Publications | Biotechnology and Bioengineering
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 276

World Academy of Science, Engineering and Technology

[Biotechnology and Bioengineering]

Online ISSN : 1307-6892

126 Statistical Optimization of the Enzymatic Saccharification of the Oil Palm Empty Fruit Bunches

Authors: Rashid S. S., Alam M. Z.

Abstract:

A statistical optimization of the saccharification process of EFB was studied. The statistical analysis was done by applying faced centered central composite design (FCCCD) under response surface methodology (RSM). In this investigation, EFB dose, enzyme dose and saccharification period was examined, and the maximum 53.45% (w/w) yield of reducing sugar was found with 4% (w/v) of EFB, 10% (v/v) of enzyme after 120 hours of incubation. It can be calculated that the conversion rate of cellulose content of the substrate is more than 75% (w/w) which can be considered as a remarkable achievement. All the variables, linear, quadratic and interaction coefficient, were found to be highly significant, other than two coefficients, one quadratic and another interaction coefficient. The coefficient of determination (R2) is 0.9898 that confirms a satisfactory data and indicated that approximately 98.98% of the variability in the dependent variable, saccharification of EFB, could be explained by this model.

Keywords: Face centered central composite design (FCCCD), Liquid state bioconversion (LSB), Palm oil mill effluent, Trichoderma reesei RUT C-30.

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125 Study on Phytochemical Properties, Antibacterial Activity and Cytotoxicity of Aloe vera L.

Authors: K. Thu, Yin Y. Mon, Tin A. Khaing, Ohn M. Tun

Abstract:

The aim of the study was to investigate phytochemical properties, antimicrobial activity and cytotoxicity of Aloe vera. The phytochemical screening of the extracts of leaves of A. vera revealed the presence of bioactive compounds such as alkaloids, tannins, flavonoids phenolic compounds, and etc. with absence of cyanogenic glycosides. Three different solvents such as methanol, ethanol and Di-Methyl sulfoxide were used to screen the antimicrobial activity of A. vera leaves against four human clinical pathogens by agar well diffusion method. The maximum antibacterial activities were observed in methanol extract followed by ethanol and Di-Methyl sulfoxide. It was also found that remarkable antibacterial activities with methanolic and ethanolic extracts of A. vera compared with the standard antibiotic, tetracycline that was not active against E. coli and S. boydii and supported the view that A. vera is a potent antimicrobial agent compared with the conventional antibiotic. Moreover, the brine shrimps (Artemia salina) toxicity test exhibited LC50 value was 569.52 ppm. The resulting data indicated that the A. vera plant have less toxic effects on brine shrimp. Hence, it is signified that Aloe vera plant extract is safe to be used as an antimicrobial agent.

Keywords: Aloe vera L., antimicrobial activity, brine shrimp, cytotoxicity, phytochemical properties.

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124 Effects of Irradiation to Morphological, Physicochemical and Biocompatibility Properties of Carrageenan

Authors: Jhalique Jane R. Fojas, Rizalinda L. De Leon, Lucille V. Abad

Abstract:

The characterization of κ-carrageenan could provide a better understanding of its functions in biological, medical and industrial applications. Chemical and physical analyses of carrageenan from seaweeds, Euchema cottonii L., were done to offer information on its properties and the effects of Co-60 γ-irradiation on its thermochemical characteristics. The structural and morphological characteristics of κ-carrageenan were determined using scanning electron microscopy (SEM) while the composition, molecular weight and thermal properties were determined using attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), gel permeation chromatography (GPC), thermal gravimetric analysis (TGA) and differential scanning calorimetry (DSC). Further chemical analysis was done using hydrogen-1 nuclear magnetic resonance (1H NMR) and functional characteristics in terms of biocompatibility were evaluated using cytotoxicity test.

Keywords: Biocompatibility, carrageenan, DSC, FTIR, GPC, irradiation, NMR, physicochemical, SEM, TGA.

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123 Molecular Docking Studies of Mycobacterium tuberculosis RNA Polymerase β Subunit (rpoB) Receptor

Authors: Virupakshaiah DBM, Madiha Ahmed, Smita T. Patil, Chandrakanth Kelmani

Abstract:

Tuberculosis (TB) is a bacterial infectious disease caused by the obligate human pathogen, Mycobacterium tuberculosis. Multidrug-resistant tuberculosis (MDR-TB) is a global reality that threatens tuberculosis control. Resistance to antibiotic Rifampicin, occurs in 95% of cases through nucleotide substitutions in an 81-bp core region of the rpoB i.e; beta subunit of DNA dependant RNA polymerase. In this paper, we studied the Rifampicin-rpoB receptor interactions In silico. First, homology modeling was performed to obtain the three dimensional structure of Mycobacterium rpoB. Sixty analogs of Rifampicin were prepared using Marvin sketch software. Both original Rifampicin and the analogs were docked with rpoB and energy values were obtained. Out of sixty analogs, 43 analogs had lesser energy values than conventional Rifampicin and hence are predicted to have greater binding affinity to rpoB. Thus, this study offers a route for the development of Rifampicin analogs against multi drug resistant Mycobacterium rpoB.

Keywords: Marvin Sketch, Mycobacterium tuberculosis, Rifampicin, rpoB.

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122 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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121 A Tool for Creation Artificial Symbiotic Associations of Wheat

Authors: Zilya R. Vershinina, Andrei K. Baymiev, Aleksei K. Baymiev, Aleksei V. Chemeris

Abstract:

This paper reports optimization of characteristics of bioballistic transformation of spring soft wheat (Triticum aestivum L. cultivar Raduga) and getting of transgenic plants, carrying pea lectin gene. This gene will let to create new associative wheat symbiosis with nodule bacteria of field pea, which has growth encouraging, fungistatic and other useful characteristics.

Keywords: transgenic wheat, pea lectin, rhizobia root colonization, symbiosis

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120 Molecular Epidemiology and Genotyping of Bovine Viral Diarrhea Virus in Xinjiang Uygur Autonomous Region of China

Authors: Yan Ren, Jun Qiao, Xianxia Liu, Pengyan Wang, Qiang Fu, Huijun Shi, Fei Guo, Yuanzhi Wang, Hui Zhang, Jinliang Sheng, Xinli Gu, Xiao-Jun Liu, Chuangfu Chen

Abstract:

As part of national epidemiological survey on bovine viral diarrhea virus (BVDV), a total of 274 dejecta samples were collected from 14 cattle farms in 8 areas of Xinjiang Uygur Autonomous Region in northwestern China. Total RNA was extracted from each sample, and 5--untranslated region (UTR) of BVDV genome was amplified by using two-step reverse transcriptase-polymerase chain reaction (RT-PCR). The PCR products were subsequently sequenced to study the genetic variations of BVDV in these areas. Among the 274 samples, 33 samples were found virus-positive. According to sequence analysis of the PCR products, the 33 samples could be arranged into 16 groups. All the sequences, however, were highly conserved with BVDV Osloss strains. The virus possessed theses sequences belonged to BVDV-1b subtype by phylogenetic analysis. Based on these data, we established a typing tree for BVDV in these areas. Our results suggested that BVDV-1b was a predominant subgenotype in northwestern China and no correlation between the genetic and geographical distances could be observed above the farm level.

Keywords: bovine viral diarrhea virus, molecular epidemiology, phylogenetic analysis.

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119 Synthesis of Sterile and Pyrogen Free Biogenic Magnetic Nanoparticles: Biotechnological Potential of Magnetotactic Bacteria for Production of Nanomaterials

Authors: Saeid Ghorbanzadeh-Mashkani, Parisa Tajer-Mohammad-Ghazvini, Ahmad Nozad-Golikand, Rouha Kasra-Kermanshahi, Mohammad-Reza Davarpanah

Abstract:

Today, biogenic magnetite nanoparticles among magnetic nanoparticles have unique attracted attention because of their magnetic characteristics and potential applications in various fields such as therapeutic and diagnostic. A well known example of these biogenic nanoparticles is magnetosomes of magnetotactic bacteria. In this research, we used two different types of technique for the isolation and purification of magnetosome nanoparticles from the isolated magnetotactic bacterial cells, heat-alkaline treatment and sonication. Also we evaluated pyrogen content and sterility of synthesized the isolated individual magnetosome by the Limulus Amoebocyte Lysate test and direct impedimetric method respectively.

Keywords: Biogenic magnetic nanoparticles, Magnetosome, Magnetotactic bacteria, Nanobiotechnology

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118 A Green Chemical Technique for the Synthesis of Magnetic Nanoparticles by Magnetotactic Bacteria

Authors: Parisa Tajer-Mohammad-Ghazvini, Rouha Kasra-Kermanshahi, Ahmad Nozad-Golikand, Majid Sadeghizadeh

Abstract:

Bacterial magnetic nanoparticles have great useful potential in biotechnological and biomedical applications. In this study, a liquid growth medium was modified for cultivation a fastidious magnetotactic bacterium that has been isolated from Anzali lagoon, Iran in our previous research. These modifications include change in vitamin, mineral, carbon sources and etcetera. In our experience, the serum bottles and designed air-tight laboratory bottles were used to create microaerobic conditions in order to development of a method for scale-up experiment. This information may serve as a guide to green chemistry based biological protocols for the synthesis of magnetic nanoparticles with control over the chemical composition, morphology and size.

Keywords: Green chemistry, Magnetosome, Magnetotactic bacteria, Magnetic nanoparticles, Nano-Biotechnology.

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117 Experimental Studies on Multiphase Flow in Porous Media and Pore Wettability

Authors: Xingxun Li, Xianfeng Fan

Abstract:

Multiphase flow transport in porous medium is very common and significant in science and engineering applications. For example, in CO2 Storage and Enhanced Oil Recovery processes, CO2 has to be delivered to the pore spaces in reservoirs and aquifers. CO2 storage and enhance oil recovery are actually displacement processes, in which oil or water is displaced by CO2. This displacement is controlled by pore size, chemical and physical properties of pore surfaces and fluids, and also pore wettability. In this study, a technique was developed to measure the pressure profile for driving gas/liquid to displace water in pores. Through this pressure profile, the impact of pore size on the multiphase flow transport and displacement can be analyzed. The other rig developed can be used to measure the static and dynamic pore wettability and investigate the effects of pore size, surface tension, viscosity and chemical structure of liquids on pore wettability.

Keywords: Enhanced oil recovery, Multiphase flow, Pore size, Pore wettability

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116 Intragenic MicroRNAs Binding Sites in MRNAs of Genes Involved in Carcinogenesis

Authors: Olga A. Berillo, Assel S. Issabekova, Anatoly T. Ivashchenko

Abstract:

MiRNAs participate in gene regulation of translation. Some studies have investigated the interactions between genes and intragenic miRNAs. It is important to study the miRNA binding sites of genes involved in carcinogenesis. RNAHybrid 2.1 and ERNAhybrid programmes were used to compute the hybridization free energy of miRNA binding sites. Of these 54 mRNAs, 22.6%, 37.7%, and 39.7% of miRNA binding sites were present in the 5'UTRs, CDSs, and 3'UTRs, respectively. The density of the binding sites for miRNAs in the 5'UTR ranged from 1.6 to 43.2 times and from 1.8 to 8.0 times greater than in the CDS and 3'UTR, respectively. Three types of miRNA interactions with mRNAs have been revealed: 5'- dominant canonical, 3'-compensatory, and complementary binding sites. MiRNAs regulate gene expression, and information on the interactions between miRNAs and mRNAs could be useful in molecular medicine. We recommend that newly described sites undergo validation by experimental investigation.

Keywords: Exon, intron, miRNA, oncogene.

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115 Novel Inhibitor of E. coli DNA Adenine Methyltransferase (Ecodam)

Authors: H. Elsawy, A. Jeltsch

Abstract:

EcoDam is an adenine-N6 DNA methyltransferase that methylates the GATC sites in the Escherichia coli genome. DNA-adenine methylation is not present in higher eukaryotes including humans. These observations raise the possibility that dam inhibitors may be used as anti-microbial agents. Polyphosphate (Poly(P)) is an important metabolite and signaling molecule in prokaryotes and eukaryotes. Here, by using gel retardation experiments to investigate the competition of DNA binding by EcoDam in the presence of polyphosphate, we found that Poly (P) strongly interferes with DNA binding by EcoDam, while same concentration of monophosphate does not. In addition, we demonstrated that Poly (P) binding inhibits the activity of EcoDam and our results suggest that Poly (P) led to strong inhibition of the EcoDam catalytic activity, while monophosphate had only moderate effect.

Keywords: Antibacterial drugs, EcoDam inhibitors, Polyphosphate.

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114 Investigation of Pre-Treatment Parameters of Rye and Triticale for Bioethanol Production

Authors: Algirdas Jasinskas, Egidijus Šarauskis, Raimondas Šarauskis, Antanas Sakalauskas

Abstract:

This paper presents the new results of energy plant – rye and triticale at yellow ripeness and ripe, pre-treatment in high pressure steam reactor and monosaccharide extraction. There were investigated the influence of steam pressure (20 to 22 bar), retention duration (180 to 240 s) and catalytic sulphuric acid concentration strength (0 to 0.5 %) on the pre-treatment process, contents of monosaccharides (glucose, arabinose, xylose, mannose) and undesirable by-compounds (furfural and HMF) in the reactor. The study has determined that the largest amount of monosaccharides (37.2 % of glucose, 2.7 % of arabinose, 8.4 % of xylose, and 1.3 % of mannose) was received in the rye at ripe, the samples of which were mixed with 0.5 % concentration of catalytic sulphuric acid, and hydrolysed in the reactor, where the pressure was 20 bar, whereas the reaction time – 240 s.

Keywords: Bioethanol, Pre-treatment, Rye, Triticale.

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113 Evolutionary Distance in the Yeast Genome

Authors: Somayyeh Azizi, Saeed Kaboli, Atsushi Yagi

Abstract:

Whole genome duplication (WGD) increased the number of yeast Saccharomyces cerevisiae chromosomes from 8 to 16. In spite of retention the number of chromosomes in the genome of this organism after WGD to date, chromosomal rearrangement events have caused an evolutionary distance between current genome and its ancestor. Studies under evolutionary-based approaches on eukaryotic genomes have shown that the rearrangement distance is an approximable problem. In the case of S. cerevisiae, we describe that rearrangement distance is accessible by using dedoubled adjacency graph drawn for 55 large paired chromosomal regions originated from WGD. Then, we provide a program extracted from a C program database to draw a dedoubled genome adjacency graph for S. cerevisiae. From a bioinformatical perspective, using the duplicated blocks of current genome in S. cerevisiae, we infer that genomic organization of eukaryotes has the potential to provide valuable detailed information about their ancestrygenome.

Keywords: Whole-genome duplication, Evolution, Double-cutand- join operation, Yeast.

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112 Study of Optical Properties of a Glutathione Capped Gold Nanoparticles Using Linker (MHDA) by Fourier Transform Infra Red Spectroscopy and Surface Enhanced Raman Scattering

Authors: A. Deręgowska, J. Depciuch, R. Wojnarowska, J. Polit, D. Broda, H. Nechai, M. Gonchar, and E. Sheregii

Abstract:

16-Mercaptohexadecanoic acid (MHDA) and tripeptide glutathione conjugated with gold nanoparticles (Au-NPs) are characterized by Fourier Transform InfaRared (FTIR) spectroscopy combined with Surface-enhanced Raman scattering (SERS) spectroscopy. Surface Plasmon Resonance (SPR) technique based on FTIR spectroscopy has become an important tool in biophysics, which is perspective for the study of organic compounds. FTIR-spectra of MHDA shows the line at 2500 cm-1 attributed to thiol group which is modified by presence of Au-NPs, suggesting the formation of bond between thiol group and gold. We also can observe the peaks originate from characteristic chemical group. A Raman spectrum of the same sample is also promising. Our preliminary experiments confirm that SERS-effect takes place for MHDA connected with Au-NPs and enable us to detected small number (less than 106 cm-2) of MHDA molecules. Combination of spectroscopy methods: FTIR and SERS – enable to study optical properties of Au- NPs and immobilized bio-molecules in context of a bio-nano-sensors.

Keywords: Glutathione; gold nanoparticles, Fourier transform infrared spectroscopy, MHDA, surface-enhanced Raman scattering.

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111 The Composition of Rice Bran Hydrolysate and Its Possibility to Use in the Ethanol Production by Zymomonas mobilis Biofilm

Authors: Tatsaporn Todhanakasem, Kamonchanok Areerat, Pornthap Thanonkeo, Roungdao KlinjapoandGlenn M. Young

Abstract:

Rice bran has been abandoned as agricultural waste for million tonnes per year in Thailand, therefore they have been proposed to be utilized as a rich carbon source in the production of bioethanol. Many toxic compounds are possibly released during the pretreatment of rice bran prior the fermentation process. This study aims to analyze on the availability of toxic compounds and the amount of glucose obtained from 2 different pretreatments using sulfuric acid and mixed cellulase enzymes (without and with delignification/ activated charcoal). The concentration of furfural, 5- hydroxymethyl furfural (5-HMF), levulinic acid, vanillin, syringaldehyde and4-hydroxybenzaldehyde (4-HB) and the percent acetic acid were found to be 0.0517 ± 0.049 mg/L, 0.032 ± 0.06 mg/L, 21074 ± 1685.62 mg/L, 126.265 ± 6.005 mg/L, 2.89 ± 0.30 mg/L, 0.37 ± 0.031mg/L and 0.72% under the pretreatment process without delignification/ activated charcoal treatment and 384.47 ± 99.02 g/L, 0.068 mg/L, 142107.62 ± 8664.6 mg/L, 0.19 mg/L, 5.43 ± 3.29 mg/L, 4.80 ± 0.76 mg/L and 0.254% under the pretreatment process with delignification/ activated charcoal treatment respectively. The presence of high concentration of acetic acid was found to impede the growth of Zymomonas mobilis strain TISTR 551 despite the present of high concentration of levulinic acid. Z. mobilis strain TISTR 551 was found to produce 8.96 ± 4.06 g/L of ethanol under 4 days fementation period in biofilm stage in which represented 40% theoretical yield.

Keywords: Rice bran, Zymomonas mobilis, biofilm, ethanol.

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110 Categorization and Estimation of Relative Connectivity of Genes from Meta-OFTEN Network

Authors: U. Kairov, T. Karpenyuk, E. Ramanculov, A. Zinovyev

Abstract:

The most common result of analysis of highthroughput data in molecular biology represents a global list of genes, ranked accordingly to a certain score. The score can be a measure of differential expression. Recent work proposed a new method for selecting a number of genes in a ranked gene list from microarray gene expression data such that this set forms the Optimally Functionally Enriched Network (OFTEN), formed by known physical interactions between genes or their products. Here we present calculation results of relative connectivity of genes from META-OFTEN network and tentative biological interpretation of the most reproducible signal. The relative connectivity and inbetweenness values of genes from META-OFTEN network were estimated.

Keywords: Microarray, META-OFTEN, gene network.

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109 Bioengineering for Customized Orthodontic Applications- Implant, Bracket and Dental Vibrator

Authors: Rajashekar Patil, S. Mohan Kumar, Shreya Ajmera

Abstract:

To understand complex living system an effort has made by mechanical engineers and dentists to deliver prompt products and services to patients concerned about their aesthetic look. Since two decades various bracket systems have designed involving techniques like milling, injection molding which are technically not flexible for the customized dental product development. The aim of this paper to design, develop a customized system which is economical and mainly emphasizes the expertise design and integration of engineering and dental fields. A custom made selfadjustable lingual bracket and customized implants are designed and developed using computer aided design (CAD) and rapid prototyping technology (RPT) to improve the smiles and to overcome the difficulties associated with conventional ones. Lengthy orthodontic treatment usually not accepted by the patients because the patient compliance is lost. Patient-s compliance can be improved by facilitating faster tooth movements by designing a localized dental vibrator using advanced engineering principles.

Keywords: Orthodontics, Prosthodontics, Lingual bracket, Implants, Dental vibrator, Computer aided design, Rapid prototyping technology.

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108 Phenotypes of B Cells Differ in EBV-positive Burkitt-s lymphoma Derived Cell Lines

Authors: Irina Spaka, Rita Birkenfelde, Svetlana Kozireva, Jevgenija Osmjana, Madara Upmane, ElenaKashuba, Irina Kholodnyuk Holodnuka

Abstract:

Epstein-Barr virus (EBV) is implicated in the pathogenesis of the endemic Burkitt-s lymphoma (BL). The EBVpositive BL-derived cell lines initially maintain the original tumor phenotype of EBV infection (latency I, LatI), but most of them drift toward a lymphoblast phenotype of EBV latency III (LatIII) during in vitro culturing. The aim of the present work was to characterize the B-cell subsets in EBV-positive BL cell lines and to verify whether a particular cell subset correlates with the type of EBV infection. The phenotype analysis of two EBV-negative and eleven EBV-positive (three of LatI and eight of LatIII) BL cell lines was performed by polychromatic flow cytomery, based on expression pattern of CD19, CD10, CD38, CD27, and CD5 markers. Two cell subsets, CD19+CD10+ and CD19+CD10-, were defined in LatIII BL cell lines. In both subsets, the CD27 and CD5 cell surface expression was detected in a proportion of the cells.

Keywords: B-cell subsets, Burkitt's lymphoma cell lines, EBV latency, phenotype profiles.

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107 Endothelial-Cell-Mediated Displacement of Extracellular Matrix during Angiogenesis

Authors: Yue Du, Sahan C. B. Herath, Qing-Guo Wang, Harry Asada, Peter C. Y. Chen

Abstract:

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.

Keywords: Angiogenesis, digital image correlation, digital volume correlation, interaction between ECs and ECM.

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106 Numbers and Biomass of Bacteria and Fungi Obtained by the Direct Microscopic Count Method

Authors: Ayuko Itsuki, Sachiyo Aburatani

Abstract:

The soil ecology of the organic and mineral soil layers of laurel-leaved and Cryptomeria japonica forest in the Kasuga-yama Hill Primeval Forest (Nara, Japan) was assessed. The number of bacteria obtained by the dilution plate count method was less than 0.05% of those counted by the direct microscopic count. We therefore found that forest soil contains large numbers of non-culturable bacteria compared with agricultural soils. The numbers of bacteria and fungi obtained by both the dilution plate count and the direct microscopic count were larger in the deeper horizons (F and H) of the organic layer than in the mineral soil layer. This suggests that active microbial metabolism takes place in the organic layer. The numbers of bacteria and the length of fungal hyphae obtained by the direct count method were greater in the H horizon than in the F horizon. The direct microscopic count revealed numerous non-culturable bacteria and fungi in the soil. The ratio of fungal to bacterial biomass was lower in the laurel-leaved forest soil. The fungal biomass was therefore relatively low in the laurel-leaved forest soil due to differences in forest vegetation.

Keywords: Bacterial number, Dilution plate count, Direct microscopic count, Forest soil.

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105 Effect of Geum Kokanicum Total Extract on Induced Nociception and Inflammation in Male Mice

Authors: M. Ramezani, S. Ghaderifard, HR. Monsef-Esfahani, S. Nasri

Abstract:

The aim of this study is evaluating the antinociceptive and anti-inflamatory activity of Geum kokanicum. After determination total extract LD50, different doses of extract were chosen for intrapritoneal injections. In inflammation test, male NMRI mice were divided into 6 groups: control (normal saline), positive control (Dexamethasone 15mg/kg), and total extract (0.025, 0.05, 0.1, and 0.2 gr/kg). The inflammation was produced by xyleneinduced edema. In order to evaluate the antinociceptive effect of total extract, formalin test was used. Mice were divided into 6 groups: control, positive control (morphine 10mg/kg), and 4 groups which received total extract. Then they received Formalin. The animals were observed for the reaction to pain. Data were analyzed using One-way ANOVA followed by Tukey-Kramer multiple comparison test. LD50 was 1 gr/kg. Data indicated that 0.5,0.1 and 0.2 gr/kg doses of total extract have particular antinociceptive and antiinflammatory effects in a comparison with control (P<0.001). The most effective dose was 0.2 gr/kg which did not show any significant difference in a comparison with positive control. Results indicated that total extract can inhibit nociception in the first and second phase. The antinociceptive effects in high doses are the same as morphine as a strong analgesic substance. TLC chromatography indicated presence of steroids and triterpenoids in this plant. The effects of extract may be related to presence of these compounds.

Keywords: Anti-inflammatory, Antinociceptive, Geum kokanicum, Mice.

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104 Metabolic Analysis of Fibroblast Conditioned Media and Comparison with Theoretical Modeling

Authors: Priyanka Gupta, Paul Verma, Kerry Hourigan, Jayesh Bellare, Sameer Jadhav

Abstract:

Understanding the consumption and production of various metabolites of fibroblast conditioned media is needed for its proper and optimized use in expansion of pluripotent stem cells. For this purpose, we have used the HPLC method to analyse the consumption of glucose and the production of lactate over time by mouse embryonic fibroblasts. The experimental data have also been compared with mathematical model fits. 0.025 moles of lactate was produced after 72 hrs while the glucose concentration decreased from 0.017 moles to 0.011 moles. The mathematical model was able to predict the trends of glucose consumption and lactate production.

Keywords: Conditioned media, HPLC, metabolite analysis, mouse embryonic fibroblast.

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103 Unnoticeable Mumps Infection in India: Does MMR Vaccine Protect against Circulating Mumps Virus Genotype C?

Authors: Jeevan Malayan, Aparna Warrier, Padmasani Venkat Ramanan, Sanjeeva Reddy N, Elanchezhiyan Manickan

Abstract:

MMR vaccine failure had been reported globally and here we report that it occurs now in India. Samples were collected from clinically suspected mumps cases were subjected for anti mumps antibodies, virus isolation, RT-PCR, sequencing and phylogenetic tree analysis. 56 samples collected from men and women belonging to various age groups. 30 had been vaccinated and the status of 26 patients was unknown. 28 out of 30 samples were found to be symptomatic and positive for Mumps IgM, indicating active mumps infection in 93.4% of the vaccinated population. A phylogenetic tree comparison of the clinical isolate is shown to be genotype C which is distinct from vaccine strain. Our study clearly sending warning signs that MMR vaccine is a failure and it needs to be revamped for the human use by increasing its efficacy and efficiency.

Keywords: Genotype C, Mumps virus, MMR vaccine, Sero types.

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102 Comparison of Anti-Shadoo Antibodies – Where is the Endogenous Shadoo protein?

Authors: Eszter Tóth, Ervin Welker

Abstract:

Shadoo protein (Sho) was described in 2003 as the newest member of Prion protein superfamily [1]. Sho has similar structural motifs like prion protein (PrP) that is known for its central role in transmissible spongiform enchephalopathies. Although a great number of functions have been proposed, the exact physiological function of PrP is not known yet. Investigation of the function and localization of Sho may help us to understand the function of the Prion protein superfamily. Analyzing the subcellular localization of YFP-tagged forms of Sho, we detected the protein in the plasma membrane and in the nucleus of various cell lines. To reveal the localization of the endogenous protein we generated antibodies against Shadoo as well as employed commercially available anti-Shadoo antibodies: i) EG62 anti-mouse Shadoo antibody generated by Eurogentec Ltd.; ii) S-12 anti-human Shadoo antibody by Santa Cruz Biotechnology Inc.; iii) R-12 anti-mouse Shadoo antibody by Santa Cruz Biotechnology Inc.; iv) SPRN antibody against human Shadoo by Abgent Inc. We carried out immunocytochemistry on non-transfected HeLa, Zpl 2-1, Zw 3-5, GT1-1, GT1-7 and SHSY5Y cells as well as on YFP-Sho, Sho-YFP, and YFP-GPI transfected HeLa cells. Their specificity (in antibody-peptide competition assay) and co-localization (with the YFP signal) were assessed.

Keywords: Shadoo, prion protein, immunocytochemistry, antibody-peptide competition assay, antibody.

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101 Promotion of Growth and Modulation of As- Induced Stress Ethylene in Maize by As- Tolerant ACC Deaminase Producing Bacteria

Authors: Charlotte C. Shagol, Tongmin Sa

Abstract:

One of the major pollutants in the environment is arsenic (As). Due to the toxic effects of As to all organisms, its remediation is necessary. Conventional technologies used in the remediation of As contaminated soils are expensive and may even compromise the structure of the soil. An attractive alternative is phytoremediation, which is the use of plants which can take up the contaminant in their tissues. Plant growth promoting bacteria (PGPB) has been known to enhance growth of plants through several mechanisms such as phytohormone production, phosphate solubilization, siderophore production and 1-aminocyclopropane-1- carboxylate (ACC) deaminase production, which is an essential trait that aids plants especially under stress conditions such as As stress. Twenty one bacteria were isolated from As-contaminated soils in the vicinity of the Janghang Smelter in Chungnam Province, South Korea. These exhibited high tolerance to either arsenite (As III) or arsenate (As V) or both. Most of these isolates possess several plant growth promoting traits which can be potentially exploited to increase phytoremediation efficiency. Among the identified isolates is Pseudomonas sp. JS1215, which produces ACC deaminase, indole acetic acid (IAA), and siderophore. It also has the ability to solubilize phosphate. Inoculation of JS1215 significantly enhanced root and shoot length and biomass accumulation of maize under normal conditions. In the presence of As, particularly in lower As level, inoculation of JS1215 slightly increased root length and biomass. Ethylene increased with increasing As concentration, but was reduced by JS1215 inoculation. JS1215 can be a potential bioinoculant for increasing phytoremediation efficiency.

Keywords: As-tolerant bacteria, plant growth promoting bacteria, As stress, phytoremediation.

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100 Molecular Dynamics Study on Laninamivir Inhibiting Neuraminidases of H5N1 and pH1N1 Influenza a Viruses

Authors: A. Meeprasert, W. Khuntawee, S. Hannongbua, T. Rungrotmongkol

Abstract:

Viral influenza A subtypes H5N1 and pandemic H1N1 (pH1N1) have worldwide emerged and transmitted. The most common anti-influenza drug for treatment of both seasonal and pandemic influenza viruses is oseltamivir that nowadays becomes resistance to influenza neuraminidase. The novel long-acting drug, laninamivir, was discovered for treatment of the patients infected with influenza B and influenza A viruses. In the present study, laninamivir complexed with wild-type strain of both H5N1 and pH1N1 viruses were comparatively determined the structures and drug-target interactions by means of molecular dynamics (MD) simulations. The results show that the hydrogen bonding interactions formed between laninamivir and its binding residues are likely similar for the two systems. Additionally, the presence of intermolecular interactions from laninamivir to the residues in the binding pocket is established through their side chains in accordance with hydrogen bond interactions.

Keywords: Laninamivir, neuraminidase, H5N1, pandemic H1N1, wild-type, MD simulation

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99 Smart Motion

Authors: Arpita Soni, Sapna Mittal

Abstract:

Austenite and Martensite indicate the phases of solids undergoing phase transformation which we usually associate with materials and not with living organisms. This article provides an overview of bacterial proteins and structures that are undergoing phase transformation and suggests its probable effect on mechanical behavior. The context is mainly within the role of phase transformations occurring in the flagellum of bacteria. The current knowledge of molecular mechanism leading to phase variation in living organisms is reviewed. Since in bacteria, each flagellum is driven by a separate motor, similarity to a Differential drive in case of four-wheeled vehicles is suggested. It also suggests the application of the mechanism in which bacteria changes its direction of movement to facilitate single point turning of a multi-wheeled vehicle. Finally, examples are presented to illustrate that the motion due to phase transformation of flagella in bacteria can start a whole new research on motion mechanisms.

Keywords: Flagella, Phase Transformation, Nanobots, Differential Drive, Single point turn, Biomimetics

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98 A Heat-Inducible Transgene Expression System for Gene Therapy

Authors: Masaki Yamaguchi, Akira Ito, Noriaki Okamoto, Yoshinori Kawabe, Masamichi Kamihira

Abstract:

Heat-inducible gene expression vectors are useful for hyperthermia-induced cancer gene therapy, because the combination of hyperthermia and gene therapy can considerably improve the therapeutic effects. In the present study, we developed an enhanced heat-inducible transgene expression system in which a heat-shock protein (HSP) promoter and tetracycline-responsive transactivator were combined. When the transactivator plasmid containing the tetracycline-responsive transactivator gene was co-transfected with the reporter gene expression plasmid, a high level of heat-induced gene expression was observed compared with that using the HSP promoter without the transactivator. In vitro evaluation of the therapeutic effect using HeLa cells showed that heat-induced therapeutic gene expression caused cell death in a high percentage of these cells, indicating that this strategy is promising for cancer gene therapy.

Keywords: Inducible gene expression, Gene therapy, Hyperthermia, Heat shock protein, Tetracycline transactivator.

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97 Biospeckle Techniques in Quality Evaluation of Indian Fruits

Authors: MD Zaheer Ansari, A.K. Nirala

Abstract:

In this study spatial-temporal speckle correlation techniques have been applied for the quality evaluation of three different Indian fruits namely apple, pear and tomato for the first time. The method is based on the analysis of variations of laser light scattered from biological samples. The results showed that crosscorrelation coefficients of biospeckle patterns change subject to their freshness and the storage conditions. The biospeckle activity was determined by means of the cross-correlation functions of the intensity fluctuations. Significant changes in biospeckle activity were observed during their shelf lives. From the study, it is found that the biospeckle activity decreases with the shelf-life storage time. Further it has been shown that biospeckle activity changes according to their respiration rates.

Keywords: Biospeckle, cross-correlation, respiration, shelf-life.

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