Enzymatic Endpoint Method Research Articles

HDL Cholesterol and Functional Scale Measured by the NIHSS in Acute Thrombotic Stroke Patients

Highlight: Stroke is one of the leading causes of death and disability in the world. There is correlation between HDL cholesterol in acute thrombotic patients and the functional degree of the NIHSS. The lower rate of HDL cholesterol is related to the higher NIHSS score or the higher level of stroke severity. ABSTRACT Introduction: Stroke is one of the world’s leading causes of death and disability. Acute thrombotic stroke accounts for 87% of all stroke cases. The study of the correlation between high-density lipoprotein (HDL) cholesterol levels and acute thrombotic stroke is still limited, especially on the relationship between HDL levels and the degree of functional stroke. Objective: This study aimed to prove a correlation between HDL cholesterol and the functional scale measured by the NIHSS (National Institutes of Health Stroke Scale) in acute thrombotic stroke. Methods: The design study was cross-sectional, with sampling by consecutive admission based on inclusion and exclusion criteria in the neurological ward (Seruni A) at Dr.Soetomo General Academic Hospital, Surabaya. The NIHSS was used to determine the degree of functional scale, and HDL levels were measured by enzymatic endpoint methods. Results: From the 40 subjects studied, 16 (40%) had normal HDL levels of ≥ 40 mg/dL with an NIHSS score between 0–10, and 24 (60%) had lower HDL levels of ≤ 40 mg/dL with an NUHSS score between 0–20. There were significant results in this study (r = 0.391 and p = 0.013). Conclusion: There was a negative correlation between HDL cholesterol levels and the functional scale measured by the NIHSS in acute thrombotic stroke patients.

1839-PUB: Baseline FFA and IR after One Year of TB Treatment in People with Prediabetes—Pilot Study from South India

Background: Association of TB and prediabetes (PreDM)/insulin resistance (IR) and FFA remains unclear. Aim was to assess FFA and IR in TB with and without PreDM and to see the changes in IR in PreDM after 1 year of TB treatment. Methods: A prospective study done from Jan 2017 to Sep 2019 in people with newly diagnosed PTB and PreDM (OGTT) in TB clinics and grouped as TB without PreDM (n=30) (group1) and TB with PreDM (n=37) (group2). Control without TB/PreDM (n=30) was included. Anthropometric, and biochemical details were noted. Insulin was measured by electro chemiluminescence and FFA by enzymatic end point method. IR was calculated by HOMA. Results: Age was similar in groups (1vs.2; 40vs44). 2 hr glucose was significantly higher in group 2 than group1 (162vs. 108;p<0.001). BMI, WC, fasting glucose and HbA1c were similar between groups. Mean IR was higher in group 2 than group1 (1.3vs.1; p=0.7). Median FFA was higher in TB PreDM (20.9) than without Pre DM (17.8) and control (14.9) (Figure1). High FFA (23vs.20) (p=0.4) and high IR (1 vs. 0.6) (p=0.2) were observed in BMI ≥vs.<18.5 among TBPreDM. In group 2, baseline IR was high (0.82) and reduced to 0.71 and also those who remained PreDM (0.52) and who reverted to normal (0.44 from 0.71) after 1 year completion of TB treatment. Conclusion: High FFA and IR was noted in TBPreDM and those with high BMI. Improvement in IR was noted 1 year after completion of TB treatment. Disclosure S.Kumpatla: None. A.Devarajan: None. H.Kornfeld: None. V.Viswanathan: None. Funding Indian Department of Biotechnology and Civilian Research; Development Foundation Global (USB1-31149-XX-13)

Association of Apolipoprotein B and Triglyceride Levels with Coronary Artery Disease

Objective: To find the association of Apolipoprotein B and triglycerides with coronary artery disease.
 Study Design: Cross-sectional study.
 Place and Duration of Study: Department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology, Rawalpindi, in collaboration with HITEC Institute of Medical Sciences, Taxila Pakistan, from Mar 2019 to Mar 2020.
 Methodology: Four hundred and forty-four individuals of either gender and 18-70 years were included in the study. According to CT angiography, participants were differentiated into those with coronary artery disease and without coronary artery disease. In addition, serum triglycerides were analysed by glycerol phosphate enzymatic endpoint method on ADVIA 1800R Clinical Chemistry Auto analyzer, and apolipoprotein B was analysed on BT1500 Clinical Chemistry turbidimetric analyzer.
 Results: The sample population had 144(32.0%) females and 300(68.0%) males. 164(37.0%) patients had coronary artery disease. 120(27.0%) participants were between 35 and 45 years of age, and 96(22.0%) were between 56 and 65 years. 126(30.0%) patients had hypertension, 352(79.3%) had apolipoprotein B levels >130 mg/dl. Positive Pearson’s correlation was found between apolipoprotein B and triglycerides with coronary artery disease.
 Conclusion: There is a strong association between apolipoprotein B and non-fasting triglyceride levels with coronary artery disease. The apolipoprotein B can be used as a routine biomarker for screening and treatment monitoring coronary artery disease along with other lipid profile parameters.

Association of TNF-α polymorphisms (-857, -863 and -1031), TNF-α serum level and lipid profile with acne vulgaris.

BackgroundAcne is an inflammatory condition principally affected by genetic and dietary factors. Investigation into functional polymorphisms of TNF-α gene and their association with acne vulgaris will be helpful in exploring genetic influence on skin immune mediated inflammatory events. In the present study, we analyzed association of TNF-α gene polymorphisms, its expression levels and lipid profiles in a large cohort of acne patients and controls.MethodsWe used PCR-RFLP to study association of TNF-α polymorphisms at −857C/T, −863C/A and −1031 T/C sites with acne vulgaris. Lipid profiles were measured using enzymatic end-point method. The serum levels of TNF-α and apolipoprotein a were measured using ELISA. NIH, LDlink was used to investigate patterns of linkage disequilibrium across south Asian reference genome (Punjabi from Lahore Pakistan).ResultsWe found that TNF-α −863 polymorphism is strongly associated with acne in overall population as well as in gender and severity based groups of acne patients. Polymorphisms at −863 and −1031 position were in linkage disequilibrium. Importantly, TNF-α serum level was significantly increased in acne patients with severe disease symptoms. Furthermore, levels of total cholesterol (TC) and triglycerides (TG) were significantly increased, whereas high density lipoprotein cholesterol (HDL-C) level was significantly decreased in acne patients. The levels of apolipoprotein a varied widely in studied populations and no significant difference was found in the analyzed groups.ConclusionIn conclusion, we found that TNF-α expression increases in acne patients affected by TNF-α polymorphisms, and that the lipid profile is specifically disrupted in acne patients.

Effect of antihyperlipidemic polyherbal formulation in high diet induced hyperlipidemia wistar albino rats. An in vitro - in vivo evaluation

Hyperlipidemia is a prevalent global health problem, and it is linked to various cardiovascular disorders. The side effects of the current lipid-lowering drugs have increased the tendency to move toward traditional and alternative remedies. The study aims to Formulate and evaluate the Antilipidemic activity of Polyherbal formulations used as a traditional medicine in the Malabar area, Kerala by in vitro and in vivo methods. Further, the present study also compares the impact of seasonal variations on chemical contents of ingredient herbs of polyherbal medicine. They were analyzed for, anti-oxidant activity by DPPH and Nitric oxide method and In-vitro anti-cholesterol activity by cholesterol enzymatic endpoint method using simavastatin as a positive control. The formulation showing highest anti-oxidant and in- vitro Antilipidemic activity was selected for in-vivo analysis. Out of four formulations, PHF 1 shows low IC50 values in DPPH and Nitric oxide methods (250.45± 0.60, 985.40±5.59), respectively. The in-vitro anti-cholesterol activity showed a maximum % of inhibition for PHF1. Based on this PHF 1was selected for in vivo analysis. Acute toxicity was performed according to OECD guidelines. The antilipidemic activity was conducted by Diet-induced hyperlipidemia model in Wistar albino rats, containing six animals in each group. All the groups except saline control received a high-fat diet for two weeks. The Polyherbal formulation (200 mg/kg & 400 mg/kg) showed significant (P<0.05) reduction in total serum cholesterol and lipid levels compared to the vehicle control group. This present study proved that Polyherbal formulation has Antilipidemic activity against the diet-induced hyperlipidemia model by reducing the total serum cholesterol (TC), triglycerides (TG), very low-density lipid (VLDL), low-density lipids(LDL) levels and increasing high-density lipid (HDL) level.

Evaluation of Anti-diabetic, Hepatoprotective and Antilipidemic Potentials of Syzygium aromaticum (Clove) on Albino Rats

Aims: The aim of this study was to evaluate the anti-diabetic, hepatorenoprotective and antilipidemic potentials of Syzygium aromaticum (clove) in albino rats.
 Study Design: This study is a non-randomized experimental study.
 Place and Duration of Study: Department of Medical Laboratory Technology, University of Port Harcourt, Nigeria, between September, 2019 and December, 2019.
 Methodology: Thirty-five male Wister rats with weight between 145 to 150 g were randomly selected into seven groups of five rats each. The first group served as Negative control (group 1). The second group was the positive control (Diabetic group). The remaining five groups being the treatment groups (3-7). Diabetes was induced intraperitoneally with 65 mg/kg of streptozotocin (STZ) single dose. Group 3 Diabetic group treated with metformin (100 mg/kg); Group 4 Diabetic group treated with low dose clove (250 mg/kg). Group 5 Diabetic group treated with low dose clove and metformin; Group 6 Diabetic group treated with high dose clove (750 mg/kg), while Group 7 were Diabetic group treated with high dose clove (750 mg/kg) and metformin for six weeks. Blood samples were collected via cardiac puncture in appropriate EDTA, heparinized and sterile bottles for standard laboratory investigations of lipid profile, glucose, liver enzymes, urea and creatinine. Plasma lipid profile, liver enzymes, urea and creatinine were determined using enzymatic end point method under standard operating procedures. Statistical analysis was done using Graph Pad Prism Version 5.03 and p values less than 0.05 were considered statistically significant.
 Results: Results revealed that the STZ-induced diabetic group exhibited highly significant increase in activity of liver enzymes AST, ALT and ALP, increase in the levels of urea, creatinine, glucose and most lipid profile parameters as compared to the negative control group (p<0.001). Histopathological examination of liver and kidney tissues of diabetic rats indicated slight changes. However, their changes were overcame by clove treatment and the majority of the cells tend to be normal. Low dose clove group 5 (250 mg/kg) with metformin decreased the levels of the analytes most when compared to the levels of the positive control group. For glucose, group 5 gave a mean glucose level of 4.40 ± 1.08 mmo/l, significantly lower than the positive control group 39.67 ± 0.67, ALT group 5 gave a mean ALT level of 56.00 ± 7.11, which was significantly lower than the positive control group 205.70 ± 14.79, for urea group 5 gave a mean urea level of 4.25 ± 0.77 which was significantly lower the control group 23.80 ± 3.56 at p < 0.001, thereby yielding a better treatment result.
 Conclusion: In conclusion, low dose clove supplementation with metformin could be excellent adjuvant support in the therapy of diabetes mellitus and its complications.

Changes in Serum Calcium, Albumin and Alkaline Phosphatase Levels of Postmenopausal Women

A case-control cross-sectional study was carried out to observe the changes in serum calcium, albumin and alkaline phosphatase levels of postmenopausal women compared to their premenopausal counterparts. This is to enable us ascertain the relative risk of developing osteoporosis in postmenopausal women in Bangladesh. One hundred and eighteen (118) apparently healthy females (59 premenopausal=control and 59 postmenopausal=case) were selected for the study. The study was carried out in the Department of Biochemistry, Mymensingh Medical College in cooperation with the Department of Gynecology, Mymensingh Medical College Hospital and Community Based Medical College Hospital Bangladesh during the period from January to December 2009. Data were collected through clinical evaluation from pre-selected questionnaires. Fasting venous blood samples were taken for laboratory investigations. Serum calcium and albumin was determined using colorimetric method and enzymatic end point method was used to determine serum alkaline phosphatase levels. Statistical significance of difference between two groups were evaluated by using Student's unpaired 't' test with the help of SPSS software package. The results showed statistically significant decrease in serum calcium and albumin (p<0.01) of menopausal women compared to reproductive age group. The postmenopausal women had significantly higher (p<0.01) concentrations of serum alkaline phosphatase than the premenopausal women with regular menstruation. Therefore, it can be concluded that menopause leads to the increased risk of osteoporosis by decreasing serum calcium and albumin and by increasing alkaline phosphatase levels. The decreased serum calcium and increased alkaline phosphatase levels indicate that menopause is an independent risk factor for developing osteoporosis in women of Bangladesh.
 CBMJ 2019 July: Vol. 08 No. 02 P: 30-35

Evaluation of Glycated Hemoglobin (HbA1c) in Acute Coronary Syndrome (ACS)

Aim: The aim of our study was to estimate the level of HbA1c in patients with Acute Coronary Syndrome (ACS). To determine any correlation between HbA1c, admission blood glucose (Random blood sugar), serum lipid profile among the cases and to evaluate the outcome during hospitalization.
 Study Design and Setting: Cohort study.
 Place and Duration of the Study: Department of Biochemistry and General Medicine including ICCU and Cardiology unit, RIMS, Imphal, between September 2015 to August 2017.
 Methodology: Data were collected from 98 patients admitted in Medicine ward who were diagnosed with ACS irrespective of their diabetes status and they were divided into three groups according to their HbA1c levels (<5.7%, 5.7-6.4%, > 6.5%). The blood samples collected by venipuncture were analyzed for HbA1c level, done by RANDOX HbA1c Rx series (latex agglutination inhibition assay), admission blood glucose (measured by Trinder’s method) and serum lipid profile (RANDOX enzymatic Endpoint Method Rx series). The data were analyzed using statistical tools like Chi Square test, Independent sample t test, Pearson’s Correlation, Fisher’s exact test through SPSS 21.0.
 Results: Majority of the patients were men (72.4%) & (27.6%) women and in the age group of 51-65 years. The mean age is 62.14 years. 54.1% of the ACS patients were already diagnosed cases of type 2 diabetes mellitus whereas 45.9% were non-diabetic. Out of 98 patients, 60 had HbA1c level in the diabetic range (≥6.5%), 25 in the pre-diabetic range (5.7-6.4%) and the remaining 13 were within normal range (<5.7%). Admission blood glucose, total cholesterol and LDL were positively correlated with HbA1c value.
 Conclusion: The mortality of the ACS patients irrespective of diabetic status during hospitalization was associated with HbA1c value irrespective of diabetic status during hospitalization. However, it was not associated with thirty days mortality.

Are serum leptin levels predicted by lipoproteins, vitamin D and body composition?

BACKGROUNDBoth obesity and vitamin D deficiency are important health issues in Pakistan. The connection between body composition, Vitamin D and leptin in young adults is important to be studied as body composition may affect bone health and therefore the possibility of osteoporosis in later life. Few studies have attempted to investigate the effect of body composition and leptin with vitamin D in adolescence.AIMTo investigate the association of serum leptin with body composition, lipids and 25-hydroxyvitamin D (25OHD) in adults.METHODSThis cross-sectional study was conducted on 167 apparently healthy adults. Demographics were recorded, bioelectrical impedance analysis was performed and clinical history noted. Serum leptin was measured using DIA source kit on ELISA and total 25OHD was measured on ADVIA-Centaur; Siemens. Total cholesterol and high density lipoprotein cholesterol were quantified using Enzymatic Endpoint Method and Cholesterol Oxidase-Phenol Aminophenazone method respectively. Biochemical analysis was done in the Departments of Pathology and Laboratory Medicine and Biological and Biomedical Sciences, Aga Khan University Hospital Karachi Pakistan.RESULTSMedian age of the group (n = 167) was 20 years (IQR 27-20); 55.7% were females. Majority (89.2%, n = 149) of the study group was 25OHD deficient, 6% (n = 10) had insufficient serum 25OHD levels and 4.8% (n = 8) had sufficient D levels. Females, had higher median leptin levels [2.71 (IQR 4.76-1.66 ng/mL)] compared to their counterparts [1.3 (3.60-0.54 ng/mL), P < 0.01]. Multiple regression analysis suggested that basal metabolic rate, muscle mass, body fat percent, bone mass and serum 25OHD were the most contributing factors to serum leptin levels. Bone mass and serum 25OHD in fact bore a negative correlation with leptin.CONCLUSIONThe results indicate that basal metabolic rate, muscle mass, body fat percent, bone mass and serum 25OHD have an impact on serum leptin. Being a cross sectional study causal relationship between leptin and other variables could not be determined.

The effect of fish oil capsule on serum total cholesterol and triglycride levels in type-2 diabetic patient

Background: Diabetes Mellitus (DM) is one of the common metabolic disorders that have effects on lipid metabolism. Fish oil supplementation may improve lipid abnormalities in diabetic patient.Objective: To observe the effect of supplementation of omega-3 fatty acid on total cholesterol (TC) and triglyceride(TG) level in type 2 DM.&#x0D; Methods: A prospective interventional study was conducted from January 2017 to December 2017. A total numbers of 52 diagnosed type 2 diabetic patients of both sexes were selected with age 40 to 50 years. Among them, 27 type 2 diabetic patient with supplementation of omega 3 fatty acid (2g/ day) for 12 weeks was considered as study group. Another 25 type 2 diabetic patients without supplementation of omega 3 fatty acid was considered as control group. The study subjects were selected from Outpatient Department of Endocrinology, Dhaka Medical College Hospital, Dhaka and personal contact from Dhaka city.Serum TC and TG were estimated by enzymatic endpoint method in autoanalyzer. The study parameters were measured at base line and after 12 weeks of study period in both groups. For statistical analysis, Paired Student’s‘t’ test and Unpaired Student’s ‘t’ test were performed as applicable using SPSS for windows version 16.0.&#x0D; Results: In this study serumTC and TG levels were significantly lower (p&lt;.001) in diabetic patients after supplementation with omega- 3 fatty acid in comparison to that of their baseline value.Again, after 12 weeks, serum TC and TG levels were significantly lower (p&lt;.001) in diabetic patients after supplementation with omega-3 fatty acid in comparison to control group. In control group, serum TC and TG levels were almost similar both at baseline and after 3 months of follow-up.&#x0D; Conclusion: After analyzing the results of the study, it can be concluded that omega-3 fatty acid may improve serum TC and TG levels in type 2 diabetic patient and may be helpful to minimize the complications of type-2 DM.&#x0D; J Bangladesh Soc Physiol. 2018, December; 13(2): 73-77

A correlative study of HbA1C and lipid profile parameters among type 2 diabetic population in a rural hospital in puducherry.

Background: Diabetes Mellitus (DM) is characterized by chronic hyperglycemia with disturbances of Carbohydrate, Lipid and Protein metabolism resulting from defects in insulin secretion, insulin action or both. Obesity is an independent risk factor for cardiovascular disease, including coronary artery disease and congestive heart failure, in both men and women. Glycated hemoglobin (HbA1C) is a routinely used marker for long-term glycemic control. Elevated HbA1c has been regarded as an independent risk factor for coronary heart disease (CHD) and stroke in subjects with or without diabetes. Aim&Objectives:To evaluate the level of HbA1C and lipid profile in type 2 diabetes mellitus patients. To find out the correlation between HbA1C and lipid profile parameters in type 2 diabetic population. Methodology:This Cross sectional study was conducted among type 2 diabetic patients and control subjects and they were divided into three groups.Group 1- control subjects = 28, Group 2– Type 2 DM with good control = 28 and Group 3– Type 2 DM with poor control = 28.The serum sample was used for the measurement of total Cholesterol, Triglycerides, HDL, LDL and VLDL.Fasting and postprandial (2 hour) blood sugar (FBS & PPBS) were estimated by Glucose Oxidase-Peroxidase (GOD-POD) enzymatic end point method. HbA1C was estimated by using Ion exchange chromatography (Crest A Coral clinical system, USA). Urine examination was done for benedicts test and proteinuria.Statistical Analysis was done by using SPSS version 75.0. Results:Our study showed significant correlation between HbA1C and lipid profile parameters between the two groups (less than 7% and more than 7% of HbA1C ). The results suggested the importance of glycemic control in order to manage dyslipidemia and risk for cardiovascular disorder in type 2 diabetes. Conclusion:This dual biomarker- HbA1C, glycemic control as well as lipid profile can be used for screening of high risk patients for early diagnosis of dyslipidemia thereby the cardiovascular and peripheral complications can be prevented by timely intervention of the disease.

Evaluation of Anti-Atherosclerotic Activity of Virgin Coconut Oil in Male Wistar Rats Against High Lipid and High Carbohydrate Diet Induced Atherosclerosis

The aim of this study was to analyze the effect of virgin coconut oil (VCO) on high lipid diet (HLD) and high carbohydrate diet (HCD) induced atherosclerosis in male Wistar rats. Spectrophotometer was used to determine the lipid parameters by enzymatic endpoint method. The plasma total cholesterol (TC), triglyceride (TG) and high-density lipoprotein (HDL) levels were measured using commercial enzymatic kits. The results showed that feeding with normal and VCO diet significantly decrease (p&lt;0.05) body weight when compared to the control group rats. Body weight and lipid profile were estimated after 8 weeks of treatment. VCO showed a significant (p&lt;0.05) reduction in TC, TG, Low density lipoprotein (LDL), Very low density lipoprotein (VLDL) levels and significant (p&lt;0.05) increase in HDL in HCD /HLD group rats. There was significant decrease in atherogenic index (AI) (p&lt;0.05) of all VCO treated groups when compared to the Control group. Hence, there was increase in the percentage of protection in VCO treated animals after 8 weeks.The decrease of lipid profiles in the experimental rats showed that VCO possesses anti-atherosclerotic activity.

Serum vaspin levels are associated with decreased insulin sensitivity in newly diagnosed type 2 diabetes mellitus in Bangladesh

Abstract Objectives The present study aimed to explore the relationship of circulating vaspin levels with insulin sensitivity and anthropometric factors. Methods This study was conducted with 65 newly diagnosed type 2 diabetes mellitus (T2DM) patients with age-matched 65 healthy controls. Serum glucose was measured using glucose-oxidase method, lipid profiles by enzymatic end-point methods, and fasting insulin and vaspin levels were assessed with ELISA techniques. Homeostasis model assessment for insulin sensitivity (HOMA%S) and insulin secretory capacity (HOMA%B) were estimated from the fasting glucose and insulin levels using HOMA-CIGMA software. Results Fasting serum insulin (μU/ml) was higher in the diabetic group than controls (16.0 ± 7.9 vs. 10.9 ±3.3, respectively, p =0.0001). The mean (±SD) HOMA%S of the diabetics was significantly lower than that of the controls (48 ±31 vs. 76±55, respectively, p = 0.001). The HOMA%B of the T2DM group was nearly 50% of that of the controls (71± 40 vs. 131 ±46, respectively, p = 0.001). The T2DM group exhibited significantly lower serum vaspin (ng/ml) levels than the controls (0.62 ±0.26 vs. 0.83± 0.28, respectively, p =0.001). Vaspin levels were negatively correlated with waist circumference (r =0.17, p =0.043) and positively correlated with HOMA%S (r =0.243, p =0.007) among all of the participants. The association of serum vaspin with diabetes remained highly significant (p =0.008) in binary logistic regression analysis performed after adjusting for the effects of confounders. Conclusions Serum vaspin level is positively associated with insulin sensitivity and negatively correlated with serum glucose, BMI and waist-height ratio.

Anti-spermatogenic and estrogenic effects of Escravos crude oil in Chinchilla rabbits

This study aimed at investigating the effects of Escravos crude oil on spermatogenesis in relation to serum concentrations of estradiol and total cholesterol. A total of 30 male Chinchilla rabbits within the age range of 12 to 14 weeks and weighing 1.2 kg to 1.45 kg were used in this study. The doses of 00, 15, 20, 25 and 30 mg kg-1 body weight of Escravos crude oil were orally administered to groups A, B, C, D and E, respectively for 28 days. Serum concentrations of estradiol and total cholesterol were estimated using the microplate enzyme immunoassay and enzymatic end point methods, respectively. The SPSS software package (version 16) was used for the statistical analyses and results expressed in mean ± Standard Deviation. The results showed dose dependent significant increases in estradiol (18.32 +/- 6.46 to 69.41 +/- 12.89 pg/ml) and total cholesterol (1.32 +/- 0.24 to 2.92 +/- 0.34 mmol/l) concentrations (p 0.05) in the relative weight of the testis (3.80 +/- 0.40 to 5.50 +/- 0.90 g). The histology of the testes revealed decreased spermatogenic activity. The findings of this study suggest that Escravos crude oil could be a potential endocrine disruptor and anti-spermatogenic agent. Keywords : Escravos crude oil, Estradiol, Anti-Spermatogenesis, Total cholesterol, Chinchilla rabbits.

Estrogenic and Hypocholesterolemic Effects of Aqueous Extract of Nicotiana tabacum (Tobacco) on the Micro-Anatomical Architecture of the Testis in Male Albino Wistar Rats

Aim: The aim of this study was to investigate the effect of aqueous extract of Nicotiana tabacum on the testicular function and micro-architectural integrity in male Albino Wistar rats in relation to the serum concentrations of estradiol, cholesterol, testis and body weights. Study Design: The experiment was carried out on Albino Wistar Rats. Methodology: A total of 18 male Albino Wistar rats aged 8 to 10 weeks and weighing 140 to 220 g was used. Aqueous extract of N. tabacum at the doses of 20 and 30 mg/kg body weight were orally administered to groups B and C, respectively while 0.5 ml of distilled water was administered to group A (Control) for 28days. Serum concentration of estradiol (E2) and total cholesterol were estimated using the microplate enzyme Original Research Article British Journal of Applied Science & Technology, 4(15): 2194-2205, 2014 2195 immunoassay and enzymatic end point methods, respectively. The SPSS software (version 20) was used for the statistical analysis and the result expressed in mean ± SEM. Result: The result showed significant increase in serum estradiol concentration (from 2.50±0.43 pg/ml in the control group to 9.33±1.87 pg/ml in the treated groups), significant decrease in serum total cholesterol concentration (from 1.92±0.04 mmol/l in the control group to 1.22±0.08 mmol/l in the treated groups) and body weight (p≤0.05), and insignificant increase in the weight of the testes (p≥0.05). Testicular microscopy showed decreased spermatogenesis, atrophic interstitial space and moderate hypertrophy of the seminiferous tubules. Conclusion: Therefore, N. tabacum could be considered a potential endocrine disruptor which can affect the micro-anatomical architecture and function of the testis.

Histopathological, Hemochromatotic, Hypercholesterolemic, and Androgenic Effects of Escravos Crude oil on the Testis in Male Chinchilla Rabbits

Aim: This study aimed to investigate the effects of Escravos crude oil on serum concentrations of testosterone, total cholesterol, body weight, relative weight and microanatomical architecture of the testis using male Chinchilla Rabbits. Place and Duration of Study: This study was carried out at the Department of Medical Laboratory Science, Nnamdi Azikiwe University, Nnewi Campus between May and June 2013 (28 days). Methodology: A total of thirty male Chinchilla Rabbits aged 12 to 14 weeks and weighing 1.2kg to 1.45kg was used. Crude oil was administered orally at the doses of 15, 20, 25 and 30mg/kg body weight to groups designated B, C, D and E respectively for 28 days, while group A was given normal saline. Serum concentration of testosterone and total Original Research Article British Biotechnology Journal, 4(6): 649-658, 2014 650 cholesterol were estimated using the microplate enzyme immunoassay and enzymatic end point methods, respectively. The SPSS software (version 20) was used for the statistical analysis and the result expressed in mean ± SD. Result: The results showed dose dependent effects on the hormone and biochemical assays, especially at the concentration of 25mg/kg body weight of the administered crude oil. Significant increases in serum concentrations of testosterone (0.32+/-0.05 to 0.46+/0.14) and total cholesterol (1.35+/-0.17 to 1.76+/-0.15) concentrations (p≤.05), and insignificant increase (p≥.05) in the relative weight of the testis (4.80+/-0.40 to 6.50+/0.90) were observed. The histology of the testes revealed hypertrophy of the seminiferous tubules, atrophy of the basal lamina and interstitial cells, and hemochromatosis. The histological findings agree with the hormonal and biochemical findings. Conclusion: These findings suggest that Escravos crude oil might be a potential endocrine disruptor and toxic substance which can affect the micro-anatomical architecture of the testis.

Association of Circulating Fasting TNF-Α with Hyperglycemia is Stronger than with Body Mass Index in Newly Diagnosed Bangladeshi Type 2 Diabetic Subjects

Background and aim: Insulin resistance and hypo-insulinemia are the major problems in Type 2 Diabetes Mellitus (T2DM). Epidemiological evidences suggest that obesity is linked with insulin resistance. Inflammatory cytokines such as TNF-α has also been associated with obesity and insulin resistance. The present study aims to evaluate fasting serum TNF-α in newly diagnosed type 2 diabetic subjects in different BMI group to observe its association with insulin sensitivity and obesity. Methods: A total number of 145 newly diagnosed T2DM subjects were recruited in this study. On a prescheduled morning fasting and postprandial (2hrs after 75g glucose load) blood was drawn. Serum glucose was measured by glucose-oxidase method, lipid profile by enzymatic end-point method, insulin and TNF-α by ELISA methods. Insulin secretory capacity (HOMA%B) and insulin sensitivity (HOMA%S) were calculated using fasting glucose and insulin by HOMA-CIGMA software. SPSS for Windows was used for statistical analysis. Results and conclusion: The subjects were divided into three groups on the basis of BMI cut-off points suggested by WHO for the Asian population (Gr1: BMI 27.5). The fasting serum insulin and TNF-α levels were significantly (p<0.005) higher in group 3 than the other groups. HOMA%B was significantly higher in Gr 3 and HOMA%S was significantly lower in group 3 compared to group 1 and group 2. From multiple linear regression analysis it can be concluded that both BMI and postprandial serum glucose had potential effects on serum TNF-α level but the later one had stronger.

Effects of 3,4-Dihydroxyacetophenone on the Hypercholesterolemia-Induced Atherosclerotic Rabbits

3,4-Dihydroxyacetophenone (3,4-DHAP) is one herbal extract from bald Mao-dong-qing leaves. We reported that 3,4-DHAP had anti-inflammatory function by decreasing tumor necrosis factor-α (TNF-α) secretion in macrophages. The aim of the study was to examine the effects of 3,4-DHAP on plasma and liver lipids, plasma alanine aminotranferase (ALT) and TNF-α level, vascular cell adhesion molecule-1 (VCAM-1) expression, plaque vulnerability and vascular inflammation in hypercholesterolemia-induced atherosclerotic rabbits. Male New Zealand white rabbits were randomized into negative control, positive control, 3,4-DHAP and simvastatin groups. From weeks 2 to 12, the rabbits were treated with 3,4-DHAP or simvastatin. At weeks 12, all the animals were sacrificed. Plasma lipids and ALT were measured using the enzymatic endpoint method. Plasma TNF-α was measured using enzyme-linked immuno sorbent assay (ELISA). Liver lipids concentrations were estimated using commercial kits. The expression of VCAM-1 was measured using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Histological analysis was used to evaluate the pathologic changes of rabbit aortas. The results showed that 3,4-DHAP markedly lowered plasma and liver lipids, lowered plasma ALT and TNF-α levels compared with the positive control group. VCAM-1 mRNA and protein were markedly inhibited by 3,4-DHAP. Decreased aortic plaque instability was evident in 3,4-DHAP-treated rabbits, as demonstrated by a thickened elastic layer, increased vascular smooth muscle cells (VSMCs) accumulation in the plaques, less neointima hyperplasia and macrophages recruitment. 3,4-DHAP may attenuate the progression of atherosclerotic lesions and stabilize plaques by lowering plasma lipids, the number of macrophages and the expression of VCAM-1, while increasing the number of VSMCs in the atherosclerotic plaques.

Promoter variants in interleukin-6 and tumor necrosis factor alpha and risk of coronary artery disease in a population from Western India

INTRODUCTION:A central component of the atherosclerotic process is inflammation. Single nucleotide polymorphisms (SNPs) present in the promoter region of various cytokines can lead to altered levels of the transcript and a state of low-grade inflammation exacerbating the risk of coronary artery disease (CAD). The present work tries to understand the role of permissive promoter variants in the interleukin-6 gene (IL-6-174G/C) and the tumor necrosis factor alpha (TNFα-308G/A) in the causation of CAD and also dyslipidemia.MATERIALS AND METHODS:Genotyping was conducted on 100 cases of CAD and 150 controls by the allele termination assay SNaPshot. Biochemical parameters were determined by routine enzymatic endpoint methods. The results were analyzed by appropriate statistical methods.RESULTS:No differences in the minor allele frequency IL-6-174G/C SNP were seen between cases and controls (0.13 vs. 0.12). The differences in the allele frequency of TNFα-308A between cases (6%) and controls (2%) have led to an odds ratio, 3.370; 95% confidence interval, 1.039-11.543; P=0.033 in the univariate analysis. In the final logistic regression analysis, however none of the variants were associated with an increased risk of CAD.CONCLUSIONS:In summary, no association of the permissive promoter variants in the IL-6 gene and the TNFα gene were seen with an increased CAD risk. These and other studies highlight the importance of doing population specific studies.

Highly selective l-threonine 3-dehydrogenase from Cupriavidus necator and its use in determination of l-threonine

l-Threonine level in blood plasma is a biomarker of some diseases and nitrogen imbalance in the body. The determination of l-threonine is interesting and is required for diagnosis and management of inherited metabolic disorder. This is the first report of the specific enzymatic determination of l-threonine by a newly discovered l-threonine 3-dehydrogenase (ThrDH, EC 1.1.1.103) from Cupriavidus necator NBRC 102504. ThrDH, a key enzyme in l-threonine catabolism in microorganisms and animals, catalyzes the NAD +-dependent oxidation of l-threonine to 2-amino-3-oxobutyrate. ThrDH from C. necator was purified to homogeneity and fully characterized. l-Threonine and d l-2-amino-3-hydroxyvalerate are the only substrates for ThrDH among other l-amino acids, alcohols, and amino alcohols. The primary amino acid structure of ThrDH belongs to the extended short-chain alcohol dehydrogenase superfamily and is related to GDP-mannose-3′,5′-epimerase (GME) from Arabidopsis thaliana. Both enzymes have a glycine-rich NAD +-binding domain at the N terminal and conserved catalytic triad of YxxxK residues, but substrate-binding residues of GME were not found in the ThrDH sequence. ThrDH significantly differs from known bacterial and archaea ThrDHs that belong to zinc-binding medium chain alcohol dehydrogenase because of low sequence similarity and the lack of a zinc-binding domain in the sequence. A specific, quantitative, and sensitive enzymatic endpoint method for l-threonine determination was developed by using a ThrDH microplate assay. The assay was successfully applied for determination of l-threonine in human serum and plasma. Our specific determination is simple, convenient, inexpensive, accurate, and suitable for mass screening determination of l-threonine in a number of samples.