Search results for: grammar-based genetic programming
255 A Biophysical Model of CRISPR/Cas9 on- and off-Target Binding for Rational Design of Guide RNAs
Authors: Iman Farasat, Howard M. Salis
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The CRISPR/Cas9 system has revolutionized genome engineering by enabling site-directed and high-throughput genome editing, genome insertion, and gene knockdowns in several species, including bacteria, yeast, flies, worms, and human cell lines. This technology has the potential to enable human gene therapy to treat genetic diseases and cancer at the molecular level; however, the current CRISPR/Cas9 system suffers from seemingly sporadic off-target genome mutagenesis that prevents its use in gene therapy. A comprehensive mechanistic model that explains how the CRISPR/Cas9 functions would enable the rational design of the guide-RNAs responsible for target site selection while minimizing unexpected genome mutagenesis. Here, we present the first quantitative model of the CRISPR/Cas9 genome mutagenesis system that predicts how guide-RNA sequences (crRNAs) control target site selection and cleavage activity. We used statistical thermodynamics and law of mass action to develop a five-step biophysical model of cas9 cleavage, and examined it in vivo and in vitro. To predict a crRNA's binding specificities and cleavage rates, we then compiled a nearest neighbor (NN) energy model that accounts for all possible base pairings and mismatches between the crRNA and the possible genomic DNA sites. These calculations correctly predicted crRNA specificity across 5518 sites. Our analysis reveals that cas9 activity and specificity are anti-correlated, and, the trade-off between them is the determining factor in performing an RNA-mediated cleavage with minimal off-targets. To find an optimal solution, we first created a scheme of safe-design criteria for Cas9 target selection by systematic analysis of available high throughput measurements. We then used our biophysical model to determine the optimal Cas9 expression levels and timing that maximizes on-target cleavage and minimizes off-target activity. We successfully applied this approach in bacterial and mammalian cell lines to reduce off-target activity to near background mutagenesis level while maintaining high on-target cleavage rate.Keywords: biophysical model, CRISPR, Cas9, genome editing
Procedia PDF Downloads 398254 Organic Geochemistry and Oil-Source Correlation of Cretaceous Sediments in the Kohat Basin, Pakistan
Authors: Syed Mamoon Siyar, Fayaz Ali, Sajjad Ahmad, Samina Jahandad, George Kontakiotis, Hammad T. Janjuhah, Assimina Antonarakou, Waqas Naseem
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The Cretaceous Chichali Formation in the Chanda-01, Chanda-02, Chanda-03 and Mela-05 wells and the oil samples from Chanda-01 and Chanda-01 wells located in the Kohat Basin, Pakistan, were analyzed with the objectives of evaluating the hydrocarbon generation potential, source, thermal maturity and depositional of organic matter, and oil-source correlation by employing geochemical screening techniques and biomarker studies. The total organic carbon (TOC) values in Chanda-02, Chanda-03 and Mela-05 indicate, in general, poor to fair, fair and fair to good source rock potential with low genetic potential, respectively. The nature of organic matter has been determined by standard cross plots of Rock Eval pyrolysis parameters, indicating that studied cuttings from the Chichali Formation dominantly contain type III kerogen at present and show maturity for oil generation in the studied wells. The organic petrographic study also confirmed the vitrinite (type III) as a major maceral in the investigated Chichali Shales and its reflectance values show maturity for oil. The different ratios of non-biomarkers and biomarkers i.e., steranes, terpenes and aromatics parameters, indicate the marine source of organic matter deposited in the anoxic environment for the Chichali Formation in Chanda-01 and Chanda-02 wells and mixed source input of organic matter deposited in suboxic conditions for oil in the same wells. The CPI, and different biomarkers parameters such as C29 S/S+R, ββ/αα+ββ), M29/H30, Ts/Ts+Tm, H31 (S/S+R) and aromatic compounds methyl phenanthrene index (MPI) and organic petrographic analysis (vitrinite reflectance) suggest mature stage of oil generation for Chichali Shales and oil samples in the study area with little high thermal maturity in case of oils. Based on source and thermal maturity biomarkers and non-biomarkers parameters, the produced oils have no correlation with the Cretaceous Chichali Formation in the studied Chanda-01 and Chanda-02 wells in Kohat Basin, Pakistan, but it has been suggested that these oils have been generated by the strata containing high terrestrial organic input compare to Chichali Shales.Keywords: Organic geochemistry, Chichali Shales and crude oils, Kohat Basin, Pakistan
Procedia PDF Downloads 78253 Evaluation of Radio Protective Potential of Indian Bamboo Leaves
Authors: Mansi Patel, Priti Mehta
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Background: Ionizing radiations have detrimental effects on humans, and the growing technological encroachment has increased human exposure to it enormously. So, the safety issues have emphasized researchers to develop radioprotector from natural resources having minimal toxicity. A substance having anti-inflammatory, antioxidant, and immunomodulatory activity can be a potential candidate for radioprotection. One such plant with immense potential i.e. Bamboo was selected for the present study. Purpose: The study aims to evaluate the potential of Indian bamboo leaves for protection against the clastogenic effect of gamma radiation. Methods: The protective effect of bamboo leaf extract against gamma radiation-induced genetic damage in human peripheral blood lymphocytes (HPBLs) was evaluated in vitro using Cytokinesis blocked micronuclei assay (CBMN). The blood samples were pretreated with varying concentration of extract 30 min before the radiation exposure (4Gy & 6Gy). The reduction in the frequency of micronuclei was observed for the irradiated and control groups. The effect of various concentration of bamboo leaf extract (400,600,800 mg/kg) on the development of radiation induced sickness and altered mortality in mice exposed to 8 Gy of whole-body gamma radiation was studied. The developed symptoms were clinically scored by multiple endpoints for 30 days. Results: Treatment of HPBLs with varying concentration of extract before exposure to a different dose of γ- radiation resulted in significant (P < 0.0001) decline of radiation induced micronuclei. It showed dose dependent and concentration driven activity. The maximum protection ~ 70% was achieved at nine µg/ml concentration. Extract treated whole body irradiated mice showed 50%, 83.3% and 100% survival for 400, 600, and 800mg/kg with 1.05, 0.43 and 0 clinical score respectively when compared to Irradiated mice having 6.03 clinical score and 0% survival. Conclusion: Our findings indicate bamboo leaf extract reduced the radiation induced cytogenetic damage. It has also increased the survival ratio and reduced the radiation induced sickness and mortality when exposed to a lethal dose of gamma radiation.Keywords: bamboo leaf extract, Cytokinesis blocked micronuclei (CBMN) assay, ionizing radiation, radio protector
Procedia PDF Downloads 140252 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 371251 Simulation of Focusing of Diamagnetic Particles in Ferrofluid Microflows with a Single Set of Overhead Permanent Magnets
Authors: Shuang Chen, Zongqian Shi, Jiajia Sun, Mingjia Li
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Microfluidics is a technology that small amounts of fluids are manipulated using channels with dimensions of tens to hundreds of micrometers. At present, this significant technology is required for several applications in some fields, including disease diagnostics, genetic engineering, and environmental monitoring, etc. Among these fields, manipulation of microparticles and cells in microfluidic device, especially separation, have aroused general concern. In magnetic field, the separation methods include positive and negative magnetophoresis. By comparison, negative magnetophoresis is a label-free technology. It has many advantages, e.g., easy operation, low cost, and simple design. Before the separation of particles or cells, focusing them into a single tight stream is usually a necessary upstream operation. In this work, the focusing of diamagnetic particles in ferrofluid microflows with a single set of overhead permanent magnets is investigated numerically. The geometric model of the simulation is based on the configuration of previous experiments. The straight microchannel is 24mm long and has a rectangular cross-section of 100μm in width and 50μm in depth. The spherical diamagnetic particles of 10μm in diameter are suspended into ferrofluid. The initial concentration of the ferrofluid c₀ is 0.096%, and the flow rate of the ferrofluid is 1.8mL/h. The magnetic field is induced by five identical rectangular neodymium−iron− boron permanent magnets (1/8 × 1/8 × 1/8 in.), and it is calculated by equivalent charge source (ECS) method. The flow of the ferrofluid is governed by the Navier–Stokes equations. The trajectories of particles are solved by the discrete phase model (DPM) in the ANSYS FLUENT program. The positions of diamagnetic particles are recorded by transient simulation. Compared with the results of the mentioned experiments, our simulation shows consistent results that diamagnetic particles are gradually focused in ferrofluid under magnetic field. Besides, the diamagnetic particle focusing is studied by varying the flow rate of the ferrofluid. It is in agreement with the experiment that the diamagnetic particle focusing is better with the increase of the flow rate. Furthermore, it is investigated that the diamagnetic particle focusing is affected by other factors, e.g., the width and depth of the microchannel, the concentration of the ferrofluid and the diameter of diamagnetic particles.Keywords: diamagnetic particle, focusing, microfluidics, permanent magnet
Procedia PDF Downloads 126250 Effect of Auraptene on the Enzymatic Glutathione Redox-System in Nrf2 Knockout Mice
Authors: Ludmila A. Gavriliuc, Jerry McLarty, Heather E. Kleiner, J. Michael Mathis
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Abstract -- Background: The citrus coumarine Auraptene (Aur) is an effective chemopreventive agent, as manifested in many models of diseases and cancer. Nuclear factor erythroid 2-related factor (Nrf2) is an important regulator of genes induced by oxidative stress, such as glutathione S-transferases, heme oxygenase-1, and peroxiredoxin 1, by activating the antioxidant response element (ARE). Genetic and biochemical evidence has demonstrated that glutathione (GSH) and glutathione-dependent enzymes, glutathione reductase (GR), glutathione peroxidases (GPs), glutathione S-transferases (GSTs) are responsible for the control of intracellular reduction-oxidation status and participate in cellular adaptation to oxidative stress. The effect of Aur on the activity of GR, GPs (Se-GP and Se-iGP), and content of GSH in the liver, kidney, and spleen is insufficiently explored. Aim: Our goal was the examination of the Aur influence on the redox-system of GSH in Nrf2 wild type and Nrf2 knockout mice via activation of Nrf2 and ARE. Methods: Twenty female mice, 10 Nrf2 wild-type (WT) and 10 Nrf2 (-/-) knockout (KO), were bred and genotyped for our study. The activity of GR, Se-GP, Se-iGP, GST, G6PD, CytP450 reductase, catalase (Cat), and content of GSH were analyzed in the liver, kidney, and spleen using Spectrophotometry methods. The results of the specific activity of enzymes and the amount of GSH were analyzed with ANOVA and Spearman statistical methods. Results: Aur (200 mg/kg) treatment induced hepatic GST, GR, Se-GP activity and inhibited their activity in the spleen of mice, most likely via activation of the ARE through Nrf2. Activation in kidney Se-GP and G6PD by Aur is also controlled, apparently through Nrf2. Results of the non-parametric Spearman correlation analysis indicated the strong positive correlation between GR and G6PD only in the liver in WT control mice (r=+0.972; p < 0.005) and in the kidney KO control mice (r=+0.958; p < 0.005). The observed low content of GSH in the liver of KO mice indicated an increase in its participation in the neutralization of toxic substances with the absence of induction of GSH-dependent enzymes, such as GST, GR, Se-GP, and Se-iGP. Activation of CytP450 in kidney and spleen and Cat in the liver in KO mice probably revealed another regulatory mechanism for these enzymes. Conclusion: Thereby, obtained results testify that Aur can modulate the activity of genes and antioxidant enzymatic redox-system of GSH, responsible for the control of intracellular reduction-oxidation status.Keywords: auraptene, glutathione, GST, Nrf2
Procedia PDF Downloads 142249 Metagenomics Analysis on Microbial Communities of Sewage Sludge from Nyeri-Kangemi Wastewater Treatment Plant, Nyeri County-Kenya
Authors: Allan Kiptanui Kimisto, Geoffrey Odhiambo Ongondo, Anastasia Wairimu Muia, Cyrus Ndungu Kimani
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The major challenge to proper sewage sludge treatment processes is the poor understanding of sludge microbiome diversities. This study applied the whole-genome. shotgun metagenomics technique to profile the microbial composition of sewage sludge in two active digestion lagoons at the Nyeri-Kangemi Wastewater Treatment Plant in Nyeri County, Kenya. Total microbial community DNA was extracted from samples using the available ZymoBIOMICS™ DNA Miniprep Kit and sequenced using Shotgun metagenomics. Samples were analyzed using MG-RAST software (Project ID: mgp100988), which allowed for comparing taxonomic diversity before β-diversities studies for Bacteria, Archaea and Eukaryotes. The study identified 57 phyla, 145 classes, 301 orders, 506 families, 963 genera, and 1980 species. Bacteria dominated the microbes and comprised 28 species, 51 classes, 110 orders, 243 families, 597 genera, and 1518 species. The Bacteroides(6.77%) were dominant, followed by Acinetobacter(1.44%) belonging to the Gammaproteobacteria and Acidororax (1.36%), Bacillus (1.24%) and Clostridium (1.02%) belonging to Betaproteobacteria. Archaea recorded 5 phyla, 13 classes, 19 orders, 29 families, 60 genera,and87 species, with the dominant genera being Methanospirillum (16.01%), methanosarcina (15.70%), and Methanoregula(14.80%) and Methanosaeta (8.74%), Methanosphaerula(5.48%) and Methanobrevibacter(5.03%) being the subdominant group. The eukaryotes were the least in abundance and comprised 24 phyla, 81 classes, 301 orders, 506 families, 963 genera, and 980 species. Arabidopsis (4.91%) and Caenorhabditis (4.81%) dominated the eukaryotes, while Dityostelium (3.63%) and Drosophila(2.08%) were the subdominant genera. All these microbes play distinct roles in the anaerobic treatment process of sewage sludge. The local sludge microbial composition and abundance variations may be due to age difference differences between the two digestion lagoons in operation at the plant and the different degradation rales played by the taxa. The information presented in this study can help in the genetic manipulation or formulation of optimal microbial ratios to improve their effectiveness in sewage sludge treatment. This study recommends further research on how the different taxa respond to environmental changes over time and space.Keywords: shotgun metagenomics, sludge, bacteria, archaea, eukaryotes
Procedia PDF Downloads 90248 Clostridium thermocellum DBT-IOC-C19, A Potential CBP Isolate for Ethanol Production
Authors: Nisha Singh, Munish Puri, Collin Barrow, Deepak Tuli, Anshu S. Mathur
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The biological conversion of lignocellulosic biomass to ethanol is a promising strategy to solve the present global crisis of exhausting fossil fuels. The existing bioethanol production technologies have cost constraints due to the involvement of mandate pretreatment and extensive enzyme production steps. A unique process configuration known as consolidated bioprocessing (CBP) is believed to be a potential cost-effective process due to its efficient integration of enzyme production, saccharification, and fermentation into one step. Due to several favorable reasons like single step conversion, no need of adding exogenous enzymes and facilitated product recovery, CBP has gained the attention of researchers worldwide. However, there are several technical and economic barriers which need to be overcome for making consolidated bioprocessing a commercially viable process. Finding a natural candidate CBP organism is critically important and thermophilic anaerobes are preferred microorganisms. The thermophilic anaerobes that can represent CBP mainly belong to genus Clostridium, Caldicellulosiruptor, Thermoanaerobacter, Thermoanaero bacterium, and Geobacillus etc. Amongst them, Clostridium thermocellum has received increased attention as a high utility CBP candidate due to its highest growth rate on crystalline cellulose, the presence of highly efficient cellulosome system and ability to produce ethanol directly from cellulose. Recently with the availability of genetic and molecular tools aiding the metabolic engineering of Clostridium thermocellum have further facilitated the viability of commercial CBP process. With this view, we have specifically screened cellulolytic and xylanolytic thermophilic anaerobic ethanol producing bacteria, from unexplored hot spring/s in India. One of the isolates is a potential CBP organism identified as a new strain of Clostridium thermocellum. This strain has shown superior avicel and xylan degradation under unoptimized conditions compared to reported wild type strains of Clostridium thermocellum and produced more than 50 mM ethanol in 72 hours from 1 % avicel at 60°C. Besides, this strain shows good ethanol tolerance and growth on both hexose and pentose sugars. Hence, with further optimization this new strain could be developed as a potential CBP microbe.Keywords: Clostridium thermocellum, consolidated bioprocessing, ethanol, thermophilic anaerobes
Procedia PDF Downloads 394247 An Emergence of Pinus taeda Needle Defoliation and Tree Mortality in Alabama, USA
Authors: Debit Datta, Jeffrey J. Coleman, Scott A. Enebak, Lori G. Eckhardt
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Pinus taeda, commonly known as loblolly pine, is a crucial timber species native to the southeastern USA. An emerging problem has been encountered for the past few years, which is better to be known as loblolly pine needle defoliation (LPND), which is threatening the ecological health of southeastern forests and economic vitality of the region’s timber industry. Currently, more than 1000 hectares of loblolly plantations in Alabama are affected with similar symptoms and have created concern among southeast landowners and forest managers. However, it is still uncertain whether LPND results from one or the combination of several fungal pathogens. Therefore, the objectives of the study were to identify and characterize the fungi associated with LPND in the southeastern USA and document the damage being done to loblolly pine as a result of repeated defoliation. Identification of fungi was confirmed using classical morphological methods (microscopic examination of the infected needles), conventional and species-specific priming (SSPP) PCR, and ITS sequencing. To date, 17 species of fungi, either cultured from pine needles or formed fruiting bodies on pine needles, were identified based on morphology and genetic sequence data. Among them, brown-spot pathogen Lecanostica acicola has been frequently recovered from pine needles in both spring and summer. Moreover, Ophistomatoid fungi such as Leptographium procerum, L. terebrantis are associated with pine decline have also been recovered from root samples of the infected stands. Trees have been increasingly and repeatedly chlorotic and defoliated from 2019 to 2020. Based on morphological observations and molecular data, emerging loblolly pine needle defoliation is due in larger part to the brown-spot pathogen L. acoicola followed by pine decline pathogens L. procerum and L. terebrantis. Root pathogens were suspected to emerge later, and their cumulative effects contribute to the widespread mortality of the trees. It is more likely that longer wet spring and warmer temperatures are favorable to disease development and may be important in the disease ecology of LPND. Therefore, the outbreak of the disease is assumed to be expanded over a large geographical area in a changing climatic condition.Keywords: brown-spot fungi, emerging disease, defoliation, loblolly pine
Procedia PDF Downloads 131246 Ultrastructural Characterization of Lipid Droplets of Rat Hepatocytes after Whole Body 60-Cobalt Gamma Radiation
Authors: Ivna Mororó, Lise P. Labéjof, Stephanie Ribeiro, Kely Almeida
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Lipid droplets (LDs) are normally presented in greater or lesser number in the cytoplasm of almost all eukaryotic and some prokaryotic cells. They are independent organelles composed of a lipid ester core and a surface phospholipid monolayer. As a lipid storage form, they provide an available source of energy for the cell. Recently it was demonstrated that they play an important role in other many cellular processes. Among the many unresolved questions about them, it is not even known how LDs is formed, how lipids are recruited to LDs and how they interact with the other organelles. Excess fat in the organism is pathological and often associated with the development of some genetic, hormonal or behavioral diseases. The formation and accumulation of lipid droplets in the cytoplasm can be increased by exogenous physical or chemical agents. It is well known that ionizing radiation affects lipid metabolism resulting in increased lipogenesis in cells, but the details of this process are unknown. To better understand the mode of formation of LDs in liver cells, we investigate their ultrastructural morphology after irradiation. For that, Wistar rats were exposed to whole body gamma radiation from 60-cobalt at various single doses. Samples of the livers were processed for analysis under a conventional transmission electron microscope. We found that when compared to controls, morphological changes in liver cells were evident at the higher doses of radiation used. It was detected a great number of lipid droplets of different sizes and homogeneous content and some of them merged each other. In some cells, it was observed diffused LDs, not limited by a monolayer of phospholipids. This finding suggests that the phospholipid monolayer of the LDs was disrupted by ionizing radiation exposure that promotes lipid peroxydation of endo membranes. Thus the absence of the phospholipid monolayer may prevent the realization of some cellular activities as follow: - lipid exocytosis which requires the merging of LDs membrane with the plasma membrane; - the interaction of LDs with other membrane-bound organelles such as the endoplasmic reticulum (ER), the golgi and mitochondria and; - lipolysis of lipid esters contained in the LDs which requires the presence of enzymes located in membrane-bound organelles as ER. All these impediments can contribute to lipid accumulation in the cytoplasm and the development of diseases such as liver steatosis, cirrhosis and cancer.Keywords: radiobiology, hepatocytes, lipid metabolism, transmission electron microscopy
Procedia PDF Downloads 305245 Sensing Endocrine Disrupting Chemicals by Virus-Based Structural Colour Nanostructure
Authors: Lee Yujin, Han Jiye, Oh Jin-Woo
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The adverse effects of endocrine disrupting chemicals (EDCs) has attracted considerable public interests. The benzene-like EDCs structure mimics the mechanisms of hormones naturally occurring in vivo, and alters physiological function of the endocrine system. Although, some of the most representative EDCs such as polychlorinated biphenyls (PCBs) and phthalates compounds already have been prohibited to produce and use in many countries, however, PCBs and phthalates in plastic products as flame retardant and plasticizer are still circulated nowadays. EDCs can be released from products while using and discarding, and it causes serious environmental and health issues. Here, we developed virus-based structurally coloured nanostructure that can detect minute EDCs concentration sensitively and selectively. These structurally coloured nanostructure exhibits characteristic angel-independent colors due to the regular virus bundle structure formation through simple pulling technique. The designed number of different colour bands can be formed through controlling concentration of virus solution and pulling speed. The virus, M-13 bacteriophage, was genetically engineered to react with specific ECDs, typically PCBs and phthalates. M-13 bacteriophage surface (pVIII major coat protein) was decorated with benzene derivative binding peptides (WHW) through phage library method. In the initial assessment, virus-based color sensor was exposed to several organic chemicals including benzene, toluene, phenol, chlorobenzene, and phthalic anhydride. Along with the selectivity evaluation of virus-based colour sensor, it also been tested for sensitivity. 10 to 300 ppm of phthalic anhydride and chlorobenzene were detected by colour sensor, and showed the significant sensitivity with about 90 of dissociation constant. Noteworthy, all measurements were analyzed through principal component analysis (PCA) and linear discrimination analysis (LDA), and exhibited clear discrimination ability upon exposure to 2 categories of EDCs (PCBs and phthalates). Because of its easy fabrication, high sensitivity, and the superior selectivity, M-13 bacteriophage-based color sensor could be a simple and reliable portable sensing system for environmental monitoring, healthcare, social security, and so on.Keywords: M-13 bacteriophage, colour sensor, genetic engineering, EDCs
Procedia PDF Downloads 236244 In Silico Analysis of Deleterious nsSNPs (Missense) of Dihydrolipoamide Branched-Chain Transacylase E2 Gene Associated with Maple Syrup Urine Disease Type II
Authors: Zainab S. Ahmed, Mohammed S. Ali, Nadia A. Elshiekh, Sami Adam Ibrahim, Ghada M. El-Tayeb, Ahmed H. Elsadig, Rihab A. Omer, Sofia B. Mohamed
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Maple syrup urine (MSUD) is an autosomal recessive disease that causes a deficiency in the enzyme branched-chain alpha-keto acid (BCKA) dehydrogenase. The development of disease has been associated with SNPs in the DBT gene. Despite that, the computational analysis of SNPs in coding and noncoding and their functional impacts on protein level still remains unknown. Hence, in this study, we carried out a comprehensive in silico analysis of missense that was predicted to have a harmful influence on DBT structure and function. In this study, eight different in silico prediction algorithms; SIFT, PROVEAN, MutPred, SNP&GO, PhD-SNP, PANTHER, I-Mutant 2.0 and MUpo were used for screening nsSNPs in DBT including. Additionally, to understand the effect of mutations in the strength of the interactions that bind protein together the ELASPIC servers were used. Finally, the 3D structure of DBT was formed using Mutation3D and Chimera servers respectively. Our result showed that a total of 15 nsSNPs confirmed by 4 software (R301C, R376H, W84R, S268F, W84C, F276C, H452R, R178H, I355T, V191G, M444T, T174A, I200T, R113H, and R178C) were found damaging and can lead to a shift in DBT gene structure. Moreover, we found 7 nsSNPs located on the 2-oxoacid_dh catalytic domain, 5 nsSNPs on the E_3 binding domain and 3 nsSNPs on the Biotin Domain. So these nsSNPs may alter the putative structure of DBT’s domain. Furthermore, we detected all these nsSNPs are on the core residues of the protein and have the ability to change the stability of the protein. Additionally, we found W84R, S268F, and M444T have high significance, and they affected Leucine, Isoleucine, and Valine, which reduces or disrupt the function of BCKD complex, E2-subunit which the DBT gene encodes. In conclusion, based on our extensive in-silico analysis, we report 15 nsSNPs that have possible association with protein deteriorating and disease-causing abilities. These candidate SNPs can aid in future studies on Maple Syrup Urine Disease type II base in the genetic level.Keywords: DBT gene, ELASPIC, in silico analysis, UCSF chimer
Procedia PDF Downloads 199243 Triploid Rainbow Trout (Oncorhynchus mykiss) for Better Aquaculture and Ecological Risk Management
Authors: N. N. Pandey, Raghvendra Singh, Biju S. Kamlam, Bipin K. Vishwakarma, Preetam Kala
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The rainbow trout (Oncorhynchus mykiss) is an exotic salmonid fish, well known for its fast growth, tremendous ability to thrive in diverse conditions, delicious flesh and hard fighting nature in Europe and other countries. Rainbow trout farming has a great potential for its contribution to the mainstream economy of Himalayan states in India and other temperate countries. These characteristics establish them as one of the most widely introduced and cultured fish across the globe, and its farming is also prominent in the cold water regions of India. Nevertheless, genetic fatigue, slow growth, early maturity, and low productivity are limiting the expansion of trout production. Moreover, farms adjacent to natural streams or other water sources are subject to escape of domesticated rainbow trout into the wild, which is a serious environmental concern as the escaped fish is subject to contaminate and disrupt the receiving ecosystem. A decline in production traits due to early maturity prolongs the culture duration and affects the profit margin of rainbow trout farms in India. A viable strategy that could overcome these farming constraints in large scale operation is the production of triploid fish that are sterile and more heterozygous. For better triploidy induction rate (TR), heat shock at 28°C for 10 minutes and pressure shock 9500 psi pressure for 5 minutes is applied to green eggs with 90-100% of triploidy success and 72-80% survival upto swim-up fry stage. There is 20% better growth in aquaculture with triploids rainbow trout over diploids. As compared to wild diploid fish, larger sized and fitter triploid rainbow trout in natural waters attract to trout anglers, and support the development of recreational fisheries by state fisheries departments without the risk of contaminating existing gene pools and disrupting local fish diversity. Overall, enhancement of productivity in rainbow trout farms and trout production in coldwater regions, development of lucrative trout angling and better ecological management is feasible with triploid rainbow trout.Keywords: rainbow trout, triploids fish, heat shock, pressure shock, trout angling
Procedia PDF Downloads 120242 ESDN Expression in the Tumor Microenvironment Coordinates Melanoma Progression
Authors: Roberto Coppo, Francesca Orso, Daniela Dettori, Elena Quaglino, Lei Nie, Mehran M. Sadeghi, Daniela Taverna
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Malignant melanoma is currently the fifth most common cancer in the white population and it is fatal in its metastatic stage. Several research studies in recent years have provided evidence that cancer initiation and progression are driven by genetic alterations of the tumor and paracrine interactions between tumor and microenvironment. Scattered data show that the Endothelial and Smooth muscle cell-Derived Neuropilin-like molecule (ESDN) controls cell proliferation and movement of stroma and tumor cells. To investigate the role of ESDN in the tumor microenvironment during melanoma progression, murine melanoma cells (B16 or B16-F10) were injected in ESDN knockout mice in order to evaluate how the absence of ESDN in stromal cells could influence melanoma progression. While no effect was found on primary tumor growth, increased cell extravasation and lung metastasis formation was observed in ESDN knockout mice compared to wild type controls. In order to understand how cancer cells cross the endothelial barrier during metastatic dissemination in an ESDN-null microenvironment, structure, and permeability of lung blood vessels were analyzed. Interestingly, ESDN knockout mice showed structurally altered and more permeable vessels compared to wild type animals. Since cell surface molecules mediate the process of tumor cell extravasation, the expression of a panel of extravasation-related ligands and receptors was analyzed. Importantly, modulations of N-cadherin, E-selectin, ICAM-1 and VAP-1 were observed in ESDN knockout endothelial cells, suggesting the presence of a favorable tumor microenvironment which facilitates melanoma cell extravasation and metastasis formation in the absence of ESDN. Furthermore, a potential contribution of immune cells in tumor dissemination was investigated. An increased recruitment of macrophages in the lungs of ESDN knockout mice carrying subcutaneous B16-F10 tumors was found. In conclusion, our data suggest a functional role of ESDN in the tumor microenvironment during melanoma progression and the identification of the mechanisms that regulate tumor cell extravasation could lead to the development of new therapies to reduce metastasis formation.Keywords: melanoma, tumor microenvironment, extravasation, cell surface molecules
Procedia PDF Downloads 325241 Tick Infestation and its Implications on Health and Welfare of Cattle under Pastoral System in Nigeria
Authors: Alabi Olufemi, Adeyanju Taiwo, Oloruntoba Oluwasegun, Adeleye Bobola, Alabi Oyekemi
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The pastoral system is a predominant form of cattle production in Nigeria, characterized by extensive grazing on communal lands. However, this system is challenged by various factors, including tick infestation, which significantly affects cattle health and welfare hence this investigation which aims to provide an in-depth understanding of tick infestation in the context of Nigerian pastoral systems, emphasizing its impact on cattle health and welfare. The country harbors a diverse array of tick species that affect cattle. These ticks belong to different genera, including Rhipicephalus, Amblyomma, and Hyalomma, among others. Each species has unique characteristics, life cycles, and host preferences, contributing to the complexity of tick infestation dynamics in pastoral settings. Tick infestation has numerous detrimental effects on cattle health. The direct effects include blood loss, anemia, skin damage due to feeding, and the transmission of pathogens that cause diseases such as anaplasmosis, babesiosis, and theileriosis. Indirectly, tick infestation can lead to reduced productivity, weight loss, and increased susceptibility to other diseases.The welfare of cattle in Nigerian pastoral systems is significantly impacted by tick infestation. Infested cattle often exhibit signs of distress, including restlessness, reduced grazing activity, and altered behavior. Furthermore, the discomfort caused by tick bites can lead to chronic stress, compromising the overall welfare of the animals. Effective tick control is crucial for mitigating the impact of infestation on cattle health and welfare. Strategies such as acaricide application, pasture management, genetic selection for tick resistance cattle, and vaccination against tick-borne diseases are commonly used. Tick infestation presents a significant challenge to cattle production under the pastoral system in Nigeria. It not only impacts cattle health but also compromises their welfare. Addressing the issue of tick infestation requires a multifaceted approach that integrates effective control strategies with sustainable management practices. Further research is needed to develop tailored interventions that account for the unique characteristics of Nigerian pastoral systems, ultimately ensuring the well-being and productivity of cattle in these settings.Keywords: tick infestation, pastoral system, welfare, cattle
Procedia PDF Downloads 40240 Marzuq Basin Palaeozoic Petroleum System
Authors: M. Dieb, T. Hodairi
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In the Southwest Libya area, the Palaeozoic deposits are an important petroleum system, with Silurian shale considered a hydrocarbon source rock and Cambro-Ordovician recognized as a good reservoir. The Palaeozoic petroleum system has the greatest potential for conventional and is thought to represent the significant prospect of unconventional petroleum resources in Southwest Libya. Until now, the lateral and vertical heterogeneity of the source rock was not well evaluated, and oil-source correlation is still a matter of debate. One source rock, which is considered the main source potential in Marzuq Basin, was investigated for its uranium contents using gamma-ray logs, rock-eval pyrolysis, and organic petrography for their bulk kinetic characteristics to determine the petroleum potential qualitatively and quantitatively. Thirty source rock samples and fifteen oil samples from the Tannezzuft source rock were analyzed by Rock-Eval Pyrolysis, microscopely investigation, GC, and GC-MS to detect acyclic isoprenoids and aliphatic, aromatic, and NSO biomarkers. Geochemistry tools were applied to screen source and age-significant biomarkers to high-spot genetic relationships. A grating heterogeneity exists among source rock zones from different levels of depth with varying uranium contents according to gamma-ray logs, rock-eval pyrolysis results, and kinetic features. The uranium-rich Tannezzuft Formations (Hot Shales) produce oils and oil-to-gas hydrocarbons based on their richness, kerogen type, and thermal maturity. Biomarker results such as C₂₇, C₂₈, and C₂₉ steranes concentrations and C₂₄ tetracyclic terpane/C₂₉ tricyclic terpane ratios, with sterane and hopane ratios, are considered the most promising biomarker information in differentiating within the Silurian Shale Tannezzuft Formation and in correlating with its expelled oils. The Tannezzuft Hot Shale is considered the main source rock for oil and gas accumulations in the Cambro-Ordovician reservoirs within the Marzuq Basin. Migration of the generated and expelled oil and gas from the Tannezzuft source rock to the reservoirs of the Cambro-Ordovician petroleum system was interpreted to have occurred along vertical and lateral pathways along the faults in the Palaeozoic Strata. The Upper Tannezzuft Formation (cold shale) is considered the primary seal in the Marzuq Basin.Keywords: heterogeneity, hot shale, kerogen, Silurian, uranium
Procedia PDF Downloads 56239 Genetic Analysis of Rust Resistance Genes in Global Wheat
Authors: Aktar-Uz-Zaman, M. Tuhina-Khatun, Mohamed Hanafi Musa
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Three rust diseases: leaf (brown) rust caused by Puccinia triticina Eriks, stripe (yellow) rust caused by Puccinia striiformis West, and stem (black) rust caused by Puccinia graminis f. sp. tritici are economically important diseases of wheat in world wide. Yield loss due to leaf rust is 40% in susceptible cultivars. Yield losses caused by the stem rust pathogens in the mid of 20 century reached 20-30% in Eastern and Central Europe and the most virulent stem rust race Ug99 emerged first in Uganda and after that in Kenya, Ethiopia, Yemen, in the Middle East and South Asia. Yield losses were estimated up to 100%, whereas, up to 80% have been reported in Kenya during 1999. In case of stripe rust, severity level has been recorded 60% - 70% as compared to 100% severity of susceptible check in disease screening nurseries in Kenya. Improvement of resistant varieties or cultivars is the sustainable, economical and environmentally friendly approaches for increasing the global wheat production to suppress the rust diseases. More than 68 leaf rust, 49 stripe rust and 53 stem rust resistance genes have been identified in the global wheat cultivars or varieties using different molecular breeding approaches. Among these, Lr1, Lr9, Lr10, Lr19, Lr21, Lr24, Lr25, Lr28, Lr29, Lr34, Lr35, Lr37, Lr39, Lr47, Lr51, Lr3bg, Lr18, Lr40, Lr46, and Lr50 leaf rust resistance genes have been identified by using molecular, enzymatic and microsatellite markers from African, Asian, European cultivars of hexaploid wheat (Triticum aestivum), durum wheat and diploid wheat species. These genes are located on 20, of the 21 chromosomes of hexaploid wheat. Similarly, Sr1, Sr2, Sr24, and Sr3, Sr31 stem rust resistance genes have been recognized from wheat cultivars of Pakistan, India, Kenya, and Uganda etc. A race of P. striiformis (stripe rust) Yr9, Yr18, and Yr29 was first observed in East Africa, Italy, Pakistan and India wheat cultivars. These stripe rust resistance genes are located on chromosomes 1BL, 4BL, 6AL, 3BS and 6BL in bread wheat cultivars. All these identified resistant genes could be used for notable improvement of susceptible wheat cultivars in the future.Keywords: hexaploid wheat, resistance genes, rust disease, triticum aestivum
Procedia PDF Downloads 476238 Development of a Bus Information Web System
Authors: Chiyoung Kim, Jaegeol Yim
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Bus service is often either main or the only public transportation available in cities. In metropolitan areas, both subways and buses are available whereas in the medium sized cities buses are usually the only type of public transportation available. Bus Information Systems (BIS) provide current locations of running buses, efficient routes to travel from one place to another, points of interests around a given bus stop, a series of bus stops consisting of a given bus route, and so on to users. Thanks to BIS, people do not have to waste time at a bus stop waiting for a bus because BIS provides exact information on bus arrival times at a given bus stop. Therefore, BIS does a lot to promote the use of buses contributing to pollution reduction and saving natural resources. BIS implementation costs a huge amount of budget as it requires a lot of special equipment such as road side equipment, automatic vehicle identification and location systems, trunked radio systems, and so on. Consequently, medium and small sized cities with a low budget cannot afford to install BIS even though people in these cities need BIS service more desperately than people in metropolitan areas. It is possible to provide BIS service at virtually no cost under the assumption that everybody carries a smartphone and there is at least one person with a smartphone in a running bus who is willing to reveal his/her location details while he/she is sitting in a bus. This assumption is usually true in the real world. The smartphone penetration rate is greater than 100% in the developed countries and there is no reason for a bus driver to refuse to reveal his/her location details while driving. We have developed a mobile app that periodically reads values of sensors including GPS and sends GPS data to the server when the bus stops or when the elapsed time from the last send attempt is greater than a threshold. This app detects the bus stop state by investigating the sensor values. The server that receives GPS data from this app has also been developed. Under the assumption that the current locations of all running buses collected by the mobile app are recorded in a database, we have also developed a web site that provides all kinds of information that most BISs provide to users through the Internet. The development environment is: OS: Windows 7 64bit, IDE: Eclipse Luna 4.4.1, Spring IDE 3.7.0, Database: MySQL 5.1.7, Web Server: Apache Tomcat 7.0, Programming Language: Java 1.7.0_79. Given a start and a destination bus stop, it finds a shortest path from the start to the destination using the Dijkstra algorithm. Then, it finds a convenient route considering number of transits. For the user interface, we use the Google map. Template classes that are used by the Controller, DAO, Service and Utils classes include BUS, BusStop, BusListInfo, BusStopOrder, RouteResult, WalkingDist, Location, and so on. We are now integrating the mobile app system and the web app system.Keywords: bus information system, GPS, mobile app, web site
Procedia PDF Downloads 213237 Meniere's Disease and its Prevalence, Symptoms, Risk Factors and Associated Treatment Solutions for this Disease
Authors: Amirreza Razzaghipour Sorkhab
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One of the most common disorders among humans is hearing impairment. This paper provides an evidence base that recovers understanding of Meniere’s disease and highlights the physical and mental health correlates of the disorder. Meniere's disease is more common in the elderly. The term idiopathic endolymphatic hydrops has been attributed to this disease by some in the previous. Meniere’s disease demonstrations a genetic tendency, and a family history is found in 10% of cases, with an autosomal dominant inheritance pattern. The COCH gene may be one of the hereditary factors contributing to Meniere’s disease, and the possibility of a COCH mutation should be considered in patients with Meniere’s disease symptoms. Should be considered Missense mutations in the COCH gene cause the autosomal dominant sensorineural hearing loss and vestibular disorder. Meniere’s disease is a complex, heterogeneous disorder of the inner ear and that is characterized by episodes of vertigo lasting from minutes to hours, fluctuating sensorineural hearing loss, tinnitus, and aural fullness. The existing evidence supports the suggestion that age and sleep disorder are risk factors for Meniere's disease. Many factors have been reported to precipitate the progress of Menier, including endolymphatic hydrops, immunology, viral infection, inheritance, vestibular migraine, and altered intra-labyrinthine fluid dynamics. Although there is currently no treatment that has a proven helpful effect on hearing levels or on the long-term evolution of the disease, however, in the primary stages, the hearing may improve among attacks, but a permanent hearing loss occurs in the majority of cases. Current publications have proposed a role for the intratympanic use of medicine, mostly aminoglycosides, for the control of vertigo. more than 85% of patients with Meniere's disease are helped by either changes in lifestyle and medical treatment or minimally aggressive surgical procedures such as intratympanic steroid therapy, intratympanic gentamicin therapy, and endolymphatic sac surgery. However, unilateral vestibular extirpation methods (intratympanic gentamicin, vestibular nerve section, or labyrinthectomy) are more predictable but invasive approaches to control the vertigo attacks. Medical therapy aimed at reducing endolymph volume, such as low-sodium diet, diuretic use, is the typical initial treatment.Keywords: meniere's disease, endolymphatic hydrops, hearing loss, vertigo, tinnitus, COCH gene
Procedia PDF Downloads 85236 Detailed Sensitive Detection of Impurities in Waste Engine Oils Using Laser Induced Breakdown Spectroscopy, Rotating Disk Electrode Optical Emission Spectroscopy and Surface Plasmon Resonance
Authors: Cherry Dhiman, Ayushi Paliwal, Mohd. Shahid Khan, M. N. Reddy, Vinay Gupta, Monika Tomar
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The laser based high resolution spectroscopic experimental techniques such as Laser Induced Breakdown Spectroscopy (LIBS), Rotating Disk Electrode Optical Emission spectroscopy (RDE-OES) and Surface Plasmon Resonance (SPR) have been used for the study of composition and degradation analysis of used engine oils. Engine oils are mainly composed of aliphatic and aromatics compounds and its soot contains hazardous components in the form of fine, coarse and ultrafine particles consisting of wear metal elements. Such coarse particulates matter (PM) and toxic elements are extremely dangerous for human health that can cause respiratory and genetic disorder in humans. The combustible soot from thermal power plants, industry, aircrafts, ships and vehicles can lead to the environmental and climate destabilization. It contributes towards global pollution for land, water, air and global warming for environment. The detection of such toxicants in the form of elemental analysis is a very serious issue for the waste material management of various organic, inorganic hydrocarbons and radioactive waste elements. In view of such important points, the current study on used engine oils was performed. The fundamental characterization of engine oils was conducted by measuring water content and kinematic viscosity test that proves the crude analysis of the degradation of used engine oils samples. The microscopic quantitative and qualitative analysis was presented by RDE-OES technique which confirms the presence of elemental impurities of Pb, Al, Cu, Si, Fe, Cr, Na and Ba lines for used waste engine oil samples in few ppm. The presence of such elemental impurities was confirmed by LIBS spectral analysis at various transition levels of atomic line. The recorded transition line of Pb confirms the maximum degradation which was found in used engine oil sample no. 3 and 4. Apart from the basic tests, the calculations for dielectric constants and refractive index of the engine oils were performed via SPR analysis.Keywords: surface plasmon resonance, laser-induced breakdown spectroscopy, ICCD spectrometer, engine oil
Procedia PDF Downloads 136235 Mitigating the Negative Health Effects from Stress - A Social Network Analysis
Authors: Jennifer A. Kowalkowski
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Production agriculture (farming) is a physically, emotionally, and cognitively stressful occupation, where workers have little control over the stressors that impact both their work and their lives. In an occupation already rife with hazards, these occupational-related stressors have been shown to increase farm workers’ risks for illness, injury, disability, and death associated with their work. Despite efforts to mitigate the negative health effects from occupational-related stress (ORS) and to promote health and well-being (HWB) among farmers in the US, marked improvements have not been attained. Social support accessed through social networks has been shown to buffer against the negative health effects from stress, yet no studies have directly examined these relationships among farmers. The purpose of this study was to use social network analysis to explore the social networks of farm owner-operators and the social supports available to them for mitigating the negative health effects of ORS. A convenience sample of 71 farm owner-operators from a Midwestern County in the US completed and returned a mailed survey (55.5% response rate) that solicited information about their social networks related to ORS. Farmers reported an average of 2.4 individuals in their personal networks and higher levels of comfort discussing ORS with female network members. Farmers also identified few connections (3.4% density) and indicated low comfort with members of affiliation networks specific to ORS. Findings from this study highlighted that farmers accessed different social networks and resources for their personal HWB than for issues related to occupational(farm-related) health and safety. In addition, farmers’ social networks for personal HWB were smaller, with different relational characteristics than reported in studies of farmers’ social networks related to occupational health and safety. Collectively, these findings suggest that farmers conceptualize personal HWB differently than farm health and safety. Therefore, the same research approaches and targets that guide occupational health and safety research may not be appropriate for personal HWB for farmers. Interventions and programming targeting ORS and HWB have largely been offered through the same platforms or mechanisms as occupational health and safety programs. This may be attributed to the significant overlap between the farm as a family business and place of residence, or that ORS stems from farm-related issues. However, these assumptions translated to health research of farmers and farm families from the occupational health and safety literature have not been directly studied or challenged. Thismay explain why past interventions have not been effective at improving health outcomes for farmers and farm families. A close examination of findings from this study raises important questions for researchers who study agricultural health. Findings from this study have significant implications for future research agendas focused on addressing ORS, HWB, and health disparities for farmersand farm families.Keywords: agricultural health, occupational-related stress, social networks, well-being
Procedia PDF Downloads 102234 Evaluating the Potential of a Fast Growing Indian Marine Cyanobacterium by Reconstructing and Analysis of a Genome Scale Metabolic Model
Authors: Ruchi Pathania, Ahmad Ahmad, Shireesh Srivastava
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Cyanobacteria is a promising microbe that can capture and convert atmospheric CO₂ and light into valuable industrial bio-products like biofuels, biodegradable plastics, etc. Among their most attractive traits are faster autotrophic growth, whole year cultivation using non-arable land, high photosynthetic activity, much greater biomass and productivity and easy for genetic manipulations. Cyanobacteria store carbon in the form of glycogen which can be hydrolyzed to release glucose and fermented to form bioethanol or other valuable products. Marine cyanobacterial species are especially attractive for countries with scarcity of freshwater. We recently identified a marine native cyanobacterium Synechococcus sp. BDU 130192 which has good growth rate and high level of polyglucans accumulation compared to Synechococcus PCC 7002. In this study, firstly we sequenced the whole genome and the sequences were annotated using the RAST server. Genome scale metabolic model (GSMM) was reconstructed through COBRA toolbox. GSMM is a computational representation of the metabolic reactions and metabolites of the target strain. GSMMs construction through the application of Flux Balance Analysis (FBA), which uses external nutrient uptake rates and estimate steady state intracellular and extracellular reaction fluxes, including maximization of cell growth. The model, which we have named isyn942, includes 942 reactions and 913 metabolites having 831 metabolic, 78 transport and 33 exchange reactions. The phylogenetic tree obtained by BLAST search revealed that the strain was a close relative of Synechococcus PCC 7002. The flux balance analysis (FBA) was applied on the model iSyn942 to predict the theoretical yields (mol product produced/mol CO₂ consumed) for native and non-native products like acetone, butanol, etc. under phototrophic condition by applying metabolic engineering strategies. The reported strain can be a viable strain for biotechnological applications, and the model will be helpful to researchers interested in understanding the metabolism as well as to design metabolic engineering strategies for enhanced production of various bioproducts.Keywords: cyanobacteria, flux balance analysis, genome scale metabolic model, metabolic engineering
Procedia PDF Downloads 153233 Evaluation of Mito-Uncoupler Induced Hyper Metabolic and Aggressive Phenotype in Glioma Cells
Authors: Yogesh Rai, Saurabh Singh, Sanjay Pandey, Dhananjay K. Sah, B. G. Roy, B. S. Dwarakanath, Anant N. Bhatt
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One of the most common signatures of highly malignant gliomas is their capacity to metabolize more glucose to lactic acid than normal brain tissues, even under normoxic conditions (Warburg effect), indicating that aerobic glycolysis is constitutively upregulated through stable genetic or epigenetic changes. However, oxidative phosphorylation (OxPhos) is also required to maintain the mitochondrial membrane potential for tumor cell survival. In the process of tumorigenesis, tumor cells during fastest growth rate exhibit both high glycolytic and high OxPhos. Therefore, metabolically reprogrammed cancer cells with combination of both aerobic glycolysis and altered OxPhos develop a robust metabolic phenotype, which confers a selective growth advantage. In our study, we grew the high glycolytic BMG-1 (glioma) cells with continuous exposure of mitochondrial uncoupler 2, 4, dinitro phenol (DNP) for 10 passages to obtain a phenotype of high glycolysis with enhanced altered OxPhos. We found that OxPhos modified BMG (OPMBMG) cells has similar growth rate and cell cycle distribution but high mitochondrial mass and functional enzymatic activity than parental cells. In in-vitro studies, OPMBMG cells showed enhanced invasion, proliferation and migration properties. Moreover, it also showed enhanced angiogenesis in matrigel plug assay. Xenografted tumors from OPMBMG cells showed reduced latent period, faster growth rate and nearly five folds reduction in the tumor take in nude mice compared to BMG-1 cells, suggesting that robust metabolic phenotype facilitates tumor formation and growth. OPMBMG cells which were found radio-resistant, showed enhanced radio-sensitization by 2-DG as compared to the parental BMG-1 cells. This study suggests that metabolic reprogramming in cancer cells enhances the potential of migration, invasion and proliferation. It also strengthens the cancer cells to escape the death processes, conferring resistance to therapeutic modalities. Our data also suggest that combining metabolic inhibitors like 2-DG with conventional therapeutic modalities can sensitize such metabolically aggressive cancer cells more than the therapies alone.Keywords: 2-DG, BMG, DNP, OPM-BMG
Procedia PDF Downloads 221232 Characterization of the Blood Microbiome in Rheumatoid Arthritis Patients Compared to Healthy Control Subjects Using V4 Region 16S rRNA Sequencing
Authors: D. Hammad, D. P. Tonge
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Rheumatoid arthritis (RA) is a disabling and common autoimmune disease during which the body's immune system attacks healthy tissues. This results in complicated and long-lasting actions being carried out by the immune system, which typically only occurs when the immune system encounters a foreign object. In the case of RA, the disease affects millions of people and causes joint inflammation, ultimately leading to the destruction of cartilage and bone. Interestingly, the disease mechanism still remains unclear. It is likely that RA occurs as a result of a complex interplay of genetic and environmental factors including an imbalance in the microorganism population inside our body. The human microbiome or microbiota is an extensive community of microorganisms in and on the bodies of animals, which comprises bacteria, fungi, viruses, and protozoa. Recently, the development of molecular techniques to characterize entire bacterial communities has renewed interest in the involvement of the microbiome in the development and progression of RA. We believe that an imbalance in some of the specific bacterial species in the gut, mouth and other sites may lead to atopobiosis; the translocation of these organisms into the blood, and that this may lead to changes in immune system status. The aim of this study was, therefore, to characterize the microbiome of RA serum samples in comparison to healthy control subjects using 16S rRNA gene amplification and sequencing. Serum samples were obtained from healthy control volunteers and from patients with RA both prior to, and following treatment. The bacterial community present in each sample was identified utilizing V4 region 16S rRNA amplification and sequencing. Bacterial identification, to the lowest taxonomic rank, was performed using a range of bioinformatics tools. Significantly, the proportions of the Lachnospiraceae, Ruminococcaceae, and Halmonadaceae families were significantly increased in the serum of RA patients compared with healthy control serum. Furthermore, the abundance of Bacteroides and Lachnospiraceae nk4a136_group, Lachnospiraceae_UGC-001, RuminococcaceaeUCG-014, Rumnococcus-1, and Shewanella was also raised in the serum of RA patients relative to healthy control serum. These data support the notion of a blood microbiome and reveal RA-associated changes that may have significant implications for biomarker development and may present much-needed opportunities for novel therapeutic development.Keywords: blood microbiome, gut and oral bacteria, Rheumatoid arthritis, 16S rRNA gene sequencing
Procedia PDF Downloads 127231 Safety Climate Assessment and Its Impact on the Productivity of Construction Enterprises
Authors: Krzysztof J. Czarnocki, F. Silveira, E. Czarnocka, K. Szaniawska
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Research background: Problems related to the occupational health and decreasing level of safety occur commonly in the construction industry. Important factor in the occupational safety in construction industry is scaffold use. All scaffolds used in construction, renovation, and demolition shall be erected, dismantled and maintained in accordance with safety procedure. Increasing demand for new construction projects unfortunately still is linked to high level of occupational accidents. Therefore, it is crucial to implement concrete actions while dealing with scaffolds and risk assessment in construction industry, the way on doing assessment and liability of assessment is critical for both construction workers and regulatory framework. Unfortunately, professionals, who tend to rely heavily on their own experience and knowledge when taking decisions regarding risk assessment, may show lack of reliability in checking the results of decisions taken. Purpose of the article: The aim was to indicate crucial parameters that could be modeling with Risk Assessment Model (RAM) use for improving both building enterprise productivity and/or developing potential and safety climate. The developed RAM could be a benefit for predicting high-risk construction activities and thus preventing accidents occurred based on a set of historical accident data. Methodology/Methods: A RAM has been developed for assessing risk levels as various construction process stages with various work trades impacting different spheres of enterprise activity. This project includes research carried out by teams of researchers on over 60 construction sites in Poland and Portugal, under which over 450 individual research cycles were carried out. The conducted research trials included variable conditions of employee exposure to harmful physical and chemical factors, variable levels of stress of employees and differences in behaviors and habits of staff. Genetic modeling tool has been used for developing the RAM. Findings and value added: Common types of trades, accidents, and accident causes have been explored, in addition to suitable risk assessment methods and criteria. We have found that the initial worker stress level is more direct predictor for developing the unsafe chain leading to the accident rather than the workload, or concentration of harmful factors at the workplace or even training frequency and management involvement.Keywords: safety climate, occupational health, civil engineering, productivity
Procedia PDF Downloads 311230 The Anesthesia Considerations in Robotic Mastectomies
Authors: Amrit Vasdev, Edwin Rho, Gurinder Vasdev
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Robotic surgery has enabled a new spectrum of minimally invasive breast reconstruction by improving visualization, surgeon posturing, and improved patient outcomes.1 The DaVinci robot system can be utilized in nipple sparing mastectomies and reconstructions. The process involves the insufflation of the subglandular space and a dissection of the mammary gland with a combination of cautery and blunt dissection. This case outlines a 35-year-old woman who has a long-standing family history of breast cancer and a diagnosis of a deleterious BRCA2 genetic mutation. She has decided to proceed with bilateral nipple sparing mastectomies with implants. Her perioperative mammogram and MRI were negative for masses, however, her left internal mammary lymph node was enlarged. She has taken oral contraceptive pills for 3-5 years and denies DES exposure, radiation therapy, human replacement therapy, or prior breast surgery. She does not smoke and rarely consumes alcohol. During the procedure, the patient received a standardized anesthetic for out-patient surgery of propofol infusion, succinylcholine, sevoflurane, and fentanyl. Aprepitant was given as an antiemetic and preoperative Tylenol and gabapentin for pain management. Concerns for the patient during the procedure included CO2 insufflation into the subcutaneous space. With CO2 insufflation, there is a potential for rapid uptake leading to severe acidosis, embolism, and subcutaneous emphysema.2To mitigate this, it is important to hyperventilate the patient and reduce both the insufflation pressure and the CO2 flow rate to the minimal acceptable by the surgeon. For intraoperative monitoring during this 6-9 hour long procedure, it has been suggested to utilize an Arterial-Line for end-tidal CO2 monitoring. However, in this case, it was not necessary as the patient had excellent cardiovascular reserve, and end-tidal CO2 was within normal limits for the duration of the procedure. A BIS monitor was also utilized to reduce anesthesia burden and to facilitate a prompt discharge from the PACU. Minimal Invasive Robotic Surgery will continue to evolve, and anesthesiologists need to be prepared for the new challenges ahead. Based on our limit number of patients, robotic mastectomy appears to be a safe alternative to open surgery with the promise of clearer tissue demarcation and better cosmetic results.Keywords: anesthesia, mastectomies, robotic, hypercarbia
Procedia PDF Downloads 100229 Pixel Façade: An Idea for Programmable Building Skin
Authors: H. Jamili, S. Shakiba
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Today, one of the main concerns of human beings is facing the unpleasant changes of the environment. Buildings are responsible for a significant amount of natural resources consumption and carbon emissions production. In such a situation, this thought comes to mind that changing each building into a phenomenon of benefit to the environment. A change in a way that each building functions as an element that supports the environment, and construction, in addition to answering the need of humans, is encouraged, the way planting a tree is, and it is no longer seen as a threat to alive beings and the planet. Prospect: Today, different ideas of developing materials that can smartly function are realizing. For instance, Programmable Materials, which in different conditions, can respond appropriately to the situation and have features of modification in shape, size, physical properties and restoration, and repair quality. Studies are to progress having this purpose to plan for these materials in a way that they are easily available, and to meet this aim, there is no need to use expensive materials and high technologies. In these cases, physical attributes of materials undertake the role of sensors, wires and actuators then materials will become into robots itself. In fact, we experience robotics without robots. In recent decades, AI and technology advances have dramatically improving the performance of materials. These achievements are a combination of software optimizations and physical productions such as multi-materials 3D printing. These capabilities enable us to program materials in order to change shape, appearance, and physical properties to interact with different situations. nIt is expected that further achievements like Memory Materials and Self-learning Materials are also added to the Smart Materials family, which are affordable, available, and of use for a variety of applications and industries. From the architectural standpoint, the building skin is significantly considered in this research, concerning the noticeable surface area the buildings skin have in urban space. The purpose of this research would be finding a way that the programmable materials be used in building skin with the aim of having an effective and positive interaction. A Pixel Façade would be a solution for programming a building skin. The Pixel Facadeincludes components that contain a series of attributes that help buildings for their needs upon their environmental criteria. A PIXEL contains series of smart materials and digital controllers together. It not only benefits its physical properties, such as control the amount of sunlight and heat, but it enhances building performance by providing a list of features, depending on situation criteria. The features will vary depending on locations and have a different function during the daytime and different seasons. The primary role of a PIXEL FAÇADE can be defined as filtering pollutions (for inside and outside of the buildings) and providing clean energy as well as interacting with other PIXEL FACADES to estimate better reactions.Keywords: building skin, environmental crisis, pixel facade, programmable materials, smart materials
Procedia PDF Downloads 84228 Investigation of the Role of Lipoprotein a rs10455872 Gene Polymorphism in Childhood Obesity
Authors: Mustafa M. Donma, Ayşen Haksayar, Bahadır Batar, Buse Tepe, Birol Topçu, Orkide Donma
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Childhood obesity is an ever-increasing health problem. The Association of obesity with severe chronic diseases such as diabetes and cardiovascular diseases makes the problem life-threatening. Aside from psychological, societal and metabolic factors, genetic polymorphisms have gained importance concerning etiology in recent years. The aim of this study was to evaluate the relationship between rs10455872 gene polymorphism in the Lipoprotein (a) locus and the development of childhood obesity. This was a prospective study carried out according to the Helsinki Declarations. The study protocol was approved by the Institutional Ethics Committee. This study was supported by Tekirdag Namik Kemal University Rectorate, Scientific Research Projects Coordination Unit. Project No: NKUBAP.02.TU.20.278. A total of 180 children (103 obese (OB) and 77 healthy), aged 6-18 years, without any acute or chronic disease, participated in the study. Two different groups were created: OB and healthy control. Each group was divided into two further groups depending on the nature of the polymorphism. Anthropometric measurements were taken during the detailed physical examination. Laboratory tests and TANITA measurements were performed. For the statistical evaluations, SPSS version 28.0 was used. A P-value smaller than 0.05 was the statistical significance degree. The distribution of lipoprotein (a) rs10455872 gene polymorphism did not differ between OB and healthy children. Children with AG genotype in both OB and control groups had lower body mass index (BMI), diagnostic obesity notation model assessment index (DONMA II), body fat ratio (BFR), C-reactive protein (CRP), and metabolic syndrome index (MetS index) values compared to children with normal AA genotype. In the OB group, serum iron, vitamin B12, hemoglobin, MCV, and MCH values were found to be higher in the AG genotype group than those of children with the normal AA genotype. A significant correlation was found between the MetS index and BFR among OB children with normal homozygous genotype. MetS index increased as BFR increased in this group. However, such a correlation was not observed in the OB group with heterozygous AG genotype. To the best of our knowledge, the association of lipoprotein (a) rs10455872 gene polymorphism with the etiology of childhood obesity has not been studied yet. Therefore, this study was the first report suggesting polymorphism with AG genotype as a good risk factor for obesity.Keywords: child, gene polymorphism, lipoprotein (a), obesity, rs10455872
Procedia PDF Downloads 66227 Population Dynamics of Cyprinid Fish Species (Mahseer: Tor Species) and Its Conservation in Yamuna River of Garhwal Region, India
Authors: Davendra Singh Malik
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India is one of the mega-biodiversity countries in the world and contributing about 11.72% of global fish diversity. The Yamuna river is the longest tributary of Ganga river ecosystem, providing a natural habitat for existing fish diversity of Himalayan region of Indian subcontinent. The several hydropower dams and barrages have been constructed on different locations of major rivers in Garhwal region. These dams have caused a major ecological threat to change existing fresh water ecosystems altering water flows, interrupting ecological connectivity, fragmenting habitats and native riverine fish species. Mahseer fishes (Indian carp) of the genus Tor, are large cyprinids endemic to continental Asia popularly known as ‘Game or sport fishes’ have continued to be decimated by fragmented natural habitats due to damming the water flow in riverine system and categorized as threatened fishes of India. The fresh water fish diversity as 24 fish species were recorded from Yamuna river. The present fish catch data has revealed that mahseer fishes (Tor tor and Tor putitora) were contributed about 32.5 %, 25.6 % and 18.2 % in upper, middle and lower riverine stretches of Yaumna river. The length range of mahseer (360-450mm) recorded as dominant size of catch composition. The CPUE (catch per unit effort) of mahseer fishes also indicated about a sharp decline of fish biomass, changing growth pattern, sex ratio and maturity stages of fishes. Only 12.5 – 14.8 % mahseer female brooders have showed only maturity phases in breeding months. The fecundity of mature mahseer female fish brooders ranged from 2500-4500 no. of ova during breeding months. The present status of mahseer fishery has attributed to the over exploitative nature in Yamuna river. The mahseer population is shrinking continuously in down streams of Yamuna river due to cumulative effects of various ecological stress. Mahseer conservation programme have implemented as 'in situ fish conservation' for enhancement of viable population size of mahseer species and restore the genetic loss of mahseer fish germplasm in Yamuna river of Garhwal Himalayan region.Keywords: conservation practice, population dynamics, tor fish species, Yamuna River
Procedia PDF Downloads 252226 Parallels between Training Parameters of High-Performance Athletes Determining the Long-Term Adaptation of the Body in Various Sports: Case Study on Different Types of Training and Their Gender Conditioning
Authors: Gheorghe Braniste
Abstract:
Gender gap has always been in dispute when comparing records and has been a major factor influencing best performances in various sports. Consequently, our study registers the evolution of the difference between men's and women’s best performances within either cyclic or acyclic sports, considering the fact that the training sessions of high performance athletes prove both similarities and differences in long-term adaptation of their body to stress and effort in breaking limits and records. Firstly, for a correct interpretation of the data and tables included in this paper, we must point out that the intense muscular activity has a considerable impact on the structural organization of the organs and systems of the performer's body through the mechanism of motor-visceral reflexes, forming a high working capacity suitable for intense muscular activity. The opportunity to obtaine high sports results during the official competitions is due, on the one hand, to the genetic characteristics of the athlete's body, and on the other hand, to the fact that playing professional sports leaves its mark on the vital morphological and functional parameters. The aim of our research is to study the landmarking differences between male and female athletes and their physical development, together with their growing capacity to stand up to the functional training during the competitive period of their annual training cycle. In order to evaluate the physical development of the athletes, the data of the anthropometric screenings obtained at the Olympic Training Center of the selected teams of the Republic of Moldova were interpreted and rated. During the study of physical development in terms of body height and weight, vital capacity, thoracic excursion, maximum force (Fmax), dynamometry of the hand and back, a further evaluation of the physical development indices that allow an evaluation of complex physical development were registered. The interdependence of the results obtained in performance sports with the morphological and functional particularities of the athletes' body is firmly determined and cannot be disputed. Nevertheless, registered data proved that with the increase of the training capacity, the morphological and functional abilities of the female body increase and, in some respects, approach and even slightly surpass the men in certain sports.Keywords: physical development, indices, parameters, active body weight, morphological maturity, physical performance
Procedia PDF Downloads 111