A Novel Strategy for Oriented Protein Immobilization

A new strategy for oriented immobilization of proteins was proposed. The strategy contains two steps. The first step is to search for a docking site away from the active site on the protein surface. The second step is trying to find a ligand that is able to grasp the targeted site of the protein. To avoid ligand binding to the active site of protein, the targeted docking site is selected to own opposite charges to those near the active site. To enhance the ligand-protein binding, both hydrophobic and electrostatic interactions need to be included. The targeted docking site should therefore contain hydrophobic amino acids. The ligand is then selected through the help of molecular docking simulations. The enzyme α-amylase derived from Aspergillus oryzae (TAKA) was taken as an example for oriented immobilization. The active site of TAKA is surrounded by negatively charged amino acids. All the possible hydrophobic sites on the surface of TAKA were evaluated by the free energy estimation through benzene docking. A hydrophobic site on the opposite side of TAKA-s active site was found to be positive in net charges. A possible ligand, 3,3-,4,4- – Biphenyltetra- carboxylic acid (BPTA), was found to catch TAKA by the designated docking site. Then, the BPTA molecules were grafted onto silica gels and measured the affinity of TAKA adsorption and the specific activity of thereby immobilized enzymes. It was found that TAKA had a dissociation constant as low as 7.0×10-6 M toward the ligand BPTA on silica gel. The increase in ionic strength has little effect on the adsorption of TAKA, which indicated the existence of hydrophobic interaction between ligands and proteins. The specific activity of the immobilized TAKA was compared with the randomly adsorbed TAKA on primary amine containing silica gel. It was found that the orderly immobilized TAKA owns a specific activity twice as high as the one randomly adsorbed by ionic interaction.

Endothelial-Cell-Mediated Displacement of Extracellular Matrix during Angiogenesis

Mechanical interaction between endothelial cells (ECs) and the extracellular matrix (or collagen gel) is known to influence the sprouting response of endothelial cells during angiogenesis. This influence is believed to impact on the capability of endothelial cells to sense soluble chemical cues. Quantitative analysis of endothelial-cell-mediated displacement of the collagen gel provides a means to explore this mechanical interaction. Existing analysis in this context is generally limited to 2D settings. In this paper, we investigate the mechanical interaction between endothelial cells and the extracellular matrix in terms of the endothelial-cellmediated displacement of the collagen gel in both 2D and 3D. Digital image correlation and Digital volume correlation are applied on confocal reflectance image stacks to analyze cell-mediated displacement of the gel. The skeleton of the sprout is extracted from phase contrast images and superimposed on the displacement field to further investigate the link between the development of the sprout and the displacement of the gel.

pH-Responsiveness Properties of a Biodigradable Hydrogels Based on Carrageenan-g-poly(NaAA-co-NIPAM)

A novel thermo-sensitive superabsorbent hydrogel with salt- and pH-responsiveness properties was obtained by grafting of mixtures of acrylic acid (AA) and N-isopropylacrylamide (NIPAM) monomers onto kappa-carrageenan, kC, using ammonium persulfate (APS) as a free radical initiator in the presence of methylene bisacrylamide (MBA) as a crosslinker. Infrared spectroscopy was carried out to confirm the chemical structure of the hydrogel. Moreover, morphology of the samples was examined by scanning electron microscopy (SEM). The effect of MBA concentration and AA/NIPAM weight ratio on the water absorbency capacity has been investigated. The swelling variations of hydrogels were explained according to swelling theory based on the hydrogel chemical structure. The hydrogels exhibited salt-sensitivity and cation exchange properties. The temperature- and pH-reversibility properties of the hydrogels make the intelligent polymers as good candidates for considering as potential carriers for bioactive agents, e.g. drugs.

Design and Analysis of a Solar Refrigeration System with a Rotating Generator

A solar refrigeration system based on the adsorptiondesorption phenomena is designed and analyzed. An annular tubular generator filled with silica gel adsorbent and with a perforated inner cylinder is integrated within a flat solar collector. The working fluid in the refrigeration cycle is water. The thermodynamic analysis and because of the temperature level that could be attained with a flat solar collector it is required that the system operates under vacuum conditions. In order to enhance the performance of the system and to get uniform temperature in the silica gel and higher desorbed mass, an apparatus for rotation of the generator is incorporated in the system. Testing is carried out and measurements are taken on the designed installation. The effect of rotation is checked on the temperature distribution and on the performance of this machine and compared to the flat solar collector with fixed generator.

Association of the p53 Codon 72 Polymorphism with Colorectal Cancer in South West of Iran

The p53 tumor suppressor gene plays two important roles in genomic stability: blocking cell proliferation after DNA damage until it has been repaired, and starting apoptosis if the damage is too critical. Codon 72 exon4 polymorphism (Arg72Pro) of the P53 gene has been implicated in cancer risk. Various studies have been done to investigate the status of p53 at codon 72 for arginine (Arg) and proline (Pro) alleles in different populations and also the association of this codon 72 polymorphism with various tumors. Our objective was to investigate the possible association between P53 Arg72Pro polymorphism and susceptibility to colorectal cancer among Isfahan and Chaharmahal Va Bakhtiari (a part of south west of Iran) population. We investigated the status of p53 at codon 72 for Arg/Arg, Arg/Pro and Pro/Pro allele polymorphisms in blood samples from 145 colorectal cancer patients and 140 controls by Nested-PCR of p53 exon 4 and digestion with BstUI restriction enzyme and the DNA fragments were then resolved by electrophoresis in 2% agarose gel. The Pro allele was 279 bp, while the Arg allele was restricted into two fragments of 160 and 119 bp. Among the 145 colorectal cancer cases 49 cases (33.79%) were homozygous for the Arg72 allele (Arg/Arg), 18 cases (12.41%) were homozygous for the Pro72 allele (Pro/Pro) and 78 cases (53.8%) found in heterozygous (Arg/Pro). In conclusion, it can be said that p53Arg/Arg genotype may be correlated with possible increased risk of this kind of cancers in south west of Iran.

Migration from Commercial to in-House Developed Learning Management Systems

The Learning Management Systems present learning environment which offers a collection of e-learning tools in a package that allows a common interface and information sharing among the tools. South East European University initial experience in LMS was with the usage of the commercial LMS-ANGEL. After a three year experience on ANGEL usage because of expenses that were very high it was decided to develop our own software. As part of the research project team for the in-house design and development of the new LMS, we primarily had to select the features that would cover our needs and also comply with the actual trends in the area of software development, and then design and develop the system. In this paper we present the process of LMS in-house development for South East European University, its architecture, conception and strengths with a special accent on the process of migration and integration with other enterprise applications.

Preparation and Characterization of M. × Piperita L. Oil Based Gel Formulation

The essential oil of M. × piperita L. was formulated into a topical gel. The prepared gel was characterized for its pH, viscosity, spreadiblity, consistency and extrudiblity, while its stability was evaluated under different temperature conditions. The prepared M. × piperita oil gel was clear and transparent. The pH value of developed gel was 6.6, while its viscosity was 1200 cP. Spreadability and consistency of the M. × piperita oil gel was 10.7 g.cm/sec and 7 mm, respectively. The prepared gel showed good extrudiblity. During the stability studies, no significant change in pH and viscosity as a function of time for gel was observed, indicating stability of prepared formulation. The gel developed in this study is expected to forward the usage of M. × piperita essential towards commercial application.