Association of insertion/deletion mutations in angiotensin converting enzyme (ACE) gene and effectiveness of Glibenclamide therapy in Iranian type 2 diabetic patients

Abstract Introduction: Type 2 diabetes mellitus (T2DM) is an expanding global health problem, closely linked to the epidemic of obesity. The incidence of diabetes is increasing because of aging, changing ethnic mix of the population and worsening obesity. Glibenclamide is used for the treatment of patients with type II diabetes mellitus. Some patients respond well to this therapy while others need to use higher doses along with other medications. Since polymorphisms in ACE gene has been associated with type 2 diabetes mellitus, in the present study we investigated the association of insertion/deletion mutations of this gene with the effectiveness of Glibenclamide in treating Iranian type 2 diabetic patients.   Methods and Results: In this experimental study, blood samples from type II diabetic patients were collected (n=99) and their genomic DNA was isolated. Specific primers for the detection of insertion/deletion mutation were used and polymerase chain reactions (PCR) were conducted using specific thermal cycles. The amplified DNA samples were detected by electrophoresis of these samples on a 0.7% agarose gel. Statistical analysis of the obtained data was performed using t-test and chi-square test. A total of 99 patients were enrolled to the study. The frequency distribution of DD, ID, and II polymorphisms were 72%, 20%, and 8%, respectively. There were no differences among genotypic groups (P = 0.146). In terms of cholesterol, there was a significant difference between DD and DI (P = 0.012). There was a significant difference between the two DD and II genotypes in terms of creatinine (P = 0.034)   Conclusions: Although the results of our study indicated no association of ACE I/D polymorphisms and Effectiveness of Glibenclamide therapy, DD genotype may play a role on effectiveness of Glibenclamide Therapy

Optimization of the expression of genes encoding poly (3-hydroxyalkanoate) synthase from Pseudomonas aeruginosa PTCC 1310 in Escherichia coli

Introduction: Over the years, the use of plastics has complicated the problem of disposal of solid wastes. One strategy to reduce plastic waste is the use of biodegradable plastics. A group of these plastics are polyhydroxyalkanoates (PHAs). To date more than 250 different microorganisms are known to synthesize and accumulate PHA. Most Pseudomonas strains are able to accumulate mcl-PHA. In previous studies, the phaC1 and phaC2 genes were identified in Pseudomonas aeruginosa (P.aeruginosa) PTCC 1310 and were cloned. The aim of this study was to express these genes and optimize the conditions for their expression.  Methods and Results: The inserts obtained from vectors pTZPHAC1 and pTZPHAC2 were subcloned into pET15b expression vector. After transformation of competent Escherichia coli (E.coli) BL21 (DE3) cells with recombinant plasmids, expression was induced using IPTG. By changing expression conditions such as IPTG concentration, time and temperature of incubation with IPTG, the expression conditions for these enzymes were optimized, and the obtained results were compared using proper statistical analysis. Conclusions: The PHA synthase genes were induced with IPTG and the expressed 62 kDa protein was observed and purified. By changing expression conditions, 1 mM IPTG, 37°C and a 2 hr incubation provided the highest level of protein production in E.coli cells. These results suggest that induction condition of PhaC genes can influence expression of PHA synthase enzymes

The study effect agency theory and signaling theory on the level of voluntary disclosure of listed companies in Tehran Stock Exchange

This study investigates the effects of agency theory and signaling theory on the level of voluntary disclosure . empirically the research data is collected from 45 firms in the Tehran Security Exchange (TSE) during 2008-2013.multiple regressions technique is used for examining the stated hypotheses. It is used for, two models. The first model is based on agency theory and the second model is based on signaling theory.The relationship between ratio of fixed assets, Leverage, ROE, Liquidity on the level of voluntary disclosure.. In order to examine the hypotheses, data is collected from the annual reports of the companies using official bulletins of the Tehran stock exchange, mainly, through Novin software, ................. Keywords:agency theory,   signaling theory,  voluntary disclosure

Molecular cloning and expression of rat µ-opioid receptor in Escherichia coli (BL21)

     The µ (mu) opioid receptors, which mediate the effects of morphine, are widely distributed in brain. The purpose of this study was to design a simple expression system for rat µ-receptor in Escherichia coli (BL21). In this laboratory study, rat µ-receptor cDNA was isolated from pcDNA3 vector using Xba1 and Hind3 restriction enzymes. pET-15b was digested by Nco1 restriction enzyme. µ-receptor cDNA and pET-15b formed a recombinant DNA that was transformed to Escherichia coli (BL21). The insert presence was proved by Rsa1 restriction enzyme and the induction of its expression was performed using IPTG. Finally, the presence of desired insert was confirmed using RSA1, and the colonies that had correct orientation in gene containing plasmid were used for further studies. On the SDS-page gel electrophoresis, a 33 kDa band was observed when IPTG was used at 0.5 and 1 mM concentrations, that is equal to calculated molecular weight of rat µ-receptor. At the end of this project, the expression of rat µ-receptor by IPTG induction was successfully performed.

Expression and activity evaluation of reteplase in Escherichia coli TOP10

   Reteplase is a part of tissue Plasminogen Activator (t-PA) used for theremoval of thrombi in blood vessels. In the present study we express the reteplase genein Escherichia coli TOP10 and then its thrombolytic activity was measured. The recombinant plasmid pBADgIIIA was transformed into the competent Escherichia coli TOP10and then transformed bacteria was seeded into bioreactor containing 1.5 L LB medium and induced by 0.02% L-Arabinoseat 37°C, pH 7, and 180 rpm until OD 600 of 0.6 was reached.Samples were analyzed by SDS-PAGE and western blotting andthe expression of reteplase was examined. Finally the activity of this recombinant protein was evaluated using Chromogenic Activity Assay Kit. The presence of reteplase in transformed Escherichia coli TOP10 wasexamined by western blotting which revealed that the target protein in form inclusion body was expressed as a unique band at39 and the refolded reteplase was 66KDa. The amount of protein produced was 90.5µg/mL and its activity was determined as 0.8 units. In this study, the expression of reteplase in Escherichia coli TOP10 wasscaled up under optimum condition. Furthermore we earned reteplase with partially suitable thrombolytic activity.

Evaluation of Leptin and Adiponectin Levels in Patients with Stable Angina Pectoris

<p class="abstract"><strong>BACKGROUND:</strong> Leptin and adiponectin are two adipose tissue hormones and their association with the incidence of cardiovascular diseases is under evaluation. The aim of this study was to determine the relationship of leptin and adiponectin with coronary artery diseases.</p> <p class="abstract"><strong>&nbsp;&nbsp; METHODS:</strong><strong> </strong>One hundred and seventy patients with angina pectoris and indications of coronary angiography underwent angiography. Serum levels of blood lipids, leptin, and adiponectin were measured. The gathered data was evaluated using SPSS software, V. 15, by multivariate variance analysis.</p> <p class="abstract"><strong>&nbsp;&nbsp; RESULTS:</strong> Analysis of the data demonstrated that 45.1% of the patients had positive angiographic findings. The serum levels of leptin and adiponectin were significantly lower than the minimum levels specified by the kit. However, the two groups, i.e., patients with positive angiographic findings and those with negative findings were not significantly different according to the serum levels of the hormones. Moreover, no significant correlation between the serum levels of the hormones and serum lipids was observed.<strong>&nbsp;</strong></p> <p class="abstract"><strong>&nbsp;&nbsp; </strong><strong>CONCLUSION:</strong> Various studies have demonstrated that high serum level of leptin and the incidence of coronary artery diseases are correlated. On the other hand, they have reported that adiponectin has cardioprotective role. Confirmation of these findings requires more detailed studies.</p> <p class="abstract">&nbsp;</p><table cellspacing="0" cellpadding="0" align="left"><tbody><tr><td width="35" height="12"><br /></td></tr> <tr><td><br /></td> <td>&nbsp;</td></tr></tbody></table><strong>Keywords:</strong> Leptin, Adiponectin, Coronary artery disease<br /

The Relationship Between Social Support and Self-care in Patients With Heart Failure: The Role of Illness Related Worries as a Mediator

Objectives: Heart failure (HF), as a chronic disease, is a progressive and debilitating problem in communities. In previous studies, the role of self-care in HF has been emphasized. Illness-related worries and social support are associated with self-care. The aim of this study is to explore the role of illness-related worries as a mediator in the relationship between social support and self-care in patients with HF. Materials and Methods: This descriptive and correlational study was conducted on 149 HF patients based on inclusion and exclusion criteria, in 2016. Patients were selected using availability sampling. For data collection, a demographic information questionnaire, self-care behavior scale, illness-related worries questionnaire, and social support scale were used. In addition, the Pearson correlation coefficient and Sobel test were also conducted. Results: Sobel test results was used for understanding the mediating role of illness related-worries (P <0.01, Sobel test result = -5.16). The correlation between social support and self-care was -0.518, P <0.01, and the correlation between illness-related worries and self-care was -0.71, P < 0.01. Conclusions: This study showed that illness related worries can serve as a mediator in the relationship between social support and self-care. There was a significant correlation between the research variables