Search results for: genome analysis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 27309

Search results for: genome analysis

26979 Influence of Food Microbes on Horizontal Transfer of β-Lactam Resistance Genes between Salmonella Strains in the Mouse Gut

Authors: M. Ottenbrite, G. Yilmaz, J. Devenish, M. Kang, H. Dan, M. Lin, C. Lau, C. Carrillo, K. Bessonov, J. Nash, E. Topp, J. Guan

Abstract:

Consumption of food contaminated by antibiotic-resistant (AR) bacteria may lead to the transmission of AR genes in the gut microbiota and cause AR bacterial infection, a significant public health concern. However, information is limited on if and how background microbes from the food matrix (food microbes) may influence resistance transmission. Thus, we assessed the colonization of a β-lactam resistant Salmonella Heidelberg strain (donor) and a β-lactam susceptible S. Typhimurium strain (recipient) and the transfer of the resistance genes in the mouse gut in the presence or absence of food microbes that were derived from washing freshly-harvested carrots. Mice were pre-treated with streptomycin and then inoculated with both donor and recipient bacteria or recipient only. Fecal shedding of the donor, recipient, and transconjugant bacteria was enumerated using selective culture techniques. Transfer of AR genes was confirmed by whole genome sequencing. Gut microbial composition was determined by 16s rRNA amplicon sequencing. Significantly lower numbers of donors and recipients were shed from mice that were inoculated with food microbes compared to those without food microbe inoculation. S. Typhimurium transconjugants were only recovered from mice without inoculation of food microbes. A significantly higher survival rate was in mice with vs. without inoculation of food microbes. The results suggest that the food microbes may compete with both the donor and recipient Salmonella, limit their growth and reduce transmission of the β-lactam resistance gene in the mouse gut.

Keywords: antibiotic resistance, gene transfer, gut microbiota, Salmonella infection

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26978 Numerical Approach of RC Structural MembersExposed to Fire and After-Cooling Analysis

Authors: Ju-young Hwang, Hyo-Gyoung Kwak, Hong Jae Yim

Abstract:

This paper introduces a numerical analysis method for reinforced-concrete (RC) structures exposed to fire and compares the result with experimental results. The proposed analysis method for RC structure under the high temperature consists of two procedures. First step is to decide the temperature distribution across the section through the heat transfer analysis by using the time-temperature curve. After determination of the temperature distribution, the nonlinear analysis is followed. By considering material and geometrical non-linearity with the temperature distribution, nonlinear analysis predicts the behavior of RC structure under the fire by the exposed time. The proposed method is validated by the comparison with the experimental results. Finally, Prediction model to describe the status of after-cooling concrete can also be introduced based on the results of additional experiment. The product of this study is expected to be embedded for smart structure monitoring system against fire in u-City.

Keywords: RC structures, heat transfer analysis, nonlinear analysis, after-cooling concrete model

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26977 Financial Analysis of Selected Private Healthcare Organizations with Special Referance to Guwahati City, Assam

Authors: Mrigakshi Das

Abstract:

The private sector investments and quantum of money required in this sector critically hinges on the financial risk and returns the sector offers to providers of capital. Therefore, it becomes important to understand financial performance of hospitals. Financial Analysis is useful for decision makers in a variety of settings. Consider the small proprietary hospitals, say, Physicians Clinic. The managers of such clinic need the information that financial statements provide. Attention to Financial Statements of healthcare Organizations can provide answers to questions like: How are they doing? What is their rate of profit? What is their solvency and liquidity position? What are their sources and application of funds? What is their Operational Efficiency? The researcher has studied Financial Statements of 5 Private Healthcare Organizations in Guwahati City.

Keywords: not-for-profit organizations, financial analysis, ratio analysis, profitability analysis, liquidity analysis, operational efficiency, capital structure analysis

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26976 Force Measurement for E-Cadherin-Mediated Intercellular Adhesion Probed by Protein Micropattern and Traction Force Microscopy

Authors: Chieh-Chung Tsou, Chun-Min Lo, Yeh-Shiu Chu

Abstract:

Cell’s mechanical forces provide important physical cues in regulation of proper cellular functions, such as cell differentiation, proliferation and migration. It is believed that adhesive forces generated by cell-cell interaction are able to transmit to the interior of cell through filamentous cortical cytoskeleton. Prominent among other membrane receptors, Cadherins are prototypical adhesive molecules able to generate remarkable forces to regulate intercellular adhesion. However, the mechanistic steps of mechano-transduction in Cadherin-mediated adhesion remain very controversial. We are interested in understanding how Cadherin protein complexes enable force generation and transmission at cell-cell contact in the initial stage of intercellular adhesion. For providing a better control of time, space, and substrate stiffness, in this study, a combination of protein micropattern, micropipette manipulation, and traction force microscopy is used. Pair micropattern with different forms confines cell spreading area and the gaps in pairs varied from 2 to 8 microns are applied for monitoring the forces that cell pairs generated, measured by traction force microscopy. Moreover, cell clones obtained from epithelial cells undergone genome editing are used to score the importance for known components of Cadherin complexes in force generation. We believe that our results from this combinatory mechanobiological method will provide deep insights on understanding the biophysical principle governing mechano- transduction of Cadherin-mediated intercellular adhesion.

Keywords: cadherin, intercellular adhesion, protein micropattern, traction force microscopy

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26975 A Cross-Dialect Statistical Analysis of Final Declarative Intonation in Tuvinian

Authors: D. Beziakina, E. Bulgakova

Abstract:

This study continues the research on Tuvinian intonation and presents a general cross-dialect analysis of intonation of Tuvinian declarative utterances, specifically the character of the tone movement in order to test the hypothesis about the prevalence of level tone in some Tuvinian dialects. The results of the analysis of basic pitch characteristics of Tuvinian speech (in general and in comparison with two other Turkic languages - Uzbek and Azerbaijani) are also given in this paper. The goal of our work was to obtain the ranges of pitch parameter values typical for Tuvinian speech. Such language-specific values can be used in speaker identification systems in order to get more accurate results of ethnic speech analysis. We also present the results of a cross-dialect analysis of declarative intonation in the poorly studied Tuvinian language.

Keywords: speech analysis, statistical analysis, speaker recognition, identification of person

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26974 A Survey of Sentiment Analysis Based on Deep Learning

Authors: Pingping Lin, Xudong Luo, Yifan Fan

Abstract:

Sentiment analysis is a very active research topic. Every day, Facebook, Twitter, Weibo, and other social media, as well as significant e-commerce websites, generate a massive amount of comments, which can be used to analyse peoples opinions or emotions. The existing methods for sentiment analysis are based mainly on sentiment dictionaries, machine learning, and deep learning. The first two kinds of methods rely on heavily sentiment dictionaries or large amounts of labelled data. The third one overcomes these two problems. So, in this paper, we focus on the third one. Specifically, we survey various sentiment analysis methods based on convolutional neural network, recurrent neural network, long short-term memory, deep neural network, deep belief network, and memory network. We compare their futures, advantages, and disadvantages. Also, we point out the main problems of these methods, which may be worthy of careful studies in the future. Finally, we also examine the application of deep learning in multimodal sentiment analysis and aspect-level sentiment analysis.

Keywords: document analysis, deep learning, multimodal sentiment analysis, natural language processing

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26973 Containment/Penetration Analysis for the Protection of Aircraft Engine External Configuration and Nuclear Power Plant Structures

Authors: Dong Wook Lee, Adrian Mistreanu

Abstract:

The authors have studied a method for analyzing containment and penetration using an explicit nonlinear Finite Element Analysis. This method may be used in the stage of concept design for the protection of external configurations or components of aircraft engines and nuclear power plant structures. This paper consists of the modeling method, the results obtained from the method and the comparison of the results with those calculated from simple analytical method. It shows that the containment capability obtained by proposed method matches well with analytically calculated containment capability.

Keywords: computer aided engineering, containment analysis, finite element analysis, impact analysis, penetration analysis

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26972 Association of Copy Number Variation of the CHKB, KLF6, GPC1, and CHRM3 Genes with Growth Traits of Datong Yak (Bos grunniens)

Authors: Habtamu Abera Goshu, Ping Yan

Abstract:

Copy number variation (CNV) is a significant marker of the genetic and phenotypic diversity among individuals that accounts for complex quantitative traits of phenotype and diseases via modulating gene dosage, position effects, alteration of downstream pathways, modification of chromosome structure, and position within the nucleus and disrupting coding regions in the genome. Associating copy number variations (CNVs) with growth and gene expression are a powerful approach for identifying genomic characteristics that contribute to phenotypic and genotypic variation. A previous study using next-generation sequencing illustrated that the choline kinase beta (CHKB), Krüpple-like factor 6 (KLF6), glypican 1(GPC1), and cholinergic receptor muscarinic 3 (CHRM3) genes reside within copy number variable regions (CNVRs) of yak populations that overlap with quantitative trait loci (QTLs) of meat quality and growth. As a result, this research aimed to determine the association of CNVs of the KLF6, CHKB, GPC1, and CHRM3 genes with growth traits in the Datong yak breed. The association between the CNV types of the KLF6, CHKB, GPC1, and CHRM3 genes and the growth traits in the Datong yak breed was determined by one-way analysis of variance (ANOVA) using SPSS software. The CNV types were classified as a loss (a copy number of 0 or 1), gain (a copy number >2), and normal (a copy number of 2) relative to the reference gene, BTF3 in the 387 individuals of Datong yak. These results indicated that the normal CNV types of the CHKB and GPC1 genes were significantly (P<0.05) associated with high body length, height and weight, and chest girth in six-month-old and five-year-old Datong yaks. On the other hand, the loss CNV types of the KLF6 gene is significantly (P<0.05) associated with body weight and length and chest girth at six-month-old and five-year-old Datong yaks. In the contrary, the gain CNV type of the CHRM3 gene is highly (P<0.05) associated with body weight, length, height, and chest girth in six-month-old and five-year-old. This work provides the first observation of the biological role of CNVs of the CHKB, KLF6, GPC1, and CHRM3 genes in the Datong yak breed and might, therefore, provide a novel opportunity to utilize data on CNVs in designing molecular markers for the selection of animal breeding programs for larger populations of various yak breeds. Therefore, we hypothesized that this study provided inclusive information on the application of CNVs of the CHKB, KLF6, GPC1, and CHRM3 genes in growth traits in Datong yaks and its possible function in bovine species.

Keywords: Copy number variation, growth traits, yak, genes

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26971 Collision Theory Based Sentiment Detection Using Discourse Analysis in Hadoop

Authors: Anuta Mukherjee, Saswati Mukherjee

Abstract:

Data is growing everyday. Social networking sites such as Twitter are becoming an integral part of our daily lives, contributing a large increase in the growth of data. It is a rich source especially for sentiment detection or mining since people often express honest opinion through tweets. However, although sentiment analysis is a well-researched topic in text, this analysis using Twitter data poses additional challenges since these are unstructured data with abbreviations and without a strict grammatical correctness. We have employed collision theory to achieve sentiment analysis in Twitter data. We have also incorporated discourse analysis in the collision theory based model to detect accurate sentiment from tweets. We have also used the retweet field to assign weights to certain tweets and obtained the overall weightage of a topic provided in the form of a query. Hadoop has been exploited for speed. Our experiments show effective results.

Keywords: sentiment analysis, twitter, collision theory, discourse analysis

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26970 Using SNAP and RADTRAD to Establish the Analysis Model for Maanshan PWR Plant

Authors: J. R. Wang, H. C. Chen, C. Shih, S. W. Chen, J. H. Yang, Y. Chiang

Abstract:

In this study, we focus on the establishment of the analysis model for Maanshan PWR nuclear power plant (NPP) by using RADTRAD and SNAP codes with the FSAR, manuals, and other data. In order to evaluate the cumulative dose at the Exclusion Area Boundary (EAB) and Low Population Zone (LPZ) outer boundary, Maanshan NPP RADTRAD/SNAP model was used to perform the analysis of the DBA LOCA case. The analysis results of RADTRAD were similar to FSAR data. These analysis results were lower than the failure criteria of 10 CFR 100.11 (a total radiation dose to the whole body, 250 mSv; a total radiation dose to the thyroid from iodine exposure, 3000 mSv).

Keywords: RADionuclide, transport, removal, and dose estimation (RADTRAD), symbolic nuclear analysis package (SNAP), dose, PWR

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26969 A Data Envelopment Analysis Model in a Multi-Objective Optimization with Fuzzy Environment

Authors: Michael Gidey Gebru

Abstract:

Most of Data Envelopment Analysis models operate in a static environment with input and output parameters that are chosen by deterministic data. However, due to ambiguity brought on shifting market conditions, input and output data are not always precisely gathered in real-world scenarios. Fuzzy numbers can be used to address this kind of ambiguity in input and output data. Therefore, this work aims to expand crisp Data Envelopment Analysis into Data Envelopment Analysis with fuzzy environment. In this study, the input and output data are regarded as fuzzy triangular numbers. Then, the Data Envelopment Analysis model with fuzzy environment is solved using a multi-objective method to gauge the Decision Making Units' efficiency. Finally, the developed Data Envelopment Analysis model is illustrated with an application on real data 50 educational institutions.

Keywords: efficiency, Data Envelopment Analysis, fuzzy, higher education, input, output

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26968 Constructivism Learning Management in Mathematics Analysis Courses

Authors: Komon Paisal

Abstract:

The purposes of this research were (1) to create a learning activity for constructivism, (2) study the Mathematical Analysis courses learning achievement, and (3) study students’ attitude toward the learning activity for constructivism. The samples in this study were divided into 2 parts including 3 Mathematical Analysis courses instructors of Suan Sunandha Rajabhat University who provided basic information and attended the seminar and 17 Mathematical Analysis courses students who were studying in the academic and engaging in the learning activity for constructivism. The research instruments were lesson plans constructivism, subjective Mathematical Analysis courses achievement test with reliability index of 0.8119, and an attitude test concerning the students’ attitude toward the Mathematical Analysis courses learning activity for constructivism. The result of the research show that the efficiency of the Mathematical Analysis courses learning activity for constructivism is 73.05/72.16, which is more than expected criteria of 70/70. The research additionally find that the average score of learning achievement of students who engaged in the learning activities for constructivism are equal to 70% and the students’ attitude toward the learning activity for constructivism are at the medium level.

Keywords: constructivism, learning management, mathematics analysis courses, learning activity

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26967 Systematic Review of Functional Analysis in Brazil

Authors: Felipe Magalhaes Lemos

Abstract:

Functional behavior analysis is a procedure that has been studied for several decades by behavior analysts. In Brazil, we still have few studies in the area, so it was decided to carry out a systematic review of the articles published in the area by Brazilians. A search was done on the following scientific article registration sites: PsycINFO, ERIC, ISI Web of Science, Virtual Health Library. The research includes (a) peer-reviewed studies that (b) have been carried out in Brazil containing (c) functional assessment as a pre-treatment through (d) experimental procedures, direct or indirect observation and measurement of behavior problems (e) demonstrating a relationship between environmental events and behavior. During the review, 234 papers were found; however, only 9 were included in the final analysis. Of the 9 articles extracted, only 2 presented functional analysis procedures with manipulation of environmental variables, while the other 7 presented different procedures for a descriptive behavior assessment. Only the two studies using "functional analysis" used graphs to demonstrate the prevalent function of the behavior. Other studies described procedures and did not make clear the causal relationship between environment and behavior. There is still confusion in Brazil regarding the terms "functional analysis", "descriptive assessment" and "contingency analysis," which are generally treated in the same way. This study shows that few articles are published with a focus on functional analysis in Brazil.

Keywords: behavior, contingency, descriptive assessment, functional analysis

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26966 Experiences of Timing Analysis of Parallel Embedded Software

Authors: Muhammad Waqar Aziz, Syed Abdul Baqi Shah

Abstract:

The execution time analysis is fundamental to the successful design and execution of real-time embedded software. In such analysis, the Worst-Case Execution Time (WCET) of a program is a key measure, on the basis of which system tasks are scheduled. The WCET analysis of embedded software is also needed for system understanding and to guarantee its behavior. WCET analysis can be performed statically (without executing the program) or dynamically (through measurement). Traditionally, research on the WCET analysis assumes sequential code running on single-core platforms. However, as computation is steadily moving towards using a combination of parallel programs and multi-core hardware, new challenges in WCET analysis need to be addressed. In this article, we report our experiences of performing the WCET analysis of Parallel Embedded Software (PES) running on multi-core platform. The primary purpose was to investigate how WCET estimates of PES can be computed statically, and how they can be derived dynamically. Our experiences, as reported in this article, include the challenges we faced, possible suggestions to these challenges and the workarounds that were developed. This article also provides observations on the benefits and drawbacks of deriving the WCET estimates using the said methods and provides useful recommendations for further research in this area.

Keywords: embedded software, worst-case execution-time analysis, static flow analysis, measurement-based analysis, parallel computing

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26965 CCR5 as an Ideal Candidate for Immune Gene Therapy and Modification for the Induced Resistance to HIV-1 Infection

Authors: Alieh Farshbaf, Tayyeb Bahrami

Abstract:

Introduction: Cc-chemokine receptor-5 (CCR5) is known as a main co-receptor in human immunodeficiency virus type-1 (HIV-1) infection. Many studies showed 32bp deletion (Δ32) in CCR5 gene, provide natural resistance to HIV-1 infection in homozygous individuals. Inducing the resistance mechanism by CCR5 in HIV-1 infected patients eliminated many problems of highly-active-anti retroviral therapy (HAART) drugs like as low safety, side-effects and virus rebounding from latent reservoirs. New treatments solved some restrictions that are based on gene modification and cell therapy. Literature review: The stories of the “Berlin and Boston patients” showed autologous hematopoietic stem cells transplantation (HSCT) could provide effective cure of HIV-1 infected patients. Furthermore, gene modification by zinc finger nuclease (ZFN) demonstrated another successful result again. Despite the other studies for gene therapy by ∆32 genotype, there is another mutation -CCR5 ∆32/m303- that provides HIV-1 resistant. It is a heterozygote genotype for ∆32 and T→A point mutation at nucleotide 303. These results approved the key role of CCR5 gene. Conclusion: Recent studies showed immune gene therapy and cell therapy could provide effective cure for refractory disease like as HIV. Eradication of HIV-1 from immune system was not observed by HAART, because of reloading virus genome from latent reservoirs after stopping them. It is showed that CCR5 could induce natural resistant to HIV-1 infection by the new approaches based on stem cell transplantation and gene modifying.

Keywords: CCR5, HIV-1, stem cell, immune gene therapy, gene modification

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26964 Differential Expression Profile Analysis of DNA Repair Genes in Mycobacterium Leprae by qPCR

Authors: Mukul Sharma, Madhusmita Das, Sundeep Chaitanya Vedithi

Abstract:

Leprosy is a chronic human disease caused by Mycobacterium leprae, that cannot be cultured in vitro. Though treatable with multidrug therapy (MDT), recently, bacteria reported resistance to multiple antibiotics. Targeting DNA replication and repair pathways can serve as the foundation of developing new anti-leprosy drugs. Due to the absence of an axenic culture medium for the propagation of M. leprae, studying cellular processes, especially those belonging to DNA repair pathways, is challenging. Genomic understanding of M. Leprae harbors several protein-coding genes with no previously assigned function known as 'hypothetical proteins'. Here, we report identification and expression of known and hypothetical DNA repair genes from a human skin biopsy and mouse footpads that are involved in base excision repair, direct reversal repair, and SOS response. Initially, a bioinformatics approach was employed based on sequence similarity, identification of known protein domains to screen the hypothetical proteins in the genome of M. leprae, that are potentially related to DNA repair mechanisms. Before testing on clinical samples, pure stocks of bacterial reference DNA of M. leprae (NHDP63 strain) was used to construct standard graphs to validate and identify lower detection limit in the qPCR experiments. Primers were designed to amplify the respective transcripts, and PCR products of the predicted size were obtained. Later, excisional skin biopsies of newly diagnosed untreated, treated, and drug resistance leprosy cases from SIHR & LC hospital, Vellore, India were taken for the extraction of RNA. To determine the presence of the predicted transcripts, cDNA was generated from M. leprae mRNA isolated from clinically confirmed leprosy skin biopsy specimen across all the study groups. Melting curve analysis was performed to determine the integrity of the amplification and to rule out primer‑dimer formation. The Ct values obtained from qPCR were fitted to standard curve to determine transcript copy number. Same procedure was applied for M. leprae extracted after processing a footpad of nude mice of drug sensitive and drug resistant strains. 16S rRNA was used as positive control. Of all the 16 genes involved in BER, DR, and SOS, differential expression pattern of the genes was observed in terms of Ct values when compared to human samples; this was because of the different host and its immune response. However, no drastic variation in gene expression levels was observed in human samples except the nth gene. The higher expression of nth gene could be because of the mutations that may be associated with sequence diversity and drug resistance which suggests an important role in the repair mechanism and remains to be explored. In both human and mouse samples, SOS system – lexA and RecA, and BER genes AlkB and Ogt were expressing efficiently to deal with possible DNA damage. Together, the results of the present study suggest that DNA repair genes are constitutively expressed and may provide a reference for molecular diagnosis, therapeutic target selection, determination of treatment and prognostic judgment in M. leprae pathogenesis.

Keywords: DNA repair, human biopsy, hypothetical proteins, mouse footpads, Mycobacterium leprae, qPCR

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26963 Predictive Analysis for Big Data: Extension of Classification and Regression Trees Algorithm

Authors: Ameur Abdelkader, Abed Bouarfa Hafida

Abstract:

Since its inception, predictive analysis has revolutionized the IT industry through its robustness and decision-making facilities. It involves the application of a set of data processing techniques and algorithms in order to create predictive models. Its principle is based on finding relationships between explanatory variables and the predicted variables. Past occurrences are exploited to predict and to derive the unknown outcome. With the advent of big data, many studies have suggested the use of predictive analytics in order to process and analyze big data. Nevertheless, they have been curbed by the limits of classical methods of predictive analysis in case of a large amount of data. In fact, because of their volumes, their nature (semi or unstructured) and their variety, it is impossible to analyze efficiently big data via classical methods of predictive analysis. The authors attribute this weakness to the fact that predictive analysis algorithms do not allow the parallelization and distribution of calculation. In this paper, we propose to extend the predictive analysis algorithm, Classification And Regression Trees (CART), in order to adapt it for big data analysis. The major changes of this algorithm are presented and then a version of the extended algorithm is defined in order to make it applicable for a huge quantity of data.

Keywords: predictive analysis, big data, predictive analysis algorithms, CART algorithm

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26962 Genome-Wide Expression Profiling of Cicer arietinum Heavy Metal Toxicity

Authors: B. S. Yadav, A. Mani, S. Srivastava

Abstract:

Chickpea (Cicer arietinum L.) is an annual, self-pollinating, diploid (2n = 2x = 16) pulse crop that ranks second in world legume production after common bean (Phaseolus vulgaris). ICC 4958 flowers approximately 39 days after sowing under peninsular Indian conditions and the crop matures in less than 90 days in rained environments. The estimated collective yield losses due to abiotic stresses (6.4 million t) have been significantly higher than for biotic stresses (4.8 million t). Most legumes are known to be salt sensitive, and therefore, it is becoming increasingly important to produce cultivars tolerant to high-salinity in addition to other abiotic and biotic stresses for sustainable chickpea production. Our aim was to identify the genes that are involved in the defence mechanism against heavy metal toxicity in chickpea and establish the biological network of heavy metal toxicity in chickpea. ICC4958 variety of chick pea was taken and grown in normal condition and 150µM concentration of different heavy metal salt like CdCl₂, K₂Cr2O₇, NaAsO₂. At 15th day leave samples were collected and stored in RNA Later solution microarray was performed for checking out differential gene expression pattern. Our studies revealed that 111 common genes that involved in defense mechanism were up regulated and 41 genes were commonly down regulated during treatment of 150µM concentration of CdCl₂, K₂Cr₂O₇, and NaAsO₂. Biological network study shows that the genes which are differentially expressed are highly connected and having high betweenness and centrality.

Keywords: abiotic stress, biological network, chickpea, microarray

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26961 PPRA Regulates DNA Replication Initiation and Cell Morphology in Escherichia coli

Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

Abstract:

PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provides better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

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26960 Heterologous Expression of a Clostridium thermocellum Proteins and Assembly of Cellulosomes 'in vitro' for Biotechnology Applications

Authors: Jessica Pinheiro Silva, Brenda Rabello De Camargo, Daniel Gusmao De Morais, Eliane Ferreira Noronha

Abstract:

The utilization of lignocellulosic biomass as source of polysaccharides for industrial applications requires an arsenal of enzymes with different mode of action able to hydrolyze its complex and recalcitrant structure. Clostridium thermocellum is gram-positive, thermophilic bacterium producing lignocellulosic hydrolyzing enzymes in the form of multi-enzyme complex, termed celulossomes. This complex has several hydrolytic enzymes attached to a large and enzymically inactive protein known as Cellulosome-integrating protein (CipA), which serves as a scaffolding protein for the complex produced. This attachment occurs through specific interactions between cohesin modules of CipA and dockerin modules in enzymes. The present work aims to construct celulosomes in vitro with the structural protein CipA, a xylanase called Xyn10D and a cellulose called CelJ from C.thermocellum. A mini-scafoldin was constructed from modules derived from CipA containing two cohesion modules. This was cloned and expressed in Escherichia coli. The other two genes were cloned under the control of the alcohol oxidase 1 promoter (AOX1) in the vector pPIC9 and integrated into the genome of the methylotrophic yeast Pichia pastoris GS115. Purification of each protein is being carried out. Further studies regarding enzymatic activity of the cellulosome is going to be evaluated. The cellulosome built in vitro and composed of mini-CipA, CelJ and Xyn10D, can be very interesting for application in industrial processes involving the degradation of plant biomass.

Keywords: cellulosome, CipA, Clostridium thermocellum, cohesin, dockerin, yeast

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26959 PPRA Controls DNA Replication and Cell Growth in Escherichia Coli

Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

Abstract:

PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provide better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

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26958 Disturbed Cellular Iron Metabolism Genes in Neurodevelopmental Disorders is Different from Neurodegenerative Disorders

Authors: O. H. Gebril, N. A. Meguid

Abstract:

Background: Iron had been a focus of interest recently as a main exaggerating factor for oxidative stresses in the central nervous system and a link to various neurological disorders is suspected. Many studies with various techniques showed evidence of disturbed iron-related proteins in the cell in human and animal models of neurodegenerative disorders. Also, linkage to significant pathological changes had been evidenced e.g. apoptosis and cell signaling. On the other hand, the role of iron in neurodevelopmental disorders is still unclear. With increasing prevalence of autism worldwide, some changes in iron parameters and its stores were documented in many studies. This study includes Haemochromatosis HFE gene polymorphisms (p.H63D and p.C282Y) and ferroportin gene (SLC40A1) Q248H polymorphism in autism and control children. Materials and Methods: Whole genome DNA was extracted; p.H63D and p.C282Y genotyping was studied using specific sequence amplification followed by restriction enzyme digestion on a sample of autism patients (25 cases) and twenty controls. Results: The p.H63D is seen more than the C282Y among both autism and control samples, with no significant association of p.H63D or p.C282Y polymorphism and autism was revealed. Also, no association with Q248H polymorphism was evidenced. Conclusion: The study results do not prove the role of cellular iron genes polymorphisms as risk factors for neurodevelopmental disorders, and in turn highlights the specificity of cellular iron related pathways in neurodegeneration. These results demand further gene expression studies to elucidate the main pathophysiological pathways that are disturbed in autism and other neurodevelopmental disorders.

Keywords: iron, neurodevelopmental, oxidative stress, haemohromatosis, ferroportin, genes

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26957 Enzymatic Repair Prior To DNA Barcoding, Aspirations, and Restraints

Authors: Maxime Merheb, Rachel Matar

Abstract:

Retrieving ancient DNA sequences which in return permit the entire genome sequencing from fossils have extraordinarily improved in recent years, thanks to sequencing technology and other methodological advances. In any case, the quest to search for ancient DNA is still obstructed by the damage inflicted on DNA which accumulates after the death of a living organism. We can characterize this damage into three main categories: (i) Physical abnormalities such as strand breaks which lead to the presence of short DNA fragments. (ii) Modified bases (mainly cytosine deamination) which cause errors in the sequence due to an incorporation of a false nucleotide during DNA amplification. (iii) DNA modifications referred to as blocking lesions, will halt the PCR extension which in return will also affect the amplification and sequencing process. We can clearly see that the issues arising from breakage and coding errors were significantly decreased in recent years. Fast sequencing of short DNA fragments was empowered by platforms for high-throughput sequencing, most of the coding errors were uncovered to be the consequences of cytosine deamination which can be easily removed from the DNA using enzymatic treatment. The methodology to repair DNA sequences is still in development, it can be basically explained by the process of reintroducing cytosine rather than uracil. This technique is thus restricted to amplified DNA molecules. To eliminate any type of damage (particularly those that block PCR) is a process still pending the complete repair methodologies; DNA detection right after extraction is highly needed. Before using any resources into extensive, unreasonable and uncertain repair techniques, it is vital to distinguish between two possible hypotheses; (i) DNA is none existent to be amplified to begin with therefore completely un-repairable, (ii) the DNA is refractory to PCR and it is worth to be repaired and amplified. Hence, it is extremely important to develop a non-enzymatic technique to detect the most degraded DNA.

Keywords: ancient DNA, DNA barcodong, enzymatic repair, PCR

Procedia PDF Downloads 390
26956 Direct Design of Steel Bridge Using Nonlinear Inelastic Analysis

Authors: Boo-Sung Koh, Seung-Eock Kim

Abstract:

In this paper, a direct design using a nonlinear inelastic analysis is suggested. Also, this paper compares the load carrying capacity obtained by a nonlinear inelastic analysis with experiment results to verify the accuracy of the results. The allowable stress design results of a railroad through a plate girder bridge and the safety factor of the nonlinear inelastic analysis were compared to examine the safety performance. As a result, the load safety factor for the nonlinear inelastic analysis was twice as high as the required safety factor under the allowable stress design standard specified in the civil engineering structure design standards for urban magnetic levitation railways, which further verified the advantages of the proposed direct design method.

Keywords: direct design, nonlinear inelastic analysis, residual stress, initial geometric imperfection

Procedia PDF Downloads 518
26955 Multilocus Phylogenetic Approach Reveals Informative DNA Barcodes for Studying Evolution and Taxonomy of Fusarium Fungi

Authors: Alexander A. Stakheev, Larisa V. Samokhvalova, Sergey K. Zavriev

Abstract:

Fusarium fungi are among the most devastating plant pathogens distributed all over the world. Significant reduction of grain yield and quality caused by Fusarium leads to multi-billion dollar annual losses to the world agricultural production. These organisms can also cause infections in immunocompromised persons and produce the wide range of mycotoxins, such as trichothecenes, fumonisins, and zearalenone, which are hazardous to human and animal health. Identification of Fusarium fungi based on the morphology of spores and spore-forming structures, colony color and appearance on specific culture media is often very complicated due to the high similarity of these features for closely related species. Modern Fusarium taxonomy increasingly uses data of crossing experiments (biological species concept) and genetic polymorphism analysis (phylogenetic species concept). A number of novel Fusarium sibling species has been established using DNA barcoding techniques. Species recognition is best made with the combined phylogeny of intron-rich protein coding genes and ribosomal DNA sequences. However, the internal transcribed spacer of (ITS), which is considered to be universal DNA barcode for Fungi, is not suitable for genus Fusarium, because of its insufficient variability between closely related species and the presence of non-orthologous copies in the genome. Nowadays, the translation elongation factor 1 alpha (TEF1α) gene is the “gold standard” of Fusarium taxonomy, but the search for novel informative markers is still needed. In this study, we used two novel DNA markers, frataxin (FXN) and heat shock protein 90 (HSP90) to discover phylogenetic relationships between Fusarium species. Multilocus phylogenetic analysis based on partial sequences of TEF1α, FXN, HSP90, as well as intergenic spacer of ribosomal DNA (IGS), beta-tubulin (β-TUB) and phosphate permease (PHO) genes has been conducted for 120 isolates of 19 Fusarium species from different climatic zones of Russia and neighboring countries using maximum likelihood (ML) and maximum parsimony (MP) algorithms. Our analyses revealed that FXN and HSP90 genes could be considered as informative phylogenetic markers, suitable for evolutionary and taxonomic studies of Fusarium genus. It has been shown that PHO gene possesses more variable (22 %) and parsimony informative (19 %) characters than other markers, including TEF1α (12 % and 9 %, correspondingly) when used for elucidating phylogenetic relationships between F. avenaceum and its closest relatives – F. tricinctum, F. acuminatum, F. torulosum. Application of novel DNA barcodes confirmed the fact that F. arthrosporioides do not represent a separate species but only a subspecies of F. avenaceum. Phylogeny based on partial PHO and FXN sequences revealed the presence of separate cluster of four F. avenaceum strains which were closer to F. torulosum than to major F. avenaceum clade. The strain F-846 from Moldova, morphologically identified as F. poae, formed a separate lineage in all the constructed dendrograms, and could potentially be considered as a separate species, but more information is needed to confirm this conclusion. Variable sites in PHO sequences were used for the first-time development of specific qPCR-based diagnostic assays for F. acuminatum and F. torulosum. This work was supported by Russian Foundation for Basic Research (grant № 15-29-02527).

Keywords: DNA barcode, fusarium, identification, phylogenetics, taxonomy

Procedia PDF Downloads 305
26954 Multidimensional Item Response Theory Models for Practical Application in Large Tests Designed to Measure Multiple Constructs

Authors: Maria Fernanda Ordoñez Martinez, Alvaro Mauricio Montenegro

Abstract:

This work presents a statistical methodology for measuring and founding constructs in Latent Semantic Analysis. This approach uses the qualities of Factor Analysis in binary data with interpretations present on Item Response Theory. More precisely, we propose initially reducing dimensionality with specific use of Principal Component Analysis for the linguistic data and then, producing axes of groups made from a clustering analysis of the semantic data. This approach allows the user to give meaning to previous clusters and found the real latent structure presented by data. The methodology is applied in a set of real semantic data presenting impressive results for the coherence, speed and precision.

Keywords: semantic analysis, factorial analysis, dimension reduction, penalized logistic regression

Procedia PDF Downloads 424
26953 Output Voltage Analysis of CMOS Colpitts Oscillator with Short Channel Device

Authors: Maryam Ebrahimpour, Amir Ebrahimi

Abstract:

This paper presents the steady-state amplitude analysis of MOS Colpitts oscillator with short channel device. The proposed method is based on a large signal analysis and the nonlinear differential equations that govern the oscillator circuit behaviour. Also, the short channel effects are considered in the proposed analysis and analytical equations for finding the steady-state oscillation amplitude are derived. The output voltage calculated from this analysis is in excellent agreement with simulations for a wide range of circuit parameters.

Keywords: colpitts oscillator, CMOS, electronics, circuits

Procedia PDF Downloads 334
26952 Model Solutions for Performance-Based Seismic Analysis of an Anchored Sheet Pile Quay Wall

Authors: C. J. W. Habets, D. J. Peters, J. G. de Gijt, A. V. Metrikine, S. N. Jonkman

Abstract:

Conventional seismic designs of quay walls in ports are mostly based on pseudo-static analysis. A more advanced alternative is the Performance-Based Design (PBD) method, which evaluates permanent deformations and amounts of (repairable) damage under seismic loading. The aim of this study is to investigate the suitability of this method for anchored sheet pile quay walls that were not purposely designed for seismic loads. A research methodology is developed in which pseudo-static, permanent-displacement and finite element analysis are employed, calibrated with an experimental reference case that considers a typical anchored sheet pile wall. A reduction factor that accounts for deformation behaviour is determined for pseudo-static analysis. A model to apply traditional permanent displacement analysis on anchored sheet pile walls is proposed. Dynamic analysis is successfully carried out. From the research it is concluded that PBD evaluation can effectively be used for seismic analysis and design of this type of structure.

Keywords: anchored sheet pile quay wall, simplified dynamic analysis, performance-based design, pseudo-static analysis

Procedia PDF Downloads 365
26951 Fault Study and Reliability Analysis of Rotative Machine

Authors: Guang Yang, Zhiwei Bai, Bo Sun

Abstract:

This paper analyzes the influence of failure mode and harmfulness of rotative machine according to FMECA (Failure Mode, Effects, and Criticality Analysis) method, and finds out the weak links that affect the reliability of this equipment. Also in this paper, fault tree analysis software is used for quantitative and qualitative analysis, pointing out the main factors of failure of this equipment. Based on the experimental results, this paper puts forward corresponding measures for prevention and improvement, and fundamentally improves the inherent reliability of this rotative machine, providing the basis for the formulation of technical conditions for the safe operation of industrial applications.

Keywords: rotative machine, reliability test, fault tree analysis, FMECA

Procedia PDF Downloads 141
26950 Load Flow Analysis of 5-IEEE Bus Test System Using Matlab

Authors: H. Abaal, R. Skouri

Abstract:

A power flow analysis is a steady-state study of power grid. The goal of power flow analysis is to determine the voltages, currents, and real and reactive power flows in a system under a given load conditions. In this paper, the load flow analysis program by Newton Raphson polar coordinates Method is developed. The effectiveness of the developed program is evaluated through a simple 5-IEEE test system bus by simulations using MATLAB.

Keywords: power flow analysis, Newton Raphson polar coordinates method

Procedia PDF Downloads 589