Search results for: protein modification.
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 716

Search results for: protein modification.

566 The Expression of a Novel Gene Encoding an Ankyrin-Repeat Protein, DRA1, is Regulated by Drought-Responsive Alternative Splicing

Authors: H. Sakamoto, Y. Nakagawara, S. Oguri

Abstract:

Drought stress is a critical environmental factor that adversely affects crop productivity and quality. Because of their immobile nature, plants have evolved mechanisms to sense and respond to drought stress. We identified a novel locus of Arabidopsis, designated DRA1 (drought responsive ankyrin1), whose disruption leads to increased drought-stress tolerance. DRA1 encodes a transmembrane protein with an ankyrin-repeat motif that has been implicated in diverse cellular processes such as signal transduction. RT-PCR analysis revealed that there were at least two splicing variants of DRA1 transcripts in wild-type plants. In response to drought stress, the levels of DRA1 transcripts retaining second and third introns were increased, whereas these introns were removed under unstressed conditions. These results suggest that DRA1 protein may negatively regulate plant drought tolerance and that the expression of DRA1is regulated in response to drought stress by alternative splicing.

Keywords: Alternative splicing, ankyrin repeat, Arabidopsis, drought tolerance.

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565 Multi-Label Hierarchical Classification for Protein Function Prediction

Authors: Helyane B. Borges, Julio Cesar Nievola

Abstract:

Hierarchical classification is a problem with applications in many areas as protein function prediction where the dates are hierarchically structured. Therefore, it is necessary the development of algorithms able to induce hierarchical classification models. This paper presents experimenters using the algorithm for hierarchical classification called Multi-label Hierarchical Classification using a Competitive Neural Network (MHC-CNN). It was tested in ten datasets the Gene Ontology (GO) Cellular Component Domain. The results are compared with the Clus-HMC and Clus-HSC using the hF-Measure.

Keywords: Hierarchical Classification, Competitive Neural Network, Global Classifier.

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564 Expression of Tissue Plasminogen Activator in Transgenic Tobacco Plants by Signal Peptides Targeting for Delivery to Apoplast, Endoplasmic Reticulum and Cytosol Spaces

Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi

Abstract:

Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA4404. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.

Keywords: Recombinant tissue plasminogen activator, plant cell comportment, leader signals, transgenic tobacco.

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563 Alignment of MG-63 Osteoblasts on Fibronectin-Coated Phosphorous Doping Lattices in Silicon

Authors: Andreas Körtge, Susanne Stählke, Regina Lange, Mario Birkholz, Mirko Fraschke, Katrin Schulz, Barbara Nebe, Patrick Elter

Abstract:

A major challenge in biomaterials research is the regulation of protein adsorption which is a key factor for controlling the subsequent cell adhesion at implant surfaces. The aim of the present study was to control the adsorption of fibronectin (FN) and the attachment of MG-63 osteoblasts with an electronic nanostructure. Shallow doping line lattices with a period of 260 nm were produced for this purpose by implantation of phosphorous in silicon wafers. Protein coverage was determined after incubating the substrate with FN by means of an immunostaining procedure and the measurement of the fluorescence intensity with a TECAN analyzer. We observed an increased amount of adsorbed FN on the nanostructure compared to control substrates. MG-63 osteoblasts were cultivated for 24h on FN-incubated substrates and their morphology was assessed by SEM. Preferred orientation and elongation of the cells in direction of the doping lattice lines was observed on FN-coated nanostructures.

Keywords: Cell adhesion, electronic nanostructures, doping lattice, fibronectin, MG-63 osteoblasts, protein adsorption.

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562 Diabetes Mellitus and Food Balance in the Kingdom of Saudi Arabia

Authors: Aljabryn Dalal Hamad

Abstract:

The present explanatory study concerns with the relation between Diabetes Mellitus and Food Balance in the Kingdom of Saudi Arabia during 2005-2010, using published data. Results illustrated that Saudi citizen daily protein consumption (DPC) during 2005-2007 (g/capita/day) is higher than the average global consumption level of protein with 15.27%, daily fat consumption (DFC) with 24.56% and daily energy consumption (DEC) with 16.93% and increases than recommended level by International Nutrition Organizations (INO) with 56% for protein, 60.49% for fat and 27.37% for energy. On the other hand, DPC per capita in Saudi Arabia decreased during the period 2008-2010 from 88.3 to 82.36 gram/ day. Moreover, DFC per capita in Saudi Arabia decreased during the period 2008-2010 from 3247.90 to 3176.43 Cal/capita/ day, and daily energy consumption (DEC) of Saudi citizen increases than world consumption with 16.93%, whereas increases with 27.37% than INO. Despite this, DPC, DFC and DEC per capita in Saudi Arabia still higher than world mean. On the other side, results illustrated that the number of diabetic patients in Saudi Arabia during the same period (2005-2010). The curve of diabetic patient’s number in Saudi Arabia during 2005-2010 is regular ascending with increasing level ranged between 7.10% in 2005 and 12.44% in 2010. It is essential to devise Saudi National programs to educate the public about the relation of food balances and diabetes so it could be avoided, and provide citizens with healthy dietary balances tables.

Keywords: Diabetes Mellitus, Food Balance, Energy, Fat, Protein, Saudi Arabia.

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561 Waste-Based Surface Modification to Enhance Corrosion Resistance of Aluminium Bronze Alloy

Authors: Wilson Handoko, Farshid Pahlevani, Isha Singla, Himanish Kumar, Veena Sahajwalla

Abstract:

Aluminium bronze alloys are well known for their superior abrasion, tensile strength and non-magnetic properties, due to the co-presence of iron (Fe) and aluminium (Al) as alloying elements and have been commonly used in many industrial applications. However, continuous exposure to the marine environment will accelerate the risk of a tendency to Al bronze alloys parts failures. Although a higher level of corrosion resistance properties can be achieved by modifying its elemental composition, it will come at a price through the complex manufacturing process and increases the risk of reducing the ductility of Al bronze alloy. In this research, the use of ironmaking slag and waste plastic as the input source for surface modification of Al bronze alloy was implemented. Microstructural analysis conducted using polarised light microscopy and scanning electron microscopy (SEM) that is equipped with energy dispersive spectroscopy (EDS). An electrochemical corrosion test was carried out through Tafel polarisation method and calculation of protection efficiency against the base-material was determined. Results have indicated that uniform modified surface which is as the result of selective diffusion process, has enhanced corrosion resistance properties up to 12.67%. This approach has opened a new opportunity to access various industrial utilisations in commercial scale through minimising the dependency on natural resources by transforming waste sources into the protective coating in environmentally friendly and cost-effective ways.

Keywords: Aluminium bronze, waste-based surface modification, Tafel polarisation, corrosion resistance.

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560 Quantitative Characteristics of Rainbow Trout, Oncorhynchus Mykiss, Neo-Males (XX Genotype) and Super-Males (YY Genotype) Sperm

Authors: Radosław K. Kowalski, Beata Sarosiek, Wiesław Demianowicz, Jędrek Judek, Krzysztof Goryczko, Stefan Dobosz, Henryk Kuźmiński, Krystyna Demska-Zakęś, Igor Babiak, Jan Glogowski

Abstract:

Rainbow trout homogametic males, (XX or YY sex genotype), can be obtained, respectively, through masculinisation of genetic females or induced androgenesis. Aim of this study was to compare reproductive potential of neo-males (XX) and super-males (YY) with heterogametic males (XY). We measured spermatozoa motility parameters, sperm concentration, osmolality and characterized protein profiles in samples of stripped and testicular sperm obtained from XY and YY males, and testicular sperm of XX males. The motile spermatozoa, as measured by both subjective method and CASA, showed no differences between testicular sperm of XX males and stripped sperm of XY and YY males whereas testicular sperm of XY and YY males had significantly lower sperm motility. Result of protein densitometry showed similarities in protein profile between seminal plasma of XY and YY males and testicular fluids of XX males. Testis of XX males showed specific histological structures of cysts consists hypertrophied Sertoli cells.

Keywords: fish, genotype, rainbow trout, sperm.

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559 Effect of Drought Stress on Nitrogen Components in Corn

Authors: Masoud Rafiee, Fatemeh Abdipoor, Hosain Lari

Abstract:

An attempt was made to study of nitrogen components response of corn (Zea mays L.) to drought stress. A farm research was done in RCBD as split-plot with four replications in Khorramabad, west Iran. Drought stress levels as irrigation regimes after 75 (control), 100, and 120 (stress) mm cumulative evaporation were in main plots, and four seed corn varieties include 500 (medium maturity), 647, 700, and 704 (long maturity) were as subplots. Soluble protein, nitrate and proline amino acid were measured in shoot and root at flowering stage, and grain yield was measured in harvesting stage. As the drought progressed, the amount of nitrate and proline followed an increasing trend, but soluble protein decreased in shoot and root. The highest amount of nitrate and proline was observed in longer maturity varieties than shorter ones, but decrease yield of long maturity varieties was higher than medium maturity varieties in drought condition, because of long duration of stress.

Keywords: Nitrate, Proline, Soluble protein, Yield

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558 Hydrolysis of Hull-Less Pumpkin Oil Cake Protein Isolate by Pepsin

Authors: Ivan Živanović, Žužana Vaštag, Senka Popović, Ljiljana Popović, Draginja Peričin

Abstract:

The present work represents an investigation of the hydrolysis of hull-less pumpkin (Cucurbita Pepo L.) oil cake protein isolate (PuOC PI) by pepsin. To examine the effectiveness and suitability of pepsin towards PuOC PI the kinetic parameters for pepsin on PuOC PI were determined and then, the hydrolysis process was studied using Response Surface Methodology (RSM). The hydrolysis was carried out at temperature of 30°C and pH 3.00. Time and initial enzyme/substrate ratio (E/S) at three levels were selected as the independent parameters. The degree of hydrolysis, DH, was mesuared after 20, 30 and 40 minutes, at initial E/S of 0.7, 1 and 1.3 mA/mg proteins. Since the proposed second-order polynomial model showed good fit with the experimental data (R2 = 0.9822), the obtained mathematical model could be used for monitoring the hydrolysis of PuOC PI by pepsin, under studied experimental conditions, varying the time and initial E/S. To achieve the highest value of DH (39.13 %), the obtained optimum conditions for time and initial E/S were 30 min and 1.024 mA/mg proteins.

Keywords: Enzymatic hydrolysis, Pepsin, Pumpkin (CucurbitaPepo L.) oil cake protein isolate, Response surface methodology.

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557 Effect of Dietary α-Cellulose Levels on the Growth Parameters of Nile Tilapia Oreochromis niloticus Fingerlings

Authors: Keri Alhadi Ighwela, Aziz Bin Ahmad, A. B. Abol-Munafi

Abstract:

Three purified diets were formulated using fish meal, soya bean, wheat flour, palm oil, minerals and maltose. The carbohydrate in the diets was increased from 5 to 15% by changing the cellulose content to study the effect of dietary carbohydrate level on the growth parameters of Nile tilapia Oreochromis niloticus. The protein and the lipid contents were kept constant in all the diets. The results showed that, weight gain, protein efficiency ratio, net protein utilisation and hepatosomatic index of fish fed the diet containing 15% cellulose were the lowest among all groups. Addition, the fish fed the diet containing 5% cellulose had the best specific growth rate, and food conversion ratio. While, there was no effect of the dietary cellulose levels on condition factor and survival rate. These results indicate that Nile tilapia fingerlings are able to utilize dietary cellulose does not exceed 10% in their feed for optimum growth.

Keywords: Dietary cellulose, growth parameters, Nile Tilapia Oreochromis niloticus, purified diets.

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556 A Stochastic Approach of Mitochondrial Dynamics

Authors: Athanasios T. Alexiou, Maria M. Psiha, John A. Rekkas, Panayiotis M. Vlamos

Abstract:

Mitochondria are dynamic organelles, capable to interact with each other. While the number of mitochondria in a cell varies, their quality and functionality depends on the operation of fusion, fission, motility and mitophagy. Nowadays, several researches declare as an important factor in neurogenerative diseases the disruptions in the regulation of mitochondrial dynamics. In this paper a stochastic model in BioAmbients calculus is presented, concerning mitochondrial fusion and its distribution in the renewal of mitochondrial population in a cell. This model describes the successive and dependent stages of protein synthesis, protein-s activation and merging of two independent mitochondria.

Keywords: Mitochondrial Dynamics, P-Calculus, StochasticModeling.

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555 Study of Mechanical Properties of Glutarylated Jute Fiber Reinforced Epoxy Composites

Authors: V. Manush Nandan, K. Lokdeep, R. Vimal, K. Hari Hara Subramanyan, C. Aswin, V. Logeswaran

Abstract:

Natural fibers have attained the potential market in the composite industry because of the huge environmental impact caused by synthetic fibers. Among the natural fibers, jute fibers are the most abundant plant fibers which are manufactured mainly in countries like India. Even though there is a good motive to utilize the natural supplement, the strength of the natural fiber composites is still a topic of discussion. In recent days, many researchers are showing interest in the chemical modification of the natural fibers to increase various mechanical and thermal properties. In the present study, jute fibers have been modified chemically using glutaric anhydride at different concentrations of 5%, 10%, 20%, and 30%. The glutaric anhydride solution is prepared by dissolving the different quantity of glutaric anhydride in benzene and dimethyl-sulfoxide using sodium formate catalyst. The jute fiber mats have been treated by the method of retting at various time intervals of 3, 6, 12, 24, and 36 hours. The modification structure of the treated fibers has been confirmed with infrared spectroscopy. The degree of modification increases with an increase in retention time, but higher retention time has damaged the fiber structure. The unmodified fibers and glutarylated fibers at different retention times are reinforced with epoxy matrix under room temperature. The tensile strength and flexural strength of the composites are analyzed in detail. Among these, the composite made with glutarylated fiber has shown good mechanical properties when compared to those made of unmodified fiber.

Keywords: Flexural properties, glutarylation, glutaric anhydride, tensile properties.

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554 Exploring Dimensionality, Systematic Mutations and Number of Contacts in Simple HP ab-initio Protein Folding Using a Blackboard-based Agent Platform

Authors: Hiram I. Beltrán, Arturo Rojo-Domínguez, Máximo Eduardo Sánchez Gutiérrez, Pedro Pablo González Pérez

Abstract:

A computational platform is presented in this contribution. It has been designed as a virtual laboratory to be used for exploring optimization algorithms in biological problems. This platform is built on a blackboard-based agent architecture. As a test case, the version of the platform presented here is devoted to the study of protein folding, initially with a bead-like description of the chain and with the widely used model of hydrophobic and polar residues (HP model). Some details of the platform design are presented along with its capabilities and also are revised some explorations of the protein folding problems with different types of discrete space. It is also shown the capability of the platform to incorporate specific tools for the structural analysis of the runs in order to understand and improve the optimization process. Accordingly, the results obtained demonstrate that the ensemble of computational tools into a single platform is worthwhile by itself, since experiments developed on it can be designed to fulfill different levels of information in a self-consistent fashion. By now, it is being explored how an experiment design can be useful to create a computational agent to be included within the platform. These inclusions of designed agents –or software pieces– are useful for the better accomplishment of the tasks to be developed by the platform. Clearly, while the number of agents increases the new version of the virtual laboratory thus enhances in robustness and functionality.

Keywords: genetic algorithms, multi-agent systems, bioinformatics, optimization, protein folding, structural biology.

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553 Effects of Boron Compounds in Rabbits Fed High Protein and Energy Diet: A Metabolomic and Transcriptomic Approach

Authors: N. Baspinar, A. Basoglu, O. Ozdemir, C. Ozel, F. Terzi, O. Yaman

Abstract:

Current research is targeting new molecular mechanisms that underlie non-alcoholic fatty liver disease (NAFLD) and associated metabolic disorders like non-alcoholic steatohepatitis (NASH). Forty New Zealand White rabbits have been used and fed a high protein (HP) and energy diet based on grains and containing 11.76 MJ/kg. Boron added to 3 experimental groups’ drinking waters (30 mg boron/L) as boron compounds. Biochemical analysis including boron levels, and nuclear magnetic resonance (NMR) based metabolomics evaluation, and mRNA expression of peroxisome proliferator-activated receptor (PPAR) family was performed. LDLcholesterol concentrations alone were decreased in all the experimental groups. Boron levels in serum and feces were increased. Content of acetate was in about 2x higher for anhydrous borax group, at least 3x higher for boric acid group. PPARα mRNA expression was significantly decreased in boric acid group. Anhydrous borax attenuated mRNA levels of PPARγ, which was further suppressed by boric acid. Boron supplementation decreased the degenerative alterations in hepatocytes. Except borax group other boron groups did not have a pronounced change in tubular epithels of kidney. In conclusion, high protein and energy diet leads hepatocytes’ degenerative changes which can be prevented by boron supplementation. Boric acid seems to be more effective in this situation.

Keywords: High protein and energy diet, boron, metabolomic, transcriptomic.

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552 ICAM-2, A Protein of Antitumor Immune Response in Mekong Giant Catfish (Pangasianodon gigas)

Authors: Jiraporn Rojtinnakorn

Abstract:

ICAM-2 (intercellular adhesion molecule 2) or CD102 (Cluster of Differentiation 102) is type I transmembrane glycoproteins, composing 2-9 immunoglobulin-like C2-type domains. ICAM-2 plays the particular role in immune response and cell surveillance. It is concerned in innate and specific immunity, cell survival signal, apoptosis, and anticancer. EST clone of ICAM-2, from P. gigas blood cell EST libraries, showed high identity to human ICAM-2 (92%) with conserve region of ICAM N-terminal domain and part of Ig superfamily. Gene and protein of ICAM-2 has been founded in mammals. This is the first report of ICAM-2 in fish

Keywords: ICAM-2, CD102, Pangasianodon gigas, antitumor.

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551 Production of the Protein-Vitamin Complex from Wheat Germ

Authors: Gulmira Kenenbay, Urishbay Chomanov, Tamara Tultabayeva, Aruzhan Shoman

Abstract:

Wheat germ has a balanced amino acid composition of the protein, which is well digested by enzymes in the gastrointestinal tract of humans, a high content of vitamins, minerals and unsaturated acids. Introduction components grain food products will enrich their biologically important substances, giving these products a number of valuable properties and reducing their caloric. A complex natural system of substances in foods will help replenish the body's need of essential nutrients, increasing its resistance to the harmful effects of the environment, prolong life. In this regard, there was a need for the development of production technology of protein complexes from wheat germ and then applying them in food, particularly in the dairy industry. Experimental studies were conducted to determine the number of herbal supplements on the sensory characteristics of the product. Studies have been conducted to determine the optimal process parameters of water activity and moisture content of the investigational product. 

Keywords: Wheat germ, sensory characteristics of the product, water activity.

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550 Physiological and Biochemical Responses to Drought Stress of Chickpea Genotypes

Authors: E. Ceyhan, A. Kahraman, M. Önder, M.K. Ateş, S. Karadaş, R. Topak, M.A. Avcı

Abstract:

The experimental design was 4 x 5 factorial with three replications in fully controlled research greenhouse in Department of Soil Sciences and Plant Nutrition, Faculty of Agriculture, University of Selcuk in the year of 2009. Determination of tolerant chickpea genotypes to drought was made in the research. Additionally, sophisticated effects of drought on plant growth and development, biochemical and physical properties or physical defense mechanisms were presented. According to the results, the primary genotypes were Ilgın YP (0.0063 g/gh) for leaf water capacity, 22235 70.44(%) for relative water content, 22159 (82.47%) for real water content, 22159 (5.03 mg/l) for chlorophyll a+b, Ilgın YP (125.89 nmol H2O2.dak-1/ mg protein-1) for peroxidase, Yunak YP (769.67 unit/ mg protein-1) for superoxide dismutase, Seydişehir YP (16.74 μg.TA-1) for proline, Gökçe (80.01 nmol H2O2.dak-1/ mg protein-1) for catalase. Consequently, all the genotypes increased their enzyme activity depending on the increasing of drought stress consider with the effects of drought stress on leaf enzyme activity. Chickpea genotypes are increasing enzyme activity against to drought stress.

Keywords: Chickpea, drought, enzyme, tolerance to drought

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549 Effect of the Polymer Modification on the Cytocompatibility of Human and Rat Cells

Authors: N. Slepickova Kasalkova, P. Slepicka, L. Bacakova, V. Svorcik

Abstract:

Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable,  semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H2O and CO2. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.

Keywords: Poly(L-lactic acid), plasma treatment, surface characterization, cytocompatibility, human osteoblasts, rat vascular smooth muscle cells, human stem cells.

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548 Synthesis and Characterization of Recycled Isotactic Polypropylene Nanocomposites Containing Date Wood Fiber

Authors: Habib Shaban

Abstract:

Nanocomposites of isotactic polypropylene (iPP) and date wood fiber were prepared after modification of the host matrix by reactive extrusion grafting of maleic anhydride. Chemical and mechanical treatment of date wood flour (WF) was conducted to obtain nanocrystalline cellulose. Layered silicates (clay) were partially intercalated with date wood fiber, and the modified layered silicate was used as filler in the PP matrix via a melt-blending process. The tensile strength of composites prepared from wood fiber modified clay was greater than that of the iPP-clay and iPP-WF composites at a 6% filler concentration, whereas deterioration of mechanical properties was observed when clay and WF were used alone for reinforcement. The dispersion of the filler in the matrix significantly decreased after clay modification with cellulose at higher concentrations, as shown by X-ray diffraction (XRD) data.

Keywords: Nanocomposites, isotactic polypropylene, date wood flour, intercalated, melt-blending.

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547 Identification of Binding Proteins That Interact with BVDV E2 Protein in Bovine Trophoblast Cell

Authors: Yan Ren, Fei Guo, Jun Qiao, Shengwei Hu, Hui Zhang, Yuanzhi Wang, Pengyan Wang, Jinliang Sheng, Xinli Gu, Xiaojun Liu, Chuangfu Chen

Abstract:

Bovine viral diarrhea virus (BVDV) can cause lifelong persistent infection. One reason for the phenomena is attributed to BVDV infection to placenta tissue. However the mechanisms that BVDV invades into placenta tissue remain unclear. To clarify the molecular mechanisms, we investigated the possible means that BVDV entered into bovine trophoblast cells (TPC). Yeast two-hybrid system was used to identify proteins extracted from TPC, which interact with BVDV envelope glycoprotein E2. A PGbkt7-E2 yeast expression vector and TPC cDNA library were constructed. Through two rounds of screening, three positive clones were identified. Sequencing analysis indicated that all the three positive clones encoded the same protein clathrin. Physical interaction between clathrin and BVDV E2 protein was further confirmed by coimmunoprecipitation experiments. This result suggested that the clathrin might play a critical role in the process of BVDV entry into placenta tissue and might be a novel antiviral target for preventing BVDV infection.

Keywords: Bovine viral diarrhea virus, clathrin, glycoprotein E2, yeast two-hybrid system.

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546 Investigations of Protein Aggregation Using Sequence and Structure Based Features

Authors: M. Michael Gromiha, A. Mary Thangakani, Sandeep Kumar, D. Velmurugan

Abstract:

The main cause of several neurodegenerative diseases such as Alzhemier, Parkinson and spongiform encephalopathies is formation of amyloid fibrils and plaques in proteins. We have analyzed different sets of proteins and peptides to understand the influence of sequence based features on protein aggregation process. The comparison of 373 pairs of homologous mesophilic and thermophilic proteins showed that aggregation prone regions (APRs) are present in both. But, the thermophilic protein monomers show greater ability to ‘stow away’ the APRs in their hydrophobic cores and protect them from solvent exposure. The comparison of amyloid forming and amorphous b-aggregating hexapeptides suggested distinct preferences for specific residues at the six positions as well as all possible combinations of nine residue pairs. The compositions of residues at different positions and residue pairs have been converted into energy potentials and utilized for distinguishing between amyloid forming and amorphous b-aggregating peptides. Our method could correctly identify the amyloid forming peptides at an accuracy of 95-100% in different datasets of peptides.

Keywords: Aggregation prone regions, amyloids, thermophilic proteins, amino acid residues, machine learning.

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545 ELISA Based hTSH Assessment Using Two Sensitive and Specific Anti-hTSH Polyclonal Antibodies

Authors: Maysam Mard-Soltani, Mohamad Javad Rasaee, Saeed Khalili, Abdol Karim Sheikhi, Mehdi Hedayati

Abstract:

Production of specific antibody responses against hTSH is a cumbersome process due to the high identity between the hTSH and the other members of the glycoprotein hormone family (FSH, LH and HCG) and the high identity between the human hTSH and host animals for antibody production. Therefore, two polyclonal antibodies were purified against two recombinant proteins. Four possible ELISA tests were designed based on these antibodies. These ELISA tests were checked against hTSH and other glycoprotein hormones, and their sensitivity and specificity were assessed. Bioinformatics tools were used to analyze the immunological properties. After the immunogen region selection from hTSH protein, c terminal of B hTSH was selected and applied. Two recombinant genes, with these cut pieces (first: two repeats of C terminal of B hTSH, second: tetanous toxin+B hTSH C terminal), were designed and sub-cloned into the pET32a expression vector. Standard methods were used for protein expression, purification, and verification. Thereafter, immunizations of the white New Zealand rabbits were performed and the serums of them were used for antibody titration, purification and characterization. Then, four ELISA tests based on two antibodies were employed to assess the hTSH and other glycoprotein hormones. The results of these assessments were compared with standard amounts. The obtained results indicated that the desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. The raised antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum. Among the four designed tests, the test in which the antibody against first protein was used as capture antibody, and the antibody against second protein was used as detector antibody did not show any hook effect up to 50 miu/l. Both proteins have the ability to induce highly sensitive and specific antibody responses against the hTSH. One of the antibody combinations of these antibodies has the highest sensitivity and specificity in hTSH detection.

Keywords: hTSH, bioinformatics, protein expression, cross reactivity.

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544 Improvement of Durability of Wood by Maleic Anhydride

Authors: Yong F. Li, Yi X. Liu, Xiang M. Wang, Feng H. Wang

Abstract:

Wood as a natural renewable material is vulnerable to degradation by microorganisms and susceptible to change in dimension by water. In order to effectively improve the durability of wood, an active reagent, maleic anhydride (Man) was selected for wood modification. Man was first dissolved into a solvent, and then penetrated into wood porous structure under a vacuum/pressure condition. After a final catalyst-thermal treatment, wood modification was finished. The test results indicate that acetone is a good solvent for transporting Man into wood matrix. SEM observation proved that wood samples treated by Man kept a good cellular structure, indicating a well penetration of Man into wood cell walls. FTIR analysis suggested that Man reacted with hydroxyl groups on wood cell walls by its ring-ether group, resulting in reduction of amount of hydroxyl groups and resultant good dimensional stability as well as fine decay resistance. Consequently, Man modifying wood to improve its durability is an effective method.

Keywords: Wood, porous structure, durability improvement, maleic anhydride

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543 Multi-Agent Systems Applied in the Modeling and Simulation of Biological Problems: A Case Study in Protein Folding

Authors: Pedro Pablo González Pérez, Hiram I. Beltrán, Arturo Rojo-Domínguez, Máximo EduardoSánchez Gutiérrez

Abstract:

Multi-agent system approach has proven to be an effective and appropriate abstraction level to construct whole models of a diversity of biological problems, integrating aspects which can be found both in "micro" and "macro" approaches when modeling this type of phenomena. Taking into account these considerations, this paper presents the important computational characteristics to be gathered into a novel bioinformatics framework built upon a multiagent architecture. The version of the tool presented herein allows studying and exploring complex problems belonging principally to structural biology, such as protein folding. The bioinformatics framework is used as a virtual laboratory to explore a minimalist model of protein folding as a test case. In order to show the laboratory concept of the platform as well as its flexibility and adaptability, we studied the folding of two particular sequences, one of 45-mer and another of 64-mer, both described by an HP model (only hydrophobic and polar residues) and coarse grained 2D-square lattice. According to the discussion section of this piece of work, these two sequences were chosen as breaking points towards the platform, in order to determine the tools to be created or improved in such a way to overcome the needs of a particular computation and analysis of a given tough sequence. The backwards philosophy herein is that the continuous studying of sequences provides itself important points to be added into the platform, to any time improve its efficiency, as is demonstrated herein.

Keywords: multi-agent systems, blackboard-based agent architecture, bioinformatics framework, virtual laboratory, protein folding.

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542 A New Approach In Protein Folding Studies Revealed The Potential Site For Nucleation Center

Authors: Nurul Bahiyah Ahmad Khairudin, Habibah A Wahab

Abstract:

A new approach to predict the 3D structures of proteins by combining the knowledge-based method and Molecular Dynamics Simulation is presented on the chicken villin headpiece subdomain (HP-36). Comparative modeling is employed as the knowledge-based method to predict the core region (Ala9-Asn28) of the protein while the remaining residues are built as extended regions (Met1-Lys8; Leu29-Phe36) which then further refined using Molecular Dynamics Simulation for 120 ns. Since the core region is built based on a high sequence identity to the template (65%) resulting in RMSD of 1.39 Å from the native, it is believed that this well-developed core region can act as a 'nucleation center' for subsequent rapid downhill folding. Results also demonstrate that the formation of the non-native contact which tends to hamper folding rate can be avoided. The best 3D model that exhibits most of the native characteristics is identified using clustering method which then further ranked based on the conformational free energies. It is found that the backbone RMSD of the best model compared to the NMR-MDavg is 1.01 Å and 3.53 Å, for the core region and the complete protein, respectively. In addition to this, the conformational free energy of the best model is lower by 5.85 kcal/mol as compared to the NMR-MDavg. This structure prediction protocol is shown to be effective in predicting the 3D structure of small globular protein with a considerable accuracy in much shorter time compared to the conventional Molecular Dynamics simulation alone.

Keywords: 3D model, Chicken villin headpiece subdomain, Molecular dynamic simulation NMR-MDavg, RMSD.

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541 Secondary Ion Mass Spectrometry of Proteins

Authors: Santanu Ray, Alexander G. Shard

Abstract:

The adsorption of bovine serum albumin (BSA), immunoglobulin G (IgG) and fibrinogen (Fgn) on fluorinated selfassembled monolayers have been studied using time of flight secondary ion mass spectrometry (ToF-SIMS) and Spectroscopic Ellipsometry (SE). The objective of the work has to establish the utility of ToF-SIMS for the determination of the amount of protein adsorbed on the surface. Quantification of surface adsorbed proteins was carried out using SE and a good correlation between ToF-SIMS results and SE was achieved. The surface distribution of proteins were also analysed using Atomic Force Microscopy (AFM). We show that the surface distribution of proteins strongly affect the ToFSIMS results.

Keywords: ToF-SIMS, Spectroscopic Ellipsometry, Protein, Atomic Force Microscopy.

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540 WLAN Positioning Based on Joint TOA and RSS Characteristics

Authors: Peerapong Uthansakul, Monthippa Uthansakul

Abstract:

WLAN Positioning has been presented by many approaches in literatures using the characteristics of Received Signal Strength (RSS), Time of Arrival (TOA) or Time Difference of Arrival (TDOA), Angle of Arrival (AOA) and cell ID. Among these, RSS approach is the simplest method to implement because there is no need of modification on both access points and client devices whereas its accuracy is terrible due to physical environments. For TOA or TDOA approach, the accuracy is quite acceptable but most researches have to modify either software or hardware on existing WLAN infrastructure. The scales of modifications are made on only access card up to the changes in protocol of WLAN. Hence, it is an unattractive approach to use TOA or TDOA for positioning system. In this paper, the new concept of merging both RSS and TOA positioning techniques is proposed. In addition, the method to achieve TOA characteristic for positioning WLAN user without any extra modification necessarily appended in the existing system is presented. The measurement results confirm that the proposed technique using both RSS and TOA characteristics provides better accuracy than using only either RSS or TOA approach.

Keywords: Received signal strength, Time of arrival, Positioning system, WLAN, Measurement.

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539 Mechanical and Morphological Properties of Polypropylene and High Density Polyethylene Matrix Composites Reinforced with Surface Modified Nano Sized TiO2 Particles

Authors: Mirigul Altan, Huseyin Yildirim

Abstract:

Plastics occupy wide place in the applications of automotive, electronics and house goods. Especially reinforced plastics become popular because of their high strength besides their advantages of low weight and easy manufacturability. In this study, mechanical and morphological properties of polypropylene (PP) and high density polyethylene (HDPE) matrix composites reinforced with surface modified nano titan dioxide (TiO2) particles were investigated. Surface modification was made by coating the nano powders with maleic anhydride grafted styrene ethylene butylene styrene (SEBS-g-MA) and silane, respectively. After surface modification, PP/TiO2 and HDPE/TiO2 composites were obtained by using twin screw extruder at titan dioxide loading of 1 wt.%, 3 wt.% and 5 wt.%. Effects of surface modification were determined by thermal and morphological analysis. SEBS-g-MA provided bridging effect between TiO2 particles and polymer matrix while silane was effective as a dispersant. Depending on that, homogenous structures without agglomeration were obtained. Mechanical tests were performed on the injection moldings of the composites for obtaining the impact strength, tensile strength, stress at break, elongation and elastic modulus. Reinforced HDPE and PP moldings gave higher tensile strength and elastic modulus due to the rigid structure of TiO2. Slight increment was seen in stress at break. Elongation and impact strength decreased due to the stiffness of the nano titan dioxide.

Keywords: High density polyethylene, mechanical properties, nano TiO2, polypropylene.

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538 Experimental and Finite Element Study of Bending Fatigue Failure: A Case Study on Main Shaft of a Gyrator Crusher

Authors: Rahim Sotoudeh Bahreini, Alireza Foroughi Nematollahi, Akbar Jafari

Abstract:

This study investigates the mechanism of a Gyratory crusher-located in Golgohar mining and industrial Co. specifically with a focus on stresses distribution and fatigue failure of its main shaft. At first step, the cross section of the fractured shaft is studied, and the crack growth is analyzed. Then, the rotational motion of the shaft and the oil temperature of oil circuit of equipment are monitored. Condition monitoring is used to help finding a better modification. Based on the results of this study, the main causes of shaft failure are identified, and corrective solution is offered to increase crusher performance, especially its main shaft life. To predict the efficiency of the proposed modification, finite element simulation is performed, and its results are compared with the similar modified cases. The comparison and interpretation of simulation results confirm the efficiency of proposed corrective method.

Keywords: Fatigue failure, finite element method, gyratory crusher, condition monitoring.

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537 Connectivity Characteristic of Transcription Factor

Authors: T. Mahalakshmi, Aswathi B. L., Achuthsankar S. Nair

Abstract:

Transcription factors are a group of proteins that helps for interpreting the genetic information in DNA. Protein-protein interactions play a major role in the execution of key biological functions of a cell. These interactions are represented in the form of a graph with nodes and edges. Studies have showed that some nodes have high degree of connectivity and such nodes, known as hub nodes, are the inevitable parts of the network. In the present paper a method is proposed to identify hub transcription factor proteins using sequence information. On a complete data set of transcription factor proteins available from the APID database, the proposed method showed an accuracy of 77%, sensitivity of 79% and specificity of 76%.

Keywords: Transcription Factor Proteins, Hub Proteins, Shannon Index, Transfer Free Energy to Surface (TFES).

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