حسن کبیری فرد

High efficiency and purity protein purification is a serious challenge in the production of recombinant proteins. This study is proposing an easy and new strategy for fabricating and describing magnetic microspheres consisting of a silica-coated magnetic core and a polyvinyl acetate shell modified with iminodiastic-acetic acid and complexed with Ni2+, (Fe3O4@SiO2@PVA@IDA-Ni2+) for the separation and purification of organophosphorus hydrolase (OPH) protein and histidine-containing outer membrane protein (OmpW).

The nanoparticles were synthesized in the following four steps: 1) synthesis of nanoparticles with vinyl groups on their surface, 2) grafting of polyvinyl alcohol (PVA) to these particles, 3) conversion of the hydroxyl groups of PVA to iminodiacetic acid, and 4) charging with nickel ions to form chelate groups. Structural properties and physicochemical parameters of nanoparticles were determined by FTIR, DLS, SEM, TEM, EDX, TGA and VSM analyzes. Recombinant protein subunits OPH and OmpW were expressed and purified as recombinant and their structure was confirmed. In this study, the genes encoding the recombinant proteins OPH and OmpW were cloned into the expression vector pet-26b (+) and expressed in E. coli BL21 (DE3) as a host and secreted into bacterial culture medium. Gene expression and purification were assessed by sodium dodecylsulfate-polyacrylamide gel (SDS-Page and Bradford test).

The results confirm that uniform and spherical magnetic polymer nanoparticles with high magnetization and superparamagnetic properties were successfully synthesized. According to the SEM images, the average diameter of the nanostructure was observed in the range of 24.56-62.19 nm. The formation of Fe3O4@SiO2@PVA/IDA-Ni2+ nanoparticles was confirmed by the peaks identified at 1405 and 1600 Cm-1 corresponding to the symmetric and asymmetric stretching vibration of the O=C−O− group and the removal of the peak at 1740 Cm-1. These microspheres showed a high degree of recovery of protein bands regardless of the nature of the protein. OmpW and OPH proteins as a His-tagged recombinant protein were expressed and purified using the final nanoparticles.

According to the SDS-PAGE results, there was a high degree of resolution in protein separation. The synthesized nanoparticles have a high protein binding capacity of about 1.25 mg of protein per mg of nanoparticles.

N-acyl hydrazones (NAH) and carbohydrazide derivatives have shown unique therapeutic properties. There is also, a growing interest in designing and evaluating the biological activity of these compounds and their derivatives to investigate their role as a novel structure in medicinal chemistry researches. Considering the importance of carbohydrazide derivatives, the indole-based structures, and isoxazole ring in drug design, The SAR of the active compounds in anti-Alzheimer, anti-cancer, and antiplatelet aggregation activity, and merits further investigation to find the optimal lead structures with maximum biological activity. In this study, the design, synthesis, and identification of the new Indole-isoxazole rings based carbohydrazides were desired. The synthesis started from 3-acetyl Indole, by using various reactants and reaction conditions (solvent, temperature), to obtain 5- (1-methyl-1-indole-3-yl) isoxazole-3-carbohydrazide as a novel compound that would be used as a fundamental structure in drug design. All structures were identified and approved at each step using spectrophotometer methods such as FT-IR, 1H NMR, and Mass.

In the present study, the concentration of some heavy metals (Fe, Zn, Pb, Ni) were measured and defined in surface waters, soil and organs of some plants: spinach, radish, cress and mint, in the spring and summer of 1390 in the Pishva area (Varamin). Heavy metals were measured in vegetables’ root, stem, leaf and their intake through consumption by human. Samplings of vegetable in five different points were accomplished. The amount of iron, zinc, lead and nickel, after chemical digestion of samples, were determined, using spectrophotometer flame atomic absorption method by 372.0, 213.9, 217.0, and 232.0 nm wavelength respectively. Four examined heavy metals in water, soil and vegetables samples in two seasons determined as 448.34±37.55, 56.68±15.53, 11.50±2.79 and 134.2±6.21 mg/kg respectively. Metals in vegetables, water and soil samples in two seasons had normal distribution and confidence level of 95%. Comparing the results with international standards for them showed that they are in acceptable range for water and soil samples in two seasons but in case of vegetables except for Zn in cress and mint roots and leave of spinach in summer which are in the acceptable range, the rest vegetable samples had heavy metal concentrations more than international levels in two seasons.

This study was conducted with the goal of investigating the amount of Chrome heavy metal in Firouz Abad river of Tehran. For this purpose, sampling from water was conducted by determining 5 measurement stations in the river path in the spring, summer and autumn of 2007. Analysis of samples was performed using Atomic Absorption method. The results of this study show that the levels of chrome metal measured in the third, fourth and fifth stations with the averages of 0.503mg/l, 1.29mg/l and 2.62mg/l, respectively are above the allowed level of chrome in the run-off determined by Environment Preservation Organization (0.5mg/l). In order to estimate the level of environmental risks of chrome in the environment of the affected area, the Environmental Failure Modes and Effects Analysis and its effects on environment was used. For this purpose, a table was first designed based on each station in which chrome concentration was graded within the scores span of 1 to 50 from the respects of risks severity and possible consequences. Then, Risk Priority Number (RPN) was calculated for each station from the product of risks severity in possible consequence. In the stations in which the level of chrome heavy metal was above the standard level, some measures were suggested for minimizing RPN. In this study, the level of environmental risks of this metal was estimated in 5 stations based on RPN as follows:RPN of first station = 2 < RPN of second station = 10 < RPN of thied station = 30 < RPN of forth station = 200 < RPN of fifth station = 450 Due to irrigation of agricultural lands by this river especially vegetable farming and growing summer crops in south of Tehran, the excess concentration of chrome has increased significant absorption of the aforesaid heavy metal in plants which its consequence is the entrance of this metal to food chain and its bad effects on human.