In Silico Characterization of Withania coagulans Bioactive Compounds as Potential Inhibitors of Hydroxymethylglutaryl (HMG-CoA) Reductase of Mus musculus.

Hypercholesterolemia is a mediator for the etiology of cardiovascular diseases, which are characterized as the global leading cause of mortality. We aimed to investigate the inhibitory activity of Withania coagulans compounds against 3-hydroxy-3-methylglutaryl-coenzyme A reductase (Hmgcr) of Mus musculus using an extensive in silico approach. The 3D structure of the Hmgcr protein is not yet known, so we performed the homology modeling using MODELLER and SWISS-MODEL tools, followed with structural validation and assessment. The PROCHECK web server showed that the top-ranked homology model from SWISS-MODEL has 93.4% of residues in the most-favorable region, the quality factor was 98%, and the Verify3D score was 91.43%, compared to the other generated models. The druggable protein-binding cavities in a 3D model of Hmgcr were investigated with the aid of commonly prescribed statin compounds using the CB-dock approach. We compiled a 3D compound library of W. coagulans, followed by drug-likeness evaluation, and found 20 eligible compounds. The pattern of consensus residues obtained from the CB-dock procedure was then used for grid-box docking of W. coagulans compounds and statin drugs using AutoDock 4.2, respectively. The results showed that withanolide R (-10.77 kcal/mol), withanolide Q (-10.56 kcal/mol), withanolide J (-10.52 kcal/mol), atorvastatin (-8.99 kcal/mol), simvastatin (-8.66 kcal/mol), and rosuvastatin (-8.58 kcal/mol) were promising candidates that bind Hmgcr protein. The key residues involved in protein-ligand (withanolide R) interactions were Y516, C526, V529, I530, M533, I535, and V537, and the formation of a H-bond was at C526, M533, and I535 residues. M533 was the consensus residue having a tendency to form a H-bond with withanolide Q, too. Molecular dynamics simulations were used to validate the top-ranked docked complexes for the stability of the modeled protein. We also predicted the pharmacokinetic properties of binding affinity-based top-ranked compounds and concluded that they could be used as potential inhibitors of Hmgcr. However, further in vitro and in vivo studies are essential to completing the drug development process.

Antioxidant and haematinic effects of methanolic and aqueous methanolic roots extracts of Rauwolfia serpentina Benth in type 2 diabetic mice.

Present investigation was carried out to evaluate the antioxidant and haematinic effects of methanolic (MREt) and aqueous methanolic (AqMREt) root extracts of R. serpentina in mice model of type 2 diabetes (T2D). Experimental mice were divided into nine groups (six per group) as: fructose-induced (T2D) diabetic group (distilled water 1ml/kg), negative control (0.05% DMSO 1ml/kg), positive control (pioglitazone 15mg/kg) and six test groups (MREt 10, 30 & 60mg/kg & AqMREt 50, 100 & 150mg/kg). Whereas tenth group was served as normal control (1ml/kg distilled water). All test doses of MREt & AqMREt significantly (p<0.05) decreases the percent inhibition of catalase (CAT) and superoxide dismutase (SOD) when compared with diabetic controls. Treatment with both extracts also improved the total hemoglobin (Hb), red blood cell (RBC), white blood cell (WBC) counts, packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) in test groups. Fourier transform infrared (FTIR) spectral analysis revealed the presence of phenols moiety in both extracts. Findings suggested that AqMREt possesses more antioxidant and haematinic potential while the MREt of R. serpentina moderately possesses the same activities, which might be due to the high content of phenols present in AqMREt.

Risk association of fasting insulin resistance index in non-diabetic heart failure patients: A case-control study at Karachi, Pakistan.

Present work investigates the risk association of insulin resistance (IResistance) in Non-Diabetic Heart Failure (NDHF) patients. Eighty (n=80) NDHF patients and same numbers of healthy controls were included to investigate with anthropometric measures, fasting blood glucose level (FBGL), serum insulin (SI), FIRI and ß-cells quantification was computed through HOMA-IR. Mean rank assessment of NDHF patients showed higher significant (p<0.0001) set of values in FBGL, SI, FIRI and HOMA-IR, when compared with controls. High (p<0.0001 & p<0.05) risk in NDHF patients was associated in SI status (OR=8.93-95% CI: 4.1-19.42) and also in HOMA-IR (OR=6.6-95% CI: 3.30-13.19), when compared for Pearson value based probability through Chi Square (χ2 Test) values estimates of probability, respectively. Area under the curve (AUC) of targeted NDHF patients showed higher set of estimation (FBGL-AUC =0.667, SI-AUC =0.763, FIRI-AUC=0.780 and HOMA-IR-AUC=0.776). Association of determinants through Pearson's (r) correlation was found significantly (p<0.0001) linked with HOMA-IR and FIRI. Regression coefficient shows that for every additional unit score in FBGL and SI can expect HOMA-IR to increase by an average of 0.883 (for FBGL) and 0.0368 (for SI), respectively. Findings concluded the association of IResistance with greater risk estimation in NDHF patients.

Therapeutic role of Rauwolfia serpentina in minimizing the risk of glycosylation and associated biomarkers in experimentally induced type 1 diabetic mice.

Present work investigates the effects of hydro-methanolic roots extract (HyMREt) of Rauwolfia serpentina in type 1 diabetic mice. Mice were divided into normal, diabetic, negative and positive controls (I-IV) and three test (HyMREt doses) groups (V-VII - 50, 100, &150mg/kg). Allocated treatment of each group was given orally for 14 days in overnight fasted state. Percent change in fasting blood glucose (FBG), body weights, body tissue weights, hepatic glycogen, total lipids, glycosylated hemoglobin (HbA1c), complete blood profile and antioxidant enzymes including catalase (CAT) and superoxide dismutase (SOD) were estimated. HyMREt doses produced meaningful (p<0.0001) reduction (-39 to -53%) in FBG. Hemoglobin (Hb) levels were raised, HbA1c were considerably decreased (4.5-3.77%) and glycosylation (HbA1c to Hb) ratio was expressively (p<0.0001) improved in test groups. Dose-wise improvement (p< 0.05) in total glycogen and decrement (p<0.05) in lipids were observed in livers of test groups. HyMREt significantly decreased (p<0.05) percent inhibition of SOD and CAT. HyMREt doses progressively (p<0.05) improved RBC and other hematological parameters while decrement was only noticed in leucocyte counts. Administration of test doses of HyMREt were significantly reduced the glycosylation, oxidative stress and anemia caused by alloxan intoxication in mice.

Persea americana seeds improve glycosylation and dyslipidemia in fructose-fed streptozotocin-injected type 2 diabetic male rats.

This work explored the in-vitro phytochemical contents and antidiabetic activity of crude seeds powder of Persea americana (CSSPa) and their in-vivo biochemical effects on glycated hemoglobin, lipid profile and other parameters in type 2 diabetic rats (fructose-STZ model). There were 2 groups of over night fasted rats, control (normal diet) and diabetic (35% Fructose for 6 weeks followed with injection (i.p.) of streptozotocin (STZ) (40mg/kg bw). Diabetic group was further divided into diabetic control, positive control (pioglitazone 15mg) and test (CSSPa 500mg) groups. After the appropriate treatments in each group for 2 weeks fasting glucose level (FGL), serum lipids, insulin, alanine aminotransferase (ALT), creatine Kinase (CK) & uric acid were determined. CSPPa showed presence of alkaloids, flavonoids, phenols etc and potent antidiabetic activity with IC50 13.23±0.76µM. CSPPa treatment showed a significant (p<0.01) decline in lipid profile, while HDL showed significant increase (p<0.01) in test group as compared with positive and diabetic control groups. The serum ALT, CK, uric acid, bilirubin & fasting glucose (fbg) showed significant improvements in test group (p<0.01). Coronary risk index (CRI), Fasting insulin resistance index (FIRI), Percent glycemic change (PGC) and HbA1c values also significantly (p<0.01) improved.

Raisins: A kitchen cabinet item can restores the liver function and structure.

The aqueous methanol extract of raisins (Vitis vinifera) was investigated in carbon tetrachloride (CCl4)-induced hepatotoxic rats model. Where it was found to revert the alteration induced by CCl4 in liver structure and function by improving the body weights, liver index, liver and bile duct specific enzymes, liver conjugative and synthetic markers, reduced glutathione and the total bilirubin/ albumin ratio while increasing the percent inhibition of lipid peroxidation in test groups treated with extract in doses of 400 and 800 mg/kg body weight as compared to negative control group only treated with CCl4 3mL/kg that showed entirely opposite picture of all these parameters. Silymarin 100 mg/kg was used as reference hepatoprotective medicine in present study. In addition, histopathological studies of liver tissues of test groups displayed the restoration of liver anatomy. Therefore, raisins' extract proved to have liver protective, regenerative and antioxidant properties. These might reside in total phenolic content particularly in gallic acid and rutin in extract estimated and detected by spectrophotometric and high performance liquid chromatographic methods.

Vitamin D and bone metabolism in breast cancer patients in Karachi, Pakistan.

Breast cancer is one of the common types of malignancy worldwide and in Pakistan. The heterogeneous disease itself and its complex treatment leads to various bone-affecting complications that make breast cancer patients more vulnerable to bone fractures. Vitamin D deficiency among these women worsens the condition and promotes breast cancer growth. Thus, the purpose of the study was to assess serum levels of 25-hydroxyvitamin D (25OHD) and bone markers in women suffering from breast cancer. Serum levels of 25OHD, alkaline phosphatase (ALP), bone specific ALP, calcium (Ca), phosphorus (P), magnesium (Mg), albumin (Alb) and beta carboxyl terminal collagen crosslink (ß-CTx) were analyzed in 201 histological diagnosed patient volunteers from breast cancer clinic. Vitamin D insufficiency was present among the total study population and deficiency was particularly observed among women with metastases. These patients had significantly increased serum levels of ß-CTx and bone specific ALP when compared with the non-metastatic group. No significant difference was observed in other biochemical parameters. A weak correlation between serum levels of 25OHD and ß-CTx was observed. Therefore, monitoring of serum levels of 25OHD and bone markers at the time of diagnosis and during the course of treatment will endeavor a better overall health status.

Risk assessment of insulin resistance with anthropometric measures in non-diabetic heart failure patients: A gender based case-control investigation at tertiary care hospitals of Karachi, Pakistan.

To assess the risk of anthropometric measures and serum markers of insulin resistance in non-diabetic heart failure (NDHF) patients and the difference among male and female subjects. 53 males, 27 females NDHF patients were enrolled and 80 healthy subjects were matched as control. Anthropometric measures, fasting blood glucose level (FBGL) and serum insulin (SI) were measured. Insulin's function (ß-cells quantification) was computed through Homeostasis Model Assessment of Insulin Resistance (HOMA-IR). Significant (p<0.05) difference in height was observed in NDHF male and female patients, whereas, weight was only significant in females. Higher mean values of FBGL, SI, and HOMA-IR in NDHF patients were observed. Highly significant (p<0.0001 & p<0.05) difference was also found in similar gender based case-control markers like FBGL, SI, and HOMA-IR. Phi coefficient for risk associations showed weak-positive correlation in both genders in FBGL, SI, and HOMA-IR. Anthropometric measures indicated lesser risk in both gender, especially in females' decreased risk in body height and waist circumference was observed. Risk measurement through odds ratios (OR) of FBGL and HOMA-IR in female subjects indicated significantly (p<0.001) double risk, whereas, in SI, three fold risks were measured in females as compared to male. Odds of exposure in cases were significantly (p<0.001) greater than in controls verified with significant Pearson and Fisher Exact Probability Test (FEPT) values with two-tailed estimates of probability in χ2 (Chi -Square) estimation test. Findings suggest increased risk of IR in female NDHF patients. Increased in FBGL IR and HOMA-IR and BMI were also found as distinguishing findings in NDHF cases.

Ethanolic seeds extract of Centratherum anthelminticum reduces oxidative stress in type 2 diabetes.

This work was accomplished to assess the in-vitro antiglycation and antioxidant activities of ethanolic seeds extract of Centratherum anthelminticum (CSEt), followed by its in-vivo examination in type 2 diabetes. Overnight fasted rabbits were divided into control and diabetic groups. Rabbits in diabetic group were fed with 35% fructose solution to develop hyperglycemia that was well-monitored by glucometer. These were divided into diabetic control (distilled water 1ml/kg), positive control group (pioglitazone 15mg/kg) and two test groups (CSEt 400 and 600mg treated). All treatments were given orally. After 14 days, rabbits were sacrificed and blood samples were used to estimate glycated haemoglobin (HbA1c) while total bilirubin (direct and indirect), uric acid, alanine aminotransferase (ALT) and creatine kinase (CK) were done in sera. In addition, antioxidant parameters viz., catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH) and lipid peroxidation (LPO) were in liver tissues. The in-vitro studies showed good antiglycation and antioxidant potential of CSEt. Similarly, in-vivo investigation showed significant reduction in glycemia and body weights in type 2 diabetic test groups. Plus values of HbA1c, ALT, CK, uric acid and bilirubin were almost back to normal along with improvement found in efficiency of antioxidant parameters.

Hypoglycemic and hypolipidemic activities of crude seeds of Centratherum anthelminticum in healthy volunteers and type 2 diabetic patients.

This study first time reports the hypoglycaemic activity of crude seeds powder (CSP) of Centratherum anthelminticum in healthy and type 2 diabetic volunteers. In addition, hypolipidemic effect of same CSP was also determined in healthy volunteers. Healthy individuals were divided into control and two test groups T1 and T2 treated with 200 & 400mg of CSP. Similarly, type 2 diabetic patients were also divided into positive control (PC) treated with metformin 600mg and two test groups DT1 (CSP 400mg + metformin 600mg) & DT2 (CSP 400mg). Each group has 6 individuals and each treatment was done orally. CSP 400mg was found more hypoglycaemic on all time intervals from 30 to 120min when oral glucose tolerance test was conducted in healthy volunteers. Both test quantities of CSP 200 & 400 mg were found successful in same healthy persons in decreasing the levels of triglycerides & total cholesterol (p<0.05), low & very low density lipoprotein cholesterols (p<0.01) and keeping the level of high density lipoprotein cholesterol as same as it was observed in control group. Similarly, CSP 400mg along with metformin and alone was also found helpful in lowering the fasting blood glucose levels in type 2 diabetic patients (DT1 & DT2) -24.99% and -20.62% respectively as compared to diabetic group only treated with metformin (PC), (p<0.01). Therefore, CSP of C antheminticum proves effective hypoglycaemic and hypolipidemic agent by possibly inducing glucose tolerance in healthy individuals and type 2 diabetic patients.

Ameliorative effect of hydro-methanolic extract from roots of Rauwolfia serpentina on some biochemical parameters of type 1 diabetic mice.

Present work seeks to investigate the biochemical parameters in terms of hypoglycemic and hypolipidemic effects of hydro-methanolic roots extract (HyMREt) of Rauwolfia serpentina in type 1 (alloxan induced) diabetic mice. Animals were divided into seven groups, four control groups, and three were test groups (HyMREt at 50, 100, & 150mg/kg). Each treatment was repeated for 14 days regularly in all seven respective groups and afterwards the body weights, fasting blood glucose (FBG), insulin, and serum lipid levels were determined. Total body weights of diabetic mice treated with HyMREt extract were dose dependently (p<0.05) improved. FBG of test groups were significantly (p<0.0001) reduced in comparison with diabetic controls which displayed elevated fasting blood glucose level. The insulin levels of HyMREt treated groups were significantly (p<0.0001) higher than those of diabetic controls. Lower triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-c) and very low density lipoprotein cholesterol (VLDL-c) whereas elevated level of high density lipoprotein cholesterol (HDL-c) were observed in test dose treated groups. In comparison with diabetic controls, the converse levels of serum lipid were observed. Significant improvement in cardio-protective indices and coronary risk index was also observed. Findings of present study support the hypoglycemic and hypolipidemic potential of HyMREt of R. serpentina.

Homeostasis model assessment of insulin resistance in non-diabetic heart failure patients: A case-control investigation at tertiary care Hospitals of Karachi.

eart failure is a progressive, chronic disorder. Insulin resistance (IR) has been more and more involved as a preliminary metabolic perturbance predisposing to hyperglycemia, hyperlipidemia and atherosclerosis with others heart diseases. To investigate the relation of insulin resistance (IR) in non-diabetic heart failure patients this case-control study was carried out to ascertain the presence of IR with the aid of Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) in non-diabetic heart failure patients (NDHF patients) compared with healthy controls. The sample size was calculated for both, cases (NDHF patients) and control (healthy subjects), which was initially consisted of 113 respondents each. The study consisted of two phase duration. In Phase I, NDHF patients were approached initially; only 80 patients with NDHF completed the study procedure. In Phase II, 80 healthy subjects were targeted and matched. Fasting blood glucose level (FBGL) and serum insulin was estimated. Mathematical model to quantify ß-cell function and insulin resistance was also computed through Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) in both groups. Data was analyzed on SPSS version 16. Mean values with ± standard deviation (SD) of insulin (10.2±4.36) and HOMA-IR (2.52±1.15) were significantly (p<0.05) higher in NDHF patients as compared to control subject (6.4±3.39, 1.45±0.80). Average insulin to glucose ratio was 0.10±0.044 in NDHF patients which was significantly (p<0.0001) lowered in controls i.e., 0.073±0.039. Marginal and matrix plot analysis revealed that a higher patients count have had the HOMA-IR values <1.5 units while opposite scenario was observed in control group. Regression analyses of HOMA-IR with FBGL (as independent indicator) also authenticate the similar pattern. The present study concludes that insulin resistance (decreased insulin sensitivity) is a characteristic finding in Pakistani population of heart failure as compared to matched healthy controls.