Search results for: sequence analysis
27906 Genome-Wide Identification and Characterization of MLO Family Genes in Pumpkin (Cucurbita maxima Duch.)
Authors: Khin Thanda Win, Chunying Zhang, Sanghyeob Lee
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Mildew resistance locus o (Mlo), a plant-specific gene family with seven-transmembrane (TM), plays an important role in plant resistance to powdery mildew (PM). PM caused by Podosphaera xanthii is a widespread plant disease and probably represents the major fungal threat for many Cucurbits. The recent Cucurbita maxima genome sequence data provides an opportunity to identify and characterize the MLO gene family in this species. Total twenty genes (designated CmaMLO1 through CmaMLO20) have been identified by using an in silico cloning method with the MLO gene sequences of Cucumis sativus, Cucumis melo, Citrullus lanatus and Cucurbita pepo as probes. These CmaMLOs were evenly distributed on 15 chromosomes of 20 C. maxima chromosomes without any obvious clustering. Multiple sequence alignment showed that the common structural features of MLO gene family, such as TM domains, a calmodulin-binding domain and 30 important amino acid residues for MLO function, were well conserved. Phylogenetic analysis of the CmaMLO genes and other plant species reveals seven different clades (I through VII) and only clade IV is specific to monocots (rice, barley, and wheat). Phylogenetic and structural analyses provided preliminary evidence that five genes belonged to clade V could be the susceptibility genes which may play the importance role in PM resistance. This study is the first comprehensive report on MLO genes in C. maxima to our knowledge. These findings will facilitate the functional analysis of the MLOs related to PM susceptibility and are valuable resources for the development of disease resistance in pumpkin.Keywords: Mildew resistance locus o (Mlo), powdery mildew, phylogenetic relationship, susceptibility genes
Procedia PDF Downloads 17627905 Remembering Route in an Unfamiliar Homogenous Environment
Authors: Ahmed Sameer, Braj Bhushan
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The objective of our study was to compare two techniques (no landmark vs imaginary landmark) of remembering route while traversing in an unfamiliar homogenous environment. We used two videos each having nine identical turns with no landmarks. In the first video participant was required to remember the sequence of turns. In the second video participant was required to imagine a landmark at each turn and associate the turn with it. In both the task the participant was asked to recall the sequence of turns as it appeared in the video. Results showed that performance in the first condition i.e. without use of landmarks was better than imaginary landmark condition. The difference, however, became significant when the participant were tested again about 30 minutes later though performance was still better in no-landmark condition. The finding is surprising given the past research in memory and is explained in terms of cognitive factors such as mental workload.Keywords: wayfinding, landmarks, unfamiliar environment, cognitive psychology
Procedia PDF Downloads 46827904 Effective Teaching Pyramid and Its Impact on Enhancing the Participation of Students in Swimming Classes
Authors: Salam M. H. Kareem
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Instructional or teaching procedures and their proper sequence are essential for high-quality learning outcomes. These actions are the path that the teacher takes during the learning process after setting the learning objectives. Teachers and specialists in the education field should include teaching procedures with putting in place an effective mechanism for the procedure’s implementation to achieve a logical sequence with the desired output of overall education process. Determining the sequence of these actions may be a strategic process outlined by a strategic educational plan or drawn by teachers with a high level of experience, enabling them to determine those logical procedures. While specific actions may be necessary for a specific form, many Physical Education (PE) teachers can work out on various sports disciplines. This study was conducted to investigate the impact of using the teaching sequence of the teaching pyramid in raising the level of enjoyment in swimming classes. Four months later of teaching swimming skills to the control and experimental groups of the study, we figured that using the tools shown in the teaching pyramid with the experimental group led to statistically significant differences in the positive tendencies of students to participate in the swimming classes by using the traditional procedures of teaching and using of successive procedures in the teaching pyramid, and in favor of the teaching pyramid, The students are influenced by enhancing their tendency to participate in swimming classes when the teaching procedures followed are sensitive to individual differences and are based on the element of pleasure in learning, and less positive levels of the tendency of students when using traditional teaching procedures, by getting the level of skills' requirements higher and more difficult to perform. The level of positive tendencies of students when using successive procedures in the teaching pyramid was increased, by getting the level of skills' requirements higher and more difficult to perform, because of the high level of motivation and the desire to challenge the self-provided by the teaching pyramid.Keywords: physical education, swimming classes, teaching process, teaching pyramid
Procedia PDF Downloads 13927903 Multi-Objective Optimization of the Thermal-Hydraulic Behavior for a Sodium Fast Reactor with a Gas Power Conversion System and a Loss of off-Site Power Simulation
Authors: Avent Grange, Frederic Bertrand, Jean-Baptiste Droin, Amandine Marrel, Jean-Henry Ferrasse, Olivier Boutin
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CEA and its industrial partners are designing a gas Power Conversion System (PCS) based on a Brayton cycle for the ASTRID Sodium-cooled Fast Reactor. Investigations of control and regulation requirements to operate this PCS during operating, incidental and accidental transients are necessary to adapt core heat removal. To this aim, we developed a methodology to optimize the thermal-hydraulic behavior of the reactor during normal operations, incidents and accidents. This methodology consists of a multi-objective optimization for a specific sequence, whose aim is to increase component lifetime by reducing simultaneously several thermal stresses and to bring the reactor into a stable state. Furthermore, the multi-objective optimization complies with safety and operating constraints. Operating, incidental and accidental sequences use specific regulations to control the thermal-hydraulic reactor behavior, each of them is defined by a setpoint, a controller and an actuator. In the multi-objective problem, the parameters used to solve the optimization are the setpoints and the settings of the controllers associated with the regulations included in the sequence. In this way, the methodology allows designers to define an optimized and specific control strategy of the plant for the studied sequence and hence to adapt PCS piloting at its best. The multi-objective optimization is performed by evolutionary algorithms coupled to surrogate models built on variables computed by the thermal-hydraulic system code, CATHARE2. The methodology is applied to a loss of off-site power sequence. Three variables are controlled: the sodium outlet temperature of the sodium-gas heat exchanger, turbomachine rotational speed and water flow through the heat sink. These regulations are chosen in order to minimize thermal stresses on the gas-gas heat exchanger, on the sodium-gas heat exchanger and on the vessel. The main results of this work are optimal setpoints for the three regulations. Moreover, Proportional-Integral-Derivative (PID) control setting is considered and efficient actuators used in controls are chosen through sensitivity analysis results. Finally, the optimized regulation system and the reactor control procedure, provided by the optimization process, are verified through a direct CATHARE2 calculation.Keywords: gas power conversion system, loss of off-site power, multi-objective optimization, regulation, sodium fast reactor, surrogate model
Procedia PDF Downloads 29827902 Genetic Analysis of the Endangered Mangrove Species Avicennia Marina in Qatar Detected by Inter-Simple Sequence Repeat DNA Markers
Authors: Talaat Ahmed, Amna Babssail
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Mangroves are evergreen trees and grow along the coastal areas of Qatar. The largest and oldest area of mangroves can be found around Al-Thakhira and Al-Khor. Other mangrove areas originate from fairly recent plantings by the government, although unfortunately the picturesque mangrove lake in Al-Wakra has now been uprooted. Avicinnia marina is the predominant mangrove species found in the region. Mangroves protect and stabilize low lying coastal land, and provide protection and food sources for estuarine and coastal fishery food chains. They also serve as feeding, breeding and nursery grounds for a variety of fish, crustaceans, reptiles, birds and other wildlife. A total of 21 individuals of A. marina, representing seven diverse Natural and artificial populations, were sampled throughout its range in Qatar. Leaves from 2-3 randomly selected trees at each location were collected. The locations are as follows: Al-Rawis, Ras-Madpak, Fuwairt, Summaseima, Al-khour, AL-Mafjar and Zekreet. Total genomic DNA was extracted using commercial DNeasy Plant System (Qiagen, Inc., Valencia, CA) kit to be used for genetic diversity analysis. Total of 12 (Inter-Simple Sequence Repeat) ISSR primers were used to amplify DNA fragments using genomic DNA. The 12 ISSR primers amplified polymorphic bands among mangrove samples in different areas as well as within each area indicating the existing of variation within each area and among the different areas of mangrove in Qatar. The results could characterize Avicinnia marina populations exist in different areas of Qatar and establish DNA fingerprint documentations for mangrove population to be used in further studies. Moreover, existing of genetic variation within and among Avicinnia marina populations is a strong indication for the ability of such populations to adapt different environmental conditions in Qatar. This study could be a warning to save mangrove in Qatar and save the environment as well.Keywords: DNA fingerprint, Avicinnia marina, genetic analysis, Qatar
Procedia PDF Downloads 38927901 Biodegradation of Malathion by Acinetobacter baumannii Strain AFA Isolated from Domestic Sewage in Egypt
Authors: Ahmed F. Azmy, Amal E. Saafan, Tamer M. Essam, Magdy A. Amin, Shaban H. Ahmed
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Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosae (PS1),andPseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazenon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 1012CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.Keywords: Acinetobacter baumannii, biodegradation, malathion, organophosphate pesticides
Procedia PDF Downloads 48427900 A Highly Efficient Broadcast Algorithm for Computer Networks
Authors: Ganesh Nandakumaran, Mehmet Karaata
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A wave is a distributed execution, often made up of a broadcast phase followed by a feedback phase, requiring the participation of all the system processes before a particular event called decision is taken. Wave algorithms with one initiator such as the 1-wave algorithm have been shown to be very efficient for broadcasting messages in tree networks. Extensions of this algorithm broadcasting a sequence of waves using a single initiator have been implemented in algorithms such as the m-wave algorithm. However as the network size increases, having a single initiator adversely affects the message delivery times to nodes further away from the initiator. As a remedy, broadcast waves can be allowed to be initiated by multiple initiator nodes distributed across the network to reduce the completion time of broadcasts. These waves initiated by one or more initiator processes form a collection of waves covering the entire network. Solutions to global-snapshots, distributed broadcast and various synchronization problems can be solved efficiently using waves with multiple concurrent initiators. In this paper, we propose the first stabilizing multi-wave sequence algorithm implementing waves started by multiple initiator processes such that every process in the network receives at least one sequence of broadcasts. Due to being stabilizing, the proposed algorithm can withstand transient faults and do not require initialization. We view a fault as a transient fault if it perturbs the configuration of the system but not its program.Keywords: distributed computing, multi-node broadcast, propagation of information with feedback and cleaning (PFC), stabilization, wave algorithms
Procedia PDF Downloads 49727899 Partial M-Sequence Code Families Applied in Spectral Amplitude Coding Fiber-Optic Code-Division Multiple-Access Networks
Authors: Shin-Pin Tseng
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Nowadays, numerous spectral amplitude coding (SAC) fiber-optic code-division-multiple-access (FO-CDMA) techniques were appealing due to their capable of providing moderate security and relieving the effects of multiuser interference (MUI). Nonetheless, the performance of the previous network is degraded due to fixed in-phase cross-correlation (IPCC) value. Based on the above problems, a new SAC FO-CDMA network using partial M-sequence (PMS) code is presented in this study. Because the proposed PMS code is originated from M-sequence code, the system using the PMS code could effectively suppress the effects of MUI. In addition, two-code keying (TCK) scheme can applied in the proposed SAC FO-CDMA network and enhance the whole network performance. According to the consideration of system flexibility, simple optical encoders/decoders (codecs) using fiber Bragg gratings (FBGs) were also developed. First, we constructed a diagram of the SAC FO-CDMA network, including (N/2-1) optical transmitters, (N/2-1) optical receivers, and one N×N star coupler for broadcasting transmitted optical signals to arrive at the input port of each optical receiver. Note that the parameter N for the PMS code was the code length. In addition, the proposed SAC network was using superluminescent diodes (SLDs) as light sources, which then can save a lot of system cost compared with the other FO-CDMA methods. For the design of each optical transmitter, it is composed of an SLD, one optical switch, and two optical encoders according to assigned PMS codewords. On the other hand, each optical receivers includes a 1 × 2 splitter, two optical decoders, and one balanced photodiode for mitigating the effect of MUI. In order to simplify the next analysis, the some assumptions were used. First, the unipolarized SLD has flat power spectral density (PSD). Second, the received optical power at the input port of each optical receiver is the same. Third, all photodiodes in the proposed network have the same electrical properties. Fourth, transmitting '1' and '0' has an equal probability. Subsequently, by taking the factors of phase‐induced intensity noise (PIIN) and thermal noise, the corresponding performance was displayed and compared with the performance of the previous SAC FO-CDMA networks. From the numerical result, it shows that the proposed network improved about 25% performance than that using other codes at BER=10-9. This is because the effect of PIIN was effectively mitigated and the received power was enhanced by two times. As a result, the SAC FO-CDMA network using PMS codes has an opportunity to apply in applications of the next-generation optical network.Keywords: spectral amplitude coding, SAC, fiber-optic code-division multiple-access, FO-CDMA, partial M-sequence, PMS code, fiber Bragg grating, FBG
Procedia PDF Downloads 38127898 Study of Microbial Diversity Associated with Tarballs and Their Exploitation in Crude Oil Degradation
Authors: Varsha Shinde, Belle Damodara Shenoy
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Tarballs are crude oil remnants found in oceans after long term weathering process and are a global concern since several decades as potential marine pollutant. Being complicated in structure microbial remediation of tarballs in natural environment is a slow process. They are rich in high molecular weight alkanes and poly aromatic hydrocarbons which are resistant to microbial attack and other environmental factors, therefore remain in environment for long time. However, it has been found that many bacteria and fungi inhabit on tarballs for nutrients and shelter. Many of them are supposed to be oil degraders, while others are supposed to be getting benefited by byproducts formed during hydrocarbon metabolism. Thus tarballs are forming special interesting ecological niche of microbes. This work aimed to study diversity of bacteria and fungi from tarballs and to see their potential application in crude oil degradation. The samples of tarballs were collected from Betul beach of south Goa (India). Different methods were used to isolate culturable fraction of bacteria and fungi from it. Those were sequenced for 16S rRNA gene and ITS for molecular level identification. The 16S rRNA gene sequence analysis revealed the presence of 13 bacterial genera/clades (Alcanivorax, Brevibacterium, Bacillus, Cellulomonas, Enterobacter, Klebsiella, Marinobacter, Nitratireductor, Pantoea, Pseudomonas, Pseudoxanthomonas, Tistrella and Vibrio), while the ITS sequence analysis placed the fungi in 8 diverse genera/ clades (Aspergillus, Byssochlamys, Monascus, Paecilomyces, Penicillium, Scytalidium/ Xylogone, Talaromyces and Trichoderma). All bacterial isolates were screened for oil degradation capacity. Potential strains were subjected to crude oil degradation experiment for quantification. Results were analyzed by GC-MS-MS.Keywords: bacteria, biodegradation, crude oil, diversity, fungi, tarballs
Procedia PDF Downloads 21627897 Isolate-Specific Variations among Clinical Isolates of Brucella Identified by Whole-Genome Sequencing, Bioinformatics and Comparative Genomics
Authors: Abu S. Mustafa, Mohammad W. Khan, Faraz Shaheed Khan, Nazima Habibi
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Brucellosis is a zoonotic disease of worldwide prevalence. There are at least four species and several strains of Brucella that cause human disease. Brucella genomes have very limited variation across strains, which hinder strain identification using classical molecular techniques, including PCR and 16 S rDNA sequencing. The aim of this study was to perform whole genome sequencing of clinical isolates of Brucella and perform bioinformatics and comparative genomics analyses to determine the existence of genetic differences across the isolates of a single Brucella species and strain. The draft sequence data were generated from 15 clinical isolates of Brucella melitensis (biovar 2 strain 63/9) using MiSeq next generation sequencing platform. The generated reads were used for further assembly and analysis. All the analysis was performed using Bioinformatics work station (8 core i7 processor, 8GB RAM with Bio-Linux operating system). FastQC was used to determine the quality of reads and low quality reads were trimmed or eliminated using Fastx_trimmer. Assembly was done by using Velvet and ABySS softwares. The ordering of assembled contigs was performed by Mauve. An online server RAST was employed to annotate the contigs assembly. Annotated genomes were compared using Mauve and ACT tools. The QC score for DNA sequence data, generated by MiSeq, was higher than 30 for 80% of reads with more than 100x coverage, which suggested that data could be utilized for further analysis. However when analyzed by FastQC, quality of four reads was not good enough for creating a complete genome draft so remaining 11 samples were used for further analysis. The comparative genome analyses showed that despite sharing same gene sets, single nucleotide polymorphisms and insertions/deletions existed across different genomes, which provided a variable extent of diversity to these bacteria. In conclusion, the next generation sequencing, bioinformatics, and comparative genome analysis can be utilized to find variations (point mutations, insertions and deletions) across different genomes of Brucella within a single strain. This information could be useful in surveillance and epidemiological studies supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.Keywords: brucella, bioinformatics, comparative genomics, whole genome sequencing
Procedia PDF Downloads 37127896 Antibacterial Activity of Salvadora persica Extracts against Oral Cavity Bacteria
Authors: Sulaiman A. Alrumman, Abd El-Latif Hesham
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Despite medical progress worldwide, dental caries are still widespread. Miswak is derived from the plant arak (Salvadora persica). It is used by Muslim people as a natural product for the cleansing of teeth, to ensure oral and dental hygiene. This study was designed to evaluate the antimicrobial effects of ethanol, methanol, and ethanol/methanol extracts of miswak against three bacterial pathogens of the oral cavity. The pathogens were isolated from the oral cavity of volunteers/patients and were identified on the basis of 16S rRNA gene amplification data. Sequence comparisons were made with 16S rRNA gene sequences available in the GenBank database. The results of sequence alignment and phylogenetic analysis identified the three pathogens as being Staphylococcus aureus strain KKU-020, Enterococcus faecalis strain KKU-021 and Klebsiella pneumoniae strain KKU-022. All miswak extracts showed powerful antimicrobial activity against the three pathogens. The maximum zone of inhibition (40.67±0.88 mm) was observed against E. faecalis with ethanolic extracts whilst methanolic extracts showed the minimum zone of inhibition (10.33±0.88 mm) against K. pneumonia KKU-022. Based on the significant effects of the miswak extracts against the oral cavity pathogens in our study, we recommend that miswak is to be used as a dental hygiene method to prevent tooth caries.Keywords: antibacterial, miswak, Salvadora persica, oral cavity pathogens
Procedia PDF Downloads 28927895 A Bilingual Didactic Sequence about Biological Control to Develop the Scientific Literacy on High School Students
Authors: André Melo Franco Lorena De Barros, Elida Geralda Campos
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The bilingual education has just started in Brazils public schools. This paper is a didactic sequence of biology bilingual lessons about biologic control in the Brazilian Savana. This sequence has been applied in the first year of a bilingual education program in the only public English and Portuguese bilingual high school in Brazil. The aim of this work is to develop and apply a didactic sequence capable of developing the scientific literacy through the bilingual education associated with Problem Based Learning. This didactic sequence was applied in a class of 30 students. It was divided in three lessons. In the first lesson the students were divided in groups and received a fiction Letter from a mayor explaining the problem and asking students for help. The organic soy plantation of the mayor’s is been attacked by caterpillars. The students read the text then raised hypothesis of how they could solve the problem. In the second lesson the students searched online to verify if theirs hypothesis were correct and to find answers for the question proposed. In the third lesson the groups got together and discussed about their results and wrote a final essay with the answers for the problem proposed. The tools used to acquire information about the didactic sequence were: researcher’s diary, survey, interview and essay developed by the students. Most of the initial hypothesis couldn’t answer the problem properly. By the second lesson most of the students could answer properly. During the third lesson all the groups figured out suitable answers. The forms of biological control, birds habits and transgenic were deeply studied by the students. This methodology was successful for developing the scientific literacy with most of the students and also concluded that the quality of learning is directly associated with the effort of each student during the process. [ARAÚJO, Denise Lino de. O que é (e como se faz) sequência didática. Entrepalavras, Fortaleza, v. 3, n. 3, p.322-334, jul. 2013.] [FRANCO, Aline Aparecida et al. Preferência alimentar de Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae) por cultivares de soja. Científica: Revista de Ciências Agrárias, Jaboticabal, v. 1, n. 42, p.32-38, 29 jan. 2014.] [RIBEIRO, Luis Roberto de Camargo. Aprendizagem baseada em problemas (PBL): Uma experiência no ensino superior. São Carlos: Editora da Universidade Federal de São Carlos Ribeiro, 2008. 151 p.] [TRIVELATO, Sílvia L. Frateschi; TONIDANDEL, Sandra M. Rudella. Ensino Por Investigação: Eixos Organizadores Para Sequências De Ensino De Biologia. Ensaio Pesquisa em Educação em Ciências, Belo Horizonte, v. 17, n. especial, p.97-114, nov. 2015.].Keywords: Bilingual Education, Environmental Education, Problem Based Learning, Science education
Procedia PDF Downloads 18027894 Mining the Proteome of Fusobacterium nucleatum for Potential Therapeutics Discovery
Authors: Abdul Musaweer Habib, Habibul Hasan Mazumder, Saiful Islam, Sohel Sikder, Omar Faruk Sikder
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The plethora of genome sequence information of bacteria in recent times has ushered in many novel strategies for antibacterial drug discovery and facilitated medical science to take up the challenge of the increasing resistance of pathogenic bacteria to current antibiotics. In this study, we adopted subtractive genomics approach to analyze the whole genome sequence of the Fusobacterium nucleatum, a human oral pathogen having association with colorectal cancer. Our study divulged 1499 proteins of Fusobacterium nucleatum, which has no homolog in human genome. These proteins were subjected to screening further by using the Database of Essential Genes (DEG) that resulted in the identification of 32 vitally important proteins for the bacterium. Subsequent analysis of the identified pivotal proteins, using the KEGG Automated Annotation Server (KAAS) resulted in sorting 3 key enzymes of F. nucleatum that may be good candidates as potential drug targets, since they are unique for the bacterium and absent in humans. In addition, we have demonstrated the 3-D structure of these three proteins. Finally, determination of ligand binding sites of the key proteins as well as screening for functional inhibitors that best fitted with the ligands sites were conducted to discover effective novel therapeutic compounds against Fusobacterium nucleatum.Keywords: colorectal cancer, drug target, Fusobacterium nucleatum, homology modeling, ligands
Procedia PDF Downloads 38027893 Genetic Divergence and Morphogenic Analysis of Sugarcane Red Rot Pathogen Colletotrichum falcatum under South Gujarat Condition
Authors: Prittesh Patel, Ramar Krishnamurthy
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In the present study, nine strains of C. falcatum obtained from different places and cultivars were characterized for sporulation, growth rate, and 18S rRNA gene sequence. All isolates had characteristic fast-growing sparse and fleecy aerial mycelia on potato dextrose agar with sickle shape conidia (length x width: varied from 20.0 X 3.89 to 25.52 X 5.34 μm) and blackish to orange acervuli with setae (length x width: varied from 112.37X 2.78 to 167.66 X 6.73 μm). They could be divided into two groups on the base of morphology; P1, dense mycelia with concentric growth and P2, sparse mycelia with uneven growth. Genomic DNA isolation followed by PCR amplification with ITS1 and ITS4 primer produced ~550bp amplicons for all isolates. Phylogeny generated by 18S rRNA gene sequence confirmed the variation in isolates and mainly grouped into two clusters; cluster 1 contained CoC671 isolates (cfNAV and cfPAR) and Co86002 isolate (cfTIM). Other isolates cfMAD, cfKAM, and cfMAR were grouped into cluster 2. Remaining isolates did not fall into any cluster. Isolate cfGAN, collected from Co86032 was found highly diverse of all the nine isolates. In a nutshell, we found considerable genetic divergence and morphological variation within C. falcatum accessions collected from different areas of south Gujarat, India and these can be used for the breeding program.Keywords: Colletotrichum falcatum, ITS, morphology, red rot, sugarcane
Procedia PDF Downloads 12127892 Assessment of DNA Sequence Encoding Techniques for Machine Learning Algorithms Using a Universal Bacterial Marker
Authors: Diego Santibañez Oyarce, Fernanda Bravo Cornejo, Camilo Cerda Sarabia, Belén Díaz Díaz, Esteban Gómez Terán, Hugo Osses Prado, Raúl Caulier-Cisterna, Jorge Vergara-Quezada, Ana Moya-Beltrán
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The advent of high-throughput sequencing technologies has revolutionized genomics, generating vast amounts of genetic data that challenge traditional bioinformatics methods. Machine learning addresses these challenges by leveraging computational power to identify patterns and extract information from large datasets. However, biological sequence data, being symbolic and non-numeric, must be converted into numerical formats for machine learning algorithms to process effectively. So far, some encoding methods, such as one-hot encoding or k-mers, have been explored. This work proposes additional approaches for encoding DNA sequences in order to compare them with existing techniques and determine if they can provide improvements or if current methods offer superior results. Data from the 16S rRNA gene, a universal marker, was used to analyze eight bacterial groups that are significant in the pulmonary environment and have clinical implications. The bacterial genes included in this analysis are Prevotella, Abiotrophia, Acidovorax, Streptococcus, Neisseria, Veillonella, Mycobacterium, and Megasphaera. These data were downloaded from the NCBI database in Genbank file format, followed by a syntactic analysis to selectively extract relevant information from each file. For data encoding, a sequence normalization process was carried out as the first step. From approximately 22,000 initial data points, a subset was generated for testing purposes. Specifically, 55 sequences from each bacterial group met the length criteria, resulting in an initial sample of approximately 440 sequences. The sequences were encoded using different methods, including one-hot encoding, k-mers, Fourier transform, and Wavelet transform. Various machine learning algorithms, such as support vector machines, random forests, and neural networks, were trained to evaluate these encoding methods. The performance of these models was assessed using multiple metrics, including the confusion matrix, ROC curve, and F1 Score, providing a comprehensive evaluation of their classification capabilities. The results show that accuracies between encoding methods vary by up to approximately 15%, with the Fourier transform obtaining the best results for the evaluated machine learning algorithms. These findings, supported by the detailed analysis using the confusion matrix, ROC curve, and F1 Score, provide valuable insights into the effectiveness of different encoding methods and machine learning algorithms for genomic data analysis, potentially improving the accuracy and efficiency of bacterial classification and related genomic studies.Keywords: DNA encoding, machine learning, Fourier transform, Fourier transformation
Procedia PDF Downloads 1127891 Cloning, Expression and Protein Purification of AV1 Gene of Okra Leaf Curl Virus Egyptian Isolate and Genetic Diversity between Whitefly and Different Plant Hosts
Authors: Dalia. G. Aseel
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Begomoviruses are economically important plant viruses that infect dicotyledonous plants and exclusively transmitted by the whitefly Bemisia tabaci. Here, replicative form was isolated from Okra, Cotton, Tomato plants and whitefly infected with Begomoviruses. Using coat protein specific primers (AV1), the viral infection was verified with amplicon at 450 bp. The sequence of OLCuV-AV1 gene was recorded and received an accession number (FJ441605) from Genebank. The phylogenetic tree of OLCuV was closely related to Okra leaf curl virus previously isolated from Cameroon and USA with nucleotide sequence identity of 92%. The protein purification was carried out using His-Tag methodology by using Affinity Chromatography. The purified protein was separated on SDS-PAGE analysis and an enriched expected size of band at 30 kDa was observed. Furthermore, RAPD and SDS-PAGE were used to detect genetic variability between different hosts of okra leaf curl virus (OLCuV), cotton leaf curl virus (CLCuV), tomato yellow leaf curl virus (TYLCuV) and the whitefly vector. Finally, the present study would help to understand the relationship between the whitefly and different economical crops in Egypt.Keywords: okra leaf curl virus, AV1 gene, sequencing, phylogenetic, cloning, purified protein, genetic diversity and viral proteins
Procedia PDF Downloads 14127890 Comprehensive Risk Analysis of Decommissioning Activities with Multifaceted Hazard Factors
Authors: Hyeon-Kyo Lim, Hyunjung Kim, Kune-Woo Lee
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Decommissioning process of nuclear facilities can be said to consist of a sequence of problem solving activities, partly because there may exist working environments contaminated by radiological exposure, and partly because there may also exist industrial hazards such as fire, explosions, toxic materials, and electrical and physical hazards. As for an individual hazard factor, risk assessment techniques are getting known to industrial workers with advance of safety technology, but the way how to integrate those results is not. Furthermore, there are few workers who experienced decommissioning operations a lot in the past. Therefore, not a few countries in the world have been trying to develop appropriate counter techniques in order to guarantee safety and efficiency of the process. In spite of that, there still exists neither domestic nor international standard since nuclear facilities are too diverse and unique. In the consequence, it is quite inevitable to imagine and assess the whole risk in the situation anticipated one by one. This paper aimed to find out an appropriate technique to integrate individual risk assessment results from the viewpoint of experts. Thus, on one hand the whole risk assessment activity for decommissioning operations was modeled as a sequence of individual risk assessment steps, and on the other, a hierarchical risk structure was developed. Then, risk assessment procedure that can elicit individual hazard factors one by one were introduced with reference to the standard operation procedure (SOP) and hierarchical task analysis (HTA). With an assumption of quantification and normalization of individual risks, a technique to estimate relative weight factors was tried by using the conventional Analytic Hierarchical Process (AHP) and its result was reviewed with reference to judgment of experts. Besides, taking the ambiguity of human judgment into consideration, debates based upon fuzzy inference was added with a mathematical case study.Keywords: decommissioning, risk assessment, analytic hierarchical process (AHP), fuzzy inference
Procedia PDF Downloads 42127889 Full Characterization of Heterogeneous Antibody Samples under Denaturing and Native Conditions on a Hybrid Quadrupole-Orbitrap Mass Spectrometer
Authors: Rowan Moore, Kai Scheffler, Eugen Damoc, Jennifer Sutton, Aaron Bailey, Stephane Houel, Simon Cubbon, Jonathan Josephs
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Purpose: MS analysis of monoclonal antibodies (mAbs) at the protein and peptide levels is critical during development and production of biopharmaceuticals. The compositions of current generation therapeutic proteins are often complex due to various modifications which may affect efficacy. Intact proteins analyzed by MS are detected in higher charge states that also provide more complexity in mass spectra. Protein analysis in native or native-like conditions with zero or minimal organic solvent and neutral or weakly acidic pH decreases charge state value resulting in mAb detection at higher m/z ranges with more spatial resolution. Methods: Three commercially available mAbs were used for all experiments. Intact proteins were desalted online using size exclusion chromatography (SEC) or reversed phase chromatography coupled on-line with a mass spectrometer. For streamlined use of the LC- MS platform we used a single SEC column and alternately selected specific mobile phases to perform separations in either denaturing or native-like conditions: buffer A (20 % ACN, 0.1 % FA) with Buffer B (100 mM ammonium acetate). For peptide analysis mAbs were proteolytically digested with and without prior reduction and alkylation. The mass spectrometer used for all experiments was a commercially available Thermo Scientific™ hybrid Quadrupole-Orbitrap™ mass spectrometer, equipped with the new BioPharma option which includes a new High Mass Range (HMR) mode that allows for improved high mass transmission and mass detection up to 8000 m/z. Results: We have analyzed the profiles of three mAbs under reducing and native conditions by direct infusion with offline desalting and with on-line desalting via size exclusion and reversed phase type columns. The presence of high salt under denaturing conditions was found to influence the observed charge state envelope and impact mass accuracy after spectral deconvolution. The significantly lower charge states observed under native conditions improves the spatial resolution of protein signals and has significant benefits for the analysis of antibody mixtures, e.g. lysine variants, degradants or sequence variants. This type of analysis requires the detection of masses beyond the standard mass range ranging up to 6000 m/z requiring the extended capabilities available in the new HMR mode. We have compared each antibody sample that was analyzed individually with mixtures in various relative concentrations. For this type of analysis, we observed that apparent native structures persist and ESI is benefited by the addition of low amounts of acetonitrile and formic acid in combination with the ammonium acetate-buffered mobile phase. For analyses on the peptide level we analyzed reduced/alkylated, and non-reduced proteolytic digests of the individual antibodies separated via reversed phase chromatography aiming to retrieve as much information as possible regarding sequence coverage, disulfide bridges, post-translational modifications such as various glycans, sequence variants, and their relative quantification. All data acquired were submitted to a single software package for analysis aiming to obtain a complete picture of the molecules analyzed. Here we demonstrate the capabilities of the mass spectrometer to fully characterize homogeneous and heterogeneous therapeutic proteins on one single platform. Conclusion: Full characterization of heterogeneous intact protein mixtures by improved mass separation on a quadrupole-Orbitrap™ mass spectrometer with extended capabilities has been demonstrated.Keywords: disulfide bond analysis, intact analysis, native analysis, mass spectrometry, monoclonal antibodies, peptide mapping, post-translational modifications, sequence variants, size exclusion chromatography, therapeutic protein analysis, UHPLC
Procedia PDF Downloads 35627888 Structural Geology along the Jhakri-Wangtu Road (Jutogh Section) Himachal Pradesh, NW Higher Himalaya, India
Authors: Rajkumar Ghosh
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The paper presents a comprehensive study of the structural analysis of the Chaura Thrust in Himachal Pradesh, India. The research focuses on several key aspects, including the activation timing of the Main Central Thrust (MCT) and the South Tibetan Detachment System (STDS), the identification and characterization of mylonitised zones through microscopic examination, and the understanding of box fold characteristics and their implications in the regional geology of the Himachal Himalaya. The primary objective of the study is to provide field documentation of the Chaura Thrust, which was previously considered a blind thrust with limited field evidence. Additionally, the research aims to characterize box folds and their signatures within the broader geological context of the Himachal Himalaya, document the temperature range associated with grain boundary migration (GBM), and explore the overprinting structures related to multiple sets of Higher Himalayan Out-of-Sequence Thrusts (OOSTs). The research methodology employed geological field observations and microscopic studies. Samples were collected along the Jhakri-Chaura transect at regular intervals of approximately 1 km to conduct strain analysis. Microstructural studies at the grain scale along the Jhakri-Wangtu transect were used to document the GBM-associated temperature range. The study reveals that the MCT activated in two parts, as did the STDS, and provides insights into the activation ages of the Main Boundary Thrust (MBT) and the Main Frontal Thrust (MFT). Under microscopic examination, the study identifies two mylonitised zones characterized by S-C fabric, and it documents dynamic and bulging recrystallization, as well as sub-grain formation. Various types of crenulated schistosity are observed in photomicrographs, including a rare occurrence where crenulation cleavage and sigmoid Muscovite are found juxtaposed. The study also notes the presence of S/SE-verging meso- and micro-scale box folds around Chaura, which may indicate structural upliftment. Kink folds near Chaura are visible, while asymmetric shear sense indicators in augen mylonite are predominantly observed under microscopic examination. Moreover, the research highlights the documentation of the Higher Himalayan Out-of-Sequence Thrust (OOST) in Himachal Pradesh, which activated the MCT and occurred within a zone south of the Main Central Thrust Upper (MCTU). The presence of multiple sets of OOSTs suggests a zigzag pattern of strain accumulation in the area. The study emphasizes the significance of understanding the overprinting structures associated with OOSTs. Overall, this study contributes to the understanding of the structural analysis of the Chaura Thrust and its implications in the regional geology of the Himachal Himalaya. The research underscores the importance of microscopic studies in identifying mylonitised zones and various types of crenulated schistosity. Additionally, the study documents the GBM-associated temperature range and provides insights into the activation of the Higher Himalayan Out-of-Sequence Thrust (OOST) in Himachal Pradesh. The findings of the study were obtained through geological field observations, microscopic studies, and strain analysis, offering valuable insights into the activation timing, mylonitization characteristics, and overprinting structures related to the Chaura Thrust and the broader tectonic framework of the region.Keywords: Main Central Thrust, Jhakri Thrust, Chaura Thrust, Higher Himalaya, Out-of-Sequence Thrust, Sarahan Thrust
Procedia PDF Downloads 9427887 Complete Genome Sequence Analysis of Pasteurella multocida Subspecies multocida Serotype A Strain PMTB2.1
Authors: Shagufta Jabeen, Faez J. Firdaus Abdullah, Zunita Zakaria, Nurulfiza M. Isa, Yung C. Tan, Wai Y. Yee, Abdul R. Omar
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Pasteurella multocida (PM) is an important veterinary opportunistic pathogen particularly associated with septicemic pasteurellosis, pneumonic pasteurellosis and hemorrhagic septicemia in cattle and buffaloes. P. multocida serotype A has been reported to cause fatal pneumonia and septicemia. Pasteurella multocida subspecies multocida of serotype A Malaysian isolate PMTB2.1 was first isolated from buffaloes died of septicemia. In this study, the genome of P. multocida strain PMTB2.1 was sequenced using third-generation sequencing technology, PacBio RS2 system and analyzed bioinformatically via de novo analysis followed by in-depth analysis based on comparative genomics. Bioinformatics analysis based on de novo assembly of PacBio raw reads generated 3 contigs followed by gap filling of aligned contigs with PCR sequencing, generated a single contiguous circular chromosome with a genomic size of 2,315,138 bp and a GC content of approximately 40.32% (Accession number CP007205). The PMTB2.1 genome comprised of 2,176 protein-coding sequences, 6 rRNA operons and 56 tRNA and 4 ncRNAs sequences. The comparative genome sequence analysis of PMTB2.1 with nine complete genomes which include Actinobacillus pleuropneumoniae, Haemophilus parasuis, Escherichia coli and five P. multocida complete genome sequences including, PM70, PM36950, PMHN06, PM3480, PMHB01 and PMTB2.1 was carried out based on OrthoMCL analysis and Venn diagram. The analysis showed that 282 CDs (13%) are unique to PMTB2.1and 1,125 CDs with orthologs in all. This reflects overall close relationship of these bacteria and supports the classification in the Gamma subdivision of the Proteobacteria. In addition, genomic distance analysis among all nine genomes indicated that PMTB2.1 is closely related with other five Pasteurella species with genomic distance less than 0.13. Synteny analysis shows subtle differences in genetic structures among different P.multocida indicating the dynamics of frequent gene transfer events among different P. multocida strains. However, PM3480 and PM70 exhibited exceptionally large structural variation since they were swine and chicken isolates. Furthermore, genomic structure of PMTB2.1 is more resembling that of PM36950 with a genomic size difference of approximately 34,380 kb (smaller than PM36950) and strain-specific Integrative and Conjugative Elements (ICE) which was found only in PM36950 is absent in PMTB2.1. Meanwhile, two intact prophages sequences of approximately 62 kb were found to be present only in PMTB2.1. One of phage is similar to transposable phage SfMu. The phylogenomic tree was constructed and rooted with E. coli, A. pleuropneumoniae and H. parasuis based on OrthoMCL analysis. The genomes of P. multocida strain PMTB2.1 were clustered with bovine isolates of P. multocida strain PM36950 and PMHB01 and were separated from avian isolate PM70 and swine isolates PM3480 and PMHN06 and are distant from Actinobacillus and Haemophilus. Previous studies based on Single Nucleotide Polymorphism (SNPs) and Multilocus Sequence Typing (MLST) unable to show a clear phylogenetic relatedness between Pasteurella multocida and the different host. In conclusion, this study has provided insight on the genomic structure of PMTB2.1 in terms of potential genes that can function as virulence factors for future study in elucidating the mechanisms behind the ability of the bacteria in causing diseases in susceptible animals.Keywords: comparative genomics, DNA sequencing, phage, phylogenomics
Procedia PDF Downloads 18227886 Identifying Protein-Coding and Non-Coding Regions in Transcriptomes
Authors: Angela U. Makolo
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Protein-coding and Non-coding regions determine the biology of a sequenced transcriptome. Research advances have shown that Non-coding regions are important in disease progression and clinical diagnosis. Existing bioinformatics tools have been targeted towards Protein-coding regions alone. Therefore, there are challenges associated with gaining biological insights from transcriptome sequence data. These tools are also limited to computationally intensive sequence alignment, which is inadequate and less accurate to identify both Protein-coding and Non-coding regions. Alignment-free techniques can overcome the limitation of identifying both regions. Therefore, this study was designed to develop an efficient sequence alignment-free model for identifying both Protein-coding and Non-coding regions in sequenced transcriptomes. Feature grouping and randomization procedures were applied to the input transcriptomes (37,503 data points). Successive iterations were carried out to compute the gradient vector that converged the developed Protein-coding and Non-coding Region Identifier (PNRI) model to the approximate coefficient vector. The logistic regression algorithm was used with a sigmoid activation function. A parameter vector was estimated for every sample in 37,503 data points in a bid to reduce the generalization error and cost. Maximum Likelihood Estimation (MLE) was used for parameter estimation by taking the log-likelihood of six features and combining them into a summation function. Dynamic thresholding was used to classify the Protein-coding and Non-coding regions, and the Receiver Operating Characteristic (ROC) curve was determined. The generalization performance of PNRI was determined in terms of F1 score, accuracy, sensitivity, and specificity. The average generalization performance of PNRI was determined using a benchmark of multi-species organisms. The generalization error for identifying Protein-coding and Non-coding regions decreased from 0.514 to 0.508 and to 0.378, respectively, after three iterations. The cost (difference between the predicted and the actual outcome) also decreased from 1.446 to 0.842 and to 0.718, respectively, for the first, second and third iterations. The iterations terminated at the 390th epoch, having an error of 0.036 and a cost of 0.316. The computed elements of the parameter vector that maximized the objective function were 0.043, 0.519, 0.715, 0.878, 1.157, and 2.575. The PNRI gave an ROC of 0.97, indicating an improved predictive ability. The PNRI identified both Protein-coding and Non-coding regions with an F1 score of 0.970, accuracy (0.969), sensitivity (0.966), and specificity of 0.973. Using 13 non-human multi-species model organisms, the average generalization performance of the traditional method was 74.4%, while that of the developed model was 85.2%, thereby making the developed model better in the identification of Protein-coding and Non-coding regions in transcriptomes. The developed Protein-coding and Non-coding region identifier model efficiently identified the Protein-coding and Non-coding transcriptomic regions. It could be used in genome annotation and in the analysis of transcriptomes.Keywords: sequence alignment-free model, dynamic thresholding classification, input randomization, genome annotation
Procedia PDF Downloads 6127885 RNA-Seq Analysis of Coronaviridae Family and SARS-Cov-2 Prediction Using Proposed ANN
Authors: Busra Mutlu Ipek, Merve Mutlu, Ahmet Mutlu
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Novel coronavirus COVID-19, which has recently influenced the world, poses a great threat to humanity. In order to overcome this challenging situation, scientists are working on developing effective vaccine against coronavirus. Many experts and researchers have also produced articles and done studies on this highly important subject. In this direction, this special topic was chosen for article to make a contribution to this area. The purpose of this article is to perform RNA sequence analysis of selected virus forms in the Coronaviridae family and predict/classify SARS-CoV-2 (COVID-19) from other selected complete genomes in coronaviridae family using proposed Artificial Neural Network(ANN) algorithm.Keywords: Coronaviridae family, COVID-19, RNA sequencing, ANN, neural network
Procedia PDF Downloads 13927884 Analysis of Exponential Nonuniform Transmission Line Parameters
Authors: Mounir Belattar
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In this paper the Analysis of voltage waves that propagate along a lossless exponential nonuniform line is presented. For this analysis the parameters of this line are assumed to be varying function of the distance x along the line from the source end. The approach is based on the tow-port networks cascading presentation to derive the ABDC parameters of transmission using Picard-Carson Method which is a powerful method in getting a power series solution for distributed network because it is easy to calculate poles and zeros and solves differential equations such as telegrapher equations by an iterative sequence. So the impedance, admittance voltage and current along the line are expanded as a Taylor series in x/l where l is the total length of the line to obtain at the end, the main transmission line parameters such as voltage response and transmission and reflexion coefficients represented by scattering parameters in frequency domain.Keywords: ABCD parameters, characteristic impedance exponential nonuniform transmission line, Picard-Carson's method, S parameters, Taylor's series
Procedia PDF Downloads 43727883 Extending Image Captioning to Video Captioning Using Encoder-Decoder
Authors: Sikiru Ademola Adewale, Joe Thomas, Bolanle Hafiz Matti, Tosin Ige
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This project demonstrates the implementation and use of an encoder-decoder model to perform a many-to-many mapping of video data to text captions. The many-to-many mapping occurs via an input temporal sequence of video frames to an output sequence of words to form a caption sentence. Data preprocessing, model construction, and model training are discussed. Caption correctness is evaluated using 2-gram BLEU scores across the different splits of the dataset. Specific examples of output captions were shown to demonstrate model generality over the video temporal dimension. Predicted captions were shown to generalize over video action, even in instances where the video scene changed dramatically. Model architecture changes are discussed to improve sentence grammar and correctness.Keywords: decoder, encoder, many-to-many mapping, video captioning, 2-gram BLEU
Procedia PDF Downloads 9327882 Effects of Evening vs. Morning Training on Motor Skill Consolidation in Morning-Oriented Elderly
Authors: Maria Korman, Carmit Gal, Ella Gabitov, Avi Karni
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The main question addressed in this study was whether the time-of-day wherein training is afforded is a significant factor for motor skill ('how-to', procedural knowledge) acquisition and consolidation into long term memory in the healthy elderly population. Twenty-nine older adults (60-75 years) practiced an explicitly instructed 5-element key-press sequence by repeatedly generating the sequence ‘as fast and accurately as possible’. Contribution of three parameters to acquisition, 24h post-training consolidation, and 1-week retention gains in motor sequence speed was assessed: (a) time of training (morning vs. evening group) (b) sleep quality (actigraphy) and (c) chronotype. All study participants were moderately morning type, according to the Morningness-Eveningness Questionnaire score. All participants had sleep patterns typical of age, with average sleep efficiency of ~ 82%, and approximately 6 hours of sleep. Speed of motor sequence performance in both groups improved to a similar extent during training session. Nevertheless, evening group expressed small but significant overnight consolidation phase gains, while morning group showed only maintenance of performance level attained at the end of training. By 1-week retention test, both groups showed similar performance levels with no significant gains or losses with respect to 24h test. Changes in the tapping patterns at 24h and 1-week post-training were assessed based on normalized Pearson correlation coefficients using the Fisher’s z-transformation in reference to the tapping pattern attained at the end of the training. Significant differences between the groups were found: the evening group showed larger changes in tapping patterns across the consolidation and retention windows. Our results show that morning-oriented older adults effectively acquired, consolidated, and maintained a new sequence of finger movements, following both morning and evening practice sessions. However, time-of-training affected the time-course of skill evolution in terms of performance speed, as well as the re-organization of tapping patterns during the consolidation period. These results are in line with the notion that motor training preceding a sleep interval may be beneficial for the long-term memory in the elderly. Evening training should be considered an appropriate time window for motor skill learning in older adults, even in individuals with morning chronotype.Keywords: time-of-day, elderly, motor learning, memory consolidation, chronotype
Procedia PDF Downloads 12927881 Molecular Characterisation and Expression of Glutathione S-Transferase of Fasciola Gigantica
Authors: J. Adeppa, S. Samanta, O. K. Raina
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Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.Keywords: fasciola gigantic, fasciola hepatica, GST, RT- PCR
Procedia PDF Downloads 18027880 Genetic Diversity Analysis in Triticum Aestivum Using Microsatellite Markers
Authors: Prachi Sharma, Mukesh Kumar Rana
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In the present study, the simple sequence repeat(SSR) markers have been used in analysis of genetic diversity of 37 genotypes of Triticum aestivum. The DNA was extracted using cTAB method. The DNA was quantified using the fluorimeter. The annealing temperatures for 27 primer pairs were standardized using gradient PCR, out of which 16 primers gave satisfactory amplification at temperature ranging from 50-62⁰ C. Out of 16 polymorphic SSR markers only 10 SSR primer pairs were used in the study generating 34 reproducible amplicons among 37 genotypes out of which 30 were polymorphic. Primer pairs Xgwm533, Xgwm 160, Xgwm 408, Xgwm 120, Xgwm 186, Xgwm 261 produced maximum percent of polymorphic bands (100%). The bands ranged on an average of 3.4 bands per primer. The genetic relationship was determined using Jaccard pair wise similarity co-efficient and UPGMA cluster analysis with NTSYS Pc.2 software. The values of similarity index range from 0-1. The similarity coefficient ranged from 0.13 to 0.97. A minimum genetic similarity (0.13) was observed between VL 804 and HPW 288, meaning they are only 13% similar. More number of available SSR markers can be useful for supporting the genetic diversity analysis in the above wheat genotypes.Keywords: wheat, genetic diversity, microsatellite, polymorphism
Procedia PDF Downloads 60227879 Rapid Discrimination of Porcine and Tilapia Fish Gelatin by Fourier Transform Infrared- Attenuated Total Reflection Combined with 2 Dimensional Infrared Correlation Analysis
Authors: Norhidayu Muhamad Zain
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Gelatin, a purified protein derived mostly from porcine and bovine sources, is used widely in food manufacturing, pharmaceutical, and cosmetic industries. However, the presence of any porcine-related products are strictly forbidden for Muslim and Jewish consumption. Therefore, analytical methods offering reliable results to differentiate the sources of gelatin are needed. The aim of this study was to differentiate the sources of gelatin (porcine and tilapia fish) using Fourier transform infrared- attenuated total reflection (FTIR-ATR) combined with two dimensional infrared (2DIR) correlation analysis. Porcine gelatin (PG) and tilapia fish gelatin (FG) samples were diluted in distilled water at concentrations ranged from 4-20% (w/v). The samples were then analysed using FTIR-ATR and 2DIR correlation software. The results showed a significant difference in the pattern map of synchronous spectra at the region of 1000 cm⁻¹ to 1100 cm⁻¹ between PG and FG samples. The auto peak at 1080 cm⁻¹ that attributed to C-O functional group was observed at high intensity in PG samples compared to FG samples. Meanwhile, two auto peaks (1080 cm⁻¹ and 1030 cm⁻¹) at lower intensity were identified in FG samples. In addition, using 2D correlation analysis, the original broad water OH bands in 1D IR spectra can be effectively differentiated into six auto peaks located at 3630, 3340, 3230, 3065, 2950 and 2885 cm⁻¹ for PG samples and five auto peaks at 3630, 3330, 3230, 3060 and 2940 cm⁻¹ for FG samples. Based on the rule proposed by Noda, the sequence of the spectral changes in PG samples is as following: NH₃⁺ amino acid > CH₂ and CH₃ aliphatic > OH stretch > carboxylic acid OH stretch > NH in secondary amide > NH in primary amide. In contrast, the sequence was totally in the opposite direction for FG samples and thus both samples provide different 2D correlation spectra ranged from 2800 cm-1 to 3700 cm⁻¹. This method may provide a rapid determination of gelatin source for application in food, pharmaceutical, and cosmetic products.Keywords: 2 dimensional infrared (2DIR) correlation analysis, Fourier transform infrared- attenuated total reflection (FTIR-ATR), porcine gelatin, tilapia fish gelatin
Procedia PDF Downloads 23627878 Modification of Escherichia coli PtolT Expression Vector via Site-Directed Mutagenesis
Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey
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Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression
Procedia PDF Downloads 36427877 Chaotic Sequence Noise Reduction and Chaotic Recognition Rate Improvement Based on Improved Local Geometric Projection
Authors: Rubin Dan, Xingcai Wang, Ziyang Chen
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A chaotic time series noise reduction method based on the fusion of the local projection method, wavelet transform, and particle swarm algorithm (referred to as the LW-PSO method) is proposed to address the problem of false recognition due to noise in the recognition process of chaotic time series containing noise. The method first uses phase space reconstruction to recover the original dynamical system characteristics and removes the noise subspace by selecting the neighborhood radius; then it uses wavelet transform to remove D1-D3 high-frequency components to maximize the retention of signal information while least-squares optimization is performed by the particle swarm algorithm. The Lorenz system containing 30% Gaussian white noise is simulated and verified, and the phase space, SNR value, RMSE value, and K value of the 0-1 test method before and after noise reduction of the Schreiber method, local projection method, wavelet transform method, and LW-PSO method are compared and analyzed, which proves that the LW-PSO method has a better noise reduction effect compared with the other three common methods. The method is also applied to the classical system to evaluate the noise reduction effect of the four methods and the original system identification effect, which further verifies the superiority of the LW-PSO method. Finally, it is applied to the Chengdu rainfall chaotic sequence for research, and the results prove that the LW-PSO method can effectively reduce the noise and improve the chaos recognition rate.Keywords: Schreiber noise reduction, wavelet transform, particle swarm optimization, 0-1 test method, chaotic sequence denoising
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