/-0.04), PCo2 (35.51+/- 5.08, 43.54+/-4.94), HCO3- (20.64+/-1.69, 21.30+-1.81) but significant difference observed in PO2 (77.26+/-37.43, 38.05+/-6.81), Ct O2 and SO2. Conclusion: We do not recommend venous blood sample as an alternate of arterial blood sample for blood gas analysis as there is poor correlation in PO2, CtO2 and SO2.

Page No. 43-46

Authors: Anju Sharma, Neha, Sahiba Kukreja, Jaskiran Kaur, Amandeep Kaur

Background: The prevalence of thalassemias and various hemoglobinopathies are a major public health problem in Punjabi population. These cases are mainly overlooked and thus only treated as a part of anaemia. Thus it is worthwhile to study its prevalence and occurrence. The main objective of our study was to find the prevalence of hemoglobinopathies in Punjab and implement strategies for preventing marriages between two carriers for effective control of these diseases. Material and Methods: This was a case control retrospective study for screening for haemoglobinopathies of patient samples collected in biochemistry clinical laboratory, SGRDIMSAR, Vallah, Amritsar from 1 June 2015 to 30 May 2016 using automated Interlab Genios S electrophoresis system. EDTA samples were collected and 30 µL of lysate was applied on Genios S electrophoresis system in alkaline haemoglobin buffer. The results were interpreted by densitometer were obtained as a graph and interpreted. Results: In this study, out of 187 samples we received for screening of haemoglobinopathies, total of 143 (78.5%) were found to have a normal electrophoretic pattern and 44 patients (21.4%) were found to have one or other form of hemoglobinopathy. Conclusion: Punjabi population has high prevalence of haemoglobinopathy screening being an affordable and accessible way to detect carriers, it should be made mandatory in high schools, before marriage and antenatal clinics. Also, public education and awareness for screening in areas of high prevalence of consanguinity should be made mandatory.

Page No. 56-59

Authors: Prakash S. Hundekar, Adinath Suryakar, Aarti Karnik, Rahul Ghone, Sonali Bhagat

Background: Sickle cell anaemia (SCA) is a hereditary disorder, associated with severe haemolytic anaemia, periodical vasooclussive pain, and premature death. Prevalence of SCA is alarmingly high in north Maharashtra comprising districts of Nandurbar, Dhule, Jalgaon and Nashik. Non heme iron and depleted erythrocytic glutathione are associated with generation of free radicals leading to oxidative stress. Oxidative stress may enhance haemolysis by damaging erythrocytic membrane. Aim and Objectives: To evaluate serum iron, erythrocytic MDA and erythrocytic Glutathione in relation with possible markers of oxidative stress in the steady state of SCA individuals. Methods: A total of 90 subjects were recruited in the study including 30 sickle cell trait (HbAS), 30 sickle cell disease (HbSS) and 30 healthy controls. In all subjects serum iron and bilirubin, erythrocytic Malondialdehyde (MDA) and glutathione (GSH) concentration were investigated simultaneously. Results: Serum iron level found significantly (p<0.001) lowered in sickle cell trait (HbAS) while significantly (p<0.001) elevated in sickle cell disease (HbSS) compared to control. Serum bilirubin and Erythrocytic MDA found significantly (p<0.001) elevated in HbAS and HbSS. Erythrocytic glutathione was found significantly (p<0.001) depleted in both HbAS and HbSS in comparison of control. There is positive correlation between erythrocytic MDA and serum iron. Conclusion: Our study suggests, elevated serum iron and bilirubin is result of excessive hemolysis. Depleted Erythrocytic glutathione and elevated malondialdehyde reveals hyphened oxidative stress, further which could be sickling and hemolytic propagator aspect in sickle cell anaemia.

Page No. 64-67

Authors: Sreekumari RG, Sandhya CV

Background: The hypermetabolic state seen in hyperthyroidism causes increased oxygen consumption resulting in an increased formation of Reactive Oxygen Species leading to tissue injury. Increased oxidative stress characterised by elevated levels of free radicals and diminished antioxidant status has been described in animal models as well as in patients with hyperthyroidism. Thyroid gland is characterized by high activity of the antioxidant enzyme Glutathione Peroxidase (GSHPx) and this has been studied much reflecting the imbalance of antioxidant enzyme systems in thyroid dysfunction. However, contradictory results have been obtained regarding plasma and erythrocyte GSHPx levels in hyperthyroidism. Studies have also shown that antithyroid drugs like Methimazole and propyl thiouracil have antioxidant effects. How the treatment with these drugs affect the antioxidant status is also a matter of debate. Hence, the present study aimed to evaluate the erythrocyte Glutathione Peroxidase levels in hyperthyroidism in comparison with normal controls and evaluate the effect of methimazole treatment. Methods: The study was carried out in patients attending the special investigation laboratory in the department of Biochemistry of our college for assessment of their thyroid function. The study groups included: Group 1 (n = 40) consisting of healthy euthyroid controls. Group 2: (n=30) clinically diagnosed hyperthyroid patients, 17 were untreated fresh cases (series1) and 13 patients were undergoing treatment with methimazole for the past one month (series2). Erythrocyte Glutathione peroxidase enzyme levels were analyzed in blood samples collected from these subjects along with thyroid function tests T3, T4 and TSH. Results & Conclusion: Comparison of the mean values showed that the elevated levels of thyroid hormones have come down in the Methimazole (MMI) treatment group as expected. There were very low TSH levels in the untreated group whereas in treated group TSH levels were at the lower limit of normal range. The mean GSH Px levels in untreated hyperthyroid patients were significantly elevated (6.970± 0.302μmoles/L) compared to normal controls (6.117 ± 0.133μmoles/L). The GSHPx levels in the treated group were closer to the GSHPx values of the control group.(6.125 ± 0.482μmoles/L)

Page No. 73-76

Authors: Shruthi Rai P, Sukanya Shetty, Roopa Bhandary, Ashalatha V Rao

Introduction: Reference Intervals are the most common decision making tools for interpretation of various biochemical reports. Age, sex, ethnicity, diet, physical and socio-economic conditions, affect the physiology of population. Hence it is necessary to setup reference values that are applicable to specific population rather than to borrow them from other population based studies or from literature from reagent kit manufacturers. So we intend to establish the reference intervals of serum bilirubin profile in southern coastal Karnataka population. Methodology: Hospital based cross sectional study was done involving 200 subjects. The subjects were randomly selected from the population, which included medical students, health professionals, and those who attended different OPDs and health camps organized by the K S Hegde hospital for general check-ups under health plan scheme. Total bilirubin and direct bilirubin was estimated by dichlorodiphenyl diazonium method. Indirect bilirubin (Total bilirubin-Direct bilirubin) was calculated. Results: The reference intervals observed for men and women are as follows: Total bilirubin (0.1-1.3)mg/dl and (0.1-1.38)mg/dl, serum direct bilirubin (0-0.5)mg/dl and (0-0.3)g/dl, and serum indirect bilirubin (0.1-0.9)mg/dl and (0.1-1.3)mg/dl respectively. With the advancement of age, a difference in the reference interval was observed among both the sex. Conclusion: The present study showed a difference in the reference intervals between males and females and also a categorical age wise difference was obtained.

Page No. 81-84

Authors: K. Vani, G. Sasirekha

Aim: To assess the biochemical parameters of renal and hepatic function in women with GDM and compare the results with those of normal pregnant women. Materials and Methods: Cases: The study group consisted of 45women with GDM. Controls: 45 healthy pregnant women without any complications of pregnancy were taken as controls. The parameters of renal function which include serum urea, creatinine, uric acid and liver function which includes serum bilirubin, albumin, alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase were estimated between 24 to 28 weeks of gestation using a fully automated clinical chemistry analyzer. Results: Serum creatinine and uric acid levels were found to be significantly elevated in cases. On the contrary, the serum albumin levels were significantly reduced in cases. No significant change was found in serum urea, bilirubin, alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase levels between the groups. Conclusion: The increased serum creatinine and uric acid levels in cases suggest that GDM may jeopardize the renal function and also increase the risk for future kidney disease in these women

Page No. 88-93

Authors: Khot Varsha V, Yadav KS, Haldankar VA

Introduction: Type 2 diabetic nephropathy (T2DN) develops inflammation in kidney which leads to failure. Study designed to evaluate risk of diabetic nephropathy (DN) by inflammatory gene expressions (IL6, IL10) and renal parameters at the early stage in type II diabetes mellitus (T2DM) in Indian population. Methods: This study includes 241 subjects (118 men, 123 women, and age ranges 30-70 years, distributed in three groups on the basis of age. Subjects recruited after screening for T2DM by measurement of blood glucose in fasting, post-prandial and glycosylated hemoglobin, microalbumin was measured from spot urine for screening of DN. Results: Analysis of data of routinely used renal parameters found within reference interval except e-GFR marginally deviated from control group (p <.00). Other parameters showed marginal significance within and between the groups. Pro-inflammatory cytokines IL-6 and IL-10 evaluated by rt-PCR, showed high degree of significance (p <.00) in both study groups. Results of this study suggest that measurements of renal parameters are not enough to diagnose early stage of diabetic nephropathy (DN) and eventually to prevent progression of the disease. Conclusion: It was concluded that early measurement of IL6, IL10 along with direct and indirect renal marker was suggested to prevent morbidity & mortality in DN.