Nephroprotective effects of b-carotene on ACE gene expression, oxidative stress and antioxidant status in thioacetamide induced renal toxicity in rats.

ß -carotene is one of carotenoid natural pigments, which are produced by plants and are accountable for the bright colors of various fruits and vegetables. These pigments have been widely studied for their ability to prevent chronic diseases and toxicities. This study was designed to evaluate the effects of ß-carotene on angiotensin converting enzyme (ACE) gene expression, oxidative stress and antioxidant status in thioacetamide induced renal toxicity. Total 24 albino wistar rats of male sex (200-250gm) were divided into 6 groups as Group-1: The control remained untreated; Group-2: Received thioacetamide (200mg/kg b.w; i.p) for 12 weeks; Group-3: Received ß-carotene orally (200mg/kg b.w), for 24 weeks; and Group-4: Received thioacetamide (200mg/kg b.w; i.p) for 12 weeks + received ß-carotene orally (200mg/kg b.w), for further 12 weeks. The expression of ACE gene in thioacetamide induced renal toxicity in rats as well as supplemented with ß-carotene was investigated and compared their level with control groups by using the quantitative RT-PCR method. The ACE gene expression was significantly increase in TAA rats as compare to control rats specifies that TAA induced changes in ACE gene of kidney, elevated renal ACE has been correlated with increase hypertensive end organ renal damage. The quantity of ACE gene were diminish in our rats who received ß-Carotene after TAA is administered, for this reason they seemed to be defended against increased ACE levels in kidney bought by TAA. In pre- and post-treatment groups, we studied the role of ß-Carotene against thioacetamide in the kidney of Wistar rats. Experimental confirmation from our study illustrates that ß-Carotene can certainly work as a successful radical-trapping antioxidant our results proved that TAA injury increased lipid peroxidation and diminish antioxidant GSH, SOD and CAT in renal tissue. Since ß-Carotene administration recover renal lipid peroxidation and antioxidants, it give the impression that ß-Carotene protects renal tissue against thioacetamide-induced oxidative damage.

Common variant within the FTO gene, rs9939609, obesity and type 2 diabetes in population of Karachi, Pakistan.

AIM: To determine the effect of genetic variants within the FTO gene (rs9939609) on obesity related traits and type 2 diabetes in South Asian population of Karachi, Pakistan. METHODS: A case-control study was conducted at Baqai Institute of Diabetology and Endocrinology (BIDE), Baqai Medical University situated in Karachi. A total of 296 patients with known type 2 diabetes and 198 controls aged greater than and equal to 45 years were recruited. The Anthropometric, clinical and biochemical data was collected on a structured questionnaire. Single nucleotide polymorphism (SNP) in FTO gene was identified by Amplification Refractory Mutation System-Polymerase Chain Reaction (ARMS-PCR). Association between the single nucleotide polymorphism and categorical variables such as type 2 diabetes and obesity category was tested through logistic regression analysis. RESULTS: We observed a strong association of the minor allele A at rs9939609 with type 2 diabetes. Significant difference was observed in frequency of FTO genotype when diabetic subjects were compared with controls in co dominant, dominant and recessive models. This association remained significant even after adjusting for body mass index (BMI) and for waist circumference. The frequency of homozygous risk Alleles (AA) was found to be higher in obese & overweight (≥ 23 kg/m(2)) and females with central obesity in our study population. The association of FTO variant with BMI and central obesity does not reach to statistical significance. CONCLUSION: In the study population of South Asian ancestry, variants of the FTO gene predispose to type 2 diabetes, but not entirely through their effect on BMI.

Association of SNP rs9939609 in FTO gene with metabolic syndrome in type 2 diabetic subjects, rectruited from a tertiary care unit of Karachi, Pakistan.

OBJECTIVE: To determine the association of SNP in FTO gene, rs9939609, with Metabolic Syndrome (MS) in type 2 diabetic subjects at a tertiary care unit of Karachi, Pakistan. METHODS: We genotyped FTO rs9939609 SNP in 296 patients with type 2 diabetes from the Out Patient Department (OPD) of Baqai Institute of Diabetology and Endocrinology (BIDE). MS was defined on the basis of International Diabetes Federation (IDF) and National Cholesterol Education program (NCEP) criterion. Association between the rs9939609 SNP and MS was tested through chi-square and Z-tests by using odds ratio (OR) with 95% confidence intervals. RESULTS: The frequency of MS as defined by IDF criterion was significantly higher in female subjects as compared to male subjects (p= 0.006). Carriers of ≥ 1 copy of the rs9939609 A allele were significantly more likely to had MS (69.6%) than non-carriers (30.4%), corresponding to a carrier odds ratio (OR) of 0.52 (95% confidence interval [CI] (0.29-0.93), with a similar trend for the ATP III-defined MS."A" allele carriers under dominant model, carry all the criterion of MS more significantly as compared to non-carriers. CONCLUSION: The FTO rs9939609 SNP was associated with an increased risk for Metabolic Syndrome in type 2 diabetic populations at a tertiary care unit of Karachi, Pakistan.

Three new carbazole alkaloids and biological activities of Murraya koenigii.

Three new carbazole alkaloids, mukoenigatin (1), bikoeniquinonine (2) and murrayadinal (3), were isolated from the aerial parts of Murraya koenigii, along with mukeonine-B (4). Their molecular structures were determined on the basis of spectral analysis including UV, IR, MS, and 2D NMR spectroscopy. The antimicrobial activity of different fractions of plant extract was also determined.

Effects of curcumin on angiotensin-converting enzyme gene expression, oxidative stress and anti-oxidant status in thioacetamide-induced hepatotoxicity.

INTRODUCTION: This study aimed to evaluate the protective effects of curcumin on angiotensin-converting enzyme (ACE) gene expression, oxidative stress and anti-oxidant status in thioacetamide (TAA)-induced hepatotoxicity in rats. MATERIALS AND METHODS: Total 32 albino Wistar rats (male, 200-250 g) were divided into six groups (n=8). Group 1: untreated controls; Group 2: received TAA (200 mg/kg body weight (b.w.); i.p.) for 12 weeks; Group 3: received curcumin (75 mg/kg b.w.) for 24 weeks; Group 4: received TAA (200 mg/kg b.w.; i.p.) for 12 weeks+curcumin (75 mg/kg b.w.) for 12 weeks. RESULTS: A significantly higher ACE gene expression was observed in TAA-induced groups as compared with control, indicating more synthesis of ACE proteins. Treatment with curcumin suppressed ACE expression in TAA liver and reversed the toxicity produced. TAA treatment results in higher lipid peroxidation and lower GSH, SOD and CAT than the normal, and this produces oxidative stress in the liver. Cirrhotic conditions were confirmed by serum enzymes (ALT, AST and ALP) as well as histopathological observations. CONCLUSION: Curcumin treatment reduced oxidative stress in animals by scavenging reactive oxygen species, protecting the anti-oxidant enzymes from being denatured and reducing the oxidative stress marker lipid peroxidation. Curcumin treatment restores hepatocytes, damaged by TAA, and protects liver tissue approaching cirrhosis.

Fresh and aged human lymphocyte metaphase slides are equally usable for GTG banding.

The identification of chromosomes for routine cytogenetic analysis is based on quality of metaphases and good banding pattern. Fresh slides of human lymphocytes have been shown to produce good bands for the identification of chromosomes morphology. G-bands by Trypsin using Giemsa (GTG) banding of aged slides is generally considered hard to get desired band pattern of chromosomes persistently. The current study is focused on GTG banding of aged slides. A total of 340 subjects including 290 primary infertile and 50 fertile were selected. The blood samples were drawn aseptically for cytogenetic analysis. Lymphocytes were cultured and GTG banding was done on 1440 glass slides. Giemsa trypsin banding of aged slides were done by adjusting average trypsin time for each month according to the slide age and metaphase concentration. Correlation analyses showed a significant and positive correlation between slide ageing and trypsin pre-treatment time. The results of this study suggest that, the fresh and aged human lymphocyte metaphases are equally usable for GTG banding.

Angiotensin converting enzyme (ACE) gene expression in experimentally induced liver cirrhosis in rats.

Angiotensin converting enzyme (ACE) is a key player of Renin Angiotensin System (RAS), involved in conversion of active product, angiotensin-II. Alterations in RAS have been implicated in the pathophysiology of various diseases involving heart, kidney, lung and liver. This study is designed to investigate the association of ACE gene expression in induction of liver cirrhosis in rats. Total 12 male albino Wistar rats were selected and divided in two groups. Control group received 0.9% NaCl, where as Test group received thioacidamide (TAA), dissolved in 0.9%NaCl, injected intraperitoneally at a dosage of 200mg/Kg of body weight, twice a week for 12 weeks. The rats were decapitated and blood sample was collected at the end of experimental period and used for liver functions, enzyme activity, antioxidant enzymes and lipid peroxidation estimations. Genomic DNA was isolated from excised tissue determine the ACE genotypes using specific primers. The ACE gene expression in liver tissue was assessed using the quantitative RT-PCR method. The activity of ALT, total and direct bilirubin, SOD and CAT levels were significantly high (p<0.05) and level of MDA was significantly low (p<0.05) in TAA treated rats as compared to control rats. The ACE gene expression after 12 weeks TAA treatment in cirrhotic rats was significantly increased (p<0.05) in comparison to controls. This study describes the importance of RAS in the development of hepatic fibrosis and the benefits of modulation of this system ACE gene expression. The finding of major up-regulation of ACE in the experimental rat liver provides further insight into the complexities of the RAS and its regulation in liver injury. The development of specific modulators of ACE activity and function, in future, will help determine the role of ACE and its genetic variants in the pathophysiology of liver disease.

Relationship between serum nitric oxide and sialic acid in coexisted diabetes, hypertension and nephropathy.

This study was designed to study the relationship between serum nitric oxide and sialic acid in patients of diabetic nephropathy. Total 210 diabetic patients including 115 males and 95 females, suffering from diabetes and nephropathy (DN) were selected followed by informed consent and approval from institutional ethical committee. Equal number of age and sex matched normal healthy subjects were selected without any known history of hyperglycemia, hypertension and renal insufficiency as controls. Fasting blood samples from patients and controls were collected and analyzed for serum nitric oxide, sialic acid, fasting blood glucose (FBG), serum urea, creatinine, HbA1c and golmerular filtration rate (GFR). The raised levels (p<0.05) of systolic and diastolic blood pressures, BMI, FBG, HbA1c, serum urea, creatinine and sialic acid were noted in DN patients as compared to controls. Significantly lower levels of GFR and serum nitric oxide (p<0.05) were observed in DN patients as compared to controls. Strong negative correlation was found between serum sialic acid and nitric oxide levels in patients diabetic nephropathy (p<0.05). The relationship between the levels of serum nitric oxide and sialic acid may be considered as a strong biochemical indicator for micro and macro vascular complications of diabetes such as hypertension and nephropathy. These parameters should be taken into account during screening procedures regarding identifications of the diabetic patients to get them rid of progressive renal impairment to ESRD.

Glycemic control, dyslipidemia and endothelial dysfunction in coexisted diabetes, hypertension and nephropathy.

Diabetes mellitus is a chronic metabolic disorder that can lead to serious cardiovascular, renal, neurologic and retinal complications. Diabetes clustered with hypertension and nephropathy has become the leading cause of end-stage renal disease globally. This study describes diabetes, hypertension and nephropathy with reference to glycemic control, dyslipidemia and endothelial dysfunction indicating the foremost basis of morbidity and mortality world wide and rapidly progressing in Pakistan. Study subjects selected and divided in four groups (60 each) followed by institutional ethical approval and informed consent. Group 1: non-diabetic, normotensive control subjects; Group 2: diabetic, normotensive patients; Group 3: diabetic, hypertensive patients and Group 4: diabetic, hypertensive patients with nephropathy. Their fasting blood samples analyzed for the estimations of blood glucose, HbA1c, serum triglyceride, cholesterol, LDL-cholesterol, HDL-cholesterol, urea, creatinine, nitric oxide and sialic acid levels. Results showed that all the groups showed significant rise in fasting blood glucose. Similarly HbA1c levels were also significantly high in all the patients as compared to controls. Group 2 showed significantly high serum cholesterol and LDL levels and low HDL levels. Group 3 and 4 showed significantly high serum triglyceride, cholesterol and LDL levels where as low HDL levels as compared to controls. Group 3 showed significantly high serum creatinine. Group 4 showed a significantly high serum urea and creatinine as compared to controls. Persistent albuminuria was characteristic in Group 4 patients. Significantly low production of serum nitric oxide with high concentration of serum sialic acid was observed in Group 3 and 4 as compared to controls. Results indicate a clear relationship of declining renal function with poor glycemic control, abnormal lipid metabolism, endothelial dysfunction and initiation of acute phase response in tissues affected from the microvascular complications of diabetes like hypertension and nephropathy. It must be taken into account while screening diabetic patients to get them rid of progressive renal impairment leading to end stage renal disease.

Electrolytes and NA(+)-K(+)-ATPase: potential risk factors for the development of diabetic nephropathy.

Diabetic nephropathy is the leading cause of death that affects more than 40% of diabetic patients. Its metabolic derangements are frequently accompanied with electrolyte imbalances. This study was aimed to evaluate the electrolyte homeostasis during the progression of diabetic nephropathy in various stages of developing nephropathy. Patients admitted in diabetic wards of various hospitals of Karachi were selected and divided into 4 groups with 50 individuals each. Group I (healthy normotensive, non-diabetics with normal renal functions as control). Group II (diabetic patients with normal blood pressure and renal functions). Group III (diabetic hypertensive patients without renal disease). Group IV (diabetic nephropathy patients with nephropathy). Their fasting blood samples were drawn and analyzed for the estimations of intra erythrocyte and serum electrolytes and NA(+)-K(+)-ATPase activity. Group II patients showed a significant increase in intra erythrocyte sodium, serum potassium and calcium levels where as intra erythrocyte potassium, NA(+)-K(+)-ATPase, serum sodium and magnesium were significantly decreased as compared to control. Group III showed a significant rise in intra erythrocyte sodium levels but intra erythrocyte potassium, NA(+)-K(+)-ATPase, serum sodium, calcium and magnesium were significantly lowered as compared to control. Group IV revealed a significant increase in intra erythrocyte sodium and significant decrease in intra erythrocyte potassium, NA(+)-K(+)-ATPase, serum sodium, calcium and magnesium levels as compared to control. The results suggest the progressive trends in electrolyte abnormalities in diabetes mellitus leading to end stage renal disease along with the abnormality of their chief transport mechanism. It points towards the potentiality of electrolytes disturbances as indicators for the progression of diabetic nephropathy and also beneficial in prognosis and treatment of the disease.

Ionic and allied variations in normotensive and hypertensive diabetic patients.

OBJECTIVE: To investigate the disturbances of serum and red cell electrolytes in association with membrane Na(+)-K(+)- ATPase activity as well as the status of serum Urea, Creatinine and osmolality in normotensive diabetic and hypertensive diabetic patients. METHODS: Thirty normotensive and thirty hypertensive patients (age and sex matched) were selected along with thirty control subjects. Erythrocytes were isolated from freshly drawn blood samples, washed and used for the estimation of sodium and potassium concentrations using flame photometer (Corning 410). Erythrocyte membranes were prepared for the estimation of Na(+)-K(+)-ATPase activity in terms of inorganic phosphate released/mg protein/hour. Serum glucose, creatinine and urea were determined by well-documented ortho toulidine, Jaffe's and diacetyl monoxime methods respectively. Osmomat 030 was used to estimate the plasma osmolality. RESULTS: The intra-erythrocyte sodium, serum glucose, urea, creatinine and osmolality were increased significantly in hypertensive diabetic patients as compared to normotensive diabetic patients whereas Na(+)-K(+)-ATPase activity, serum sodium, potassium, magnesium and calcium were decreased significantly in hypertensive diabetic patients as compared to normotensive diabetic patients. CONCLUSION: Results confirmed that there is a significant difference between normotensive and hypertensive diabetic patients with respect to their electrolyte metabolism and associated pathways. These results will notably help the physicians to treat diabetic patientswith associated morbidity like hypertension.