Search results for: gene selection
3188 Analysis of Saudi Breast Cancer Patients’ Primary Tumors using Array Comparative Genomic Hybridization
Authors: L. M. Al-Harbi, A. M. Shokry, J. S. M. Sabir, A. Chaudhary, J. Manikandan, K. S. Saini
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Breast cancer is the second most common cause of cancer death worldwide and is the most common malignancy among Saudi females. During breast carcinogenesis, a wide-array of cytogenetic changes involving deletions, or amplification, or translocations, of part or whole of chromosome regions have been observed. Because of the limitations of various earlier technologies, newer tools are developed to scan for changes at the genomic level. Recently, Array Comparative Genomic Hybridization (aCGH) technique has been applied for detecting segmental genomic alterations at molecular level. In this study, aCGH was performed on twenty breast cancer tumors and their matching non-tumor (normal) counterparts using the Agilent 2x400K. Several regions were identified to be either amplified or deleted in a tumor-specific manner. Most frequent alterations were amplification of chromosome 1q, chromosome 8q, 20q, and deletions at 16q were also detected. The amplification of genetic events at 1q and 8q were further validated using FISH analysis using probes targeting 1q25 and 8q (MYC gene). The copy number changes at these loci can potentially cause a significant change in the tumor behavior, as deletions in the E-Cadherin (CDH1)-tumor suppressor gene as well as amplification of the oncogenes-Aurora Kinase A. (AURKA) and MYC could make these tumors highly metastatic. This study validates the use of aCGH in Saudi breast cancer patients and sets the foundations necessary for performing larger cohort studies searching for ethnicity-specific biomarkers and gene copy number variations.Keywords: breast cancer, molecular biology, ecology, environment
Procedia PDF Downloads 3703187 Managing the Local Manager: A Comparative Study of Core HRM Functions in Multinationals
Authors: Maria Khan
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Framing good core Human Resource Management (HRM) functions like recruitment, selection, training and development, which if executed effectively, can become a strategic advantage for a company. HRM policies related to mid-level managers can depend on the type of top management. This may be due to the difference in perception of effective HRM policies of an expatriate and local leadership. This comparative case study assesses how local mid-level managers are managed in leading multinational telecom companies in Pakistan. Core HRM functions related to managers were analysed through field research based on semi-structured interviews with relevant Human Resource Managers. Results suggest that recruitment and selection practices are not too different and are in compliance with best HRM practices. However, there is a difference in the effective implementation of Training and Development policies. Changing global management trends and skill development dictate that MNCs continuously develop the local talent effectively for local and international success.Keywords: recruitment, selection, training, development, core HRM, human resource management, subsidiary, international staffing, managers, MNC, expatriate
Procedia PDF Downloads 3173186 Evaluation of QSRR Models by Sum of Ranking Differences Approach: A Case Study of Prediction of Chromatographic Behavior of Pesticides
Authors: Lidija R. Jevrić, Sanja O. Podunavac-Kuzmanović, Strahinja Z. Kovačević
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The present study deals with the selection of the most suitable quantitative structure-retention relationship (QSRR) models which should be used in prediction of the retention behavior of basic, neutral, acidic and phenolic pesticides which belong to different classes: fungicides, herbicides, metabolites, insecticides and plant growth regulators. Sum of ranking differences (SRD) approach can give a different point of view on selection of the most consistent QSRR model. SRD approach can be applied not only for ranking of the QSRR models, but also for detection of similarity or dissimilarity among them. Applying the SRD analysis, the most similar models can be found easily. In this study, selection of the best model was carried out on the basis of the reference ranking (“golden standard”) which was defined as the row average values of logarithm of retention time (logtr) defined by high performance liquid chromatography (HPLC). Also, SRD analysis based on experimental logtr values as reference ranking revealed similar grouping of the established QSRR models already obtained by hierarchical cluster analysis (HCA).Keywords: chemometrics, chromatography, pesticides, sum of ranking differences
Procedia PDF Downloads 3693185 An Efficient Strategy for Relay Selection in Multi-Hop Communication
Authors: Jung-In Baik, Seung-Jun Yu, Young-Min Ko, Hyoung-Kyu Song
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This paper proposes an efficient relaying algorithm to obtain diversity for improving the reliability of a signal. The algorithm achieves time or space diversity gain by multiple versions of the same signal through two routes. Relays are separated between a source and destination. The routes between the source and destination are set adaptive in order to deal with different channels and noises. The routes consist of one or more relays and the source transmits its signal to the destination through the routes. The signals from the relays are combined and detected at the destination. The proposed algorithm provides a better performance than the conventional algorithms in bit error rate (BER).Keywords: multi-hop, OFDM, relay, relaying selection
Procedia PDF Downloads 4403184 Oncogenic Functions of Long Non-Coding RNA XIST in Human Nasopharyngeal Carcinoma by Targeting MiR-34a-5p
Authors: Cheng-Cao Sun, Shu-Jun Li, De-Jia Li
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Long non-coding RNA (lncRNA) X inactivate-specific transcript (XIST) has been verified as an oncogenic gene in several human malignant tumors, and its dysregulation was closed associated with tumor initiation, development and progression. Nevertheless, whether the aberrant expression of XIST in human nasopharyngeal carcinoma (NPC) is corrected with malignancy, metastasis or prognosis has not been elaborated. Here, we discovered that XIST was up-regulated in NPC tissues and higher expression of XIST contributed to a markedly poorer survival time. In addition, multivariate analysis demonstrated XIST was an independent risk factor for prognosis. XIST over-expression enhanced, while XIST silencing hampered the cell growth in NPC. Additionally, mechanistic analysis revealed that XIST up-regulated the expression of miR-34a-5p targeted gene E2F3 through acting as a competitive ‘sponge’ of miR-34a-5p. Taking all into account, we concluded that XIST functioned as an oncogene in NPC through up-regulating E2F3 in part through ‘spongeing’ miR-34a-5p.Keywords: X inactivate-specific transcript; hsa-miRNA-34a-5p, miR-34a-5p; E2F3, nasopharyngeal carcinoma, tumorigenesis
Procedia PDF Downloads 2353183 Machine Learning Assisted Prediction of Sintered Density of Binary W(MO) Alloys
Authors: Hexiong Liu
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Powder metallurgy is the optimal method for the consolidation and preparation of W(Mo) alloys, which exhibit excellent application prospects at high temperatures. The properties of W(Mo) alloys are closely related to the sintered density. However, controlling the sintered density and porosity of these alloys is still challenging. In the past, the regulation methods mainly focused on time-consuming and costly trial-and-error experiments. In this study, the sintering data for more than a dozen W(Mo) alloys constituted a small-scale dataset, including both solid and liquid phases of sintering. Furthermore, simple descriptors were used to predict the sintered density of W(Mo) alloys based on the descriptor selection strategy and machine learning method (ML), where the ML algorithm included the least absolute shrinkage and selection operator (Lasso) regression, k-nearest neighbor (k-NN), random forest (RF), and multi-layer perceptron (MLP). The results showed that the interpretable descriptors extracted by our proposed selection strategy and the MLP neural network achieved a high prediction accuracy (R>0.950). By further predicting the sintered density of W(Mo) alloys using different sintering processes, the error between the predicted and experimental values was less than 0.063, confirming the application potential of the model.Keywords: sintered density, machine learning, interpretable descriptors, W(Mo) alloy
Procedia PDF Downloads 753182 Obesity, Leptin Levels and Leptin Receptor Gene Polymorphisms in Afro-Caribbean Subjects
Authors: Lydia Foucan, Christine Rambhojan, Rachel Billy, Christophe Armand, Carl-Thony Michel, Jean-Marc Lacorte, Laurent Larifla
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Leptin, an adipocyte-derived hormone, modulates insulin secretion and action via the leptin receptor (LEPR) that is expressed in pancreatic beta cells, adipose tissue, and muscle. Several polymorphisms have been described in the human LEPR gene including p.K109R (rs1137100), p.Q223R (rs1137101) and p.K656N (rs1805094) polymorphisms. The role of these polymorphisms is not yet studied in Guadeloupian population. Our aim was to explore the association of LEPR polymorphisms (K109R, Q223R and K656N) with leptin levels and obesity in non-diabetic Afro-Caribbean subjects. Genotypic analysis of the three polymorphisms was performed in 425 subjects using TaqMan and KASPar Assays. Serum leptin was measured with ELISA kits Biovendor® (RD191001100). Logistic regressions were used for assessment of statistical associations. Mean age was 47.6 ± 12.7 years. Among the participants, 238 (56 %) were women, 124 (30%) were obese and 155 (36.5%) had abdominal obesity. Carriers of LEPR K656N rs1805094 rare allele had significant higher frequencies of obesity (P = 0.007), abdominal obesity (P = 0.004) and metabolic syndrome (P = 0.021) but mean leptin level was not significantly different between both groups (P = 0.075). Odds ratios, adjusted for age and sex associated with presence of rs1805094 rare allele were 1.8 (1.1-2.9), P = 0.012 for obesity, 2.0 (1.2-3.3), P = 0.008 for abdominal obesity and 1.8 (1.1-3.0), P = 0.031 for MetS. No significant association was found with K109R, Q223R. These findings suggest that the K656N polymorphism (but not the K109R or Q223R polymorphism) of LEPR is associated with obesity, abdominal obesity and metabolic syndrome in this Afro-Caribbean non-diabetic population.Keywords: Afro-Caribbean, leptin levels, leptin receptor gene polymorphisms, obesity
Procedia PDF Downloads 3713181 Impact of CYP3A5 Polymorphism on Tacrolimus to Predict the Optimal Initial Dose Requirements in South Indian Renal Transplant Recipients
Authors: S. Sreeja, Radhakrishnan R. Nair, Noble Gracious, Sreeja S. Nair, M. Radhakrishna Pillai
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Background: Tacrolimus is a potent immunosuppressant clinically used for the long term treatment of antirejection of transplanted organs in liver and kidney transplant recipients though dose optimization is poorly managed. However, So far no study has been carried out on the South Indian kidney transplant patients. The objective of this study is to evaluate the potential influence of a functional polymorphism in CYP3A5*3 gene on tacrolimus physiological availability/dose ratio in South Indian renal transplant patients. Materials and Methods: Twenty five renal transplant recipients receiving tacrolimus were enrolled in this study. Their body weight, drug dosage, and therapeutic concentration of Tacrolimus were observed. All patients were on standard immunosuppressive regime of Tacrolimus-Mycophenolate mofetil along with steroids on a starting dose of Tac 0.1 mg/kg/day. CYP3A5 genotyping was performed by PCR followed with RFLP. Conformation of RFLP analysis and variation in the nucleotide sequence of CYP3A5*3 gene were determined by direct sequencing using a validated automated generic analyzer. Results: A significant association was found between tacrolimus per dose/kg/d and CYP3A5 gene (A6986G) polymorphism in the study population. The CYP3A5 *1/*1, *1/*3 and *3/*3 genotypes were detected in 5 (20 %), 5 (20 %) and 15 (60 %) of the 25 graft recipients, respectively. CYP3A5*3 genotypes were found to be a good predictor of tacrolimus Concentration/Dose ratio in kidney transplant recipients. Significantly higher L/D was observed among non-expressors 9.483 ng/mL(4.5- 14.1) as compared with the expressors 5.154 ng/mL (4.42-6.5 ) of CYP3A5. Acute rejection episodes were significantly higher for CYP3A5*1 homozygotes compared to patients with CYP3A5*1/*3 and CYP3A5*3/*3 genotypes (40 % versus 20 % and 13 %, respectively ). The dose normalized TAC concentration (ng/ml/mg/kg) was significantly lower in patients having CYP3A5*1/*3 polymorphism. Conclusion: This is the first study to extensively determine the effect of CYP3A5*3 genetic polymorphism on tacrolimus pharmacokinetics in South Indian renal transplant recipients and also shows that majority of our patients carry mutant allele A6986G in CYP3A5*3 gene. Identification of CYP3A5 polymorphism prior to transplantation could contribute to evaluate the appropriate initial dosage of tacrolimus for each patient.Keywords: kidney transplant patients, CYP3A5 genotype, tacrolimus, RFLP
Procedia PDF Downloads 2943180 hsa-miR-1204 and hsa-miR-639 Prominent Role in Tamoxifen's Molecular Mechanisms on the EMT Phenomenon in Breast Cancer Patients
Authors: Mahsa Taghavi
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In the treatment of breast cancer, tamoxifen is a regularly prescribed medication. The effect of tamoxifen on breast cancer patients' EMT pathways was studied. In this study to see if it had any effect on the cancer cells' resistance to tamoxifen and to look for specific miRNAs associated with EMT. In this work, we used continuous and integrated bioinformatics analysis to choose the optimal GEO datasets. Once we had sorted the gene expression profile, we looked at the mechanism of signaling, the ontology of genes, and the protein interaction of each gene. In the end, we used the GEPIA database to confirm the candidate genes. after that, I investigated critical miRNAs related to candidate genes. There were two gene expression profiles that were categorized into two distinct groups. Using the expression profile of genes that were lowered in the EMT pathway, the first group was examined. The second group represented the polar opposite of the first. A total of 253 genes from the first group and 302 genes from the second group were found to be common. Several genes in the first category were linked to cell death, focal adhesion, and cellular aging. Two genes in the second group were linked to cell death, focal adhesion, and cellular aging. distinct cell cycle stages were observed. Finally, proteins such as MYLK, SOCS3, and STAT5B from the first group and BIRC5, PLK1, and RAPGAP1 from the second group were selected as potential candidates linked to tamoxifen's influence on the EMT pathway. hsa-miR-1204 and hsa-miR-639 have a very close relationship with the candidates genes according to the node degrees and betweenness index. With this, the action of tamoxifen on the EMT pathway was better understood. It's important to learn more about how tamoxifen's target genes and proteins work so that we can better understand the drug.Keywords: tamoxifen, breast cancer, bioinformatics analysis, EMT, miRNAs
Procedia PDF Downloads 1263179 FDX1, a Cuproptosis-Related Gene, Identified as a Potential Target for Human Ovarian Aging
Authors: Li-Te Lin, Chia-Jung Li, Kuan-Hao Tsui
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Cuproptosis, a newly identified cell death mechanism, has attracted attention for its association with various diseases. However, the genetic interplay between cuproptosis and ovarian aging remains largely unexplored. This study aims to address this gap by analyzing datasets related to ovarian aging and cuproptosis. Spatial transcriptome analyses were conducted in the ovaries of both young and aged female mice to elucidate the role of FDX1. Comprehensive bioinformatics analyses, facilitated by R software, identified FDX1 as a potential cuproptosis-related gene with implications for ovarian aging. Clinical infertility biopsies were examined to validate these findings, showing consistent results in elderly infertile patients. Furthermore, pharmacogenomic analyses of ovarian cell lines explored the intricate association between FDX1 expression levels and sensitivity to specific small molecule drugs. Spatial transcriptome analyses revealed a significant reduction in FDX1 expression in aging ovaries, supported by consistent findings in biopsies from elderly infertile patients. Pharmacogenomic investigations indicated that modulating FDX1 could influence drug responses in ovarian-related therapies. This study pioneers the identification of FDX1 as a cuproptosis-related gene linked to ovarian aging. These findings not only contribute to understanding the mechanisms of ovarian aging but also position FDX1 as a potential diagnostic biomarker and therapeutic target. Further research may establish FDX1's pivotal role in advancing precision medicine and therapies for ovarian-related conditions.Keywords: cuproptosis, FDX1, ovarian aging, biomarker
Procedia PDF Downloads 343178 Association between a Serotonin Re-Uptake Transporter Gene Polymorphism and Mucosal Serotonin Level in Women Patients with Irritable Bowel Syndrome and Healthy Control: A Pilot Study from Northern India
Authors: Sunil Kumar, Uday C. Ghoshal
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Background and aims: Serotonin (5-hydroxtryptamine, 5-HT) is an important factor in gut function, playing key roles in intestinal peristalsis and secretion, and in sensory signaling in the brain-gut axis. Removal from its sites of action is mediated by a specific protein called the serotonin reuptake transporter (SERT). Polymorphisms in the promoter region of the SERT gene have effects on transcriptional activity, resulting in altered 5-HT reuptake efficiency. Functional polymorphisms may underlie disturbance in gut function in individuals suffering with disorders such as irritable bowel syndrome (IBS). The aim of this study was to assess the potential association between SERT polymorphisms and the diarrhea predominant IBS (D-IBS) phenotype Subjects: A total of 36 northern Indian female patients and 55 female northern Indian healthy controls (HC) were subjected to genotyping. Methods: Leucocyte DNA of all subjects was analyzed by polymerase chain reaction based technologies for SERT polymorphisms, specifically the insertion/deletion polymorphism in the promoter (SERT-P). Statistical analysis was performed to assess association of SERT polymorphism allele with the D-IBS phenotype. Results: The frequency of distribution of SERT-P gene was comparable between female patients with IBS and HC (p = 0.086). However, frequency of SERT-P deletion/deletion genotype was significantly higher in female patients with D-IBS compared to C-IBS and A-IBS [17/19 (89.5%) vs. 4/12 (33.3%) vs. 1/5 (20%), p=0.001, respectively]. The mucosal level of serotonin was higher in D-IBS compared to C-IBS and A-IBS [Median, range (159.26, 98.78–212.1) vs. 110.4, 67.87–143.53 vs. 92.34, 78.8–166.3 pmol/mL, p=0.001, respectively]. The mucosal level of serotonin was higher in female patients with IBS with SERT-P deletion/deletion genotype compared deletion/insertion and insertion/insertion [157.65, 67.87–212.1 vs. 110.4, 78.1–143.32 vs. 100.5, 69.1–132.03 pmol/mL, p=0.001, respectively]. Patients with D-IBS with deletion/deletion genotype more often reported symptoms of abdominal pain, discomfort (p=0.025) and bloating (p=0.039). Symptoms development following lactose ingestion was strongly associated with D-IBS and SERT-P deletion/deletion genotype (p=0.004). Conclusions: Significant association was observed between D-IBS and the SERT-P deletion/deletion genotype, suggesting that the serotonin transporter is a potential candidate gene for D-IBS in women.Keywords: serotonin, SERT, inflammatory bowel disease, genetic polymorphism
Procedia PDF Downloads 3313177 Identification of the Target Genes to Increase the Immunotherapy Response in Bladder Cancer Patients using Computational and Experimental Approach
Authors: Sahar Nasr, Lin Li, Edwin Wang
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Bladder cancer (BLCA) is known as the 13th cause of death among cancer patients worldwide, and ~575,000 new BLCA cases are diagnosed each year. Urothelial carcinoma (UC) is the most prevalent subtype among BLCA patients, which can be categorized into muscle-invasive bladder cancer (MIBC) and non-muscle-invasive bladder cancer (NMIBC). Currently, various therapeutic options are available for UC patients, including (1) transurethral resection followed by intravesical instillation of chemotherapeutics or Bacillus Calmette-Guérin for NMIBC patients, (2) neoadjuvant platinum-based chemotherapy (NAC) plus radical cystectomy is the standard of care for localized MIBC patients, and (3) systematic chemotherapy for metastatic UC. However, conventional treatments may lead to several challenges for treating patients. As an illustration, some patients may suffer from recurrence of the disease after the first line of treatment. Recently, immune checkpoint therapy (ICT) has been introduced as an alternative treatment strategy for the first or second line of treatment in advanced or metastatic BLCA patients. Although ICT showed lucrative results for a fraction of BLCA patients, ~80% of patients were not responsive to it. Therefore, novel treatment methods are required to augment the ICI response rate within BLCA patients. It has been shown that the infiltration of T-cells into the tumor microenvironment (TME) is positively correlated with the response to ICT within cancerous patients. Therefore, the goal of this study is to enhance the infiltration of cytotoxic T-cells into TME through the identification of target genes within the tumor that are responsible for the non-T-cell inflamed TME and their inhibition. BLCA bulk RNA-sequencing data from The Cancer Genome Atlas (TCGA) and immune score for TCGA samples were used to determine the Pearson correlation score between the expression of different genes and immune score for each sample. The genes with strong negative correlations were selected (r < -0.2). Thereafter, the correlation between the expression of each gene and survival in BLCA patients was calculated using the TCGA data and Cox regression method. The genes that are common in both selected gene lists were chosen for further analysis. Afterward, BLCA bulk and single-cell RNA-sequencing data were ranked based on the expression of each selected gene and the top and bottom 25% samples were used for pathway enrichment analysis. If the pathways related to the T-cell infiltration (e.g., antigen presentation, interferon, or chemokine pathways) were enriched within the low-expression group, the gene was included for downstream analysis. Finally, the selected genes will be used to calculate the correlation between their expression and the infiltration rate of the activated CD+8 T-cells, natural killer cells and the activated dendric cells. A list of potential target genes has been identified and ranked based on the above-mentioned analysis and criteria. SUN-1 got the highest score within the gene list and other identified genes in the literature as benchmarks. In conclusion, inhibition of SUN1 may increase the tumor-infiltrating lymphocytes and the efficacy of ICI in BLCA patients. BLCA tumor cells with and without SUN-1 CRISPR/Cas9 knockout will be injected into the syngeneic mouse model to validate the predicted SUN-1 effect on increasing tumor-infiltrating lymphocytes.Keywords: data analysis, gene expression analysis, gene identification, immunoinformatic, functional genomics, transcriptomics
Procedia PDF Downloads 1503176 Multi-Class Text Classification Using Ensembles of Classifiers
Authors: Syed Basit Ali Shah Bukhari, Yan Qiang, Saad Abdul Rauf, Syed Saqlaina Bukhari
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Text Classification is the methodology to classify any given text into the respective category from a given set of categories. It is highly important and vital to use proper set of pre-processing , feature selection and classification techniques to achieve this purpose. In this paper we have used different ensemble techniques along with variance in feature selection parameters to see the change in overall accuracy of the result and also on some other individual class based features which include precision value of each individual category of the text. After subjecting our data through pre-processing and feature selection techniques , different individual classifiers were tested first and after that classifiers were combined to form ensembles to increase their accuracy. Later we also studied the impact of decreasing the classification categories on over all accuracy of data. Text classification is highly used in sentiment analysis on social media sites such as twitter for realizing people’s opinions about any cause or it is also used to analyze customer’s reviews about certain products or services. Opinion mining is a vital task in data mining and text categorization is a back-bone to opinion mining.Keywords: Natural Language Processing, Ensemble Classifier, Bagging Classifier, AdaBoost
Procedia PDF Downloads 2263175 Phylogenetic Relationships of the Malaysian Primates Cercopithecine Based on COI Gene Sequences
Authors: B. M. Md-Zain, N. A. Rahman, M. A. B. Abdul-Latiff, W. M. R. Idris
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We conducted molecular research to portray phylogenetic relationships of Malaysian primates particularly in the genus of Macaca. We have sequenced cytochrome C oxidase subunit I (COI) of mitochondrial DNA of several individuals from M. fascicularis and M. arctoides. PCR amplifications were performed and COI DNA sequences were aligned using ClustalW. Phylogenetic trees were constructed using distance analyses by employing neighbor-joining algorithm (NJ). We managed to sequence 700 bp of COI DNA sequences. The tree topology showed that M. fascicularis did not clump based on phyleogeography division in Peninsular Malaysia. Individuals from Negeri Sembilan merged together with samples from Perak and Penang into one clade. In addition, phylogenetic analyses indicated that M. arctoides was classified into sinica group instead of fascicularis group supported by genetic distance data. COI gene is an effective locus to clarify phylogenetic position of M. arctoides but not in discriminating M. fascicularis population in Peninsular Malaysia.Keywords: cercopithecine, long-tailed macaque, Macaca fascicularis, Macaca arctoides
Procedia PDF Downloads 3513174 Metaheuristic to Align Multiple Sequences
Authors: Lamiche Chaabane
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In this study, a new method for solving sequence alignment problem is proposed, which is named ITS (Improved Tabu Search). This algorithm is based on the classical Tabu Search (TS). ITS is implemented in order to obtain results of multiple sequence alignment. Several ideas concerning neighbourhood generation, move selection mechanisms and intensification/diversification strategies for our proposed ITS is investigated. ITS have generated high-quality results in terms of measure of scores in comparison with the classical TS and simple iterative search algorithm.Keywords: multiple sequence alignment, tabu search, improved tabu search, neighbourhood generation, selection mechanisms
Procedia PDF Downloads 2973173 Criterion-Referenced Test Reliability through Threshold Loss Agreement: Fuzzy Logic Analysis Approach
Authors: Mohammad Ali Alavidoost, Hossein Bozorgian
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Criterion-referenced tests (CRTs) are designed to measure student performance against a fixed set of predetermined criteria or learning standards. The reliability of such tests cannot be based on internal reliability. Threshold loss agreement is one way to calculate the reliability of CRTs. However, the selection of master and non-master in such agreement is determined by the threshold point. The problem is if the threshold point witnesses a minute change, the selection of master and non-master may have a drastic change, leading to the change in reliability results. Therefore, in this study, the Fuzzy logic approach is employed as a remedial procedure for data analysis to obviate the threshold point problem. Forty-one Iranian students were selected; the participants were all between 20 and 30 years old. A quantitative approach was used to address the research questions. In doing so, a quasi-experimental design was utilized since the selection of the participants was not randomized. Based on the Fuzzy logic approach, the threshold point would be more stable during the analysis, resulting in rather constant reliability results and more precise assessment.Keywords: criterion-referenced tests, threshold loss agreement, threshold point, fuzzy logic approach
Procedia PDF Downloads 3623172 Genetic Diversity of Exon-20 of the IIS6 of the Voltage Gated Sodium Channel Gene from Pyrethroid Resistant Anopheles Mosquitoes in Sudan Savannah Region of Jigawa State
Authors: Asma'u Mahe, Abdullahi A. Imam, Adamu J. Alhassan, Nasiru Abdullahi, Sadiya A. Bichi, Nura Lawal, Kamaluddeen Babagana
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Malaria is a disease with global health significance. It is caused by parasites and transmitted by Anopheles mosquitoes. Increase in insecticide resistance threatens the disease vector control. The strength of selection pressure acting on a mosquito population in relation to insecticide resistance can be assess by determining the genetic diversity of a fragment spanning exon- 20 of IIS6 of the voltage gated sodium channel (VGSC). Larval samples reared to adulthood were identified and kdr (knock down resistance) profile was determined. The DNA sequences were used to assess the patterns of genetic differentiation by determining the levels of genetic variability between the Anopheles mosquitoes. Genetic differentiation of the Anopheles mosquitoes based on a portion of the voltage gated sodium channel gene was obtained. Polymorphisms were detected; sequence variation and analysis were presented as a phylogenetic tree. Phylogenetic tree of VGSC haplotypes was constructed for samples of the Anopheles mosquitoes using the maximum likelihood method in MEGA 6.0 software. DNA sequences were edited using BioEdit sequence editor. The edited sequences were aligned with reference sequence (Kisumu strain). Analyses were performed as contained in dnaSP 5.10. Results of genetic parameters of polymorphism and haplotype reconstruction were presented in count. Twenty sequences were used for the analysis. Regions selected were 1- 576, invariable (monomorphic) sites were 460 while variable (polymorphic) sites were 5 giving the number of total mutations observed in this study. Mutations obtained from the study were at codon 105: TTC- Phenylalanine replaces TCC- Serine, codon 513: TAG- Termination replaces TTG- Leucine, codon 153, 300 and 553 mutations were non-synonymous. From the constructed phylogenetic tree, some groups were shown to be closer with Exon20Gambiae Kisumu (Reference strain) having some genetic distance, while 5-Exon20Gambiae-F I13.ab1, 18-Exon20Gambiae-F C17.ab1, and 2-Exon20Gambiae-F C13.ab1 clustered together genetically differentiated away from others. Mutations observed in this study can be attributed to the high insecticide resistance profile recorded in the study areas. Haplotype networks of pattern of genetic variability and polymorphism for the fragment of the VGSC sequences of sampled Anopheles mosquitoes revealed low haplotypes for the present study. Haplotypes are set of closely linked DNA variation on X-chromosome. Haplotypes were scaled accordingly to reflect their respective frequencies. Low haplotype number, four VGSC-1014F haplotypes were observed in this study. A positive association was previously established between low haplotype number of VGSC diversity and pyrethroid resistance through kdr mechanism. Significant values at (P < 0.05) of Tajima D and Fu and Li D’ were observed for some of the results indicating possible signature of positive selection on the fragment of VGSC in the study. This is the first report of VGSC-1014F in the study site. Based on the results, the mutation was present in low frequencies. However, the roles played by the observed mutations need further investigation. Mutations, environmental factors among others can affect genetic diversity. The study area has recorded increase in insecticide resistance that can affect vector control in the area. This finding might affect the efforts made against malaria. Sequences were deposited in GenBank for Accession Number.Keywords: anopheles mosquitoes, insecticide resistance, kdr, malaria, voltage gated sodium channel
Procedia PDF Downloads 523171 Association of 105A/C IL-18 Gene Single Nucleotide Polymorphism with House Dust Mite Allergy in an Atopic Filipino Population
Authors: Eisha Vienna M. Fernandez, Cristan Q. Cabanilla, Hiyasmin Lim, John Donnie A. Ramos
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Allergy is a multifactorial disease affecting a significant proportion of the population. It is developed through the interaction of allergens and the presence of certain polymorphisms in various susceptibility genes. In this study, the correlation of the 105A/C single nucleotide polymorphism (SNP) of the IL-18 gene and house dust mite-specific IgE among Filipino allergic and non-allergic population was investigated. Atopic status was defined by serum total IgE concentration of ≥100 IU/mL, while house dust mite allergy was defined by specific IgE value ≥ +1SD of IgE of nonatopic participants. Two hundred twenty match-paired Filipino cases and controls aged 6-60 were the subjects of this investigation. The level of total IgE and Specific IgE were measured using Enzyme-Linked Immunosorbent Assay (ELISA) while Polymerase Chain Reaction – Restriction Fragment Length Polymorphism (PCR-RFLP) analysis was used in the SNP detection. Sensitization profiles of the allergic patients revealed that 97.3% were sensitized to Blomia tropicalis, 40.0% to Dermatophagoides farinae, and 29.1% to Dermatophagoides pteronyssinus. Multiple sensitization to HDMs was also observed among the 47.27% of the atopic participants. Any of the allergy classes of the atopic triad were exhibited by the cases (allergic asthma: 48.18%; allergic rhinitis: 62.73%; atopic dermatitis: 19.09%), and two or all of these atopic states are concurrently occurring in 26.36% of the cases. A greater proportion of the atopic participants with allergic asthma and allergic rhinitis were sensitized to D. farinae, and D. pteronyssinus, while more of those with atopic dermatitis were sensitized to D. pteronyssinus than D. farinae. Results show that there is overrepresentation of the allele “A” of the 105A/C IL-18 gene SNP in both cases and control groups of the population. The genotype that predominate the population is the heterozygous “AC”, followed by the homozygous wild “AA”, and the homozygous variant “CC” being the least. The study confirmed a positive association between serum specific IgE against B. tropicalis and D. pteronyssinus and the allele “C” (Bt P=0.021, Dp P=0.027) and “AC” (Bt P=0.003, Dp P=0.026) genotype. Findings also revealed that the genotypes “AA” (OR:1.217; 95% CI: 0.701-2.113) and “CC” (OR, 3.5; 95% CI: 0.727-16.849) increase the risk of developing allergy. This indicates that the 105A/C IL-18 gene SNP is a candidate genetic marker for HDM allergy among Filipino patients.Keywords: house dust mite allergy, interleukin-18 (IL-18), single nucleotide polymorphism,
Procedia PDF Downloads 4553170 A Comparative Study of Additive and Nonparametric Regression Estimators and Variable Selection Procedures
Authors: Adriano Z. Zambom, Preethi Ravikumar
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One of the biggest challenges in nonparametric regression is the curse of dimensionality. Additive models are known to overcome this problem by estimating only the individual additive effects of each covariate. However, if the model is misspecified, the accuracy of the estimator compared to the fully nonparametric one is unknown. In this work the efficiency of completely nonparametric regression estimators such as the Loess is compared to the estimators that assume additivity in several situations, including additive and non-additive regression scenarios. The comparison is done by computing the oracle mean square error of the estimators with regards to the true nonparametric regression function. Then, a backward elimination selection procedure based on the Akaike Information Criteria is proposed, which is computed from either the additive or the nonparametric model. Simulations show that if the additive model is misspecified, the percentage of time it fails to select important variables can be higher than that of the fully nonparametric approach. A dimension reduction step is included when nonparametric estimator cannot be computed due to the curse of dimensionality. Finally, the Boston housing dataset is analyzed using the proposed backward elimination procedure and the selected variables are identified.Keywords: additive model, nonparametric regression, variable selection, Akaike Information Criteria
Procedia PDF Downloads 2603169 Unravelling the Relationship Between Maternal and Fetal ACE2 Gene Polymorphism and Preeclampsia Risk
Authors: Sonia Tamanna, Akramul Hassan, Mohammad Shakil Mahmood, Farzana Ansari, Gowhar Rashid, Mir Fahim Faisal, M. Zakir Hossain Howlader
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Background: Preeclampsia (PE), a pregnancy-specific hypertensive disorder, significantly impacts maternal and fetal health. It is particularly prevalent in underdeveloped countries and is linked to preterm delivery and fetal growth. The renin-angiotensin system (RAS) plays a crucial role in ensuring a successful pregnancy outcome, with Angiotensin-Converting Enzyme 2 (ACE2) being a key component. ACE2 converts ANG II to Ang-(1-7), offering protection against ANG II-induced stress and inflammation while regulating blood pressure and osmotic balance during pregnancy. The reduced maternal plasma angiotensin-converting enzyme 2 (ACE2) seen in preeclampsia might contribute to its pathogenesis. However, there has been a dearth of comprehensive research into the association between ACE2 gene polymorphism and preeclampsia. In the South Asian population, hypertension is strongly linked to two SNPs: rs2285666 and rs879922. This genotype was therefore considered, and the possible association of maternal and fetal ACE2 gene polymorphism with preeclampsia within the Bangladeshi population was evaluated. Method: DNA was extracted from peripheral white blood cells (WBCs) using the organic method, and SNP genotyping was done via PCR-RFLP. Odds ratios (OR) with 95% confidence intervals (95% CI) were calculated using logistic regression to determine relative risk. Result: A comprehensive case-control study was conducted on 51 PE patients and their infants, along with 56 control subjects and their infants. Maternal single nuvleotide polymorphisms (SNP) (rs2285666) analysis revealed a strong association between the TT genotype and preeclampsia, with a four-fold increased risk in mothers (P=0.024, OR=4.00, 95% CI=1.36-11.37) compared to their ancestral genotype CC. However, the CT genotype (rs2285666) showed no significant difference (P=0.46, OR=1.54, 95% CI=0.57-4.14). Notably, no significant correlation was found in infants, regardless of their gender. For rs879922, no significant association was observed in both mothers and infants. This pioneering study suggests that mothers carrying the ACE2 gene variant rs2285666 (TT allele) may be at higher risk for preeclampsia, potentially influencing hypertension characteristics, whereas rs879922 does not appear to be associated with developing preeclampsia. Conclusion: This study sheds light on the role of ACE2 gene polymorphism, particularly the rs2285666 TT allele, in maternal susceptibility to preeclampsia. However, rs879922 does not appear to be linked to the risk of PE. This research contributes to our understanding of the genetic underpinnings of preeclampsia, offering insights into potential avenues for prevention and management.Keywords: ACE2, PCR-RFLP, preeclampsia, single nuvleotide polymorphisms (SNPs)
Procedia PDF Downloads 563168 Ground Motion Modeling Using the Least Absolute Shrinkage and Selection Operator
Authors: Yildiz Stella Dak, Jale Tezcan
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Ground motion models that relate a strong motion parameter of interest to a set of predictive seismological variables describing the earthquake source, the propagation path of the seismic wave, and the local site conditions constitute a critical component of seismic hazard analyses. When a sufficient number of strong motion records are available, ground motion relations are developed using statistical analysis of the recorded ground motion data. In regions lacking a sufficient number of recordings, a synthetic database is developed using stochastic, theoretical or hybrid approaches. Regardless of the manner the database was developed, ground motion relations are developed using regression analysis. Development of a ground motion relation is a challenging process which inevitably requires the modeler to make subjective decisions regarding the inclusion criteria of the recordings, the functional form of the model and the set of seismological variables to be included in the model. Because these decisions are critically important to the validity and the applicability of the model, there is a continuous interest on procedures that will facilitate the development of ground motion models. This paper proposes the use of the Least Absolute Shrinkage and Selection Operator (LASSO) in selecting the set predictive seismological variables to be used in developing a ground motion relation. The LASSO can be described as a penalized regression technique with a built-in capability of variable selection. Similar to the ridge regression, the LASSO is based on the idea of shrinking the regression coefficients to reduce the variance of the model. Unlike ridge regression, where the coefficients are shrunk but never set equal to zero, the LASSO sets some of the coefficients exactly to zero, effectively performing variable selection. Given a set of candidate input variables and the output variable of interest, LASSO allows ranking the input variables in terms of their relative importance, thereby facilitating the selection of the set of variables to be included in the model. Because the risk of overfitting increases as the ratio of the number of predictors to the number of recordings increases, selection of a compact set of variables is important in cases where a small number of recordings are available. In addition, identification of a small set of variables can improve the interpretability of the resulting model, especially when there is a large number of candidate predictors. A practical application of the proposed approach is presented, using more than 600 recordings from the National Geospatial-Intelligence Agency (NGA) database, where the effect of a set of seismological predictors on the 5% damped maximum direction spectral acceleration is investigated. The set of candidate predictors considered are Magnitude, Rrup, Vs30. Using LASSO, the relative importance of the candidate predictors has been ranked. Regression models with increasing levels of complexity were constructed using one, two, three, and four best predictors, and the models’ ability to explain the observed variance in the target variable have been compared. The bias-variance trade-off in the context of model selection is discussed.Keywords: ground motion modeling, least absolute shrinkage and selection operator, penalized regression, variable selection
Procedia PDF Downloads 3243167 Activity of Malate Dehydrogenase in Cell Free Extracts from S. proteamaculans, A. hydrophila, and K. pneumoniae
Authors: Mohamed M. Bumadian, D. James Gilmour
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Three bacterial species were isolated from the River Wye (Derbyshire, England) and identified using 16S rRNA gene sequencing as Serratia proteamaculans, Aeromonas hydrophila and Klebsiella pneumoniae. Respiration rates of the strains were measured in order to determine the metabolic activity under salt stress. The highest respiration rates of all three strains were found at 0.17 M and 0.5 M NaCl and then the respiration rate decreased with increasing concentrations of NaCl. In addition, the effect of increasing concentrations of NaCl on malate dehydrogenase activity was determined using cell-free extracts of the three strains. Malate dehydrogenase activity was stimulated at NaCl concentrations up to 0.5 M, and a small level of activity remained even at 3.5 M NaCl. The pH optimum of the malate dehydrogenase in cell-free extracts of all strains was higher than pH 7.5.Keywords: fresh water, halotolerant pathogenic bacteria, 16S rRNA gene, cell-free extracts, respiration rates, malate dehydrogenase
Procedia PDF Downloads 4563166 Adaptative Metabolism of Lactic Acid Bacteria during Brewers' Spent Grain Fermentation
Authors: M. Acin-Albiac, P. Filannino, R. Coda, Carlo G. Rizzello, M. Gobbetti, R. Di Cagno
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Demand for smart management of large amounts of agro-food by-products has become an area of major environmental and economic importance worldwide. Brewers' spent grain (BSG), the most abundant by-product generated in the beer-brewing process, represents an example of valuable raw material and source of health-promoting compounds. To the date, the valorization of BSG as a food ingredient has been limited due to poor technological and sensory properties. Tailored bioprocessing through lactic acid bacteria (LAB) fermentation is a versatile and sustainable means for the exploitation of food industry by-products. Indigestible carbohydrates (e.g., hemicelluloses and celluloses), high phenolic content, and mostly lignin make of BSG a hostile environment for microbial survival. Hence, the selection of tailored starters is required for successful fermentation. Our study investigated the metabolic strategies of Leuconostoc pseudomesenteroides and Lactobacillus plantarum strains to exploit BSG as a food ingredient. Two distinctive BSG samples from different breweries (Italian IT- and Finish FL-BSG) were microbially and chemically characterized. Growth kinetics, organic acid profiles, and the evolution of phenolic profiles during the fermentation in two BSG model media were determined. The results were further complemented with gene expression targeting genes involved in the degradation cellulose, hemicelluloses building blocks, and the metabolism of anti-nutritional factors. Overall, the results were LAB genus dependent showing distinctive metabolic capabilities. Leuc. pseudomesenteroides DSM 20193 may degrade BSG xylans while sucrose metabolism could be furtherly exploited for extracellular polymeric substances (EPS) production to enhance BSG pro-technological properties. Although L. plantarum strains may follow the same metabolic strategies during BSG fermentation, the mode of action to pursue such strategies was strain-dependent. L. plantarum PU1 showed a great preference for β-galactans compared to strain WCFS1, while the preference for arabinose occurred at different metabolic phases. Phenolic compounds profiling highlighted a novel metabolic route for lignin metabolism. These findings will allow an improvement of understanding of how lactic acid bacteria transform BSG into economically valuable food ingredients.Keywords: brewery by-product valorization, metabolism of plant phenolics, metabolism of lactic acid bacteria, gene expression
Procedia PDF Downloads 1223165 Application of Data Mining Techniques for Tourism Knowledge Discovery
Authors: Teklu Urgessa, Wookjae Maeng, Joong Seek Lee
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Application of five implementations of three data mining classification techniques was experimented for extracting important insights from tourism data. The aim was to find out the best performing algorithm among the compared ones for tourism knowledge discovery. Knowledge discovery process from data was used as a process model. 10-fold cross validation method is used for testing purpose. Various data preprocessing activities were performed to get the final dataset for model building. Classification models of the selected algorithms were built with different scenarios on the preprocessed dataset. The outperformed algorithm tourism dataset was Random Forest (76%) before applying information gain based attribute selection and J48 (C4.5) (75%) after selection of top relevant attributes to the class (target) attribute. In terms of time for model building, attribute selection improves the efficiency of all algorithms. Artificial Neural Network (multilayer perceptron) showed the highest improvement (90%). The rules extracted from the decision tree model are presented, which showed intricate, non-trivial knowledge/insight that would otherwise not be discovered by simple statistical analysis with mediocre accuracy of the machine using classification algorithms.Keywords: classification algorithms, data mining, knowledge discovery, tourism
Procedia PDF Downloads 2863164 The Discussion on the Composition of Feng Shui by the Environmental Planning Viewpoint
Authors: Jhuang Jin-Jhong, Hsieh Wei-Fan
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Climate change causes natural disasters persistently. Therefore, nowadays environmental planning objective tends to the issues of respecting nature and coexisting with nature. As a result, the natural environment analysis, e.g., the analysis of topography, soil, hydrology, climate, vegetation, is highly emphasized. On the other hand, Feng Shui has been a criterion of site selection for residence in Eastern since the ancient times and has had farther influence on site selection for castles and even for temples and tombs. The primary criterion of site selection is judging the quality of Long: mountain range, Sha: nearby mountains, Shui: hydrology, Xue: foundation, Xiang: aspect, which are similar to the environmental variables of mountain range, topography, hydrology and aspect. For the reason, a lot researchers attempt to probe into the connection between the criterion of Feng Shui and environmental planning factors. Most researches only discussed with the composition and theory of space of Feng Shui, but there is no research which explained Feng Shui through the environmental field. Consequently, this study reviewed the theory of Feng Shui through the environmental planning viewpoint and assembled essential composition factors of Feng Shui. The results of this study point. From literature review and comparison of theoretical meanings, we find that the ideal principles for planning the Feng Shui environment can also be used for environmental planning. Therefore, this article uses 12 ideal environmental features used in Feng Shui to contrast the natural aspects of the environment and make comparisons with previous research and classifies the environmental factors into climate, topography, hydrology, vegetation, and soil.Keywords: the composition of Feng Shui, environmental planning, site selection, main components of the Feng Shui environment
Procedia PDF Downloads 5043163 Investment Decision among Public Sector Retirees: A Behavioural Finance View
Authors: Bisi S. Olawoyin
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This study attempts an exploration into behavioural finance in which the traditional assumptions of expected utility maximization with rational investors in efficient markets are dropped. It reviews prior research and evidence about how psychological biases affect investors behaviour and stock selection. This study examined the relationship between demographic variables and financial behaviour biases among public sector retirees who invested in the Nigerian Stock Exchange prior to their retirement. By using questionnaire survey method, a total of 214 valid convenient samples were collected in order to determine how specific demographic and psychological trait affect stock selection between dividend paying and non-dividend paying stocks. Descriptive statistics and OLS were used to analyse the results. Findings showed that most of the retirees prefer dividend paying stocks in few years preceding their retirement but still hold on to their non-dividend paying stock on retirement. A significant difference also exists between senior and junior retirees in preference for non-dividend paying stocks. These findings are consistent with the clientele theories of dividend.Keywords: behavioural finance, clientele theories, dividend paying stocks, stock selection
Procedia PDF Downloads 1343162 Disturbed Cellular Iron Metabolism Genes in Neurodevelopmental Disorders is Different from Neurodegenerative Disorders
Authors: O. H. Gebril, N. A. Meguid
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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
Procedia PDF Downloads 3573161 Harnessing Deep-Level Metagenomics to Explore the Three Dynamic One Health Areas: Healthcare, Domiciliary and Veterinary
Authors: Christina Killian, Katie Wall, Séamus Fanning, Guerrino Macori
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Deep-level metagenomics offers a useful technical approach to explore the three dynamic One Health axes: healthcare, domiciliary and veterinary. There is currently limited understanding of the composition of complex biofilms, natural abundance of AMR genes and gene transfer occurrence in these ecological niches. By using a newly established small-scale complex biofilm model, COMBAT has the potential to provide new information on microbial diversity, antimicrobial resistance (AMR)-encoding gene abundance, and their transfer in complex biofilms of importance to these three One Health axes. Shotgun metagenomics has been used to sample the genomes of all microbes comprising the complex communities found in each biofilm source. A comparative analysis between untreated and biocide-treated biofilms is described. The basic steps include the purification of genomic DNA, followed by library preparation, sequencing, and finally, data analysis. The use of long-read sequencing facilitates the completion of metagenome-assembled genomes (MAG). Samples were sequenced using a PromethION platform, and following quality checks, binning methods, and bespoke bioinformatics pipelines, we describe the recovery of individual MAGs to identify mobile gene elements (MGE) and the corresponding AMR genotypes that map to these structures. High-throughput sequencing strategies have been deployed to characterize these communities. Accurately defining the profiles of these niches is an essential step towards elucidating the impact of the microbiota on each niche biofilm environment and their evolution.Keywords: COMBAT, biofilm, metagenomics, high-throughput sequencing
Procedia PDF Downloads 493160 Fuzzy Population-Based Meta-Heuristic Approaches for Attribute Reduction in Rough Set Theory
Authors: Mafarja Majdi, Salwani Abdullah, Najmeh S. Jaddi
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One of the global combinatorial optimization problems in machine learning is feature selection. It concerned with removing the irrelevant, noisy, and redundant data, along with keeping the original meaning of the original data. Attribute reduction in rough set theory is an important feature selection method. Since attribute reduction is an NP-hard problem, it is necessary to investigate fast and effective approximate algorithms. In this paper, we proposed two feature selection mechanisms based on memetic algorithms (MAs) which combine the genetic algorithm with a fuzzy record to record travel algorithm and a fuzzy controlled great deluge algorithm to identify a good balance between local search and genetic search. In order to verify the proposed approaches, numerical experiments are carried out on thirteen datasets. The results show that the MAs approaches are efficient in solving attribute reduction problems when compared with other meta-heuristic approaches.Keywords: rough set theory, attribute reduction, fuzzy logic, memetic algorithms, record to record algorithm, great deluge algorithm
Procedia PDF Downloads 4493159 Expression of Tissue Plasminogen Activator in Transgenic Tobacco Plants by Signal Peptides Targeting for Delivery to Apoplast, Endoplasmic Reticulum and Cytosol Spaces
Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi
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Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA4404. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.Keywords: tPA, recombinant, transgenic, tobacco
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