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

Experimental and theoretical investigations of thermal transport in graphene

Graphene has been actively investigated because its unique structural, electronic, and thermal properties are desirable for a number of technological applications ranging from electronic to energy devices. The thermal transport properties of graphene can influence the device performances. Because of the high surface to volume ratio and confinement of phonons and electrons, the thermal transport properties of graphene can differ considerably from those in graphite. Developing a better understanding of thermal transport in graphene is necessary for rational design of graphene-based functional devices and materials. It is known that the thermal conductivity of single-layer graphene is considerably suppressed when it is in contact with an amorphous material compared to when it is suspended. However, the effects of substrate interaction in phonon transport in both single and multi-layer graphene still remains elusive. This work presents sensitive in-plane thermal transport measurements of few-layer and multi-layer graphene samples on amorphous silicon dioxide with the use of suspended micro-thermometer devices. It is shown that full recovery to the thermal conductivity of graphite has yet to occur even after the thickness of the supported multi-layer graphene sample is increased to 34 layers, which is considerably thicker than previously thought. This surprising finding is explained by the long intrinsic scattering mean free paths of phonons in graphite along both the basal-plane and cross-plane directions, as well as partially diffuse scattering of phonons by the graphene-amorphous support interface, which is treated by an interface scattering model developed for highly anisotropic materials. In addition, an experimental method is introduced to investigate electronic thermal transport in graphene and other layered materials through the measurement of longitudinal and transverse thermal and electrical conductivities and Seebeck coefficient under applied electric and magnetic fields. Moreover, this work includes an investigation of quantitative scanning thermal microscopy measurements of electrically biased graphene supported on a flexible polyimide substrate. Based on a triple scan technique and another zero heat flux measurement method, the temperature rise in flexible devices is found to be higher by more than one order of magnitude, and shows much more significant lateral heat spreading than graphene devices fabricated on silicon.Mechanical Engineerin

Complications of Orthognathic Surgery

Orthognathic surgery is a common approach for treatment of maxillofacial deformities. Sagittal split ramus osteotomy (SSRO) is one of the most common techniques used to treat various mandibular deformities. A LeFort I osteotomy is suggested in deformities of the maxilla and can be used along with SSRO or intra‐oral vertical ramus osteotomy (IVRO).The aim of orthognathic surgery is to improve function and facial appearance; this benefits the patient psychologically and socially. Common complications which may occur in orthognathic surgery include vascular disease, temporomandibular joints (TMJ) problems, nerve damage, infection, bone necrosis, periodontal disease, vision impairment, hearing problems, hair loss, and neuropsychiatric problems. Rarely complications could be fatal. Because of the wide range of complications the surgeon should keep prevention protocols in mind and be prepared to treat them should they occur. In this chapter, common complications of various osteotomies in the mandible and maxilla are discussed

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

Optimization Programming for Stormwater Control Measures: Methods for Sizing and Selection

The Study Problem and Research Objectives. Fouling of water quality in receiving urban storm runoff is chronic in metropolitan areas across the USA and large cities worldwide. These urban areas have well-known problems of polluted storm runoff and urban flooding. Urban storm runoff exhibits deleterious physical-chemical-biological characteristics, such as bacteria, trash content, large biochemical-oxygen-demand (BOD), oil & grease, toxic sediments, water-borne pathogens, suspended solids (SS) and total dissolved solids (TDS), heavy metals, and nutrient content that degrade water quality of receiving waters. The contamination of urban runoff is the result of a number of natural and anthropogenic processes: changing and rapid expanding population and land use within urban areas; vulnerable receiving water bodies that become contaminated from degraded storm runoff that hinders their hydrologic, ecologic, and socioeconomic functions, and intense rainfall events with pronounced seasonal and inter-annual variability of storm intensity (typical in the western United States). State and federal regulations on Total Maximum Daily Loads (TMDLs) of pollutants to natural waters from urban storm runoff are not met by most cities in the USA. This thesis presents a modeling and experimental study using one of the best available data sets on urban land use, soils, groundwater, streets, storm conveyance infrastructure, non-point and point sources of pollution, rainfall, and Stormwater Control Measures (SCMs) technologies. This thesis proposes a novel approach to (1) model, screen and or evaluate urban areas using Geographic Information Systems (GIS) with the purpose of selecting appropriate SCMs in watershed hot spots, (2) select suitable SCMs to be deployed for the capture of and treatment or retention of urban runoff using Optimization Programming, (3) implement and test the proposed research in a large urban area with pervasive urban runoff pollution, and (4) field test SCMs for which there is limited or no information on treatment efficiency to assess their runoff-cleaning potential. Optimization methods are developed and presented in this research work to minimize the total cost of SCM deployment while satisfying constraints on (i) the total cost of deployment, (ii) SCM capacities, (iii) volumetric balance at SCM sites, (iv) stormwater volumes at arbitrary sites, (v) unit Operational, Maintenance, and Replacement (OMR) cost, and (vi) water-quality and quantity at monitoring locations. Two alternative optimization models for SCM siting and sizing are presented in this thesis:• Linear Programming (LP) for optimal sizing of SCMs relies with a linear programming formulation. In addition, the Binary Linear Integer Programming (BLIP) for optimal selection of SCMs based on a binary (0,1) linear integer programming formulation.• Nonlinear Programming (NLP) for optimal sizing and selection of SCMs uses mixed (binary-real) nonlinear integer programming formulation.Summary of the Research Method. The key tasks accomplished in this thesis are: • Modeling and determination of priority catchments for SCM deployment based on SCM;• Development and application of a SCM Optimization programing model to select from within the GIS-evaluated SCMs those that are most cost effective in reducing the pollutant loads and concentration in urban runoff; • Nonlinear model’s results have been field tested by implementing its SCMs selection with site-specific data;• Conducted field experiment to evaluate the pollution removal efficiency of percolation wells and vegetated swales. The implementation of GIS-based electronic maps to classify areas according to their conditions: heavy traffic, impervious soils, high rainfall intensity for design storms, high urban density, and steep topography. The selected SCMs are tested in chosen priority catchments to assess model-predicted performance with field performance. The research task consists of field-experimenting with dry wells and vegetated swales. These types of SCMs appear to have good performance potential in permeable soils, and swales exhibits high aesthetic distinction and value. These SCMs are field tested to determine their pollutant removal efficiency for selected indicator pollutants (for example, total suspended solids). Broad Impacts of the Proposed Research. This thesis overarching hypothesis is that the sequential application of (1) computer based modeling in screening of high priority urban catchments and cost effective SCMs, and (2) optimization programming, can be used successfully in (i) identifying catchments with high SCM indices to urban runoff pollution and (ii) selecting the most cost-effective SCMs to reduce runoff pollution. This novel research concept represents a trend setting approach in combating urban runoff pollution in the United States, and in other places where the resources and know-how for urban runoff pollution control are in high demand and required. Intellectual Merit of the Research. This is a novel research attempt to develop and integrate novel analysis of pollution and suitability for SCM deployment with optimization programming method for the selection of the types and sizes of SCMs in urban catchments

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.

Two novel methods for synthesizing poly (propylene fumarate): Technical aspects and role of vacuum and N2 purging effects‏

Poly (propylene fumarate) (PPF), a linear unsaturated polyester consisting of alternating propylene glycol and fumaric acid units, can be cured in vivo to fill the skeletal defects with minimal surgical intervention. Many different methods have been reported for synthesizing PPF, but none of them gives a clear method. The present paper introduces two new methods in PPF synthesis: Modified reflux system (MRS) and mixed reflux-distillation system (MRDS). Similarly, the effects of applying vacuum (vacuum sequence, time, vacuum applying position, and the distance between vacuum applying position and reactor) as well as nitrogen gas (used continuously or only as an N2 blanket) on the PPF synthesis have been studied. The PPF obtained using optimum reaction condition has been characterized by using NMR, FTIR, and GPC analyses. It is demonstrated that the efficiency of MRDS in synthesizing PPF is higher than that of MRS. Nitrogen gas, vacuum applying position, continuously/stepwise-continuously applying vacuum and other parameters show an important role in the polymerization of PPF in both the MRDS and MRS systems

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.