Search results for: whole genome sequence
1005 Approaching a Tat-Rev Independent HIV-1 Clone towards a Model for Research
Authors: Walter Vera-Ortega, Idoia Busnadiego, Sam J. Wilson
Abstract:
Introduction: Human Immunodeficiency Virus type 1 (HIV-1) is responsible for the acquired immunodeficiency syndrome (AIDS), a leading cause of death worldwide infecting millions of people each year. Despite intensive research in vaccine development, therapies against HIV-1 infection are not curative, and the huge genetic variability of HIV-1 challenges to drug development. Current animal models for HIV-1 research present important limitations, impairing the progress of in vivo approaches. Macaques require a CD8+ depletion to progress to AIDS, and the maintenance cost is high. Mice are a cheaper alternative but need to be 'humanized,' and breeding is not possible. The development of an HIV-1 clone able to replicate in mice is a challenging proposal. The lack of human co-factors in mice impedes the function of the HIV-1 accessory proteins, Tat and Rev, hampering HIV-1 replication. However, Tat and Rev function can be replaced by constitutive/chimeric promoters, codon-optimized proteins and the constitutive transport element (CTE), generating a novel HIV-1 clone able to replicate in mice without disrupting the amino acid sequence of the virus. By minimally manipulating the genomic 'identity' of the virus, we propose the generation of an HIV-1 clone able to replicate in mice to assist in antiviral drug development. Methods: i) Plasmid construction: The chimeric promoters and CTE copies were cloned by PCR using lentiviral vectors as templates (pCGSW and pSIV-MPCG). Tat mutants were generated from replication competent HIV-1 plasmids (NHG and NL4-3). ii) Infectivity assays: Retroviral vectors were generated by transfection of human 293T cells and murine NIH 3T3 cells. Virus titre was determined by flow cytometry measuring GFP expression. Human B-cells (AA-2) and Hela cells (TZMbl) were used for infectivity assays. iii) Protein analysis: Tat protein expression was determined by TZMbl assay and HIV-1 capsid by western blot. Results: We have determined that NIH 3T3 cells are able to generate HIV-1 particles. However, they are not infectious, and further analysis needs to be performed. Codon-optimized HIV-1 constructs are efficiently made in 293T cells in a Tat and Rev independent manner and capable of packaging a competent genome in trans. CSGW is capable of generating infectious particles in the absence of Tat and Rev in human cells when 4 copies of the CTE are placed preceding the 3’LTR. HIV-1 Tat mutant clones encoding different promoters are functional during the first cycle of replication when Tat is added in trans. Conclusion: Our findings suggest that the development of an HIV-1 Tat-Rev independent clone is challenging but achievable aim. However, further investigations need to be developed prior presenting our HIV-1 clone as a candidate model for research.Keywords: codon-optimized, constitutive transport element, HIV-1, long terminal repeats, research model
Procedia PDF Downloads 3011004 Inbreeding Study Using Runs of Homozygosity in Nelore Beef Cattle
Authors: Priscila A. Bernardes, Marcos E. Buzanskas, Luciana C. A. Regitano, Ricardo V. Ventura, Danisio P. Munari
Abstract:
The best linear unbiased predictor (BLUP) is a method commonly used in genetic evaluations of breeding programs. However, this approach can lead to higher inbreeding coefficients in the population due to the intensive use of few bulls with higher genetic potential, usually presenting some degree of relatedness. High levels of inbreeding are associated to low genetic viability, fertility, and performance for some economically important traits and therefore, should be constantly monitored. Unreliable pedigree data can also lead to misleading results. Genomic information (i.e., single nucleotide polymorphism – SNP) is a useful tool to estimate the inbreeding coefficient. Runs of homozygosity have been used to evaluate homozygous segments inherited due to direct or collateral inbreeding and allows inferring population selection history. This study aimed to evaluate runs of homozygosity (ROH) and inbreeding in a population of Nelore beef cattle. A total of 814 animals were genotyped with the Illumina BovineHD BeadChip and the quality control was carried out excluding SNPs located in non-autosomal regions, with unknown position, with a p-value in the Hardy-Weinberg equilibrium lower than 10⁻⁵, call rate lower than 0.98 and samples with the call rate lower than 0.90. After the quality control, 809 animals and 509,107 SNPs remained for analyses. For the ROH analysis, PLINK software was used considering segments with at least 50 SNPs with a minimum length of 1Mb in each animal. The inbreeding coefficient was calculated using the ratio between the sum of all ROH sizes and the size of the whole genome (2,548,724kb). A total of 25.711 ROH were observed, presenting mean, median, minimum, and maximum length of 3.34Mb, 2Mb, 1Mb, and 80.8Mb, respectively. The number of SNPs present in ROH segments varied from 50 to 14.954. The longest ROH length was observed in one animal, which presented a length of 634Mb (24.88% of the genome). Four bulls were among the 10 animals with the longest extension of ROH, presenting 11% of ROH with length higher than 10Mb. Segments longer than 10Mb indicate recent inbreeding. Therefore, the results indicate an intensive use of few sires in the studied data. The distribution of ROH along the chromosomes showed that chromosomes 5 and 6 presented a large number of segments when compared to other chromosomes. The mean, median, minimum, and maximum inbreeding coefficients were 5.84%, 5.40%, 0.00%, and 24.88%, respectively. Although the mean inbreeding was considered low, the ROH indicates a recent and intensive use of few sires, which should be avoided for the genetic progress of breed.Keywords: autozygosity, Bos taurus indicus, genomic information, single nucleotide polymorphism
Procedia PDF Downloads 1431003 Speech Detection Model Based on DNN Classifier for Speech Emotions Recognition
Authors: Aisultan Shoiynbek, Darkhan Kuanyshbay, Paulo Menezes, Akbayan Bekarystankyzy, Assylbek Mukhametzhanov, Temirlan Shoiynbek
Abstract:
Speech emotion recognition has received increasing research interest in recent years. Most research has utilized emotional speech collected under controlled conditions, which was recorded by actors imitating and artificially producing emotions in front of a microphone. There are four issues related to that approach: (1) emotions are not natural, meaning that machines are learning to recognize fake emotions; (2) emotions are very limited in quantity and poor in variety of speaking; (3) there is some language dependency in SER; (4) consequently, each time researchers want to start work with SER, they need to find a good emotional database in their language. This paper proposes an approach to create an automatic tool for speech emotion extraction based on facial emotion recognition and describes the sequence of actions involved in the proposed approach. One of the first objectives in the sequence of actions is the speech detection issue. The paper provides a detailed description of the speech detection model based on a fully connected deep neural network for Kazakh and Russian. Despite the high results in speech detection for Kazakh and Russian, the described process is suitable for any language. To investigate the working capacity of the developed model, an analysis of speech detection and extraction from real tasks has been performed.Keywords: deep neural networks (DNN), speech detection, speech emotion recognition (SER), Mel-frequency cepstrum coefficients (MFCC), collecting speech emotion corpus, collecting speech emotion dataset, Kazakh speech dataset
Procedia PDF Downloads 41002 PCR Based DNA Analysis in Detecting P53 Mutation in Human Breast Cancer (MDA-468)
Authors: Debbarma Asis, Guha Chandan
Abstract:
Tumor Protein-53 (P53) is one of the tumor suppressor proteins. P53 regulates the cell cycle that conserves stability by preventing genome mutation. It is named so as it runs as 53-kilodalton (kDa) protein on Polyacrylamide gel electrophoresis although the actual mass is 43.7 kDa. Experimental evidence has indicated that P53 cancer mutants loses tumor suppression activity and subsequently gain oncogenic activities to promote tumourigenesis. Tumor-specific DNA has recently been detected in the plasma of breast cancer patients. Detection of tumor-specific genetic materials in cancer patients may provide a unique and valuable tumor marker for diagnosis and prognosis. Commercially available MDA-468 breast cancer cell line was used for the proposed study.Keywords: tumor protein (P53), cancer mutants, MDA-468, tumor suppressor gene
Procedia PDF Downloads 4721001 Pale, Firm and Non-Exudative (PFN): An Emerging Major Broiler Breast Meat Group
Authors: Cintia Midori Kaminishikawahara, Fernanda Jéssica Mendonça, Moisés Grespan, Elza Iouko Ida, Massami Shimokomaki, Adriana Lourenço Soares
Abstract:
The quality of broiler breast meat is changing as a result of continuing emphasis on genetically bird’s selection for efficiently higher meat production. The consumer is experiencing a cooked product that is drier and less juicy when consumed. Breast meat has been classified as PSE (pale, soft, exudative), DFD (dark, firm, dry) and normal color meat. However, recently variations of this color have been observed and they are not in line with the specificity of the meat functional properties. Thus, the objective of this work was to report the finding of a new pale meat color group characterized as Pale, Firm and Non-exudative (PFN) based on its pH, color, meat functional properties and micro structural evaluation. Breast meat fillets samples (n=1045) from commercial line were classified into PSE (pH ≤5.8, L* ≥ 53.0), PFN (pH > 5.8 and L* ≥ 53.0) and Normal (pH >5.8 and L* < 53.0), based on pH and L* values. In sequence, a total of 30 samples of each group were analyzed for the water holding capacity (WHC) and shear force (SF). The incidence was 9.1% for PSE meat, 85.7% for PFN and 5.2% for Normal meat. The PSE meat presented lower values of WHC (P ≤ 0.05) followed in sequence by PFN and Normal samples and also the SF values of fresh PFN was higher than PSE meat (P ≤ 0.05) and similar to Normal samples. Under optical microscopy, the cell diameter was 10% higher for PFN in relation to PSE meat and similar to Normal meat. These preliminary results indicate an emerging group of breast meat and it should be considered that the Pale, Firm and Non-exudative should be considered as an ideal broiler breast meat quality.Keywords: broiler PSE meat, light microscopy, texture, water holding capacity
Procedia PDF Downloads 3401000 Sequence Polymorphism and Haplogroup Distribution of Mitochondrial DNA Control Regions HVS1 and HVS2 in a Southwestern Nigerian Population
Authors: Ogbonnaya O. Iroanya, Samson T. Fakorede, Osamudiamen J. Edosa, Hadiat A. Azeez
Abstract:
The human mitochondrial DNA (mtDNA) is about 17 kbp circular DNA fragments found within the mitochondria together with smaller fragments of 1200 bp known as the control region. Knowledge of variation within populations has been employed in forensic and molecular anthropology studies. The study was aimed at investigating the polymorphic nature of the two hypervariable segments (HVS) of the mtDNA, i.e., HVS1 and HVS2, and to determine the haplogroup distribution among individuals resident in Lagos, Southwestern Nigeria. Peripheral blood samples were obtained from sixty individuals who are not related maternally, followed by DNA extraction and amplification of the extracted DNA using primers specific for the regions under investigation. DNA amplicons were sequenced, and sequenced data were aligned and compared to the revised Cambridge Reference Sequence (rCRS) GenBank Accession number: NC_012920.1) using BioEdit software. Results obtained showed 61 and 52 polymorphic nucleotide positions for HVS1 and HVS2, respectively. While a total of three indels mutation were recorded for HVS1, there were seven for HVS2. Also, transition mutations predominate nucleotide change observed in the study. Genetic diversity (GD) values for HVS1 and HVS2 were estimated to be 84.21 and 90.4%, respectively, while random match probability was 0.17% for HVS1 and 0.89% for HVS2. The study also revealed mixed haplogroups specific to the African (L1-L3) and the Eurasians (U and H) lineages. New polymorphic sites obtained from the study are promising for human identification purposes.Keywords: hypervariable region, indels, mitochondrial DNA, polymorphism, random match probability
Procedia PDF Downloads 107999 Strain Based Failure Criterion for Composite Notched Laminates
Authors: Ibrahim A. Elsayed, Mohamed H. Elalfy, Mostafa M. Abdalla
Abstract:
A strain-based failure criterion for composite notched laminates is introduced where the most critical stress concentration factor for the anisotropic notched laminates could be related to the failure of the corresponding quasi-isotropic laminate and the anisotropy ratio of the laminate. The proposed criterion will simplify the design of composites to meet notched failure requirements by eliminating the need for the detailed specifications of the stacking sequence at the preliminary design stage. The designer will be able to design based on the stiffness of the laminate, then at a later stage, select an appropriate stacking sequence to meet the stiffness requirements. The failure strains for the notched laminates are computed using the material’s Omni-strain envelope. The concept of Omni-strain envelope concerns the region of average strain where the laminate is safe regardless of ply orientation. In this work, we use Hashin’s failure criteria and the strains around the hole are computed using Savin’s analytic solution. A progressive damage analysis study has been conducted where the failure loads for the notched laminates are computed using finite element analysis. The failure strains are computed and used to estimate the concentration factor. It is found that the correlation found using Savin’s analytic solution predicts the same ratio of concentration factors between anisotropic and quasi-isotropic laminates as the more expensive progressive failure analysis.Keywords: anisotropy ratio, failure criteria, notched laminates, Omni-strain envelope, savin’s solution
Procedia PDF Downloads 109998 A Study on the Treatment of Municipal Waste Water Using Sequencing Batch Reactor
Authors: Bhaven N. Tandel, Athira Rajeev
Abstract:
Sequencing batch reactor process is a suspended growth process operating under non-steady state conditions which utilizes a fill and draw reactor with complete mixing during the batch reaction step (after filling) and where the subsequent steps of aeration and clarification occur in the same tank. All sequencing batch reactor systems have five steps in common, which are carried out in sequence as follows, (1) fill (2) react (3) settle (sedimentation/clarification) (4) draw (decant) and (5) idle. The study was carried out in a sequencing batch reactor of dimensions 44cmx30cmx70cm with a working volume of 40 L. Mechanical stirrer of 100 rpm was used to provide continuous mixing in the react period and oxygen was supplied by fish tank aerators. The duration of a complete cycle of sequencing batch reactor was 8 hours. The cycle period was divided into different phases in sequence as follows-0.25 hours fill phase, 6 hours react period, 1 hour settling phase, 0.5 hours decant period and 0.25 hours idle phase. The study consisted of two runs, run 1 and run 2. Run 1 consisted of 6 hours aerobic react period and run 2 consisted of 3 hours aerobic react period followed by 3 hours anoxic react period. The influent wastewater used for the study had COD, BOD, NH3-N and TKN concentrations of 308.03±48.94 mg/L, 100.36±22.05 mg/L, 14.12±1.18 mg/L, and 24.72±2.21 mg/L respectively. Run 1 had an average COD removal efficiency of 41.28%, BOD removal efficiency of 56.25%, NH3-N removal efficiency of 86.19% and TKN removal efficiency of 54.4%. Run 2 had an average COD removal efficiency of 63.19%, BOD removal efficiency of 73.85%, NH3-N removal efficiency of 90.74% and TKN removal efficiency of 65.25%. It was observed that run 2 gave better performance than run 1 in the removal of COD, BOD and TKN.Keywords: municipal waste water, aerobic, anoxic, sequencing batch reactor
Procedia PDF Downloads 534997 Genetic Diversity and Molecular Basis of Carbapenem Resistance in Acinetobacter Baumannii Isolates from Cattle
Authors: Minhas Alam, Muhammad Hidayat Rasool, Mohsin Khurshid, Bilal Aslam
Abstract:
Acinetobacter baumannii is a notorious bacterial pathogen that is an emerging nightmare in clinical settings and is mainly involved in severe nosocomial infections. However, the data related to carbapenem-resistant A. baumannii (CRAB) from veterinary settings is limited, especially in developing countries like Pakistan. To investigate the genetic diversity and molecular basis of carbapenem resistance in Acinetobacter baumannii isolates from Cattle, a total of 1960 samples were collected from cattle from Punjab, Pakistan. The isolates were analyzed by routine microbiological procedures and confirmed by polymerase chain reaction (PCR). The isolates were further screened for antimicrobial susceptibility and the presence of multiple antimicrobial-resistant determinants by PCR. Multilocus sequence typing (MLST) was performed. The results of the current study revealed that the overall prevalence of A. baumannii in cattle was 3.28% (65/1980). Among cattle 27.7% (18/65) were found CRAB strains. The CRAB isolates harbor class D β- lactamases genes, e-g, blaOXA-23 and blaOXA-51, 94.4% (17/18). CRAB isolates carry class B β- lactamases gene blaIMP, and only one isolate carries the blaNDM-1 gene. The MLST results of CRAB isolates from cattle demonstrated 5 STs and one new ST. The commonly found sequence types in CRAB isolates were ST2 (n=10, 55.5%), followed by ST642 (n=5, 27.8%) and ST600 & ST889 (n=1, 5.55%). The presence of CRAB isolates in cattle indicates an alarming situation in Punjab, Pakistan. Immediate control measures should be taken to stop the transmission of CRAB isolates within cattle, to the environment, and to clinical settings.Keywords: acinetobacter baumannii, carbapenemases, veterinary, drug resistance
Procedia PDF Downloads 47996 Association of Nuclear – Mitochondrial Epistasis with BMI in Type 1 Diabetes Mellitus Patients
Authors: Agnieszka H. Ludwig-Slomczynska, Michal T. Seweryn, Przemyslaw Kapusta, Ewelina Pitera, Katarzyna Cyganek, Urszula Mantaj, Lucja Dobrucka, Ewa Wender-Ozegowska, Maciej T. Malecki, Pawel Wolkow
Abstract:
Obesity results from an imbalance between energy intake and its expenditure. Genome-Wide Association Study (GWAS) analyses have led to discovery of only about 100 variants influencing body mass index (BMI), which explain only a small portion of genetic variability. Analysis of gene epistasis gives a chance to discover another part. Since it was shown that interaction and communication between nuclear and mitochondrial genome are indispensable for normal cell function, we have looked for epistatic interactions between the two genomes to find their correlation with BMI. Methods: The analysis was performed on 366 T1DM patients using Illumina Infinium OmniExpressExome-8 chip and followed by imputation on Michigan Imputation Server. Only genes which influence mitochondrial functioning (listed in Human MitoCarta 2.0) were included in the analysis – variants of nuclear origin (MAF > 5%) in 1140 genes and 42 mitochondrial variants (MAF > 1%). Gene expression analysis was performed on GTex data. Association analysis between genetic variants and BMI was performed with the use of Linear Mixed Models as implemented in the package 'GENESIS' in R. Analysis of association between mRNA expression and BMI was performed with the use of linear models and standard significance tests in R. Results: Among variants involved in epistasis between mitochondria and nucleus we have identified one in mitochondrial transcription factor, TFB2M (rs6701836). It interacted with mitochondrial variants localized to MT-RNR1 (p=0.0004, MAF=15%), MT-ND2 (p=0.07, MAF=5%) and MT-ND4 (p=0.01, MAF=1.1%). Analysis of the interaction between nuclear variant rs6701836 (nuc) and rs3021088 localized to MT-ND2 mitochondrial gene (mito) has shown that the combination of the two led to BMI decrease (p=0.024). Each of the variants on its own does not correlate with higher BMI [p(nuc)=0.856, p(mito)=0.116)]. Although rs6701836 is intronic, it influences gene expression in the thyroid (p=0.000037). rs3021088 is a missense variant that leads to alanine to threonine substitution in the MT-ND2 gene which belongs to complex I of the electron transport chain. The analysis of the influence of genetic variants on gene expression has confirmed the trend explained above – the interaction of the two genes leads to BMI decrease (p=0.0308). Each of the mRNAs on its own is associated with higher BMI (p(mito)=0.0244 and p(nuc)=0.0269). Conclusıons: Our results show that nuclear-mitochondrial epistasis can influence BMI in T1DM patients. The correlation between transcription factor expression and mitochondrial genetic variants will be subject to further analysis.Keywords: body mass index, epistasis, mitochondria, type 1 diabetes
Procedia PDF Downloads 167995 In silico and in vitro Investigation of the Role of Acinetobacter baumannii in the Pathogenesis of Multiple Sclerosis
Authors: Kieren Luellman, Makenzi Rockwell, Eduardo Callegari, Nichole Haag, Chun Wu
Abstract:
Multiple sclerosis (MS) is an autoimmune disorder that damages the myelin sheath of neurons in the central nervous system. The presence of Acinetobacter bacteria and anti-Acinetobacter antibodies in MS patients has led to the hypothesis that the bacteria may contribute to MS pathogenesis. In this study, the protein sequences of Acinetobacter baumannii were compared to five peptides from three mammalian myelin proteins, i.e., Proteolipid Protein (PLP): PLP 139-151, PLP 178-191, Myelin Basic Protein (MBP): MBP 84-104 and Myelin Oligodendrocyte Glycoprotein (MOG): MOG 35-55 and MOG 92-106 respectively, known to induce experimental autoimmune encephalomyelitis (EAE), a condition similar to MS. We found 11 hits (i.e., with five or more amino acid sequence similarity) in Acinetobacter baumannii, which are identical or similar to PLP139-151, 32 hits to PLP178-191, 35 to MBP 84-104, 41 hits to MOG 35-55 and 26 hits to MOG92-106. In addition, Western blotting was used to assess possible interaction between the bacterial proteins and human anti-MBP, anti-MOG, and anti-PLP antibodies produced in rabbits, corresponding to MBP 84-104, MOG 35-55, and PLP 139-151, respectively. We found that both human Polyclonal anti-MOG antibody and anti-PLP antibody recognized a protein or more proteins of the same molecular mass of around 25 kDa. in Acinetobacter baumannii. The results suggested that this/these protein(s) might potentially serve as antigen(s) to induce anti-MOG antibody and anti-PLP antibody production in mammalian B cells. The proteomic study identified 433 hits, among which the sequence of Acinetobacter baumannii protein 491 subunit A matches a previously published enzyme Acinetobacter 3-Oxoadipate CoA-Transferase, in which a fragment of its peptide was observed to recognize MS patient serum via ELISA method. Our findings might pave the road to understanding one of the pathogenesis mechanisms of MS.Keywords: multiple sclerosis, pathogenesis, Acinetobacter baumannii, antibody recognition
Procedia PDF Downloads 107994 Palynology of the Cretaceous Deposits of the Southeast Sirt Basin, Libya
Authors: Khaled Mahmud Gaddah
Abstract:
Cretaceous deposits in the southeast of the Sirt Basin of Libya occur in several grabens that formed during a major phase of rifting related to the opening of the Tethys. They include continental deposits of Early Cretaceous age that belong to the Nubian Formation and marginal to fully marine deposits of Late Cretaceous age that belong to the Lidam Formation and transitional beds. The sequence was extensively sampled from twenty-two boreholes and palynologically analysed. Much of the sequence is barren. However, subordinate shales in all formations yield diverse assemblages of poorly to well preserved and thermally middle to post mature palynomorphs. Most of the assemblages contain non-marine palynomorphs (spores, pollen, and freshwater algae), although some contain rare marine elements (dinoflagellate cysts and acritarchs). The palynofloras enabled the recognition of six assemblage zones of the late Barremian-Turonian age based on the dominant and base/top occurrences of stratigraphically useful palynomorphs: AI (Afropollisspp.-Aequitriraditesspinulosus) of late Barremian age; AIIa (Scenedesmusbifidus-S. sp.) of late Barremian?-early Aptian age; AIIb (Afropollisoperculatus-A. zonatus) of Aptian age; AIII (Crybelosporitespannuceus-Afropollisjardinus) of early Albian age; AIV (Subtilisphaera sp.-Lophosphaeridiumspp.) of Cenomanian-?Turonian age; AIV (Pediastrumbifidites-Leiosphaeridiaspp.) of Cenomanian?-Turonian age. These assemblages are comparable to others from Northern Gondwana (particularly from Libya and Egypt) and correspond to established Cretaceous palynofloral provinces. Palynofacies analysis is used to interpret the depositional environments, and five palynofacies types are recognised that reflect increasing marine influence up section.Keywords: palynology, palynomorphs, palynofacies, tethys, sirt basin
Procedia PDF Downloads 85993 In vivo Estimation of Mutation Rate of the Aleutian Mink Disease Virus
Authors: P.P. Rupasinghe, A.H. Farid
Abstract:
The Aleutian mink disease virus (AMDV, Carnivore amdoparvovirus 1) causes persistent infection, plasmacytosis, and formation and deposition of immune complexes in various organs in adult mink, leading to glomerulonephritis, arteritis and sometimes death. The disease has no cure nor an effective vaccine, and identification and culling of mink positive for anti-AMDV antibodies have not been successful in controlling the infection in many countries. The failure to eradicate the virus from infected farms may be caused by keeping false-negative individuals on the farm, virus transmission from wild animals, or neighboring farms. The identification of sources of infection, which can be performed by comparing viral sequences, is important in the success of viral eradication programs. High mutation rates could cause inaccuracies when viral sequences are used to trace back an infection to its origin. There is no published information on the mutation rate of AMDV either in vivo or in vitro. The in vivo estimation is the most accurate method, but it is difficult to perform because of the inherent technical complexities, namely infecting live animals, the unknown numbers of viral generations (i.e., infection cycles), the removal of deleterious mutations over time and genetic drift. The objective of this study was to determine the mutation rate of AMDV on which no information was available. A homogenate was prepared from the spleen of one naturally infected American mink (Neovison vison) from Nova Scotia, Canada (parental template). The near full-length genome of this isolate (91.6%, 4,143 bp) was bidirectionally sequenced. A group of black mink was inoculated with this homogenate (descendant mink). Spleen sampled were collected from 10 descendant mink after 16 weeks post-inoculation (wpi) and from anther 10 mink after 176 wpi, and their near-full length genomes were bi-directionally sequenced. Sequences of these mink were compared with each other and with the sequence of the parental template. The number of nucleotide substitutions at 176 wpi was 3.1 times greater than that at 16 wpi (113 vs 36) whereas the estimates of mutation rate at 176 wpi was 3.1 times lower than that at 176 wpi (2.85×10-3 vs 9.13×10-4 substitutions/ site/ year), showing a decreasing trend in the mutation rate per unit of time. Although there is no report on in vivo estimate of the mutation rate of DNA viruses in animals using the same method which was used in the current study, these estimates are at the higher range of reported values for DNA viruses determined by various techniques. These high estimates are logical based on the wide range of diversity and pathogenicity of AMDV isolates. The results suggest that increases in the number of nucleotide substitutions over time and subsequent divergence make it difficult to accurately trace back AMDV isolates to their origin when several years elapsed between the two samplings.Keywords: Aleutian mink disease virus, American mink, mutation rate, nucleotide substitution
Procedia PDF Downloads 118992 A Comprehensive Analysis of LACK (Leishmania Homologue of Receptors for Activated C Kinase) in the Context of Visceral Leishmaniasis
Authors: Sukrat Sinha, Abhay Kumar, Shanthy Sundaram
Abstract:
The Leishmania homologue of activated C kinase (LACK) is known T cell epitope from soluble Leishmania antigens (SLA) that confers protection against Leishmania challenge. This antigen has been found to be highly conserved among Leishmania strains. LACK has been shown to be protective against L. donovani challenge. A comprehensive analysis of several LACK sequences was completed. The analysis shows a high level of conservation, lower variability and higher antigenicity in specific portions of the LACK protein. This information provides insights for the potential consideration of LACK as a putative candidate in the context of visceral Leishmaniasis vaccine target.Keywords: bioinformatics, genome assembly, leishmania activated protein kinase c (lack), next-generation sequencing
Procedia PDF Downloads 330991 Symmetric Key Encryption Algorithm Using Indian Traditional Musical Scale for Information Security
Authors: Aishwarya Talapuru, Sri Silpa Padmanabhuni, B. Jyoshna
Abstract:
Cryptography helps in preventing threats to information security by providing various algorithms. This study introduces a new symmetric key encryption algorithm for information security which is linked with the "raagas" which means Indian traditional scale and pattern of music notes. This algorithm takes the plain text as input and starts its encryption process. The algorithm then randomly selects a raaga from the list of raagas that is assumed to be present with both sender and the receiver. The plain text is associated with the thus selected raaga and an intermediate cipher-text is formed as the algorithm converts the plain text characters into other characters, depending upon the rules of the algorithm. This intermediate code or cipher text is arranged in various patterns in three different rounds of encryption performed. The total number of rounds in the algorithm is equal to the multiples of 3. To be more specific, the outcome or output of the sequence of first three rounds is again passed as the input to this sequence of rounds recursively, till the total number of rounds of encryption is performed. The raaga selected by the algorithm and the number of rounds performed will be specified at an arbitrary location in the key, in addition to important information regarding the rounds of encryption, embedded in the key which is known by the sender and interpreted only by the receiver, thereby making the algorithm hack proof. The key can be constructed of any number of bits without any restriction to the size. A software application is also developed to demonstrate this process of encryption, which dynamically takes the plain text as input and readily generates the cipher text as output. Therefore, this algorithm stands as one of the strongest tools for information security.Keywords: cipher text, cryptography, plaintext, raaga
Procedia PDF Downloads 278990 A Quinary Coding and Matrix Structure Based Channel Hopping Algorithm for Blind Rendezvous in Cognitive Radio Networks
Authors: Qinglin Liu, Zhiyong Lin, Zongheng Wei, Jianfeng Wen, Congming Yi, Hai Liu
Abstract:
The multi-channel blind rendezvous problem in distributed cognitive radio networks (DCRNs) refers to how users in the network can hop to the same channel at the same time slot without any prior knowledge (i.e., each user is unaware of other users' information). The channel hopping (CH) technique is a typical solution to this blind rendezvous problem. In this paper, we propose a quinary coding and matrix structure-based CH algorithm called QCMS-CH. The QCMS-CH algorithm can guarantee the rendezvous of users using only one cognitive radio in the scenario of the asynchronous clock (i.e., arbitrary time drift between the users), heterogeneous channels (i.e., the available channel sets of users are distinct), and symmetric role (i.e., all users play a same role). The QCMS-CH algorithm first represents a randomly selected channel (denoted by R) as a fixed-length quaternary number. Then it encodes the quaternary number into a quinary bootstrapping sequence according to a carefully designed quaternary-quinary coding table with the prefix "R00". Finally, it builds a CH matrix column by column according to the bootstrapping sequence and six different types of elaborately generated subsequences. The user can access the CH matrix row by row and accordingly perform its channel, hoping to attempt rendezvous with other users. We prove the correctness of QCMS-CH and derive an upper bound on its Maximum Time-to-Rendezvous (MTTR). Simulation results show that the QCMS-CH algorithm outperforms the state-of-the-art in terms of the MTTR and the Expected Time-to-Rendezvous (ETTR).Keywords: channel hopping, blind rendezvous, cognitive radio networks, quaternary-quinary coding
Procedia PDF Downloads 80989 Improved Traveling Wave Method Based Fault Location Algorithm for Multi-Terminal Transmission System of Wind Farm with Grounding Transformer
Authors: Ke Zhang, Yongli Zhu
Abstract:
Due to rapid load growths in today’s highly electrified societies and the requirement for green energy sources, large-scale wind farm power transmission system is constantly developing. This system is a typical multi-terminal power supply system, whose structure of the network topology of transmission lines is complex. What’s more, it locates in the complex terrain of mountains and grasslands, thus increasing the possibility of transmission line faults and finding the fault location with difficulty after the faults and resulting in an extremely serious phenomenon of abandoning the wind. In order to solve these problems, a fault location method for multi-terminal transmission line based on wind farm characteristics and improved single-ended traveling wave positioning method is proposed. Through studying the zero sequence current characteristics by using the characteristics of the grounding transformer(GT) in the existing large-scale wind farms, it is obtained that the criterion for judging the fault interval of the multi-terminal transmission line. When a ground short-circuit fault occurs, there is only zero sequence current on the path between GT and the fault point. Therefore, the interval where the fault point exists is obtained by determining the path of the zero sequence current. After determining the fault interval, The location of the short-circuit fault point is calculated by the traveling wave method. However, this article uses an improved traveling wave method. It makes the positioning accuracy more accurate by combining the single-ended traveling wave method with double-ended electrical data. What’s more, a method of calculating the traveling wave velocity is deduced according to the above improvements (it is the actual wave velocity in theory). The improvement of the traveling wave velocity calculation method further improves the positioning accuracy. Compared with the traditional positioning method, the average positioning error of this method is reduced by 30%.This method overcomes the shortcomings of the traditional method in poor fault location of wind farm transmission lines. In addition, it is more accurate than the traditional fixed wave velocity method in the calculation of the traveling wave velocity. It can calculate the wave velocity in real time according to the scene and solve the traveling wave velocity can’t be updated with the environment and real-time update. The method is verified in PSCAD/EMTDC.Keywords: grounding transformer, multi-terminal transmission line, short circuit fault location, traveling wave velocity, wind farm
Procedia PDF Downloads 256988 LIZTOXD: Inclusive Lizard Toxin Database by Using MySQL Protocol
Authors: Iftikhar A. Tayubi, Tabrej Khan, Mansoor M. Alsubei, Fahad A. Alsaferi
Abstract:
LIZTOXD provides a single source of high-quality information about proteinaceous lizard toxins that will be an invaluable resource for pharmacologists, neuroscientists, toxicologists, medicinal chemists, ion channel scientists, clinicians, and structural biologists. We will provide an intuitive, well-organized and user-friendly web interface that allows users to explore the detail information of Lizard and toxin proteins. It includes common name, scientific name, entry id, entry name, protein name and length of the protein sequence. The utility of this database is that it can provide a user-friendly interface for users to retrieve the information about Lizard, toxin and toxin protein of different Lizard species. These interfaces created in this database will satisfy the demands of the scientific community by providing in-depth knowledge about Lizard and its toxin. In the next phase of our project we will adopt methodology and by using A MySQL and Hypertext Preprocessor (PHP) which and for designing Smart Draw. A database is a wonderful piece of equipment for storing large quantities of data efficiently. The users can thus navigate from one section to another, depending on the field of interest of the user. This database contains a wealth of information on species, toxins, toxins, clinical data etc. LIZTOXD resource that provides comprehensive information about protein toxins from lizard toxins. The combination of specific classification schemes and a rich user interface allows researchers to easily locate and view information on the sequence, structure, and biological activity of these toxins. This manually curated database will be a valuable resource for both basic researchers as well as those interested in potential pharmaceutical and agricultural applications of lizard toxins.Keywords: LIZTOXD, MySQL, PHP, smart draw
Procedia PDF Downloads 149987 A Sequence of Traumatic Pain: Feminist Issues within Laila Al-Othman’s Ṣamt al-Farāshāt (Silence of the Butterflies)
Authors: Khaled Igbaria
Abstract:
Laila Al-Othman is a well-known feminist writer in Kuwait and the entire Arab world. She was born in 1943 in Kuwait to a large and wealthy family. The author has written several short stories, as well as novels, such as The Woman and the Cat (1985) and Wasumayya Comes out of the Sea (1986), which was chosen as one of the best 100 Arab novels of the 21st century. Another prominent novel of hers is Ṣamt al-Farāshāt [Silence of the Butterflies] (2007), which was highly controversial in her native Kuwait upon publication. For this study, her engagement in feminism was achieved by exploring the different ways in which her novel, Ṣamt al-Farāshāt [Silence of the Butterflies], addresses several feminist issues, mainly forced marriage, rape and sexual abuse, gender-based physical, sexual violence, and enforced silence. This paper focuses on demonstrating social obstacles and continuous trauma caused by a sequence of pain experienced by Arab females in their patriarchal society. This study argues that the novel reveals a sustained effort to raise the banner of feminism and a strong desire to liberate Arab women from patriarchal domination. Al-Othman successfully and uniquely represents women as gender-based traumatic victims of sexual and physical violence, forced silence, and general oppression in the patriarchal Arab society, as those needing help, support, protection, and liberation. They are not represented as independent or free. Methodologically, the study employs a qualitative literary analysis method in addition to trauma theory psychoanalysis, concentrating on feminist issues highlighted in the novel.Keywords: Al-Othman, Arab women pain, trauma within narration., Silence of the Butterflies
Procedia PDF Downloads 46986 Telomerase, a Biomarker in Oral Cancer Cell Proliferation and Tool for Its Prevention at Initial Stage
Authors: Shaista Suhail
Abstract:
As cancer populations is increasing sharply, the incidence of oral squamous cell carcinoma (OSCC) has also been expected to increase. Oral carcinogenesis is a highly complex, multistep process which involves accumulation of genetic alterations that lead to the induction of proteins promoting cell growth (encoded by oncogenes), increased enzymatic (telomerase) activity promoting cancer cell proliferation. The global increase in frequency and mortality, as well as the poor prognosis of oral squamous cell carcinoma, has intensified current research efforts in the field of prevention and early detection of this disease. The advances in the understanding of the molecular basis of oral cancer should help in the identification of new markers. The study of the carcinogenic process of the oral cancer, including continued analysis of new genetic alterations, along with their temporal sequencing during initiation, promotion and progression, will allow us to identify new diagnostic and prognostic factors, which will provide a promising basis for the application of more rational and efficient treatments. Telomerase activity has been readily found in most cancer biopsies, in premalignant lesions or germ cells. Activity of telomerase is generally absent in normal tissues. It is known to be induced upon immortalization or malignant transformation of human cells such as in oral cancer cells. Maintenance of telomeres plays an essential role during transformation of precancer to malignant stage. Mammalian telomeres, a specialized nucleoprotein structures are composed of large conctamers of the guanine-rich sequence 5_-TTAGGG-3_. The roles of telomeres in regulating both stability of genome and replicative immortality seem to contribute in essential ways in cancer initiation and progression. It is concluded that activity of telomerase can be used as a biomarker for diagnosis of malignant oral cancer and a target for inactivation in chemotherapy or gene therapy. Its expression will also prove to be an important diagnostic tool as well as a novel target for cancer therapy. The activation of telomerase may be an important step in tumorgenesis which can be controlled by inactivating its activity during chemotherapy. The expression and activity of telomerase are indispensable for cancer development. There are no drugs which can effect extremely to treat oral cancers. There is a general call for new emerging drugs or methods that are highly effective towards cancer treatment, possess low toxicity, and have a minor environment impact. Some novel natural products also offer opportunities for innovation in drug discovery. Natural compounds isolated from medicinal plants, as rich sources of novel anticancer drugs, have been of increasing interest with some enzyme (telomerase) blockage property. The alarming reports of cancer cases increase the awareness amongst the clinicians and researchers pertaining to investigate newer drug with low toxicity.Keywords: oral carcinoma, telomere, telomerase, blockage
Procedia PDF Downloads 163985 Genetic Diversity Analysis in Ecological Populations of Persian Walnut
Authors: Masoud Sheidai, Fahimeh Koohdar, Hashem Sharifi
Abstract:
Juglans regia (L.) commonly known as Persian walnut of the genus Juglans L. (Juglandaceae) is one of the most important cultivated plant species due to its high-quality wood and edible nuts. The genetic diversity analysis is essential for conservation and management of tree species. Persian walnut is native from South-Eastern Europe to North-Western China through Tibet, Nepal, Northern India, Pakistan, and Iran. The species like Persian walnut, which has a wide range of geographical distribution, should harbor extensive genetic variability to adapt to environmental fluctuations they face. We aimed to study the population genetic structure of seven Persian walnut populations including three wild and four cultivated populations by using ISSR (Inter simple sequence repeats) and SRAP (Sequence related amplified polymorphism) molecular markers. We also aimed to compare the genetic variability revealed by ISSR neutral multilocus marker and rDNA ITS sequences. The studied populations differed in morphological features as the samples in each population were clustered together and were separate from the other populations. Three wild populations studied were placed close to each other. The mantel test after 5000 times permutation performed between geographical distance and morphological distance in Persian walnut populations produced significant correlation (r = 0.48, P = 0.002). Therefore, as the populations become farther apart, they become more divergent in morphological features. ISSR analysis produced 47 bands/ loci, while we obtained 15 SRAP bands. Gst and other differentiation statistics determined for these loci revealed that most of the ISSR and SRAP loci have very good discrimination power and can differentiate the studied populations. AMOVA performed for these loci produced a significant difference (< 0.05) supporting the above-said result. AMOVA produced significant genetic difference based on ISSR data among the studied populations (PhiPT = 0.52, P = 0.001). AMOVA revealed that 53% of the total variability is due to among population genetic difference, while 47% is due to within population genetic variability. The results showed that both multilocus molecular markers and ITS sequences can differentiate Persian walnut populations. The studied populations differed genetically and showed isolation by distance (IBD). ITS sequence based MP and Bayesian phylogenetic trees revealed that Iranian walnut cultivars form a distinct clade separated from the cultivars studied from elsewhere. Almost all clades obtained have high bootstrap value. The results indicated that a combination of multilpcus and sequencing molecular markers can be used in genetic differentiation of Persian walnut.Keywords: genetic diversity, population, molecular markers, genetic difference
Procedia PDF Downloads 156984 Approach to Honey Volatiles' Profiling by Gas Chromatography and Mass Spectrometry
Authors: Igor Jerkovic
Abstract:
Biodiversity of flora provides many different nectar sources for the bees. Unifloral honeys possess distinctive flavours, mainly derived from their nectar sources (characteristic volatile organic components (VOCs)). Specific or nonspecific VOCs (chemical markers) could be used for unifloral honey characterisation as addition to the melissopalynologycal analysis. The main honey volatiles belong, in general, to three principal categories: terpenes, norisoprenoids, and benzene derivatives. Some of these substances have been described as characteristics of the floral source, and other compounds, like several alcohols, branched aldehydes, and furan derivatives, may be related to the microbial purity of honey processing and storage conditions. Selection of the extraction method for the honey volatiles profiling should consider that heating of the honey produce different artefacts and therefore conventional methods of VOCs isolation (such as hydrodistillation) cannot be applied for the honey. Two-way approach for the isolation of the honey VOCs was applied using headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE). The extracts were analysed by gas chromatography and mass spectrometry (GC-MS). HS-SPME (with the fibers of different polarity such as polydimethylsiloxane/ divinylbenzene (PDMS/DVB) or divinylbenzene/carboxene/ polydimethylsiloxane (DVB/CAR/PDMS)) enabled isolation of high volatile headspace VOCs of the honey samples. Among them, some characteristic or specific compounds can be found such as 3,4-dihydro-3-oxoedulan (in Centaurea cyanus L. honey) or 1H-indole, methyl anthranilate, and cis-jasmone (in Citrus unshiu Marc. honey). USE with different solvents (mainly dichloromethane or the mixture pentane : diethyl ether 1 : 2 v/v) enabled isolation of less volatile and semi-volatile VOCs of the honey samples. Characteristic compounds from C. unshiu honey extracts were caffeine, 1H-indole, 1,3-dihydro-2H-indol-2-one, methyl anthranilate, and phenylacetonitrile. Sometimes, the selection of solvent sequence was useful for more complete profiling such as sequence I: pentane → diethyl ether or sequence II: pentane → pentane/diethyl ether (1:2, v/v) → dichloromethane). The extracts with diethyl ether contained hydroquinone and 4-hydroxybenzoic acid as the major compounds, while (E)-4-(r-1’,t-2’,c-4’-trihydroxy-2’,6’,6’-trimethylcyclo-hexyl)but-3-en-2-one predominated in dichloromethane extracts of Allium ursinum L. honey. With this two-way approach, it was possible to obtain a more detailed insight into the honey volatile and semi-volatile compounds and to minimize the risks of compound discrimination due to their partial extraction that is of significant importance for the complete honey profiling and identification of the chemical biomarkers that can complement the pollen analysis.Keywords: honey chemical biomarkers, honey volatile compounds profiling, headspace solid-phase microextraction (HS-SPME), ultrasonic solvent extraction (USE)
Procedia PDF Downloads 190983 A Basic Modeling Approach for the 3D Protein Structure of Insulin
Authors: Daniel Zarzo Montes, Manuel Zarzo Castelló
Abstract:
Proteins play a fundamental role in biology, but their structure is complex, and it is a challenge for teachers to conceptually explain the differences between their primary, secondary, tertiary, and quaternary structures. On the other hand, there are currently many computer programs to visualize the 3D structure of proteins, but they require advanced training and knowledge. Moreover, it becomes difficult to visualize the sequence of amino acids in these models, and how the protein conformation is reached. Given this drawback, a simple and instructive procedure is proposed in order to teach the protein structure to undergraduate and graduate students. For this purpose, insulin has been chosen because it is a protein that consists of 51 amino acids, a relatively small number. The methodology has consisted of the use of plastic atom models, which are frequently used in organic chemistry and biochemistry to explain the chirality of biomolecules. For didactic purposes, when the aim is to teach the biochemical foundations of proteins, a manipulative system seems convenient, starting from the chemical structure of amino acids. It has the advantage that the bonds between amino acids can be conveniently rotated, following the pattern marked by the 3D models. First, the 51 amino acids were modeled, and then they were linked according to the sequence of this protein. Next, the three disulfide bonds that characterize the stability of insulin have been established, and then the alpha-helix structure has been formed. In order to reach the tertiary 3D conformation of this protein, different interactive models available on the Internet have been visualized. In conclusion, the proposed methodology seems very suitable for biology and biochemistry students because they can learn the fundamentals of protein modeling by means of a manipulative procedure as a basis for understanding the functionality of proteins. This methodology would be conveniently useful for a biology or biochemistry laboratory practice, either at the pre-graduate or university level.Keywords: protein structure, 3D model, insulin, biomolecule
Procedia PDF Downloads 42982 Mutational and Evolutionary Analysis of Interleukin-2 Gene in Four Pakistani Goat Breeds
Authors: Tanveer Hussain, Misbah Hussain, Masroor Ellahi Babar, Muhammad Traiq Pervez, Fiaz Hussain, Sana Zahoor, Rashid Saif
Abstract:
Interleukin 2 (IL-2) is a cytokine which is produced by activated T cells, play important role in immune response against antigen. It act in both autocrine and paracrine manner. It can stimulate B cells and various other phagocytic cells like monocytes, lymphokine-activated killer cells and natural killer cells. Acting in autocrine fashion, IL-2 protein plays a crucial role in proliferation of T cells. IL-2 triggers the release of pro and anti- inflammatory cytokines by activating several pathways. In present study, exon 1 of IL-2 gene of four local Pakistani breeds (Dera Din Panah, Beetal, Nachi and Kamori) from two provinces was amplified by using reported Ovine IL-2 primers, yielding PCR product of 501 bp. The sequencing of all samples was done to identify the polymorphisms in amplified region of IL-2 gene. Analysis of sequencing data resulted in identification of one novel nucleotide substitution (T→A) in amplified non-coding region of IL-2 gene. Comparison of IL-2 gene sequence of all four breeds with other goat breeds showed high similarity in sequence. While phylogenetic analysis of our local breeds with other mammals showed that IL-2 is a variable gene which has undergone many substitutions. This high substitution rate can be due to the decreased or increased changed selective pressure. These rapid changes can also lead to the change in function of immune system. This pioneering study of Pakistani goat breeds urge for further studies on immune system of each targeted breed for fully understanding the functional role of IL-2 in goat immunity.Keywords: interleukin 2, mutational analysis, phylogeny, goat breeds, Pakistan
Procedia PDF Downloads 600981 Development of One-Pot Sequential Cyclizations and Photocatalyzed Decarboxylative Radical Cyclization: Application Towards Aspidospermatan Alkaloids
Authors: Guillaume Bélanger, Jean-Philippe Fontaine, Clémence Hauduc
Abstract:
There is an undeniable thirst from organic chemists and from the pharmaceutical industry to access complex alkaloids with short syntheses. While medicinal chemists are interested in the fascinating wide range of biological properties of alkaloids, synthetic chemists are rather interested in finding new routes to access these challenging natural products of often low availability from nature. To synthesize complex polycyclic cores of natural products, reaction cascades or sequences performed one-pot offer a neat advantage over classical methods for their rapid increase in molecular complexity in a single operation. In counterpart, reaction cascades need to be run on substrates bearing all the required functional groups necessary for the key cyclizations. Chemoselectivity is thus a major issue associated with such a strategy, in addition to diastereocontrol and regiocontrol for the overall transformation. In the pursuit of synthetic efficiency, our research group developed an innovative one-pot transformation of linear substrates into bi- and tricyclic adducts applied to the construction of Aspidospermatan-type alkaloids. The latter is a rich class of indole alkaloids bearing a unique bridged azatricyclic core. Despite many efforts toward the synthesis of members of this family, efficient and versatile synthetic routes are still coveted. Indeed, very short, non-racemic approaches are rather scarce: for example, in the cases of aspidospermidine and aspidospermine, syntheses are all fifteen steps and over. We envisaged a unified approach to access several members of the Aspidospermatan alkaloids family. The key sequence features a highly chemoselective formamide activation that triggers a Vilsmeier-Haack cyclization, followed by an azomethine ylide generation and intramolecular cycloaddition. Despite the high density and variety of functional groups on the substrates (electron-rich and electron-poor alkenes, nitrile, amide, ester, enol ether), the sequence generated three new carbon-carbon bonds and three rings in a single operation with good yield and high chemoselectivity. A detailed study of amide, nucleophile, and dipolarophile variations to finally get to the successful combination required for the key transformation will be presented. To complete the indoline fragment of the natural products, we developed an original approach. Indeed, all reported routes to Aspidospermatan alkaloids introduce the indoline or indole early in the synthesis. In our work, the indoline needs to be installed on the azatricyclic core after the key cyclization sequence. As a result, typical Fischer indolization is not suited since this reaction is known to fail on such substrates. We thus envisaged a unique photocatalyzed decarboxylative radical cyclization. The development of this reaction as well as the scope and limitations of the methodology, will also be presented. The original Vilsmeier-Haack and azomethine ylide cyclization sequence as well as the new photocatalyzed decarboxylative radical cyclization will undoubtedly open access to new routes toward polycyclic indole alkaloids and derivatives of pharmaceutical interest in general.Keywords: Aspidospermatan alkaloids, azomethine ylide cycloaddition, decarboxylative radical cyclization, indole and indoline synthesis, one-pot sequential cyclizations, photocatalysis, Vilsmeier-Haack Cyclization
Procedia PDF Downloads 72980 Biodegradation of Malathion by Acinetobacter baumannii Strain AFA Isolated from Domestic Sewage in Egypt
Authors: Ahmed F. Azmy, Amal E. Saafan, Tamer M. Essam, Magdy A. Amin, Shaban H. Ahmed
Abstract:
Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosae (PS1),andPseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazenon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 1012CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.Keywords: Acinetobacter baumannii, biodegradation, malathion, organophosphate pesticides
Procedia PDF Downloads 481979 Study of Microbial Diversity Associated with Tarballs and Their Exploitation in Crude Oil Degradation
Authors: Varsha Shinde, Belle Damodara Shenoy
Abstract:
Tarballs are crude oil remnants found in oceans after long term weathering process and are a global concern since several decades as potential marine pollutant. Being complicated in structure microbial remediation of tarballs in natural environment is a slow process. They are rich in high molecular weight alkanes and poly aromatic hydrocarbons which are resistant to microbial attack and other environmental factors, therefore remain in environment for long time. However, it has been found that many bacteria and fungi inhabit on tarballs for nutrients and shelter. Many of them are supposed to be oil degraders, while others are supposed to be getting benefited by byproducts formed during hydrocarbon metabolism. Thus tarballs are forming special interesting ecological niche of microbes. This work aimed to study diversity of bacteria and fungi from tarballs and to see their potential application in crude oil degradation. The samples of tarballs were collected from Betul beach of south Goa (India). Different methods were used to isolate culturable fraction of bacteria and fungi from it. Those were sequenced for 16S rRNA gene and ITS for molecular level identification. The 16S rRNA gene sequence analysis revealed the presence of 13 bacterial genera/clades (Alcanivorax, Brevibacterium, Bacillus, Cellulomonas, Enterobacter, Klebsiella, Marinobacter, Nitratireductor, Pantoea, Pseudomonas, Pseudoxanthomonas, Tistrella and Vibrio), while the ITS sequence analysis placed the fungi in 8 diverse genera/ clades (Aspergillus, Byssochlamys, Monascus, Paecilomyces, Penicillium, Scytalidium/ Xylogone, Talaromyces and Trichoderma). All bacterial isolates were screened for oil degradation capacity. Potential strains were subjected to crude oil degradation experiment for quantification. Results were analyzed by GC-MS-MS.Keywords: bacteria, biodegradation, crude oil, diversity, fungi, tarballs
Procedia PDF Downloads 215978 Optical and Near-UV Spectroscopic Properties of Low-Redshift Jetted Quasars in the Main Sequence in the Main Sequence Context
Authors: Shimeles Terefe Mengistue, Ascensión Del Olmo, Paola Marziani, Mirjana Pović, María Angeles Martínez-Carballo, Jaime Perea, Isabel M. Árquez
Abstract:
Quasars have historically been classified into two distinct classes, radio-loud (RL) and radio-quiet (RQ), taking into account the presence and absence of relativistic radio jets, respectively. The absence of spectra with a high S/N ratio led to the impression that all quasars (QSOs) are spectroscopically similar. Although different attempts were made to unify these two classes, there is a long-standing open debate involving the possibility of a real physical dichotomy between RL and RQ quasars. In this work, we present new high S/N spectra of 11 extremely powerful jetted quasars with radio-to-optical flux density ratio > 1000 that concomitantly cover the low-ionization emission of Mgii𝜆2800 and Hbeta𝛽 as well as the Feii blends in the redshift range 0.35 < z < 1, observed at Calar Alto Observatory (Spain). This work aims to quantify broad emission line differences between RL and RQ quasars by using the four-dimensional eigenvector 1 (4DE1) parameter space and its main sequence (MS) and to check the effect of powerful radio ejection on the low ionization broad emission lines. Emission lines are analysed by making two complementary approaches, a multicomponent non-linear fitting to account for the individual components of the broad emission lines and by analysing the full profile of the lines through parameters such as total widths, centroid velocities at different fractional intensities, asymmetry, and kurtosis indices. It is found that broad emission lines show large reward asymmetry both in Hbeta𝛽 and Mgii2800A. The location of our RL sources in a UV plane looks similar to the optical one, with weak Feii UV emission and broad Mgii2800A. We supplement the 11 sources with large samples from previous work to gain some general inferences. The result shows, compared to RQ, our extreme RL quasars show larger median Hbeta full width at half maximum (FWHM), weaker Feii emission, larger 𝑀BH, lower 𝐿bol/𝐿Edd, and a restricted space occupation in the optical and UV MS planes. The differences are more elusive when the comparison is carried out by restricting the RQ population to the region of the MS occupied by RL quasars, albeit an unbiased comparison matching 𝑀BH and 𝐿bol/𝐿Edd suggests that the most powerful RL quasars show the highest redward asymmetries in Hbeta.Keywords: galaxies, active, line, profiles, quasars, emission lines, supermassive black holes
Procedia PDF Downloads 53977 Lucilia Sericata Netrin-A: Secreted by Salivary Gland Larvae as a Potential to Neuroregeneration
Authors: Hamzeh Alipour, Masoumeh Bagheri, Tahereh Karamzadeh, Abbasali Raz, Kourosh Azizi
Abstract:
Netrin-A, a protein identified for conducting commissural axons, has a similar role in angiogenesis. In addition, studies have shown that one of the netrin-A receptors is expressed in the growing cells of small capillaries. It will be interesting to study this new group of molecules because their role in wound healing will become clearer in the future due to angiogenesis. The greenbottle blowfly Luciliasericata (L. sericata) larvae are increasingly used in maggot therapy of chronic wounds. This aim of this was the identification of moleculareatures of Netrin-A in L. sericata larvae. Larvae were reared under standard maggotarium conditions. The nucleic acid sequence of L. sericataNetrin-A (LSN-A) was then identified using Rapid Amplification of cDNA Ends (RACE) and Rapid Amplification of Genomic Ends (RAGE). Parts of the Netrin-A gene, including the middle, 3′-, and 5′-ends were identified, TA cloned in pTG19 plasmid, and transferred into DH5ɑ Escherichia coli. Each part was sequenced and assembled using SeqMan software. This gene structure was further subjected to in silico analysis. The DNA of LSN-A was identified to be 2407 bp, while its mRNA sequence was recognized as 2115 bp by Oligo0.7 software. It translated the Netrin-A protein with 704 amino acid residues. Its molecular weight is estimated to be 78.6 kDa. The 3-D structure ofNetrin-A drawn by SWISS-MODEL revealed its similarity to the Netrin-1 of humans with 66.8% identity. The LSN-A protein conduces to repair the myelin membrane in neuronal cells. Ultimately, it can be an effective candidate in neural regeneration and wound healing. Furthermore, our next attempt is to deplore recombinant proteins for use in medical sciences.Keywords: maggot therapy, netrin-A, RACE, RAGE, lucilia sericata
Procedia PDF Downloads 99976 Carbon Based Classification of Aquaporin Proteins: A New Proposal
Authors: Parul Johri, Mala Trivedi
Abstract:
Major Intrinsic proteins (MIPs), actively involved in the passive transport of small polar molecules across the membranes of almost all living organisms. MIPs that specifically transport water molecules are named aquaporins (AQPs). The permeability of membranes is actively controlled by the regulation of the amount of different MIPs present but also in some cases by phosphorylation and dephosphorylation of the channel. Based on sequence similarity, MIPs have been classified into many categories. All of the proteins are made up of the 20 amino acids, the only difference is there in their orientations. Again all the 20 amino acids are made up of the basic five elements namely: carbon, hydrogen, oxygen, sulphur and nitrogen. These elements are responsible for giving the amino acids the properties of hydrophilicity/hydrophobicity which play an important role in protein interactions. The hydrophobic amino acids characteristically have greater number of carbon atoms as carbon is the main element which contributes to hydrophobic interactions in proteins. It is observed that the carbon level of proteins in different species is different. In the present work, we have taken a sample set of 150 aquaporins proteins from Uniprot database and a dynamic programming code was written to calculate the carbon percentage for each sequence. This carbon percentage was further used to barcode the aqauporins of animals and plants. The protein taken from Oryza sativa, Zea mays and Arabidopsis thaliana preferred to have carbon percentage of 31.8 to 35, whereas on the other hand sequences taken from Mus musculus, Saccharomyces cerevisiae, Homo sapiens, Bos Taurus, and Rattus norvegicus preferred to have carbon percentage of 31 to 33.7. This clearly demarks the carbon range in the aquaporin proteins from plant and animal origin. Hence the atom level analysis of protein sequences can provide us with better results as compared to the residue level comparison.Keywords: aquaporins, carbon, dynamic prgramming, MIPs
Procedia PDF Downloads 360