Association of a single-nucleotide polymorphism in C12orf43 region with the risk of coronary artery disease.

The genetics of organisms play a vital role in the development of coronary artery disease (CAD), with its heritability estimated at approximately 50-60%. For this purpose, we examined the relationship between CAD risk and C12orf43/rs2258287 polymorphisms in the Pakistani population. In this study based on the genetic approach to dyslipidemia, a total of 200 subjects were included from the southern Punjab. The biochemical analysis of parameters (total cholesterol, triglycerides, blood glucose, high-density lipoprotein, and low-density lipoprotein) was carried out along with molecular analysis using an ARMS-PCR-based assay for single-nucleotide polymorphism (SNP) C12orf43/rs2258287 to identify the genotype. Genotypes showed a substantial correlation with both family history and metabolic markers. The cholesterol, low-density lipoprotein cholesterol (LDL-C), triglycerides and blood glucose levels were higher while the high-density lipoprotein cholesterol (HDL-C) level was lower significantly (p<0.05) in cases than in controls. Age, pulse rate, diabetes, physical activity, smoking, family history, and dietary habits were also significantly associated (p<0.05) with CAD individuals. The SNP C12orf43/rs2258287 also showed an association with CAD in the population of southern Punjab. Based upon this study, it could be concluded that CAD is characterized by an unfavorable lipid profile in association with SNP C12orf43/rs2258287.

Elevated serum level of human alkaline phosphatase in obesity.

OBJECTIVE: To investigate a correlation between serum alkaline phosphatase level and body mass index in human subjects. METHODS: The comparative cross-sectional study was carried out at the National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan, from April 2012 to June 2013. Blood serum alkaline phosphatase levels were estimated and the subjects were divided into three sub-groups on the basis of their body mass. INDEX: normal weight (<25kg/m2), overweight (25-27kg/m2) and obese (>27kg/m2) subjects. The serum samples were used for the estimation of clinically important biochemical parameters, using commercial kits on clinical chemistry analyser. RESULTS: Of the 197 subjects, 97(49%) were obese and 100(51%) were non-obese. The serum alkaline phosphatase level increased in obese (214±6.4 IU/L) compared to the non-obese subjects (184.5±5 IU/L). Furthermore, a significant linear relationship (r=0.3;p-0.0001) was found between serum alkaline phosphatase and body mass index. Other biochemical variables were not correlated to the body mass index. CONCLUSIONS: Over activity and higher amounts of alkaline phosphatase were linked to the development of obesity.

Renin Angiotensin Aldosterone System (RAAS): its biology and drug targets for treating diabetic nephropathy.

Diabetes mellitus is a multifactorial disorder of hyperglycemia caused by a combination of biochemical, molecular and genetic factors, which leads to the dysfunction of various organs including kidneys. Diabetic nephropathy (DN) is one of the microvascular complications of diabetes that results due to poor glycemic control. Several molecular and biochemical pathways have been implicated in the pathogenesis of DN. Of these, the Renin Angiotensin Aldosterone System (RAAS) is considered as a key pathway. RAAS involves various subsystems which contribute to the development of DN. Mutations in several genes of the RAAS pathway have been associated with the development of DN. These genes or their products present them as therapeutic targets for potent drugs to control or prevent DN, and development of new drugs for targeting the RAAS. Drugs in use for DN are mainly the Angiotensin Converting Enzyme (ACE) inhibitors, Angiotensin Receptors Blockers (ARB) and renin inhibitors which play important roles in reducing DN. Hence, the present review is focused on the pathophysiology and genetic factors for DN by exploring the RAAS pathway and emphasizing the benefits of blocking this pathway to control and prevent DN.

Biogenic Synthesis, Characterization, and In Vitro Biological Evaluation of Silver Nanoparticles Using Cleome brachycarpa.

The therapeutical attributes of silver nanoparticles (Ag-NPs) in both conditions (in vitro and in vivo) have been investigated using different plants. This study focused on the green chemistry approach that was employed to optimize the synthesis of silver nanoparticles (AgNPs) using Cleome brachycarpa aqueous extract as a reducing and stabilizing agent. The characterization of obtained CB-AgNPs was undertaken using UV-visible spectroscopy, Atomic-force microscopy (AFM), Fourier-Transform Infrared Spectroscopy (FTIR), scanning electron microscopy (SEM), and Energy-Dispersive X-ray (EDX) analysis. Results suggest that CB-AgNPs synthesized via stirring produced small-sized particles with more even distribution. The synthesized silver nanoparticles were spherical with a 20 to 80 nm size range. In vitro studies were used to analyze antioxidant, antidiabetic, and cytotoxic potential under different conditions. The results also indicated that CB-AgNPs may have significant potential as an antidiabetic in low concentrations, but also exhibited potential antioxidant activity at different concentrations. Moreover, the anticancer activity against the breast cell line (MCF-7) with IC50 reached up to 18 µg/mL. These results suggest that green synthesized silver nanoparticles provide a promising phytomedicine for the management of diabetes and cancer therapeutics.

Discovering the anti-diabetic potential of pomegranate peel metabolites by examining molecular interplay with the thioredoxin-interacting protein.

Introduction: Medicinal plants like Punica granatum (pomegranate) have traditional uses against diabetes, inflammation and other diseases. The study was initiated to get an insight into the interaction tendency of P. granatum derived compounds with diabetes associated human thioredoxin-interacting protein (TXNIP). High glucose in diabetes induces production of TXNIP resulting in ß-cells apoptosis. Its inhibition might reduce the diabetes incidence. Methods: To elucidate the therapeutic potential of P. granatum peel against diabetes through GC-MS based identification of extracted compounds followed by application of computational algorithms. P. granatum peel extracts were screened for antioxidant, anti-inflammatory, anti-diabetic, antimicrobial and wound healing properties. Phytochemical and GC-MS based analysis were performed to identify the bioactive compounds. Molecular docking analysis was performed by Auto Dock Vina to predict the binding tendency of P. granatum derived compounds with TXNIP. Results and Discussion: The peel exhibited antioxidant, anti-inflammatory and anti-diabetic activities, which were attributed to phytochemicals like phenols, tannins and steroids. GC-MS analysis identified 3,5-octadien-2-one, 1H-pyrrole -2,5-dione, Beta-D-lyxofuranoside, 5-O-(beta-D-lyxofuranosyl)-decyl, diethyl phthalate, 9-octadecenoic acid (Z)-, methyl ester, hexadecanoic acid, methyl ester, n-hexadecanoic acid, tetradecane, 2,6,10-trimethyl, bis (2-ethylhexyl) phthalate, decane, 3,8-dimethyl, 9-octadecenoic acid (Z)-, methyl ester and bis (2-ethylhexyl) phthalate in P. granatum peel extracts. Docking analysis revealed high binding affinities of bis (2-ethylhexyl) phthalate and 9-octadecenoic acid with TXNIP i.e., -4.5 and -5.0 kcal/mol, respectively, reflecting these compounds as potent antidiabetic agents. This study validates the traditional uses of P. granatum peel and demonstrates how computational approaches can uncover pharmacologically active phytochemicals. The results suggest P. granatum peel is a promising source of novel therapeutics against diabetes, inflammation, and oxidation. Further studies on the optimization of identified ligands are warranted.

Isolation and Characterization of Plant Growth Promoting Antagonistic Bacteria from Cotton and Sugarcane Plants for Suppression of Phytopathogenic Fusarium Species.

BACKGROUND: Plant Growth Promoting Rhizobacteria (PGPR) may be utilized to augment plant growth and suppress the plant pathogens. Objective: The present study was conducted to isolate and characterize the antagonistic bacteria indigenous to cotton and sugarcane rhizosphere in Pakistan, and to evaluate their ability to suppress phytopathogenic Fusarium spp. Out of 63 isolates 37 different morphotypes were studied for their antagonistic activity against Fusarium monoliformae, Fusarium oxysporum and Fusarium solani. Among these 31 strains showed the percentage suppression ranging from 40 to 66% against Fusarium spp. OBJECTIVES: The antagonistic bacteria having antifungal activity were studied for different morphological and physiological characteristics using Gram staining and light microscopy. Most of them were Gram negative and tentatively identified as Pseudomonas spp. The selected strains were screened in vitro for plant growth regulation and antifungal traits. MATERIAL AND METHODS: Our study included 1000 premature CAD patients that classified into two groups with history of MI (n = 461) and without of MI (n = 539). The polymorphism variants in 10% of samples were determined by PCR-RFLP technique and genotyping of the polymorphism in all subjects was conducted by High Resolution Melting method. Given the two conditions of patients residing in Tehran and also faced with their first episode of MI, 640 out of 1000 study samples that had been previously followed-up were assessed in a retrospective cohort phase regarding long-term major adverse cardiac events (MACE). RESULTS: Four bacterial strains were able to produce the chitinase enzyme while four other bacterial strains showed protease production. Ten strains were positive for HCN production. Out of 37, eight strains showed phosphate solubilization ranging from 13 to 24 µg/ml. eighteen strains produced indole acetic acid ranging from 5 to 19 µg/ml. CONCLUSIONS: This study identified specific traits in the isolated rhizobacteria which make them good candidates as PGPR and might contribute to enhance growth of crop plants. This information is of general interest and also helpful for devising strategies to manage diseases caused by Fusarium in cotton and sugarcane.

Implications of advanced oxidation protein products (AOPPs), advanced glycation end products (AGEs) and other biomarkers in the development of cardiovascular diseases.

OBJECTIVE: To study the putative effects of Advanced Oxidation Protein Products (AOPPs) and Advanced Glycation End Products (AGEs) in the development and progression of cardiovascular disease (CVD). METHODOLOGY: AGEs, AOPPs, e-NOS, lipid profile, circulating stress and inflammatory biomarkers were evaluated among fifty cardiovascular patients and fifty controls. Independent student's t-test was done for statistical analysis. RESULTS: The malondialdehyde mean level in CVD patients (5.45 nmol/ml) was significantly higher than control (1.36 nmol/ml) (p value = 0.018). Nitric oxide in CVD patients (55.72 ng/ml) was remarkably increased as compared to normal subjects (19.19 ng/ml). A significant change in the mean serum level of AGEs in CVD patients (2.74 ng/ml) and normal individuals (0.85 ng/ml) was recorded (p value = 0.000). The AOPPs also showed significant increased levels in CVD group (132.07 ng/ml) in comparison with normal subjects (83.05 ng/ml) (p value = 0.011). The mean eNOS serum level in CVD group (15.50 U/L) was higher than control group (11.28 U/L) (p value = 0.004). Cardiovascular disease patients, in comparison with healthy controls, showed increased level of total cholesterol (5.48 mmol/L vs 4.45 mmol/L), triglycerides (2.59 mmol/L vs 1.24 mmol/L), and low density lipoprotein (2.47 mmol/L vs 2.31 mmol/L) along with decrease in high density lipoprotein (1.39 mmol/L vs 1.74 mmol/L). The mean MMP-11 serum levels in CVD group (98.69 ng/ml) was almost double of control group (45.60 ng/ml) (p value = 0.017). The mean serum level of TNF-α and IL1-α were 32.16 pg/ml and 6.64 pg/ml in CVD patient. The significant decreasing trend of SOD (p value = 0.041), CAT (p value = 0.018), GSH (p value = 0.036) and GRx (p value = 0.029) but increasing drift of GPx (0.023) level was observed in CVD patients. CONCLUSION: This study provides strong evidence that CVD patients presented with elevated oxidative stress, enhanced inflammation and lipid profile in their serum. Therefore, the study strongly approves that AGEs, AOPPs, inflammatory and lipoxidative biomarkers hold predictive potential in causing and aggravating the disease, thus by controlling these factors CVD progression can be inhibited.

Role of diagnostic factors associated with antioxidative status and expression of matrix metalloproteinases (MMPs) in patients with cancer therapy induced ocular disorders.

BACKGROUND: Cancer patients when treated with different chemotherapeutic drugs often develop mild to severe sight threatening diseases during or after chemotherapy. The mechanism involved in the pathogenesis of ocular toxicities is poorly understood. Oxidative stress, inflammation and MMPs (angiogenic factor) are involved in the progression of chemotherapy related ocular disorders. MATERIALS AND METHODS: The concentration of oxidative stress markers such as MDA, NO and levels of different antioxidant molecules such as SOD, CAT, GSH, GPx, GPr, VIT A, VIT E and VIT C present in the serum of chemotherapy treated patients (n = 50) and in normal persons (n = 20) were estimated by the direct spectrophotometric method while the concentration of TNF-α and MMP-9 activity were determined using human TNF-α and MMP-9 ELISA kits. RESULTS: The concentration of SOD and CAT (0.356 ±â€¯0.05 µg/dl and 1.26 ±â€¯0.01 µmol/mol of protein) was significantly lower as compared to that (1.09 ±â€¯0.03 µg/dl and 3.99 ±â€¯0.04 µmol/mol of protein) in controls. The levels of GPx (0.06 ±â€¯0.01 mmol/dl) in the cancer patients were much lower than those in the controls (0.78 ±â€¯0.06 mmol/dl). Lower level of GSH (0.96 ±â€¯0.003 µg/dl) in serum of the diseased group was observed as compared to healthy group (7.26 ±â€¯1.40 µg/dl). The level of Vit A, Vit C and Vit E was lower in systemic circulation of cancer patients (109.99 ±â€¯6.35 µg/ml, 1.26 ±â€¯0.36 µg/ml and 1.29 ±â€¯0.191 µg/ml) as compared to control subjects (166.35 ±â€¯14.26 µg/ml, 3.25 ±â€¯0.099 µg/ml and 6.354 ±â€¯2.26 µg/ml) respectively. The concentration of nitric oxide was significantly higher in the cancer patients (45.26 ±â€¯6.35 ng/ml) than that in the normal subjects (16.35 ±â€¯3.26 ng/ml). The higher concentration of MDA (8.65 ±â€¯3.26 nmol/ml) was observed in the patients than normal ones (1.254 ±â€¯0.065 nmol/ml). The quantity of TNF-α was significantly higher in chemotherapy treated patients (32.68 ±â€¯4.33 pg/ml) as compared to the control group (20.979 ±â€¯1.98 pg/ml). Significantly higher concentration of MMP-9 (40.26 ±â€¯3.26 ng/ml) was observed in the cancer patients than the controls (7.256 ±â€¯1.95 ng/ml). CONCLUSION: Lower levels of antioxidant enzymes and non-enzymatic small molecules and higher levels of oxidative stress and inflammatory clinical parameters such as NO, MDA, TNF-α and MMP-9 may be involved in the pathogenesis of systemic chemotherapy related ocular complications such as cataract, glaucoma, blepharitis, retinitis pigmentosa, macular degeneration, pterygium and retinal degeneration.

A case control association study of COMT gene polymorphism (I/D) with type 2 diabetes and its related factors in Pakistani Punjabi population.

BACKGROUND: The Catechol-O-Methyl Transferase (COMT) gene polymorphism (I/D of C nucleotide at base position 900) has been previously implicated in the development of type 2 diabetes (T2D) and kidney disease. So, aim of this study was to find association of I/D polymorphism with T2D, and its associated factors like family history and nephropathy (End Stage Renal Disease, ESRD) patients in a cohort of Pakistani Punjabis. METHODS: Genomic DNA was extracted from human subjects divided as four study groups: controls (n = 46), diabetics (n = 46), diabetics with nephropathy/ESRD (n = 53), and non-diabetics without nephropathy/ESRD (n = 43). The 900 I/D C polymorphism in the COMT gene was tested by PCR-RFLP method. Genotype and allele frequencies as well as Odds Ratios were calculated for these groups. Groups were compared statistically for the analysis of genotypes, alleles, biochemical parameters as well as disease status. RESULTS: In comparison with control group (non-diabetic, non-nephropathy), there was no significant difference in rest of the three groups for allele or genotype frequencies of COMT gene. However, Chi square (χ(2)) analysis identified a significant (p = 0.02) correlation of the 900 I/D C polymorphism with family history of diabetes, as it was found that greater number (74%) of patients having I allele had a positive family history of T2D. CONCLUSIONS: A significant correlation of the COMT polymorphism (900 I/D C) with the family history of T2D has been observed, which has not been previously reported in Pakistani Punjabi population, however, this preliminary finding requires further validation studies.

Association of TLL1 gene polymorphism (rs1503298, T > C) with coronary heart disease in PREDICT, UDACS and ED cohorts.

OBJECTIVE: To determine the sequence variant of TLL1 gene (rs1503298, T > C) in three British cohorts (PREDICT, UDACS and ED) of patients with type-2 Diabetes mellitus (T2DM) in order to assess its association with coronary heart disease (CHD). STUDY DESIGN: Analytical study. PLACE AND DURATION OF STUDY: UCL, London, UK. Participants were genotyped in 2011-2012 for TLL1 SNP. Samples and related information were previously collected in 2001-2003 for PREDICT, and in 2001-2002 for UDACS and ED groups. METHODOLOGY: Patients included in PREDICT (n=600), UDACS (n=1020) and ED (n=1240) had Diabetes. TLL1 SNP (rs1503298, T > C) was genotyped using TaqMan technology. Allele frequencies were compared using c2 test, and tested for Hardy-Weinberg equilibrium. The risk of disease was assessed from Odds ratios (OR) with 95% Confidence Intervals (95% CI). Moreover, for the PREDICT cohort, the SNP association was tested with Coronary Artery Calcification (CAC) scores. RESULTS: No significant association was found for this SNP with CHD or CAC scores in these cohorts. CONCLUSION: This SNP could not be confirmed as a risk factor for CHD in T2DM patients. However, the low power of thesmall sample size available is a limitation to the modest effect on risk. Further studies in larger samples would be useful.