International journal of Clinical Biochemistry and Research

73-76

Authors: Seema Verma, Arunima Karkun, Huma Naz Siddiqui

The present study was carried out to evaluate the antimicrobial activity of clove oil extracted from clove. Screening for antibacterial and antifungal of clove oil was done using agar well diffusion. Four Bacterial and ten fungal species were taken. According to the result clove oil was more effective against Bacteria as compared to Fungi. We can conclude by this study that clove oil works better as antibacterial. It was also observed that by increasing concentration of clove oil the growth of fungi was decreased.

83-88

Authors: Radhika Krishnaswamy, Soniya Lukose

Background: The Glomerular filtration rate (GFR) is considered to be the most reliable measure of the functional capacity of the kidneys and is it has proved to be the most sensitive and specific marker of changes in overall renal function. The accurate measurement of the GFR is time consuming, expensive, and not practical for routine clinical use.1 Though Inulin clearance is considered to be the gold standard test, it has not been used widely because it time consuming, expensive, requires continuous infusion, and constraining for the patient. 2, 3, 4 Creatinine clearance is most commonly used for the estimation of GFR. The need to collect a urine sample remains a major limitation of the creatinine clearance test.5 Many formulae have been developed to transform serum creatinine so that it may accurately reflect, GFR.6 Variations in creatinine production owing to age and sex related differences in muscle mass have been measured and incorporated into the formulas to improve the GFR.7 In adults, the most commonly used formulae are those derived from the Modification of Diet in Renal Disease (MDRD) and the Cockcroft-Gault (CG), which have not been validated in Indian population. 8 This study was aimed to compare the diagnostic performance of the MDRD Cockcroft-Gault formulae, and Measured Creatinine Clearance in Indian population. Materials and Methods: In 370 subjects Creatinine Clearance was calculated by Routine Creatinine Clearance test using serum and urine Creatinine and also through MDRD and the Cockcroft-Gault formulae. Subjects were divided into two groups based on the Measured Creatinine Clearance values. Group A consisted of subjects with Measured Creatinine Clearance >60 ml/min. Group B consisted of subjects with Measured Creatinine Clearance <60 ml/min Statistical analysis: Correlation (Pearsons correlation) between the values of Clearance obtained through the three methods in the above two groups was analyzed using SPSS-16 version. Results: Group A (Measured Creatinine Clearance >60 ml/min) The Cockcroft-Gault formula showed strong positive correlation (r value 0.756, p value <0.001) with the Measured Creatinine Clearance than the MDRD formula. (r value 0.684,p value <0. 001) Group B (Measured Creatinine Clearance <60 ml/min) both Cockcroft and MDRD showed weak positive correlation with Measured Creatinine Clearance. Cockcroft ((r value 0.617, p value <0.001) MDRD (r value 0.613, p value <0.001) Conclusion: The present study shows weak positive correlation between the Measured Creatinine Clearance, MDRD and Cockcroft formulae at Clearance values<60 ml/min and strong positive correlation between Measured Creatinine Clearance and Cockcroft formula at Clearance values >60 ml/min. Since MDRD formula includes age, gender, and race and developed for the western population with chronic renal insufficiency, further research is needed in our population, to establish and incorporate adjustment& Correction factors in the formula, so that it can be used for a wider range of renal function, in our population to give reliable results and therefore can replace the traditional Measured Creatinine Clearance test.

97-109

Authors: Sanjay D Gabhale, Pallavi Taparia, Dharamveer Yadav, S.P. Agnihotri

Background: Pleural effusion is an abnormal collection of fluid in pleural space resulting from excess production or disruptions of homeostatic forces that regulate the flow of fluid in and out of the area. It is a frequent manifestation of serious thoracic disease whose specific diagnosis is a challenging task. Pleural effusion can be due to Infectious, malignant, parapneumonic disease and tubercular or other causes. Diagnosis of PE is not straight forward due to associated heart disease, malignancy or infection. A specific biomarker is therefore required for differential diagnosis of pleural effusion. C- Reactive Protein is an acute phase protein synthesized by hepatocytes and widely used to assess severity of infection and is directly associated to degree of inflammation in diseased state. The aim of this study is to assess specificity and sensitivity of Pleural Fluid CRP and other routine biochemical indices in diagnosis of exudative Pleural effusion arising due to varying etiologies. Material and Method: The study involved 187 adult patients diagnosed with exudative pleural effusionand classified into 5 groups as follows: 1. malignant pleural effusion (MPE), 2. Chronic non-specific inflammation (CNI), 3. Parapneumonic pleural effusion (PPE), 4.Tubercular pleural effusion (TBPE) and 5. Others. After complete clinical evaluation, routine Pleural fluid analysis and CRP was analysed. Reciever Operating Characterstics (ROC) analysis established the cutoffs of CRP for discriminating between groups. Result and Discussion: Pleural Fluid CRP level was significantly higher in infectious parapneumonic group, followed by Chronic non specific inflammatory group and Tubercular cases with lowest value in malignant group. ROC analysis of Pleural fluid CRP provided good sensitivity (97.05%), specificity (71.76%), NPV of 95.31% and PPV of 80.48 %for the differentiation of tubercular vs. non tubercular effusions. In ROC analysis of CRP for differentiation of Parapneumonic from non Parapneumonic pleural effusion (tubercular, chronic non-specific inflammation, malignant and others), sensitivity of 100%, specificity 98.88%, NPV of 100% and PPV of 81.82% was seen at cut off level of 90.8 mg/l. It provides largest Area under Curve I.e 1.000. CONCLUSION: Pleural fluid CRP can be used as a diagnostic aid specially differentiating between acute and chronic inflammation and also between infectious and non infectious inflammation with CRP value more than 30 mg/l almost excludes malignancy. Pleural fluid CRP can be used as specific biomarker for differential diagnosis of Parapneumonic pleural effusion with excellent sensitivity and acceptable specificity.

120-122

Authors: Ramesh Chandra Thanna, Rekha Choudhary, Swati Pathak, Amrita Vamne, Shreya Nigoskar

Background: Uric acid is the final product of the purine metabolism in humans. The 2 final reactions in its production which catalyze the conver-sion of hypoxanthine to xanthine and the latter to uric acid are catalyzed by the enzyme xanthine oxidoreductase. The role of uric acid in the progression of prediabetes to diabetes has been known. Serum uric acid has been shown to be associated with cardiovascular disease, hypertension, and chronic kidney disease. Aims & objectives: The present study was done to see the level of uric acid in preeclampsia and regnancy induced hypertension (PIH) patients. Material and Methods: The study involves 50 cases of preeclamsia of age group between 20-35 year and 50 controls with similar age group.Srum uric acid was done from all subjects by uricase method in semi-automated biochemistry analyser after centrifugation of samples. Results: The observed mean serum uric acid levels in preeclampsia was 7.48±0.59 mg/dl as compare to 4.12±0.59 in control group. The difference between them is highly significant because p-value is less than 0.05. Conclusion: High serum uric acid level could be a useful indicator of the maternal and fetal complication.

5-12

Authors: Sunaiba M, Avinash S.S, Arunkumar K, Malathi M, Shivashankara A. R, Ashwini K

Introduction: AOPP (Advanced oxidation protein products) are the dityrosine containing protein cross linking products indicating the oxidised tyrosine residues of the plasma protein albumin, fibrinogen and lipoproteins. AOPP are elevated in oxidative stress and inflammation. Unlike AOPP the mAOPP estimation is not interfered by triglyeride levels. Objective: Analytical method comparison of AOPP with mAOPP. Materials and Methods: Measurement of AOPP involves the principle of spectrophotometric determination of oxidation of I- plasma AOPP under acidic condition by Witko Sarsat method. To remove the interference by the triglycerides especially seen in hypertriglyceridemia, the plasma was treated with lipid precipitating agent and the mAOPP were measured. In this study using pooled serum samples we compared the performance characteristics like precision, accuracy, linearity limit and reference range of AOPP assay with the mAOPP assay under our laboratory conditions as per CLIA guidelines of method validation. Results and Discussion: The intrarun, intraday, interday and overall precision of AOPP (4.25%, 5.18%, 3.89%, 5%) was better than that of mAOPP (5.76%, 5.5%, 5.8%, and 5.8%). But on linear regression, mAOPP = 0.5×AOPP + 9.8 chloramine T equivalents (r2-0.62; p < 0.001) and the AOPP was 18% higher than mAOPP. In eutriglyceridemia however, mAOPP = 0.7xAOPP + 4.8 chloramine T equivalents (r2 = 0.75; p < 0.001) and the AOPP was only 14.5% higher. Also the correlation (r = 0.74) of AOPP – mAOPP levels with TG showed that the difference was significantly higher (p < 0.001) in the persons with higher TG levels clearly establishing the overestimation of AOPP. Conclusion: Hence mAOPP may prove to be a better estimate of protein oxidation status and a better diagnostic and prognostic indicator compared to AOPP especially in hypertriglyceridemic patients.

18-21

Authors: Dinesh Kumar Sharma

Background: The present study concentrate to get significant association of serum uric acid levels with pre-eclampsia and PIH and also between serum uric acid and severity of disease. Material and Methods: 80 Patients diagnosed as having Pre-eclampsia and 80 controls with similar age group were studied after taking their consent. Blood samples were collected under aseptic precautions in plain vacutainer for serum uric acid estimations. Uric acid estimation was done by Uricase Peroxidase Method on Fully Automated Biochemistry Analyzer Miura 300. Results: The observed mean serum uric acid levels in preeclampsia was 7.52±0.77 mg/dl as compared to 3.70±0.94 mg/dl in controls.

27-32

Authors: Nishtha Saral, Mamta Rathore, V. D. Bohra, Madhuri Gupta

Background: β-Thalassemia results from a defect in β-globin chain production and ranges from clinically silent heterogeneous thalassemia minor to severe transfusion dependent homozygous thalassemia major. After approximately one year of transfusion, iron begins to deposit in parenchymal tissues, which might cause damage to the parenchyma and organ function impairment. So, iron overload may be develop liver, kidney, heart and endocrine abnormalities. Objectives: The aim of study was to correlate the early hepatic and renal complications in condition of β-thalassemia with serum iron and total iron binding capacity (TIBC) levels. Method: In study 50 β-thalassemia major patients were compared with 50 normal healthy subjects as controls. Fasting serum iron, TIBC, LFT and RFT were estimated using standard kit methods. Data were statistically evaluated by Student’s t-test and Pearson’s correlation coefficient (r). Result: A significant increase in the levels of serum iron and a significant decrease in TIBC in β-thalassemia patients were observed. Serum levels of liver enzymes (Aspartate transaminase, Alanine transaminase and alkaline phosphatase) were significantly higher in β-thalassemia patients as compared to controls. A significant decrease in total proteins and increase in serum bilirubin level was found in β-thalassemia patients. Serum urea and creatinine levels were significantly higher, but no change in levels of serum uric acid was found in β-thalassemia patients when compared to controls. There was significant negative correlation between serum iron and TIBC, total protein among patient group. Conclusion: Iron overload due to frequent transfusions in β-thalassemia results in abnormal organ function tests. Proper and timely screening of these parameters can help in early diagnosis & prevention of iron overload.

41-47

Authors: Viyatprajna Acharya, B D Toora

Background: Obesity is now occurring in pandemic proportions and has earned a new name as “Globesity” for it. Unlike earlier times developing countries too are facing the challenge of obesity and the modern epidemics have a common root cause tapering to obesity. Obesity during adolescent and young adulthood usually persists to adulthood in almost 50% cases and gives rise to early onset of type 2 diabetes mellitus, cardiovascular disorders and metabolic syndrome for which insulin resistance being the common link to all. Objective: The present study was taken up to study the prevalence of insulin resistance in apparently healthy young adult obese population and study its correlation with different cardiovascular risk factors like lipid profile and sdLDL-C. Material and Methods: In a randomized control study 106 apparently healthy young adults in the age group of 21-34 years were chosen from the community out of which 45 were obese and 61were age and gender matched non-obese controls. They were divided in obese and non-obese groups based on cut-off BMI of 25Kg/m2. Along with physical parameters fasting plasma glucose, lipid profile and routine biochemical parameters were assayed by standard kit methods and plasma insulin was measured by sandwich ELISA method. Different lipid ratios, atherogenic index and insulin resistance were calculated. Atherogenic index was calculated and insulin resistance was measured by HOMA-IR model and QUICKI index. Small dense LDL-C (sdLDL-C) was quantified by modified Tsutomu –Hirano method. Results: In the obese group BMI, waist circumference (WC) and waist-hip ratio (WHR) elevated significantly (p=0.0001) and TG, VLDL-C and sdLDL-C as well as atherogenic index elevated significantly (p<0.001).Significant Hyperinsulinaemia (p<0.0001) was found in the obese group and 50% of obese cases had hyperinsulinaemia. Insulin resistance calculated by HOMA-IR and QUICKI index was statistically significant (p<0.0001) in obese. Linear regression analysis showed sdLDL-C (R2=0.08, p=0.05 at 95% C.I.), hyperinsulinaemia (R2=0.089, p=0.054 at 95% C.I.) and insulin resistance (R2=0.099, p=0.03 at 95% C.I.) significantly dependent on WC and atherogenic index was significantly dependent on TG (R2=0.0036, p=0.05 at 95% C.I.) rather than any other lipid factors. On ROC analysis either method of insulin resistance showed equal efficacy (AUC for HOMA-IR= 80.3% and QUICKI = 80.14%; C.I. 95%) and atherogenic index turned out to be a better predictor than sdLDL-C (AUC for Atherogenic index= 76.14% and QUICKI = 71.46%). Conclusions: For Indian subpopulation WC and WHR should also be evaluated along with BMI. Insulin resistance should be identified early and interventional measures should be taken in terms of low carbohydrate protein-rich diet, physical exercise and insulin receptor sensitizers for a short-term. sdLDL-C rises earlier than total cholesterol and hence can be accepted as CVS risk predictor. Not many studies have been done in India on young adult health which is the group that can be targeted for early prevention of the modern epidemics like DM, Metabolic syndrome, CVS disorders and cancers.

54-58

Authors: Smita S. Swami, Sanjay C. Swami, Vinayak W. Patil, Anuradha M. Kanhere

Hypertension (HT) Type 2 diabetes mellitus (DM) are among the most common chronic non-communicable diseases and multifactorial disorders affecting both developed and developing countries, including India. Although DM and HT are not among the top leading causes of death, such as cancer and stroke, these two diseases draw attention from the public due to their increasing trends. The presence of hypertension in diabetic patients substantially increases the risks of coronary heart disease, stroke, nephropathy and retinopathy. When HT coexists with DM, the risk of CVD is increased by 75%, which further contributes to the overall morbidity and mortality of already high risk population. India contributes a lion’s share in the diabetic and hypertensive population. This needs to be tackled with proper awareness and proper government policies. The current review article throws light on comorbidity of HT and DM and its current scenario in India.

61-64

Authors: Dhiraj Trivedi, K Sankara Narayana

Background: Estimating the time since death is one of the prime objectives of conducting Medico-legal autopsies. The exact time of death can be known from hospital records, police investigation report etc. But at times, it is difficult to predict when such records are not available. Various methods have been experimented using biological fluids like blood, vitreous humor, synovial fluid, urine etc. to determine time lapse with varying degree of success. After somatic death, blood components undergo biochemical changes at every tissue. This principle can be utilized effectively to predict time since death in routine forensic practice where time of death is uncertain. Study purpose/Objectives: The present study was conducted to evaluate the relationship of postmortem blood glucose level with time elapsed from death on thirty-nine postmortem blood samples of time registered deaths of an adult occurred at our hospital. Methods: The 10 ml of jugular venous blood was collected from each autopsy case. Blood glucose level was estimated by glucometer. The results were correlated with time since death. Results and Mean Findings: Significant difference between blood glucose level before death (125.32 ± 489.29) and after death (56.73 ± 42.00) was observed. When the time interval since death (884.11 ± 489.29 min)were compared with difference in blood glucose level (68.59 ± 57.76mg/dl) shows high significance (p<0.0001). Similar significance (p<0.0001) were noted when the time interval since death were compared with rate of fall (0.103 ± 0.093 mg/dl min) of glucose level. Conclusions: Estimation of postmortem blood glucose level doesn’t provide any specific pattern of fall. Hence, it cannot be recommended for estimation of time since death as individual parameter.

7-9

Authors: Dinesh Kumar Sharma.

Background: It has been suggested that there is an association between calcium intake and pregnancy-induced hypertension (PIH).The present study concentrate to get significant association of serum calcium levels with pre-eclampsia. Material and Methods: 80 Patients diagnosed as having Pre-eclampsia and 80 controls with similar age group were studied after taking their consent. Blood samples were collected under aseptic precautions in plain vacutainer for serum total calcium estimations. Total calcium estimation was done by Arsenazo III end point method on Fully Automated Biochemistry Analyzer Miura 300. Results: The observed mean serum calcium level in preeclampsia patients was 7.09± 0.37 mg/dl as compared to 7.91±0.911 mg/dl in controls. Conclusion: Low serum calcium levels could be a useful indicator of the maternal and fetal complication in hypertensive patients.

16-24

Authors: Chandrasekar R, Dhanarajan MS, Lalitha prajna S and Srinivasan M

Cataract is one of change in human aging process, generally the lens become clouding and disturb normal vision of every individuals. The routine biochemical analysis shows, there is significant changes in blood parameters between normal and cataract population that were reported by many previous experiments but study related to routine biochemical parameters and its phenomenon were not known among different types of cataract. Hence, a study was planned to understand the relationship among the different types cataract population among Indian ethnics to fill-up the lacuna that could facilitate the prevention of those types by simply identifying them using routine biochemical analysis. About 25 samples in each group were planned. Biochemical data were retrieved from medical records and analyzed by statistical tools, One way ANOVA followed by an ad hoc Tukey's Multiple Comparison Test (P < 0.05) to compare variances among the four experimental groups, they are Cortical, Nuclear Sclerotic, Posterior Subcapsular and Mixed types. Results indicated that there is no characteristics significance among different types of cataract in serum biochemical parameters analysed. This study suggest that, there is a need to identify some simple markers from blood samples and exploring the fact behind the appearance of these markers which may be useful for diagnostic and prevention systems of health care market.

30-40

Authors: Panduranga Murthy,G., Puttaramaiah,G., Ravishankar,H.G., Mokshith,M.C., Leelaja,B.C and B.Shivalinga.

About 30 years back rural areas in Tumkur district were getting drinking water from open wells. Due to extensive digging of bore wells, the ground water table receded alarmingly. Investigations have shown that ground water 60mts at below ground level is unfit for human consumption. This is a serious matter since ground water table has gone down to more than 200mts below ground level. Hence there is an urgent need to provide safe drinking water to the rural population. Water samples from bore wells/Hand pumps were collected from 9 villages belonging to Chikkadalavata Gram panchayath, Madhugiri Taluk, Tumkur district, Karnataka to study the water quality parameters. It is found that ground water in this area contains excess fluorides, Iron, and Alkalinity in general. Results of analysis for water quality parameters are presented in this paper.

46-53

Authors: Dr. Vilas U. Chavan, Dr. Pushpa P. Durgawale, Dr. Ajit V Sontakke, Dr. Hasit D Lad.

Background: Microalbuminuria (MAU) has been considered the first indication of renal injury in patients with diabetes and associated with a higher cardiovascular risk. It is evident from epidemiological studies that urine albumin concentration (UAC) or albumin: creatinine ratio (ACR) in spot urine sample gives similar result like 24-hour urinary albumin excretion (UAE). Objectives: The study was aimed to find out whether UAC and/or ACR calculated in 24-hour urine sample correlate or not with same 24-hour UAE and what is diagnostic outcome by various measures of albumin excretion calculated in 24-hour urine samples in type 2 diabetic patients? Methodology: We collected 24-hour urine sample from 125 type 2 diabetic patients and urinary albumin and creatinine were measured. We calculated different parameter of albumin excretion in 24-hour urine sample and interpretation of each parameter was derived. Data was expressed as median and range. Results: The median ACR was 187.30 (mg/g), UAC was 135.5 (mg/L) and UAE was 270.92(mg/24 hours) in 24-hours urine sample. Clinical outcome of nephropathy by different calculated measures of albumin excretion in 24-hour sample was different. 72 patients had similar interpretation while 53 patients showed different diagnostic interpretation by different measures of albumin excretion in the same 24-hour urine sample. Conclusion: Various measures of urine albumin excretion like UAC, ACR calculated in 24-hour urine sample in type 2 diabetic patients showed different diagnostic interpretation. We recommend when the urinary albumin concentration exceeds 10 mg/L in spot or random urine sample, another 24-hour sample should be collected for confirmed diagnosis of the stage of the albuminuria / nephropathy. We conclude that 24-hour urine sample is gold standard for diagnosis of nephropathy in diabetic patients. Further prospective studies are needed to confirm our findings.

59-66

Authors: Kaja Urbańska, Chandi C. Mandal.

Glioblastoma multiforme (GBM) is the most common primary tumor of a central nervous system, with the IV grade of histological malignancy. Despite the advancement of GBM treatment strategies the overall survival in GBM patients is still less than one year and it is probably due to location of the tumor and its high proliferative as well as metastasis activity. Thus, there is an urgent need to intensify basic as well as clinical research on GBM for understanding the molecular biology of the diseases and for the treatment of brain tumor. Most of therapeutic and molecular studies are performed with using of commercially available GBM cell lines. Use of the right cell line, which is based on the precise knowledge of its characteristics, may not only facilitate to design the experiments but it also provides more authentic information from the study. Thus, the aim of this study is to present the characteristics of the U-87glioblastoma cell line, which is the most frequently used in brain tumor research. In the presented article there are the information indicating the similarities in U-87 cells and spontaneous GBM biology, description of current studies with using this cell line, as well as benefits of choosing U-87 cell line in researches.