Influence of Bcl-1 Gene Polymorphism of Glucocorticoid Receptor Gene (NR3C1, rs41423247) on Blood Pressure,Glucose in Northern Indians

Glucocorticoids and its receptor are known to be involved in the dysregulation of hormone and lipid levels. Therefore, we evaluated the association of Bcl1 gene polymorphism of glucocorticoids receptor (GCR) gene variant with hormone and lipid levels in Northern Indians obese. A total of 435 obese and non-obese age matched subjects were included in the case–control study. Lipid and hormonal levels were estimated using standard protocols. Analysis of +646 C>G NR3C1 gene polymorphism was done using PCR–RFLP. The frequencies of GR Bcl1, C>G genotypes and alleles did not differ significantly (P > 0.05) between obese and non-obese. The +646 G allele carriers had higher waist to hip ratio, blood pressure, insulin and glucose levels than non-carriers in obese subjects while diastolic blood pressure and glucose in non-obese. The NR3C1, +646 C>G polymorphism did not associate with obesity. However, the GG genotype may modulate blood pressure, blood glucose and hormonal levels in northern Indians.     

Seminal plasma levels of 15-F2α-isoprostane, malondialdehyde and total homocysteine in normozoospermic and asthenozoospermic males

It has been proposed that oxidative stress plays an important role in male infertility. The aims of this study were to compare seminal plasma levels of 15-F2t-isoprostane (8-iso-PGF2α), malondialdehyde (MDA), and total (sum of free and bound) homocysteine (tHcy) from normozoospermic vs. asthenozoospermic men, and to examine the relationships between tHcy and lipid peroxidation products. The study was a case-control study with a simple random sampling. The case group was consisted of 15 asthenozoospermic males. This group was compared with 15 normozoospermic men. Seminal plasma levels of 15-F2α-isoprostane and tHcy were measured using commercially available enzyme immunoassay (EIA) kits. MDA levels were determined by the thiobarbituric acid (TBA) assay. The Mann-Whitney U test was used to compare two groups. Coefficients of correlation were calculated using Spearman’s correlation analysis. All hypothesis tests were two-tailed with statistical significance assessed at the p value <0.05 level. MDA levels were higher in asthenozoospermic subjects than in control subjects (0.72±0.06 μM vs. 0.40±0.06 μM; p<0.05). No differences were seen in 15-F2α-isoprostane levels in asthenozoospermic subjects and controls (65.00±3.20 pg/ml vs. 58.17±4.12 pg/ml; p>0.05). Interestingly, tHcy levels were to be slightly higher in asthenozoospermic subjects than in controls (6.18±1.17 μM vs. 4.8±0.52 μM). Sperm motility was inversely correlated with seminal plasma 15-F2α-isoprostane and MDA levels, respectively (p<0.05). In summary, seminal plasma levels of 15-F2α-isoprostane and tHcy showed no significant difference between normozoospermic and asthenozoospermic men. Sperm motility was not correlated with seminal plasma levels of tHcy. No relationship was found between tHcy and lipid peroxidation.     

Serum dehydroepiandrosterone and lipid peroxides in human volunteers of different age groups

The objective of the present study was to evaluate the changes in serum dehydroepiandrosterone (DHEA) and lipid peroxide levels during ageing in human subjects. Random blood samples were collected from a total of 128 apparently normal human volunteers of both sexes, whose age ranged between 21–70 years. The subjects were divided into groups of a decade years of age difference. Serum levels of dehydroepiandrosterone (DHEA), lipid peroxides as malondialdehyde (MDA), and insulin were analysed in all the subjects recruited in the study. In the present study there was decrease in serum DHEA levels with age (11.30 to 7.99 ng/ml). However the differences were significant only after 50 years up to 70 years. Serum MDA levels of these subjects were higher (3.91 to 4.74 ng/ml) as compared to the values reported earlier (2.64 to 3.94 ng/ml). The serum MDA levels also showed an increasing trend with age but the increase was significant only in the 40–50 years age group and values plateaud off at later ages. The MDA and DHEA levels in women in general were lower than in men and this could be due to female hormones, which are known to protect lipid against peroxidation. There was a significant negative correlation between age and DHEA (r=−0.311 P<0.05) and positive correction between MDA and age (r=+0.405 P<0.01). No significant differences were seen in serum insulin, albumin and total protein levels. These preliminary findings support the possible utility of DHEA and MDA as markers for chronological ageing.     

Depleted nitrite and enhanced oxidative stress in urolithiasis

Crystal aggregation and retention are critical events for the formation of kidney stones. There is a close association between crystal development and free radical activity in vivo. In the present study 30 subjects presenting with urolithiasis were included. Serum levels of total lipid peroxides, nitric oxide (as nitrite), α-tocopherol, plasma ascorbic acid (vitamin C) and erythrocyte superoxide dismutase activity were measured. These findings were compared with 30 age matched control subjects irrespective of sex. Student's ‘t’ test was applied for statistical analysis. There was a significant increase in lipid peroxides (p<0.001), where as significant decrease in nitrite (p<0.01) and α-tocopherol (p<0.001) levels were observed. Plasma ascorbate (p>0.05) and erythrocyte superoxide dismutase activity (p>0.05) was also found to be decreased but the difference was not statistically significant which suggests that oxidative stress is evident in urolithiasis with depletion in antioxidant status where as decrease in nitric oxide may be less abetting in disease condition.     

Hyperlipidemia, increased lipid peroxidation and changes in antioxidant enzymes, Na+−K+-ATPase in erythrocytes of type 2 diabetic patients in andhra pradesh

Plasma levels of lipids, lipoproteins and lipid peroxides and erythrocyte Na⁺−K⁺ ATPase, Mg²⁺ATPase and antioxidant enzymes were measured in type-2 diabetic patients. A significant decrease in Na⁺-K⁺-ATPase activity was observed in diabetic patients which was negatively correlated with blood glucose and lipid peroxides, while the Mg²⁺-ATPase activity was increased. In the diabetic subjects the plasma concentrations of Na⁺ and K⁺ were increased where as erythrocyte levels of Na⁺ were increased and K⁺ were decreased. Hyperlipidaemia and increased levels of lipid peroxides were observed in the diabetic subjects. There was a significant increase in erythrocyte catalase activity in diabetics which positively correlated with their lipid peroxides. There was no change in GPx activities between controls and diabetics.     

Oxidative stress and serum α<Subscript>1</Subscript> — Antitrypsin in smokers

The present study was undertaken to evaluate the levels of serum lipid peroxide, nitric oxide end poducts, erythrocytic superoxide dismutase activity and serum α ₁-antitrypsin in smokers. Total 90 active cigarette smokers were subdivided into Group I (subjects with smoking habit of less than 10 cigarettes per day) and Group II (with smoking habit of more than 10 cigarettes per day). In both groups lipid peroxide and nitric oxide end products were significantly increased with significantly decrease in erythrocytic superoxide dismutase activity and serum α ₁-antitrypsin as compared to controls. Our findings show enhanced oxidative stress and reduced α ₁-antitrypsin in cigarette smokers. Further increase in number of cigarettes per day exacerbates the oxidative stress with decrease in α ₁-antitrypsin.     

Effect of pH on spectral characteristics of P5C-ninhydrin derivative: Application in the assay of ornithine amino transferase activity from tissue lysate

Currently available method(s) for assaying pyrroline-5-carboxylate (P5C), an important intermediate metabolite of ornithine, proline and glutamate metabolic pathways, are cumbersome or not sensitive enough for microanalysis. The present study involving the synthesis of P5C followed by purity check, molecular mass (amu =113.1) determination by mass spectrometry and spectral characterization of P5C-ninhydrin derivative (λ max: 510 nm) confirmed the authenticity of the preparation. Studies on the effect of pH on spectral characteristics of P5C ninhydrin derivative demonstrated a significant change with respect to λ max (620 nm) and several ∼ 12 fold increase in molar extinction coefficient (ε: 1.96 × 10⁵) in alkaline conditions (pH:7.0–8.0) as compared to the reported Molar ε of 1.65 × 10⁴ at max λ 510 nm in ethanolic solution. The modified method, with the improved sensitivity, is adopted for the assay of ornithine amino transferase activity in WBC’s/platelets lysate(s) from human blood.     

Oxidants and antioxidants in hypoxic ischaemic encephalopathy

In the present study, the role of serum lipid peroxide and serum nitric oxide as oxidants and erythrocytic superoxide dismutase & serum vitamin E as antioxidants were determined in the 50 neonates with hypoxic ischaemic encephalopathy. (HIE) as against 25 healthy neonates as controls 50 patients of HIE were further divided into two groups i. e. mild and moderate HIE patients. All subjects were in the age group of 37–41 weeks of gestation. The levels of serum lipid peroxide, serum nitric oxide and erythrocytic superoxide dismutase were significantly elevated in both groups of neonates with HIE than those of controls (P<0.001), whereas serum vitamin E levels were significantly decreased in both groups of HIE patients than those of controls (P<0.001). A positive correlation was obtained between serum lipid peroxide and erythrocytic superoxide dismutase (r=+0.86). Alterations in the status of oxidants and antioxidants indicate role of free radicals in the development of HIE.     

Circadian periodicity of plasma lipid peroxides and anti-oxidant enzymes in pulmonary tuberculosis

The circadian periodicity of plasma lipid peroxide levels and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were studied in 50 clinically, bacteriologically and radiologically proven fresh cases of pulmonary tuberculosis (age: 21–45 years) and 60 age-matched healthy volunteers with diurnal activity from 06∶00 to about 22∶00 and nocturnal rest. A marked circadian variation in plasma lipid peroxide level was recorded in healthy subjects and pulmonary tuberculosis patients with significant amplitude and acrophase around 16∶21 and 17∶12 respectively. The acrophase tended to be delayed in tuberculosis patients. Furthermore, a statistically significant circadian rhythm was found in SOD, CAT and GPx activities in normal volunteers and pulmonary tuberculopsis patients. SOD and CAT enzyme activity was noted to be maximum at 06∶00 and minimum at 00∶00 in tuberculosis patients. The circadian acrophase for GPx activity was recorded at 16∶15 in normals and around 22∶45 in patients. Moreover, the activity was found to be decreased at all sampling hours during 24-hours sleep-awake period in patients in comparison to healthy counterparts. The MESOR and circadian amplitude also decreased markedly. The decreased activity of measured antioxidant enzymes in pulmonary tuberculosis patients could probably be associated with oxidative stress and/or decreased anti-oxidant defensive mechanism in such patients.     

Lipoprotein (A): Better assessor of coronary heart disease risk in south Indian population

In an attempt to search for risk factors which can explain the increasing prevalence of coronary heart disease (CHD) in Indian population, we conducted a case-control study to assess the association of Lipoprotein (a)(Lp(a)) with CHD. One hundred and fifty one consecutive patients with clinical and angiographic evidence of CHD and forty-nine healthy controls were drawn for the study. Triglycerides, very low density cholesterol (VLDL-C), total cholesterol (total-C)/high density cholesterol (HDL-C) ratio, low density cholesterol (LDL-C)/HDL cholesterol ratio and Lp(a) were found to be higher in patients than controls. In female sex and in those with family history of CHD, higher total and LDL cholesterol levels were observed to be associated with higher Lp(a) levels. Lp(a) levels were also found to be higher in triple vessel disease than other vessel disease patients. Significant difference in Lp(a) levels were observed between normal coronaries vs. single and triple vessel disease(P<0.05) and also between single vs. double and triple vessel disease (P<0.01).Lp(a) levels correlated positively with vessel severity(P<0.005). Lp(a) levels >25 mg/dl were associated with coronary heart disease (Odds ratio 1.98 P<0.05 95% CI 0.007–1.18). Our findings suggest a cut-off level of 25mg/dl for determination of risk of CHD. Studies from different areas involving larger sample size are needed to confirm the findings of the present study.     

Inflammatory Cytokines,Apoptotic, Tissue Injury and Remodeling Biomarkers in Children with Congenital Heart Disease

The present study aims to evaluate specific biomarkers involved in congenital heart disease (CHD), and whether there is a significant differences between the levels of these biomarkers in the cyanotic CHD (CCHD) and acyanotic CHD (ACHD). We prospectively measured tumor necrosis factor (TNF-α), interleukin-6 (IL-6), C-reactive protein (CRP), vasoendothelial growth factor (VEGF), troponin T, creatin kinase MB (CKMB), and Caspase 3 levels in 120 consecutive children with CHD (60 cyanotic and 60 a cyanotic with age 1:4 years), and 30 healthy control children. Significant elevated levels of inflammatory markers; TNF-α, IL-6 and CRP was detected in CHD, with percentage increase in cyanotic than a cyanotic subjects as compared to the normal one. Apoptotic biomarker; caspase 3 showed also significant increases in CCHD than ACHD. In addition, tissue injury mechanisms included troponin T and CKMB, exhibited significant increase in cyanotic than a cyanotic CHD. The present results demonstrate also, significant enhancement in remodeling process (VEGF), in cyanotic than a cyanotic patients. Thus, it could be concluded that, the children with CCHD were shown to have elevated levels of inflammatory cytokines, caspase 3, troponin T, and CKMB as these biomarkers may implicated in cardiac functional status.     

Pro-oxidant and anti-oxidant status in patients of sickle cell anaemia

The role of oxidant damage to red cells in sickle cell anaemia has been of interest in recent years. Although, available reports suggest that sickle cell erythrocytes are susceptible to endogenous free radical mediated oxidant damage there remains discrepancy in the status of antioxidant enzymes and antioxidant vitamins in these patients. In view of this, 107 cases of sickle cell anaemia (36 ‘SS’ and 71 ‘AS’ pattern—as confirmed by haemoglobin electrophoresis) were subjected to analysis of malondialdehyde, ascorbic acid, superoxide dismutase and albumin. The results were compared with 54 age and sex matched healthy controls. The results indicate a marked increase in lipid peroxidation and superoxide dismutase levels in both ‘SS’ and ‘AS’ types of sickle cell anaemia as compared to controls. Although no difference was observed in the levels of albumin in these groups the levels of ascorbic acid were significantly depleted in sickle cell anaemia patients. The results are indicative of enhanced lipid peroxidation along with imbalance in the pro-oxidant and antioxidant status in patients of sickle cell anaemia.     

Stimation of reference interval of lipid profile in Assamese population

12hrs fasting samples of 1485 apparently healthy, Assamese population in the age group of 20–80 yrs., mostly from the urban area of Assam were tested for serum lipid profile that includes total cholesterol, triacylglycerol, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and very low density lipoprotein cholesterol (VLDL-C) over a period of three years. The values (median and range in mg/dl) obtained were 170 (93–263); 110 (40–256); 40 (23–73); 103 (40–173) and 22 (8–51) respectively. After grouping these subjects according to the age and sex no significant difference were observed between most of the groups. Median and upper range of total cholesterol, HDL-C and LDL-C were found to be higher in women than in men in all the age groups. But triacylglycerol and VLDL-C concentrations were observed to be higher in the men than women except in age group of 61–70 yrs. It can be suggested that lipid values obtained in this study can be used as the reference value, based on which clinical correlation can be made.     

A rabbit model for studying hypocholesterolemic effect of drugs and hypocholesterolemic effect of extracts of garlic (Allium sativum)

An experimental model of hypercholesterolemic rabbits suitable for studying the hypocholesterolemic effect of compounds was developed. Rabbits were made hypercholesterolemic by oral administration of cholesterol (100mg/kg body weight/day) suspended in groud nut oil by gastric intubation (Ryle's tube). Cholesterol can be given to rabbits from 10 days to 6 months depending on the degree of hypercholesterolemia required and duration of study of hypocholesterolemic effect. In one month cholesterol feeding experiment, the serum cholesterol level in normal controls (not given cholesterol) was 67±11.3 mg/dl and in cholesterol fed animals 191.3±70.6 mg/dl. In 2 months experiment, besides hypercholesterolemia, abnormal serum lipid profile and increase in lipid content in liver, heart and aorta were also seen. To such animals when water and methanol extracts of garlic were given along with cholesterol, there was significant decrease in serum cholesterol level. The advantage of this method over the method in which cholesterol is mixed with diet to induce hypercholesterolemia is that exactly same and calculated amount of cholesterol can be given for each animal. This minimises the variations in serum cholesterol levels in different animals.     

Antioxidant status and peroxidative stress in mother and newborn —A pilot study

Serum antioxidant status and peroxidative stress was estimated in 30 pregnant women, their newborn infants and 25 non-pregnant women. Serum Ascorbic acid, α-tocopherol, retinol & β-carotene levels were found to be significantly reduced and peroxidative stress was significantly higher in mothers as compared to matched non-pregnant women. Newborn had significantly higher levels of ascorbic acid as compared to their mother but had significantly lower levels of lipid soluble vitamins. The peroxidative stress in newborn was found to be significantly less as compared to their mothers. A positive correlation of ascorbic acid and α-tocopherol levels between mother and newborn reflects that their status in mother does influence the newborn status.     

Oxidative stress in Parkinson’s disease

Oxidative stress contributes to the cascade, leading to dopamine cell degeneration in Parkinson’s disease. However, oxidative stress is intimately linked to other components of the degenerative process, such as mitochondrial dysfunction, excitotoxicity, nitric oxide toxicity and inflammation. It is therefore difficult to determine whether oxidative stress leads to or is a consequence of, these events. Oxidative stress was assessed by estimating lipid peroxidation product in the form of thiobarbituric acid reactive substances, nitric oxide in the form of nitrite & nitrate. Enzymatic antioxidants in the form of superoxide dismutase, glutathione peroxidase, catalase, ceruloplasmin and non enzymatic antioxidant vitamins e.g. vitamin E and C in either serum or plasma or erythrocyte in 40 patients of Parkinson’s disease in the age group 40–80 years. Trace elements e.g. copper, zinc and selenium were also estimated. Plasma thiobarbituric acid reactive substances and nitric oxide levels were Significantly high but superoxide dismutase, glutathione peroxidase, catalase, ceruloplasmin, vitamin-E, vitamin-C, copper, zinc and selenium levels were significantly low in Parkinson’s disease when compared with control subjects. Present study showed that elevated oxidative stress may be playing a role in dopaminergic neuronal loss in substentia nigra pars compacta and involved in pathogenesis of the Parkinson’s disease.     

Role of Cytokines in the Pathogenesis of Non-Alcoholic Fatty Liver Disease

A number of factors are linked with non-alcoholic fatty liver diseases (NAFLD), a condition that ranges from clinically benign fatty liver to its more severe form, non alcoholic steatohepatitis (NASH). In this study, we evaluated the role of cytokines secreted from adipose tissue in the pathogenesis and progression of NAFLD. We also compared anthropometric profile, lipid profile and insulin resistance data in 105 NAFLD patients with 77 normal subjects. These subjects showed a normal serum albumin level, prothrombin time and renal function but elevated aminotransferases. Predisposing factors were diabetes mellitus (35%), overweight (56%) and hyperlipidemia (44%). Insulin resistance (IR), determined by homeostasis model assessment (HOMA) was confirmed in 70% patients with NAFLD and 42% patients fulfilled the minimum criteria for insulin resistance syndrome (IRS). NAFLD patients showed elevated levels of pro-inflammatory cytokines tumor necrosis factor (TNF)-α, and interleukin (IL)-6, while anti-inflammatory cytokines IL-4 level decreased and IL-10 level remain unchanged; however, TGF-β1 level elevated significantly compared to normal subjects. While insulin level and HOMA-IR both were significantly positively correlated with BMI, waist-to-hip ratio, total cholesterol, VLDL-cholesterol, triglyceride and TGF-β1; glucose, IL-6 and TNF-α levels were significantly positively correlated with HOMA-IR only. In conclusion, pro-inflammatory cytokines play an important link between metabolic and liver disorders in the fat accumulation, and thereby cause IR, inflammation and liver fibrosis.     

hs-CRP: A potential marker for hypertension in Kashmiri population

Hypertension is the most important public health problem in developing countries and one of the major risk factors for cardiovascular diseases, and it has been reported that hypertension is in part an inflammatory disorder and several workers have reported elevated levels of CRP in hypertensive individuals. The main aim of the present study was to evaluate the association between blood pressure and serum CRP levels across the range of blood pressure categories including prehypertension. A total of 104 patients and 63 control subjects were included in the present study. The level of CRP in the serum samples was estimated by a high sensitivity immunoturbidometric assay. Standard unpaired student’s ‘t’ test was used for comparison of hs-CRP levels between hypertensive patients and normotensive control subjects and between patient groups with different grades of hypertension and different durations of hypertensive histories. The mean serum hs-CRP level in hypertensive patients was 3.26 mg/L compared with 1.36 mg/L among normotensive control subjects (P<0.001). On comparison with normotensive control subjects, the hs-CRP levels vary significantly both with grades and duration of hypertension, with most significant difference found in patients with prehypertension (P<0.001), followed by Stage-I (P=0.01) and Stage-II(P=0.02) hypertensives. Significant difference in hs-CRP levels was also found in patients with shorter duration of hypertensive history (≤ 1year) when compared with those with ≥5 years of hypertensive history (P<0.01). Our study reveals a graded association between blood pressure and CRP elevation in people with hypertension. Individuals with prehypertension or with shorter duration of hypertension (≤1 Year) had significantly a greater likelihood of CRP elevation in comparison to chronic stage-I or stage-II hypertensives.     

Hypocholesterolemic and hepatoprotective effects of flaxseed chutney: Evidence from animal studies

Rats fed with hypercholesterolemic diet showed a significant increase in serum total—cholesterol, liver homogenate total-cholesterol, HDL-cholesterol and changed LDL-cholesterol, and HDL/LDL ratio in comparison to control. Flaxseedchutney (FC) supplemented diet (15%, w/w) was found to be more effective in restoring lipid profile changes in rats fed with cholesterol, (1.0%). The activities of serum marker enzymes glutamate oxaloacetate transminase (GOT), glutamate pyruvate transaminase (GPT) and alkaline phosphatase (ALP) were elevated significantly in carbon tetrachloride induced rats. Administration of flaxseedchutney (15%, w/w) resulted in depletion of serum marker enzymes and exhibited recoupment thus showing significant hepatoprotective effect. It was observed that flaxseedchutney supplemented diet could lower the serum cholesterol and as a potential source of antioxidants it could exert protection against hepatotoxic damage induced by carbon tetrachloride (CCl₄) in rats.     

Vitamin E supplementation on biochemical changes observed in agricultural field workers exposed to different classes of pesticides

A total of 41 healthy male pesticide sprayers exposed to different clases of pesticides for 3-5 years were compared with 21 controls matched for age and economic status with respect to free radical generation, lipid peroxidation, antioxidant status, cholesterol, lipoprotein status and haematological profile. Plasma lipid peroxidation was estimated in the form of thiobarbituric acid reactive substances (TBARS) produced. Significant increase in TBARS was observed in sprayers population when compared with control subjects and the level of TBARS increased with increase in the duration of exposure. The levels of antioxidants such as glutathione (GSH) were significantly depleted, whereas those of superoxide dismutase (SOD) were remarkably increased than control population. Significant reduction in total cholesetrol, alteration in lipoprotein fractions and nonsignificant changes in hematological parameters were observed. These results suggested that exposure to pesticidal residual drift augments the free radical generation, and lipid peroxidation. Decline in non-enzymatic antioxidant and elevation of enzymatic antioxidant were observed. Supplementation of α-tocopherol for 45 days resulted in the partial restoration of these biochemical changes produced by pesticides.