Search results for: gene module
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2157

Search results for: gene module

2097 Wt1 and FoxL2 Genes Expression Pattern in Mesonephros-Gonad Complexes of Green Sea Turtle (Chelonia mydas) Embryos Incubated in Feminization and Masculinization Temperature

Authors: Fitria D. Ayuningtyas, Anggraini Barlian

Abstract:

Green turtle (Chelonia mydas) is one of TSD (Temperature-dependent Sex Determination, TSD) animals which sex is determined by the egg’s incubation temperature. GSD (Genotypic Sex Determination) homologous genes such as Wilms’ Tumor (Wt1) and Forkhead Box L2 (FoxL2) play a role in TSD animal sex determination process. Wt1 plays a role in both male pathway, as a transcription factor for Sf1 gene and in female pathway, as a transcription factor for Dax1. FoxL2 plays a role specifically in female sex determination, and known as transcriptional factor for Aromatase gene. Until now, research on the pattern of Wt1 and FoxL2 genes expression in C.mydas has not been conducted yet. The aim of this research is to know the pattern of Wt1 and FoxL2 genes expression in Mesonephros-Gonad (MG) complexes of Chelonia mydas embryos incubated in masculinizing temperature (MT) and feminizing temperature (FT). Eggs of C.mydas incubated in 3 different stage of TSP (Thermosensitive Period) at masculinizing temperature (26±10C, MT) and feminizing temperature (31±10C FT). Mesonefros-gonad complexes were isolated at Pre-TSP stage (FT at days 14th, MT at days 24th), TSP stage (FT at days 24th, MT at days 36th) and differentiated stage (FT at days 40th, MT at days 58th). RNA from mesonephros-gonad (MG) complexes were converted into cDNA by RT-PCR process, and the pattern of Wt1 and FoxL2 genes expression is analyzed by quantitative Real Time PCR (qPCR) method, β-actin gene is used as an internal control. The pattern of Wt1 gene expression in Pre-TSP stage was almost the same between MG complexes incubated at MT or FT, while TSP and differentiation stage, the pattern of Wt1 gene expression in MG complexes incubated at MT or FT was increased. Wt1 gene expression of MG complexes that incubated at FT was higher than at MT. There was a difference pattern between Wt1 gene expression in this research compared to the previous research in protein level. It could be assumed that the difference caused by post-transcriptional regulation mechanisms before mRNA of Wt1 gene translated into protein structure. The pattern of FoxL2 gene expression in Pre-TSP stage was almost the same between MG complexes that incubated at MT and FT, and increased in both TSP and differentiated stage. The FoxL2 gene expression in MG complexes that incubated in FT is higher than MT on TSP and differentiated stage. Based on the results of this research, it can be assumed that Wt1 and FoxL2 gene were expressed in MG complexes that incubated both at MT and FT since Pre-TSP stage. The pattern of Wt1 gene expression was increased in every stage of gonadal development, and so do the pattern of FoxL2 gene expression. Wt1 and FoxL2 gene expressions were higher in MG complexes incubated at FT than MT.

Keywords: chelonia mydas, FoxL2, gene expression, TSD, Wt1

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2096 A New Design of Vacuum Membrane Distillation Module for Water Desalination

Authors: Adnan Alhathal Alanezi

Abstract:

The performance of vacuum membrane distillation (VMD) process for water desalination was investigated utilizing a new design membrane module using two commercial polytetrafluoroethylene (PTFE) and polyvinylidene fluoride (PVDF) flat sheet hydrophobic membranes. The membrane module's design demonstrated its suitability for achieving a high heat transfer coefficient of the order of 103 (W/m2K) and a high Reynolds number (Re). The heat and mass transport coefficients within the membrane module were measured using VMD experiments. The permeate flux has been examined in relation to process parameters such as feed temperature, feed flow rate, vacuum degree, and feed concentration. Because the feed temperature, feed flow rate, and vacuum degree all play a role in improving the performance of the VMD process, optimizing all of these parameters is the best method to achieve a high permeate flux. In VMD desalination, the PTFE membrane outperformed the PVDF membrane. When compared to previous studies, the obtained water flux is relatively high, reaching 43.8 and 52.6 (kg/m2h) for PVDF and PTFE, respectively. For both membranes, the salt rejection of NaCl was greater than 99%.

Keywords: desalination, vacuum membrane distillation, PTFE and PVDF, hydrophobic membranes, O-ring membrane module

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2095 Cloning and Expression of the ansZ Gene from Bacillus sp. CH11 Isolated from Chilca salterns in Peru

Authors: Stephy Saavedra, Annsy C. Arredondo, Gisele Monteiro, Adalberto Pessoa Jr, Carol N. Flores-Fernandez, Amparo I. Zavaleta

Abstract:

L-asparaginase from bacterial sources is used in leukemic treatment and food industry. This enzyme is classified based on its affinity towards L-asparagine and L-glutamine. Likewise, ansZ genes express L-asparaginase with higher affinity to L-asparagine. The aim of this work was to clone and express of ansZ gene from Bacillus sp. CH11 isolated from Chilca salterns in Peru. The gene encoding L-asparaginase was cloned into pET15b vector and transformed in Escherichia coli BL21 (DE3) pLysS. The expression was carried out in a batch culture using LB broth and 0.5 mM IPTG. The recombinant L-asparaginase showed a molecular weight of ~ 39 kDa by SDS PAGE and a specific activity of 3.19 IU/mg of protein. The cloning and expression of ansZ gene from this halotolerant Bacillus sp. CH11 allowed having a biological input to improve a future scaling-up.

Keywords: ansZ gene, Bacillus sp, Chilca salterns, recombinant L-asparaginase

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2094 Bioinformatics Analysis of DGAT1 Gene in Domestic Ruminnants

Authors: Sirous Eydivandi

Abstract:

Diacylglycerol-O-acyltransferase (DGAT1) gene encodes diacylglycerol transferase enzyme that plays an important role in glycerol lipid metabolism. DGAT1 is considered to be the key enzyme in controlling the synthesis of triglycerides in adipocytes. This enzyme catalyzes the final step of triglyceride synthesis (transform triacylglycerol (DAG) into triacylglycerol (TAG). A total of 20 DGAT1 gene sequences and corresponding amino acids belonging to 4 species include cattle, goats, sheep and yaks were analyzed, and the differentiation within and among the species was also studied. The length of the DGAT1 gene varies greatly, from 1527 to 1785 bp, due to deletion, insertion, and stop codon mutation resulting in elongation. Observed genetic diversity was higher among species than within species, and Goat had more polymorphisms than any other species. Novel amino acid variation sites were detected within several species which might be used to illustrate the functional variation. Differentiation of the DGAT1 gene was obvious among species, and the clustering result was consistent with the taxonomy in the National Center for Biotechnology Information.

Keywords: DGAT1gene, bioinformatic, ruminnants, biotechnology information

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2093 Pathway and Differential Gene Expression Studies for Colorectal Cancer

Authors: Ankita Shukla, Tiratha Raj Singh

Abstract:

Colorectal cancer (CRC) imposes serious mortality burden worldwide and it has been increasing for past consecutive years. Continuous efforts have been made so far to diagnose the disease condition and to identify the root cause for it. In this study, we performed the pathway level as well as the differential gene expression studies for CRC. We analyzed the gene expression profile GSE24514 from Gene Expression Omnibus (GEO) along with the gene pathways involved in the CRC. This analysis helps us to understand the behavior of the genes that have shown differential expression through their targeted pathways. Pathway analysis for the targeted genes covers the wider area which therefore decreases the possibility to miss the significant ones. This will prove to be beneficial to expose the ones that have not been given attention so far. Through this analysis, we attempt to understand the various neighboring genes that have close relationship to the targeted one and thus proved to be significantly controlling the CRC. It is anticipated that the identified hub and neighboring genes will provide new directions to look at the pathway level differently and will be crucial for the regulatory processes of the disease.

Keywords: mismatch repair, microsatellite instability, carcinogenesis, morbidity

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2092 Developing Soil Accumulation Effect Correction Factor for Solar Photovoltaic Module

Authors: Kelebaone Tsamaase, Rapelang Kemoabe, Japhet Sakala, Edward Rakgati, Ishmael Zibani

Abstract:

Increasing demand for energy, depletion of non-renewable energy, effects of climate change, the abundance of renewable energy such as solar energy have increased the interest in investing in renewable energies, in particular solar photovoltaic (PV) energy. Solar photovoltaic energy systems as part of clean technology are considered to be environmentally friendly, freely available, offer clean production systems, long term costs benefits as opposed to conventional sources, and are the attractive power source for a wide range of applications in remote areas where there is no easy access to the national grid. To get maximum electrical power, maximum solar power should penetrate the module and be converted accordingly. However, some environmental and other geographical related factors reduce the electrical power. One of them is dust which accumulates on the surface of the module and forming a dust layer and in the process obstructing the solar power from penetrating PV module. This study intends to improve the performance of solar photovoltaic (PV) energy modules by establishing soil accumulation effects correction factor from dust characteristics and properties, and also from dust accumulation and retention pattern on PV module surface. The non-urban dry deposition flux model was adapted to determine monthly and yearly dust accumulation pattern. Consideration was done on prevailing environmental and other geographical conditions. Preliminary results showed that cumulative dust settlement increased during the months of July to October leading to a higher drop in module electrical output power.

Keywords: dust, electrical power output, PV module, soil correction factor

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2091 ZBTB17 Gene rs10927875 Polymorphism in Slovak Patients with Dilated Cardiomyopathy

Authors: I. Boroňová, J. Bernasovská, J. Kmec, E. Petrejčíková

Abstract:

Dilated cardiomyopathy (DCM) is a severe cardiovascular disorder characterized by progressive systolic dysfunction due to cardiac chamber dilatation and inefficient myocardial contractility often leading to chronic heart failure. Recently, a genome-wide association studies (GWASs) on DCM indicate that the ZBTB17 gene rs10927875 single nucleotide polymorphism is associated with DCM. The aim of the study was to identify the distribution of ZBTB17 gene rs10927875 polymorphism in 50 Slovak patients with DCM and 80 healthy control subjects using the Custom Taqman®SNP Genotyping assays. Risk factors detected at baseline in each group included age, sex, body mass index, smoking status, diabetes and blood pressure. The mean age of patients with DCM was 52.9±6.3 years; the mean age of individuals in control group was 50.3±8.9 years. The distribution of investigated genotypes of rs10927875 polymorphism within ZBTB17 gene in the cohort of Slovak patients with DCM was as follows: CC (38.8%), CT (55.1%), TT (6.1%), in controls: CC (43.8%), CT (51.2%), TT (5.0%). The risk allele T was more common among the patients with dilated cardiomyopathy than in normal controls (33.7% versus 30.6%). The differences in genotype or allele frequencies of ZBTB17 gene rs10927875 polymorphism were not statistically significant (p=0.6908; p=0.6098). The results of this study suggest that ZBTB17 gene rs10927875 polymorphism may be a risk factor for susceptibility to DCM in Slovak patients with DCM. Studies of numerous files and additional functional investigations are needed to fully understand the roles of genetic associations.

Keywords: ZBTB17 gene, rs10927875 polymorphism, dilated cardiomyopathy, cardiovascular disorder

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2090 Inheritance of Protein Content and Grain Yield in Half Diallel Maize (Zea mays L.) Populations

Authors: Gül Ebru Orhun

Abstract:

A half diallel crossing design was carried out during 2011 and 2012 growing seasons under Çanakkale-Turkey ecological conditions. In this research, 20 F1 maize hybrids obtained by 6x6 half diallel crossing were used. Gene action for protein content and grain yield traits were explored in half set involving six elite inbred lines. According to the results diallel analysis dominance and additive gene variances were determined for protein content. Variance/Co-variance graphs revealed for grain yield and protein content traits. In this study, inheritance of grain yield and protein content demonstrated over-dominance type of gene action.

Keywords: protein, maize, inheritance, gene action

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2089 Evaluation of Promoter Hypermethylation in Tissue and Blood of Non-Small Cell Lung Cancer Patients and Association with Survival

Authors: Ashraf Ali, Kriti Upadhyay, Puja Sohal, Anant Mohan, Randeep Guleria

Abstract:

Background: Gene silencing by aberrant promoter hypermethylation is common in lung cancer and is an initiating event in its development. Aim: To evaluate the gene promoter hypermethylation frequency in serum and tissue of lung cancer patients. Method: 95 newly diagnosed untreated advance stage lung cancer patients and 50 cancer free matched controls were studied. Bisulfite modification of tissue and serum DNA was done; modified DNA was used as a template for methylation-specific PCR analysis. Survival was assessed for one year. Results: Of 95 patients, 82% were non-small cell lung cancer (34% squamous cell carcinoma, 34% non-small cell lung cancer and 14% adenocarcinoma) and 18% were small cell lung cancer. Biopsy revealed that tissue of 89% and 75% of lung cancer patients and 85% and 52% of controls had promoter hypermethylated for MGMT (p=0.35) and p16(p<0.001) gene, respectively. In serum, 33% and 49% of lung cancer patients and 28% and 43% controls were positive for MGMT and p16 gene. No significant correlation was found between survival and clinico-pathological parameters. Conclusion: High gene promoter methylation frequency of p16 gene in tissue biopsy may be linked with early stages of carcinogenesis. Appropriate follow-up is required for confirmation of this finding.

Keywords: lung cancer, MS- PCR, methylation, molecular biology

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2088 Transcriptomic Response of Calmodulin Encoding Gene (CaM) in Pesticide Utilizing Talaromyces Fungal Strains

Authors: M. D. Asemoloye, S. G. Jonathan, A. Rafiq, O. J. Olawuyi, D. O. Adejoye

Abstract:

Calmodulin is one of the intracellular calcium proteins that regulates large spectrum of enzymes and cellular functions including metabolism of cyclic nucleotides and glycogen. The potentials of calmodulin gene in fungi necessitates their genetic response and their strong cassette of enzyme secretions for pesticide degradation. Therefore, this study was carried out to investigate the ‘Transcriptomic’ response of calmodulin encoding genes in Talaromyces fungi in response to 2, 2-dichlorovinyl dimethyl phosphate (DDVP or Dichlorvos) an organophosphate pesticide and γ-Hexachlorocyclohexane (Lindane) an organochlorine pesticide. Fungi strains isolated from rhizosphere from grasses rhizosphere in pesticide polluted sites were subjected to percentage incidence test. Two most frequent fungi were further characterized using ITS gene amplification (ITS1 and ITS4 combinations), they were thereafter subjected to In-vitro DDVP and lindane tolerance tests at different concentrations. They were also screened for presence and expression of calmodulin gene (caM) using RT-PCR technique. The two Talaromyces strains had the highest incidence of 50-72% in pesticide polluted site, they were both identified as Talaromyces astroroseus asemoG and Talaromyces purpurogenum asemoN submitted in NCBI gene-bank with accession numbers KY488464 and KY488468 respectively. T. astroroseus KY488464 tolerated DDVP (1.23±0.023 cm) and lindane (1.11±0.018 cm) at 25 % concentration while T. purpurogenum KY488468 tolerated DDVP (1.33±0.061 cm) and lindane (1.54±0.077 cm) at this concentration. Calmodulin gene was detected in both strains, but RT-PCR expression of caM gene revealed at 900-1000 bp showed an under-expression of caM in T. astrorosues KY488464 but overexpressed in T. purpurogenum KY488464. Thus, the calmodulin gene response of these fungal strains to both pesticides could be considered in monitoring the potentials of fungal strains to pesticide tolerance and bioremediation of pesticide in polluted soil.

Keywords: Calmodulin gene, pesticide, RT-PCR, talaromyces, tolerance

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2087 The Instablity of TetM Gene Encode Tetracycline Resistance Gene in Lactobacillus casei FNCC 0090

Authors: Sarah Devi Silvian, Hanna Shobrina Iqomatul Haq, Fara Cholidatun Nabila, Agustin Krisna Wardani

Abstract:

Bacteria ability to survive in antibiotic is controlled by the presence of gene that encodes the antibiotic resistance protein. The instability of the antibiotic resistance gene can be observed by exposing the bacteria under the lethal dose of antibiotic. Low concentration of antibiotic can induce mutation, which may take a role in bacterial adaptation through the antibiotic concentration. Lactobacillus casei FNCC 0090 is one of the probiotic bacteria that has an ability to survive in tetracycline by expressing the tetM gene. The aims of this study are to observe the possibilities of mutation happened in L.casei FNCC 0090 by exposing in sub-lethal dose of tetracycline and also observing the instability of the tetM gene by comparing the sequence between the wild type and mutant. L.casei FNCC 0090 has a lethal dose in 60 µg/ml, low concentration is applied to induce the mutation, the range from 10 µg/ml, 15 µg/ml, 30 µg/ml, 45 µg/ml, and 50 µg/ml. L.casei FNCC 0090 is exposed to the low concentration from lowest to the highest concentration to induce the adaptation. Plasmid is isolated from the highest concentration culture which is 50 µg/ml by using modified alkali lysis method with the addition of lysozyme. The tetM gene is isolated by using PCR (Polymerase Chain Reaction) method, then PCR amplicon is purified and sequenced. Sequencing is done on both samples, wild type and mutant. Both sequences are compared and the mutations can be traced in the presence of nucleotides changes. The changing of the nucleotides means that the tetM gene is instable.

Keywords: L. casei FNCC 0090, probiotic, tetM, tetracycline

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2086 Characterization of (GRAS37) Gibberellin Acid Insensitive (GAI), Repressor (RGA), and Scarecrow (SCR) Gene by Using Bioinformatics Tools

Authors: Yusra Tariq

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The Grass 37 gene is presently known in tomatoes, which are the source of healthy substances such as ascorbic acid, polyphenols, carotenoids and nutrients. It has a significant impact on the growth and development of humans. The GRASS 37 gene is a plant Transcription factor group assuming significant parts in various reactions of different Abiotic stresses such as (drought, salinity, thermal stresses, temperature, and bright waves) which could highly affect the growth. Tomatoes are very sensitive to temperature, and their growth or production occurs optimally in a temperature range from 21 C to 29.5 C during the daytime and from 18.5 C to 21 C during the night. This protein acts as a positive regulator of salt stress response and abscisic acid signaling. This study summarizes the structure characterized by molecular formula and protein-binding domains by different bioinformatics tools such as Expasy translate tool, Expasy Portparam, Swiss Prot and Inter Pro Scan, Clustal W tool regulatory procedure of GRASS gene components, also their reactions to both biotic and Abiotic stresses.

Keywords: GRAS37, gene, bioinformatics, tool

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2085 Paper-Based Detection Using Synthetic Gene Circuits

Authors: Vanessa Funk, Steven Blum, Stephanie Cole, Jorge Maciel, Matthew Lux

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Paper-based synthetic gene circuits offer a new paradigm for programmable, fieldable biodetection. We demonstrate that by freeze-drying gene circuits with in vitro expression machinery, we can use complimentary RNA sequences to trigger colorimetric changes upon rehydration. We have successfully utilized both green fluorescent protein and luciferase-based reporters for easy visualization purposes in solution. Through several efforts, we are aiming to use this new platform technology to address a variety of needs in portable detection by demonstrating several more expression and reporter systems for detection functions on paper. In addition to RNA-based biodetection, we are exploring the use of various mechanisms that cells use to respond to environmental conditions to move towards all-hazards detection. Examples include explosives, heavy metals for water quality, and toxic chemicals.

Keywords: cell-free lysates, detection, gene circuits, in vitro

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2084 Induction of HIV-1 Resistance: The New Approaches Based on Gene Modification and Stem Cell Engineering

Authors: Alieh Farshbaf

Abstract:

Introduction: Current anti-retroviral drugs have some restrictions for treatment of HIV-1 infection. The efficacy of retroviral drugs is not same in different infected patients and the virus rebound from latent reservoirs after stopping them. Recently, the engineering of stem cells and gene therapy provide new approaches to eliminate some drug problems by induction of resistance to HIV-1. Literature review: Up to now, AIDS-restriction genes (ARGs) were suitable candidate for gene and cell therapies, such as cc-chemokine receptor-5 (CCR5). In this manner, CCR5 provide effective cure in Berlin and Boston patients by inducing of HIV-1 resistance with allogeneic stem cell transplantation. It is showed that Zinc Finger Nuclease (ZFN) could induce HIV-1 resistance in stem cells of infected patients by homologous recombination or non-end joining mechanism and eliminate virus loading after returning the modified cells. Then, gene modification by HIV restriction factors, as TRIM5, introduced another gene candidate for HIV by interfering in infection process. These gene modifications/editing provided by stem cell futures that improve treatment in refractory disease such as HIV-1. Conclusion: Although stem cell transplantation has some complications, but in compare to retro-viral drugs demonstrated effective cure by elimination of virus loading. On the other hand, gene therapy is cost-effective for an infected patient than retroviral drugs payment in a person life-long. The results of umbilical cord blood stem cell transplantation showed that gene and cell therapy will be applied easier than previous treatment of AIDS with high efficacy.

Keywords: stem cell, AIDS, gene modification, cell engineering

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2083 Mutations in the GJB2 Gene Are the Cause of an Important Number of Non-Syndromic Deafness Cases

Authors: Habib Onsori, Somayeh Akrami, Mohammad Rahmati

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Deafness is the most common sensory disorder with the frequency of 1/1000 in many populations. Mutations in the GJB2 (CX26) gene at the DFNB1 locus on chromosome 13q12 are associated with congenital hearing loss. Approximately 80% of congenital hearing loss cases are recessively inherited and 15% dominantly inherited. Mutations of the GJB2 gene, encoding gap junction protein Connexin 26 (Cx26), are the most common cause of hereditary congenital hearing loss in many countries. This report presents two cases of different mutations from Iranian patients with bilateral hearing loss. DNA studies were performed for the GJB2 gene by PCR and sequencing methods. In one of them, direct sequencing of the gene showed a heterozygous T→C transition at nucleotide 604 resulting in a cysteine to arginine amino acid substitution at codon 202 (C202R) in the fourth extracellular domain (TM4) of the protein. The analyses indicate that the C202R mutation appeared de novo in the proband with a possible dominant effect (GenBank: KF 638275). In the other one, DNA sequencing revealed a compound heterozygous mutation (35delG, 363delC) in the Cx26 gene that is strongly associated with congenital non-syndromic hearing loss (NSHL). So screening the mutations for hearing loss individuals referring to genetics counseling centers before marriage and or pregnancy is recommended.

Keywords: CX26, deafness, GJB2, mutation

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2082 Robustness Conditions for the Establishment of Stationary Patterns of Drosophila Segmentation Gene Expression

Authors: Ekaterina M. Myasnikova, Andrey A. Makashov, Alexander V. Spirov

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First manifestation of a segmentation pattern in the early Drosophila development is the formation of expression domains (along with the main embryo axis) of genes belonging to the trunk gene class. Highly variable expression of genes from gap family in early Drosophila embryo is strongly reduced by the start of gastrulation due to the gene cross-regulation. The dynamics of gene expression is described by a gene circuit model for a system of four gap genes. It is shown that for the formation of a steep and stationary border by the model it is necessary that there existed a nucleus (modeling point) in which the gene expression level is constant in time and hence is described by a stationary equation. All the rest genes expressed in this nucleus are in a dynamic equilibrium. The mechanism of border formation associated with the existence of a stationary nucleus is also confirmed by the experiment. An important advantage of this approach is that properties of the system in a stationary nucleus are described by algebraic equations and can be easily handled analytically. Thus we explicitly characterize the cross-regulation properties necessary for the robustness and formulate the conditions providing this effect through the properties of the initial input data. It is shown that our formally derived conditions are satisfied for the previously published model solutions.

Keywords: drosophila, gap genes, reaction-diffusion model, robustness

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2081 Simulation of Photovoltaic Array for Specified Ratings of Converter

Authors: Smita Pareek, Ratna Dahiya

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The power generated by solar photovoltaic (PV) module depends on surrounding irradiance, temperature, shading conditions, and shading pattern. This paper presents a simulation of photovoltaic module using Matlab/Simulink. PV Array is also simulated by series and parallel connections of modules and their characteristics curves are given. Further PV module topology/configuration are proposed for 5.5kW inverter available in the literature. Shading of a PV array either complete or partial can have a significant impact on its power output and energy yield; therefore, the simulated model characteristics curves (I-V and P-V) are drawn for uniform shading conditions (USC) and then output power, voltage and current are calculated for variation in insolation for shading conditions. Additionally the characteristics curves are also given for a predetermined shadowing condition.

Keywords: array, series, parallel, photovoltaic, partial shading

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2080 A Novel PfkB Gene Cloning and Characterization for Expression in Potato Plants

Authors: Arfan Ali, Idrees Ahmad Nasir

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Potato (Solanum tuberosum) is an important cash crop and popular vegetable in Pakistan and throughout the world. Cold storage of potatoes accelerates the conversion of starch into reduced sugars (glucose and fructose). This process causes dry mass and bitter taste in the potatoes that are not acceptable to end consumers. In the current study, the phosphofructokinase B gene was cloned into the pET-30 vector for protein expression and the pCambia-1301 vector for plant expression. Amplification of a 930bp product from an E. coli strain determined the successful isolation of the phosphofructokinase B gene. Restriction digestion using NcoI and BglII along with the amplification of the 930bp product using gene specific primers confirmed the successful cloning of the PfkB gene in both vectors. The protein was expressed as a His-PfkB fusion protein. Western blot analysis confirmed the presence of the 35 Kda PfkB protein when hybridized with anti-His antibodies. The construct Fani-01 was evaluated transiently using a histochemical gus assay. The appearance of blue color in the agroinfiltrated area of potato leaves confirmed the successful expression of construct Fani-01. Further, the area displaying gus expression was evaluated for PfkB expression using ELISA. Moreover, PfkB gene expression evaluated through transient expression determined successful gene expression and highlighted its potential utilization for stable expression in potato to reduce sweetening due to long-term storage.

Keywords: potato, Solanum tuberosum, transformation, PfkB, anti-sweetening

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2079 Association of Xeroderma pigmentosum Group D Gene Polymorphism with Colorectal Cancer Risk in Kashmiri Population

Authors: Syed Sameer Aga, Saniya Nissar

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The Xeroderma pigmentosum group D gene (XPD) plays a key role in nucleotide excision repair (NER) pathway of the damaged DNA. Genetic polymorphisms in the coding region of the XPD gene may alter DNA repair capacity of the protein and hence can modulate the risk of colorectal cancer (CRC) risk. The aim of the study was to determine the genetic association of XPD Lys751Gln polymorphism with the risk of colorectal cancer (CRC) development. 120 CRC patients and 160 normal controls were assessed for genotype frequencies of XPD Lys751Gln polymorphism using PCR-RFLP technique. We observed a significant association (p < 0.05) between the XPD Lys751Gln polymorphism and the risk of developing CRC (p < 0.05). Additionally, Gln/Gln genotype of the XPD gene doubled the risk for the development of CRC [p < 0.05; OR=2.25 95% CI (1.07-4.7)]. Our results suggest that there is a significant association between the XPD Lys751Gln polymorphism and the risk of CRC.

Keywords: colorectal cancer, polymorphism, RFLP, DNA Repair, NER, XPD

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2078 Identification of Two Novel Carbapenemase Gene Variants from a Carbapenem-Resistant Aeromonas Veronii Environmental Isolate

Authors: Rafael Estrada, Cristian Ruiz Rueda

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Carbapenems are last-resort antibiotics used in clinical settings to treat antibiotic-resistant bacterial infections. Thus, the emergence and spread of resistance to carbapenems is a major public health concern. Here, we have studied a carbapenem-resistant Aeromonas veronii strain previously isolated from a water sample from Sam Simeon Creek (Hearst San Simeon State Park, CA). Analysis of this isolate using disk-diffusion, CarbaNP, eCIM and mCIM assays revealed that it was resistant to amoxicillin-clavulanic acid and all carbapenems tested and that this isolate produced a potentially novel carbapenemase of the Metallo-β-lactamase family. Whole genome sequencing analysis revealed that this A. veronii isolate carries a novel variant of the blacₚₕₐ class β-carbapenemase gene that was closely related to the blacₚₕₐ₇ gene of Aeromonas jandaei. This isolate also carried a novel variant of the blaₒₓₐ class D carbapenemase gene that was most closely related to the blaₒₓₐ-₉₁₂ gene found in other Aeromonas veronii isolates. Finally, we also identified a novel class C β-lactamase gene moderately related to the blaFₒₓ-₁₇ gene of Providencia stuartii and other blaFₒₓ variants identified in Klebsiella pneumoniae, Escherichia coli and other Enterobacteriaceae. Overall, our findings reveal that environmental isolates are an important reservoir of multiple carbapenemases and other β-lactamases of clinical significance.

Keywords: β-lactamases, carbapenem, antibiotic-resistant, aeromonas veronii

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2077 Comparison of Efficient Production of Small Module Gears

Authors: Vaclav Musil, Robert Cep, Sarka Malotova, Jiri Hajnys, Frantisek Spalek

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The new designs of satellite gears comprising a number of small gears pose high requirements on the precise production of small module gears. The objective of the experimental activity stated in this article was to compare the conventional rolling gear cutting technology with the modern wire electrical discharge machining (WEDM) technology for the production of small module gear m=0.6 mm (thickness of 2.5 mm and material 30CrMoV9). The WEDM technology lies in copying the profile of gearing from the rendered trajectory which is then transferred to the track of a wire electrode. During the experiment, we focused on the comparison of these production methods. Main measured parameters which significantly influence the lifetime and noise was chosen. The first parameter was to compare the precision of gearing profile in respect to the mathematic model. The second monitored parameter was the roughness and surface topology of the gear tooth side. The experiment demonstrated high accuracy of WEDM technology, but a low quality of machined surface.

Keywords: precision of gearing, small module gears, surface topology, WEDM technology

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2076 The Effectiveness of the Counselling Module in Counseling Interventions for Low Performance Employees

Authors: Hazaila Hassan

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This research aims and discusses about the effectiveness of the Psynnova i-Behaviour Modification Technique (iBMT) module towards the change in behaviour of low-performing employees. The purpose of the study is to examine the effectiveness of the Psynnova Module on changing behaviour through five factors among low-performing employees in the public sector. The five main factors/constructs were cognitive enhancement and rationality, emotional stability, attitude alignment and adjustment, social skills development and psycho-spirituality enhancement. In this research, 5 main constructs will be using to indicate behaviour changing performance of the employees after attending The Psynnova Program that using this Psynnova IBMT Module. The respondents are among those who have low scores in terms of annual performance through annual performance value reports and have gone through various stages before being required to attend Psynnova Program. Besides that, the research plan was also to critically examine and understand the change in behaviour among the low-performing employees through the five dimensions in the Psynnova Module. A total of 50 respondent will purposively sampled to be the respondents of this research. This study will use the Experimental Method to One Group Purposively Pre and Post Test using the Time Series Design. Experimental SPSS software version 22.0 will be used to analyse this data. Hopefully this research can see the changing of their behaviour in five factors as an indicator to the respondent after attending the Psynnova Programme. Findings from this study are also used to propose to assisting psychologist to see the changes that occurred to the respondents with the best framework of behaviour changing for them.

Keywords: five dimension of behaviour changing, among adult, low performance, modul effectiveness

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2075 Introducing a Video-Based E-Learning Module to Improve Disaster Preparedness at a Tertiary Hospital in Oman

Authors: Ahmed Al Khamisi

Abstract:

The Disaster Preparedness Standard (DPS) is one of the elements that is evaluated by the Accreditation Canada International (ACI). ACI emphasizes to train and educate all staff, including service providers and senior leaders, on emergency and disaster preparedness upon the orientation and annually thereafter. Lack of awareness and deficit of knowledge among the healthcare providers about DPS have been noticed in a tertiary hospital where ACI standards were implemented. Therefore, this paper aims to introduce a video-based e-learning (VB-EL) module that explains the hospital’s disaster plan in a simple language which will be easily accessible to all healthcare providers through the hospital’s website. The healthcare disaster preparedness coordinator in the targeted hospital will be responsible to ensure that VB-EL is ready by 25 April 2019. This module will be developed based on the Kirkpatrick evaluation method. In fact, VB-EL combines different data forms such as images, motion, sounds, text in a complementary fashion which will suit diverse learning styles and individual learning pace of healthcare providers. Moreover, the module can be adjusted easily than other tools to control the information that healthcare providers receive. It will enable healthcare providers to stop, rewind, fast-forward, and replay content as many times as needed. Some anticipated limitations in the development of this module include challenges of preparing VB-EL content and resistance from healthcare providers.

Keywords: Accreditation Canada International, Disaster Preparedness Standard, Kirkpatrick evaluation method, video-based e-learning

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2074 Analysis of Expression Data Using Unsupervised Techniques

Authors: M. A. I Perera, C. R. Wijesinghe, A. R. Weerasinghe

Abstract:

his study was conducted to review and identify the unsupervised techniques that can be employed to analyze gene expression data in order to identify better subtypes of tumors. Identifying subtypes of cancer help in improving the efficacy and reducing the toxicity of the treatments by identifying clues to find target therapeutics. Process of gene expression data analysis described under three steps as preprocessing, clustering, and cluster validation. Feature selection is important since the genomic data are high dimensional with a large number of features compared to samples. Hierarchical clustering and K Means are often used in the analysis of gene expression data. There are several cluster validation techniques used in validating the clusters. Heatmaps are an effective external validation method that allows comparing the identified classes with clinical variables and visual analysis of the classes.

Keywords: cancer subtypes, gene expression data analysis, clustering, cluster validation

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2073 An Analysis on Clustering Based Gene Selection and Classification for Gene Expression Data

Authors: K. Sathishkumar, V. Thiagarasu

Abstract:

Due to recent advances in DNA microarray technology, it is now feasible to obtain gene expression profiles of tissue samples at relatively low costs. Many scientists around the world use the advantage of this gene profiling to characterize complex biological circumstances and diseases. Microarray techniques that are used in genome-wide gene expression and genome mutation analysis help scientists and physicians in understanding of the pathophysiological mechanisms, in diagnoses and prognoses, and choosing treatment plans. DNA microarray technology has now made it possible to simultaneously monitor the expression levels of thousands of genes during important biological processes and across collections of related samples. Elucidating the patterns hidden in gene expression data offers a tremendous opportunity for an enhanced understanding of functional genomics. However, the large number of genes and the complexity of biological networks greatly increase the challenges of comprehending and interpreting the resulting mass of data, which often consists of millions of measurements. A first step toward addressing this challenge is the use of clustering techniques, which is essential in the data mining process to reveal natural structures and identify interesting patterns in the underlying data. This work presents an analysis of several clustering algorithms proposed to deals with the gene expression data effectively. The existing clustering algorithms like Support Vector Machine (SVM), K-means algorithm and evolutionary algorithm etc. are analyzed thoroughly to identify the advantages and limitations. The performance evaluation of the existing algorithms is carried out to determine the best approach. In order to improve the classification performance of the best approach in terms of Accuracy, Convergence Behavior and processing time, a hybrid clustering based optimization approach has been proposed.

Keywords: microarray technology, gene expression data, clustering, gene Selection

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2072 Application of Genetic Algorithm with Multiobjective Function to Improve the Efficiency of Photovoltaic Thermal System

Authors: Sonveer Singh, Sanjay Agrawal, D. V. Avasthi, Jayant Shekhar

Abstract:

The aim of this paper is to improve the efficiency of photovoltaic thermal (PVT) system with the help of Genetic Algorithms with multi-objective function. There are some parameters that affect the efficiency of PVT system like depth and length of the channel, velocity of flowing fluid through the channel, thickness of the tedlar and glass, temperature of inlet fluid i.e. all above parameters are considered for optimization. An attempt has been made to the model and optimizes the parameters of glazed hybrid single channel PVT module when two objective functions have been considered separately. The two objective function for optimization of PVT module is overall electrical and thermal efficiency. All equations for PVT module have been derived. Using genetic algorithms (GAs), above two objective functions of the system has been optimized separately and analysis has been carried out for two cases. Two cases are: Case-I; Improvement in electrical and thermal efficiency when overall electrical efficiency is optimized, Case-II; Improvement in electrical and thermal efficiency when overall thermal efficiency is optimized. All the parameters that are used in genetic algorithms are the parameters that could be changed, and the non-changeable parameters, like solar radiation, ambient temperature cannot be used in the algorithm. It has been observed that electrical efficiency (14.08%) and thermal efficiency (19.48%) are obtained when overall thermal efficiency was an objective function for optimization. It is observed that GA is a very efficient technique to estimate the design parameters of hybrid single channel PVT module.

Keywords: genetic algorithm, energy, exergy, PVT module, optimization

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2071 Exploring Academic Writing Challenges of First Year English as an Additional Language Students at an ODeL Institution in South Africa

Authors: Tumelo Jaquiline Ntsopi

Abstract:

This study explored the academic writing challenges of first-year students who use English as an Additional Language (EAL) registered in the EAW101 module at an ODeL institution. Research shows that academic writing is a challenge for EAL teaching and learning contexts across the globe in higher education institutions (HEIs). Academic writing is an important aspect of academic literacy in any institution of higher learning, more so in an ODeL institution. This has probed research that shows that academic writing is and continues to pose challenges for EAL teaching and learning contexts in higher education institutions. This study stems from the researcher’s experience in teaching academic writing to first-year students in the EAW101 module. The motivation for this study emerged from the fact that EAW101 is a writing module that has a high number of students in the Department of English Studies with an average of between 50-80 percent pass rate. These statistics elaborate on the argument that most students registered in this module struggle with academic writing, and they need intervention to assist and support them in achieving competence in the module. This study is underpinned by Community of Inquiry (CoI) framework and Transactional distance theory. This study adopted a qualitative research methodology and utilised a case study approach as a research design. Furthermore, the study gathered data from first year students and the EAW101 module’s student support initiatives. To collect data, focus group discussions, structured open-ended evaluation questions, and an observation schedule were used to gather data. The study is vital towards exploring academic writing challenges that first-year students in EAW101 encounter so that lecturers in the module may consider re-evaluating their methods of teaching to improve EAL students’ academic writing skills. This study may help lecturers towards enhancing academic writing in a ODeL context by assisting first year students through using student support interventions.

Keywords: academic writing, academic writing challenge, ODeL, EAL

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2070 The Development Learning Module Physics based on Guided Inquiry Approach on Model Cooperative Learning Type STAD (Student Team Achievement Division) in the Main Subject of Temperature and Heat

Authors: Fani Firmahandari

Abstract:

The development learning module physics based on guided inquiry approach on model cooperative learning type STAD (Student Team Achievement Division) in the main subject of temperature and heat. The research development aimed to produce physics learning module based on guided cooperative learning type STAD (Student Team Achievement Division) in the main subject of temperature and heat to the student in X class. The research method used Research and Development approach. The development procedure of this module includes potential problems, data collection to meet the need, product design, and feasibility of this module. The impact of learning can be seen or observed clearly when the learning process takes place, the teachers or the students already implemented measures cooperative learning model type STAD, so that the learning process goes well, the interaction of teachers and students, students with student looks good, besides that students can interact and work together in group.

Keywords: cooperative learning type STAD (student team achievement division), development, inquiry, interaction students

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2069 The Role of MAOA Gene in the Etiology of Autism Spectrum Disorder in Males

Authors: Jana Kisková, Dana Gabriková

Abstract:

Monoamine oxidase A gene (MAOA) is suggested to be a candidate gene implicated in many neuropsychiatric disorders, including autism spectrum disorder (ASD). This meta-analytic review evaluates the relationship between ASD and MAOA markers such as 30 bp variable number tandem repeats in the promoter region (uVNTR) and single nucleotide polymorphisms (SNPs) by using findings from recently published studies. It seems that in Caucasian males, the risk of developing ASD increase with the presence of 4-repeat allele in the promoter region of MAOA gene whereas no differences were found between autistic patients and controls in Egyptian, West Bengal and Korean population. Some studies point to the importance specific haplotype groups of SNPs and interaction of MAOA with others genes (e.g. FOXP2 or SRY). The results of existing studies are insufficient and further research is needed.

Keywords: autism spectrum disorder, MAOA, uVNTR, single nucleotide polymorphism

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2068 THRAP2 Gene Identified as a Candidate Susceptibility Gene of Thyroid Autoimmune Diseases Pedigree in Tunisian Population

Authors: Ghazi Chabchoub, Mouna Feki, Mohamed Abid, Hammadi Ayadi

Abstract:

Autoimmune thyroid diseases (AITDs), including Graves’ disease (GD) and Hashimoto’s thyroiditis (HT), are inherited as complex traits. Genetic factors associated with AITDs have been tentatively identified by candidate gene and genome scanning approaches. We analysed three intragenic microsatellite markers in the thyroid hormone receptor associated protein 2 gene (THRAP2), mapped near D12S79 marker, which have a potential role in immune function and inflammation [THRAP2-1(TG)n, THRAP2-2 (AC)n and THRAP2-3 (AC)n]. Our study population concerned 12 patients affected with AITDs belonging to a multiplex Tunisian family with high prevalence of AITDs. Fluorescent genotyping was carried out on ABI 3100 sequencers (Applied Biosystems USA) with the use of GENESCAN for semi-automated fragment sizing and GENOTYPER peak-calling software. Statistical analysis was performed using the non parametric Lod score (NPL) by Merlin software. Merlin outputs non-parametric NPLall (Z) and LOD scores and their corresponding asymptotic P values. The analysis for three intragenic markers in the THRAP2 gene revealed strong evidence for linkage (NPL=3.68, P=0.00012). Our results suggested the possible role of THRAP2 gene in AITDs susceptibility in this family.

Keywords: autoimmunity, autoimmune disease, genetic, linkage analysis

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