Search results for: gene bank
1683 EnumTree: An Enumerative Biclustering Algorithm for DNA Microarray Data
Authors: Haifa Ben Saber, Mourad Elloumi
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In a number of domains, like in DNA microarray data analysis, we need to cluster simultaneously rows (genes) and columns (conditions) of a data matrix to identify groups of constant rows with a group of columns. This kind of clustering is called biclustering. Biclustering algorithms are extensively used in DNA microarray data analysis. More effective biclustering algorithms are highly desirable and needed. We introduce a new algorithm called, Enumerative tree (EnumTree) for biclustering of binary microarray data. is an algorithm adopting the approach of enumerating biclusters. This algorithm extracts all biclusters consistent good quality. The main idea of EnumLat is the construction of a new tree structure to represent adequately different biclusters discovered during the process of enumeration. This algorithm adopts the strategy of all biclusters at a time. The performance of the proposed algorithm is assessed using both synthetic and real DNA micryarray data, our algorithm outperforms other biclustering algorithms for binary microarray data. Biclusters with different numbers of rows. Moreover, we test the biological significance using a gene annotation web tool to show that our proposed method is able to produce biologically relevent biclusters.Keywords: DNA microarray, biclustering, gene expression data, tree, datamining.
Procedia PDF Downloads 3721682 The Fadama Initiative: Implications for Human Security and Sustainable Development in Nigeria
Authors: Albert T. Akume, Yahya M. Abdullahi
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The impact of poverty on individual and society is grave, hence the efforts by the government to eradicate or alleviate. In Nigeria the various efforts to reduce rural poverty by empowering them and making the process of their development self-sustaining have ended dismally. That notwithstanding, government determination to conquer poverty has not diminish as in the early 1990s the government with financial collaboration from the World Bank and African Development Bank introduced the fadama project. It is against this backdrop that this paper uses the documentary and analytical research methods to examine the implication the fadama development project has for community capacity development and human security in Nigeria. From the analysis it was discovered the fadama project improved household income of fadama farmers, community empowerment, participatory development planning and support for demand driven productive investment in farm and non-farm activities including community infrastructures. Despite this impressive result the fadama project is challenged by conflict especially in northern Nigeria and late delivery of necessary farm consumables that aid improved productivity. It was therefore recommended that the government should strengthen her various state security institutions to proactively mitigate conflicts and to ensure that farm consumables and other support services reach farmers timely.Keywords: capacity development, empowerment, fadama, human security, poverty reduction, theory of change, sustainable development
Procedia PDF Downloads 4961681 TNFRSF11B Gene Polymorphisms A163G and G11811C in Prediction of Osteoporosis Risk
Authors: I. Boroňová, J.Bernasovská, J. Kľoc, Z. Tomková, E. Petrejčíková, D. Gabriková, S. Mačeková
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Osteoporosis is a complex health disease characterized by low bone mineral density, which is determined by an interaction of genetics with metabolic and environmental factors. Current research in genetics of osteoporosis is focused on identification of responsible genes and polymorphisms. TNFRSF11B gene plays a key role in bone remodeling. The aim of this study was to investigate the genotype and allele distribution of A163G (rs3102735) osteoprotegerin gene promoter and G1181C (rs2073618) osteoprotegerin first exon polymorphisms in the group of 180 unrelated postmenopausal women with diagnosed osteoporosis and 180 normal controls. Genomic DNA was isolated from peripheral blood leukocytes using standard methodology. Genotyping for presence of different polymorphisms was performed using the Custom Taqman®SNP Genotyping assays. Hardy-Weinberg equilibrium was tested for each SNP in the groups of participants using the chi-square (χ2) test. The distribution of investigated genotypes in the group of patients with osteoporosis were as follows: AA (66.7%), AG (32.2%), GG (1.1%) for A163G polymorphism; GG (19.4%), CG (44.4%), CC (36.1%) for G1181C polymorphism. The distribution of genotypes in normal controls were follows: AA (71.1%), AG (26.1%), GG (2.8%) for A163G polymorphism; GG (22.2%), CG (48.9%), CC (28.9%) for G1181C polymorphism. In A163G polymorphism the variant G allele was more common among patients with osteoporosis: 17.2% versus 15.8% in normal controls. Also, in G1181C polymorphism the phenomenon of more frequent occurrence of C allele in the group of patients with osteoporosis was observed (58.3% versus 53.3%). Genotype and allele distributions showed no significant differences (A163G: χ2=0.270, p=0.605; χ2=0.250, p=0.616; G1181C: χ2= 1.730, p=0.188; χ2=1.820, p=0.177). Our results represents an initial study, further studies of more numerous file and associations studies will be carried out. Knowing the distribution of genotypes is important for assessing the impact of these polymorphisms on various parameters associated with osteoporosis. Screening for identification of “at-risk” women likely to develop osteoporosis and initiating subsequent early intervention appears to be most effective strategy to substantially reduce the risks of osteoporosis.Keywords: osteoporosis, real-time PCR method, SNP polymorphisms
Procedia PDF Downloads 3291680 Phylogenetic Relationships between the Whole Sets of Individual Flow Sorted U, M, S and C Chromosomes of Aegilops and Wheat as Revealed by COS Markers
Authors: András Farkas, István Molnár, Jan Vrána, Veronika Burešová, Petr Cápal, András Cseh, Márta Molnár-Láng, Jaroslav Doležel
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Species of Aegilops played a central role in the evolution of wheat and are sources of traits related to yield quality and tolerance against biotic and abiotic stresses. These wild genes and alleles are desirable to use in crop improvement programs via introgressive hybridization. However, the success of chromosome mediated gene transfer to wheat are hampered by the pour knowledge on the genome structure of Aegilops relative to wheat and by the low number of cost-effective molecular markers specific for Aegilops chromosomes. The COS markers specific for genes conserved throughout evolution in both sequence and copy number between Triticeae/Aegilops taxa and define orthologous regions, thus enabling the comparison of regions on the chromosomes of related species. The present study compared individual chromosomes of Aegilops umbellulata (UU), Ae. comosa (MM), Ae. speltoides (SS) and Ae. caudata (CC) purified by flourescent labelling with oligonucleotid SSR repeats and biparametric flow cytometry with wheat by identifying orthologous chromosomal regions by COS markers. The linear order of bin-mapped COS markers along the wheat D chromosomes was identified by the use of chromosome-specific sequence data and virtual gene order. Syntenic regions of wheat identifying genome rearrangements differentiating the U, M, S or C genomes from the D genome of wheat were detected. The conserved orthologous set markers assigned to Aegilops chromosomes promise to accelerate gene introgression by facilitating the identification of alien chromatin. The syntenic relationships between the Aegilops species and wheat will facilitate the targeted development of new markers specific for U, M, S and C genomic regions and will contribute to the understanding of molecular processes related to the evolution of Aegilops.Keywords: Aegilops, cos-markers, flow-sorting, wheat
Procedia PDF Downloads 5021679 The Impact of Corruption on Exports and Innovation in Small and Medium-Sized Enterprises: The Case of Tunisia
Authors: Moujib Bahri, Rahim Kallel, Ouafa Sakka
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Corruption is a phenomenon that increases uncertainty and risk of SMEs as it undermines the quality of the business environment and the easy access to public services. Our research builds on existing research on corruption's effects on economic growth at the firm level. Several papers have analyzed the effect of firms’ payments of bribes on their performance; however, only limited research has investigated the link between corruption, innovation, and exports. Drawing on principal-agent theory, we explore how corruption weakens the institutional context and makes the business environment unsound and not conducive to innovation and exports. This study employs data from The Enterprise Surveys conducted in Tunisia between March 2013 and July 2014 by the World Bank, the European Bank for Reconstruction and Development (EBRD) and the European Investment Bank (EIB). The main objective of this survey was to gain a better understanding of Tunisian firms’ perception of the environment in which they operate. Since 2011, the country's political situation has become fragile and unstable, and public services are perceived as inefficient and corrupt. We test our hypotheses on a sample of 537 Tunisian manufacturing SMEs using structural equation modeling and path analysis. We find that political instability leads to higher level of corruption, and that excessive business licensing regulations create a fertile ground for bribery. Our findings do not support the greasing hypothesis suggesting that corruption can reduce the negative effect of bureaucratic delays and the hard access of companies to public services related to innovation and exports. Instead, our results support the sanding hypothesis according to which corruption hinders innovation activities and exports. Furthermore, corruption is found to, negatively and significantly, impact firms’ ownership of quality certificates. Our results suggest that, in an environment with a high level of corruption, governments and policymakers interested in assisting SMEs with their innovation and export activities should have a better control on corruption to allow them developing those activities without being forced to bribe government officers.Keywords: corruption, innovation, exports, SMEs
Procedia PDF Downloads 1801678 Altered Gene Expression: Induction/Suppression of some Pathogenesis Related Protein Genes in an Egyptian Isolate of Potato Leafroll Virus (PLRV)
Authors: Dalia G. Aseel
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The potato (Solanum tubersum, L.) has become one of the major vegetable crops in Egypt and all over the world. Potato leafroll virus(PLRV) was observed on potato plants collected from different governorates in Egypt. Three cultivars, Spunta, Diamont, and Cara, infected with PLRV were collected; RNA was extracted and subjected to Real-Time PCR using the coat protein gene primers. The results showed that the expression of the coat protein was 39.6-fold, 12.45-fold, and 47.43-fold, respectively, for Spunta, Diamont, and Cara cultivars. Differential Display Polymerase Chain Reaction (DD-PCR) using pathogenesis-related protein 1 (PR-1), β-1,3-glucanases (PR-2), chitinase (PR-3), peroxidase (POD), and polyphenol oxidase (PPO) forward primers for pathogenesis-related proteins (PR). The obtained data revealed different banding patterns depending on the viral type and the region of infection. Regarding PLRV, 58 up-regulated and 19 down-regulated genes were detected. Sequence analysis of the up-and down-regulated genes revealed that infected plants were observed in comparison with the healthy control. Sequence analysis of the up-regulated gene was performed, and the encoding sequence analysis showed that the obtained genes include: induced stolen tip protein. On the other hand, two down-regulated genes were identified: disease resistance RPP-like protein and non-specific lipid-transfer protein. In this study, the expressions of PR-1, PR-2, PR-3, POD, and PPO genes in the infected leaves of three potato cultivars were estimated by quantitative real-time PCR. We can conclude that the PLRV-infection of potato plants inhibited the expression of the five PR genes. On the contrary, infected leaves by PLRV elevated the expression of some defense genes. This interaction may also induce and/or suppress the expression of some genes responsible for the plant's defense mechanisms.Keywords: PLRV, pathogenesis-related proteins (PRs), DD-PCR, sequence, real-time PCR
Procedia PDF Downloads 1421677 Fatty Acid Binding Protein 3 Gene Polymorphisms and Their Associations with Growth Traits and Blood Parameters in Two Iranian Sheep Breeds
Authors: Sahar Javadi-Novashnagh, Mohammad Moradi-Shahrbabak, Mostafa Sadeghi, Katarzyna Ropka-Molik, Hossein Moradi-Shahrbabak, Maria Consuelo Mura
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The objective of this study was to investigate two single nucleotide polymorphisms located in exon 2 (g.939A > G) and intron 3 (g.4349A > G) of fatty acid binding protein 3 (FABP3) gene in two Iranian sheep breeds, Lori-Bakhtiari and Zel, using polymerase chain reaction -restriction fragment length polymorphism (PCR-RFLP) approach. The association of the polymorphisms with growth traits and blood parameters was also examined. Results revealed a g.939A > G SNP (single nucleotide polymorphism) in the exon 2 exhibiting three genotypes: AA, AG, and GG. Statistical analysis indicated that this polymorphism significantly influenced blood triglyceride (P < 0.05) and cholesterol (P < 0.08) levels as well as weaning weight (P < 0.05). Animals with AG genotype had the highest blood triglyceride level and weaning weight while the highest amount of blood cholesterol was observed in animals with GG genotype. On the other hand, no significant effect was observed on birth and fat-tail weight traits. The intron 3 (g.4349A > G) was monomorphic across the studied samples. Lori-Bakhtiari breed showed significantly higher blood triglyceride and cholesterol levels, as also birth and weaning weight compared to Zel breed (P < 0.01). Considering that the literature is bereft of any report on the association study between FABP3 SNPs and sheep growth traits and blood parameters, our findings suggest that the investigated polymorphism might be one of the main genetic factors affecting growth and physiological traits in sheep.Keywords: FABP3 gene, fatness, weaning weight, blood triglyceride, cholesterol, Zel, Lori-Bakhtiari
Procedia PDF Downloads 6991676 DNA Polymorphism Studies of β-Lactoglobulin Gene in Native Saudi Goat Breeds
Authors: Amr A. El Hanafy, Muhammad I. Qureshi, Jamal Sabir, Mohamed Mutawakil, Mohamed M. Ahmed, Hassan El Ashmaoui, Hassan Ramadan, Mohamed Abou-Alsoud, Mahmoud Abdel Sadek
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β-Lactoglobulin (β-LG) is the dominant non-casein whey protein found in bovine milk and of most ruminants. The amino acid sequence of β-LG along with its 3-dimensional structure illustrates linkage with the lipocalin superfamily. Preliminary studies in goats indicated that milk yield can be influenced by polymorphism in genes coding for whey proteins. The aim of this study is to identify and evaluate the incidence of functional polymorphisms in the exonic and intronic portions of β-LG gene in native Saudi goat breeds (Ardi, Habsi, and Harri). Blood samples were collected from 300 animals (100 for each breed) and genomic DNA was extracted using QIAamp DNA extraction Kit. A fragment of the β-LG gene from exon 7 to 3’ flanking region was amplified with pairs of specific primers. Subsequent digestion with Sac II restriction endonuclease revealed two alleles (A and B) and three different banding patterns or genotypes i.e. AA, AB and BB. The statistical analysis showed that β-LG AA genotype had higher milk yield than β-LG AB and β-LG BB genotypes. Nucleotide sequencing of the selected β-LG fragments was done and submitted to GenBank NCBI (Accession No. KJ544248, KJ588275, KJ588276, KJ783455, KJ783456 and KJ874959). Two already established SNPs in exon 7 (+4601 and +4603) and one fresh SNP in the 3’ UTR region were detected in the β-LG fragments with designated AA genotype. The polymorphisms in exon 7 did not produce any amino acid change. Phylogenetic analysis on the basis of nucleotide sequences of native Saudi goats indicated evolutional similarity with the GenBank reference sequences of goat, Bubalus bubalis and Bos taurus.Keywords: β-Lactoglobulin, Saudi goats, PCR-RFLP, functional polymorphism, nucleotide sequencing, phylogenetic analysis
Procedia PDF Downloads 5011675 Hypoxia Tolerance, Longevity and Cancer-Resistance in the Mole Rat Spalax – a Liver Transcriptomics Approach
Authors: Hanno Schmidt, Assaf Malik, Anne Bicker, Gesa Poetzsch, Aaron Avivi, Imad Shams, Thomas Hankeln
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The blind subterranean mole rat Spalax shows a remarkable tolerance to hypoxia, cancer-resistance and longevity. Unravelling the genomic basis of these adaptations will be important for biomedical applications. RNA-Seq gene expression data were obtained from normoxic and hypoxic Spalax and rat liver tissue. Hypoxic Spalax broadly downregulates genes from major liver function pathways. This energy-saving response is likely a crucial adaptation to low oxygen levels. In contrast, the hypoxiasensitive rat shows massive upregulation of energy metabolism genes. Candidate genes with plausible connections to the mole rat’s phenotype, such as important key genes related to hypoxia-tolerance, DNA damage repair, tumourigenesis and ageing, are substantially higher expressed in Spalax than in rat. Comparative liver transcriptomics highlights the importance of molecular adaptations at the gene regulatory level in Spalax and pinpoints a variety of starting points for subsequent functional studies.Keywords: cancer, hypoxia, longevity, transcriptomics
Procedia PDF Downloads 1571674 Salt Tolerance of Potato: Genetically Engineered with Atriplex canescens BADH Gene Driven by 3 Copies of CAMV35s Promoter
Authors: Arfan Ali, Muhammad Shahzad Iqbal, Idrees Ahmad Nasir
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Potato (Solanum tuberosum L.) is ranked among the top leading staple foods in the world. Salinity adversely affects potato crop yield and quality. Therefore, increased level of salt tolerance is a key factor to ensure high yield. The present study focused on the Agrobacterium-mediated transformation of Atriplex canescens betaine aldehyde dehydrogenase (BADH) gene, using single, double and triple CAMV35s promoter to improve salt tolerance in potato. Detection of seven potato lines harboring BADH gene, followed by identification of T-DNA insertions, determination of transgenes copies no through Southern Hybridization and quantification of BADH protein through Enzyme Linked Immunosorbent Assay were considered in this study. The results clearly depict that the salt tolerance of potato was found to be promoter-dependent, as the potato transgenic lines with triple promoter showed 4.4 times more glycine betaine production which consequently leads towards high resistance to salt stress as compared to transgenic potato lines with single and double promoters having least production of glycine betaine. Moreover, triple promoter transgenic potato lines have also shown lower levels of H2O2, malondialdehyde (MDA), relative electrical conductivity, high proline and chlorophyll content as compared other two lines having a single and double promoter. Insilco analysis also confirmed that Atriplex canescens BADH has the tendency to interact with sodium ions and water molecules. Taken together these facts it can be concluded that over-expression of BADH under triple CAMV35s promoter with more glycine betaine, chlorophyll & MDA contents, high relative quantities of other metabolites results in an enhanced level of salt tolerance in potato.Keywords: Atriplex canescens, BADH, CAMV35s promotor, potato, Solanum tubersum
Procedia PDF Downloads 2771673 Bioconversion of Antifungal Antibiotic Derived from Aspergillus Nidulans
Authors: Savitha Janakiraman, Shivakumar M. C
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Anidulafungin, an advanced class of antifungal agent used for the treatment of chronic fungal infections, is derived from Echinocandin B nucleus, an intermediate metabolite of Echinocandin B produced by Aspergillus nidulans. The enzyme acylase derived from the fermentation broth of Actinoplanes utahensis (NRRL 12052) plays a key role in the bioconversion of echinocandin B to echinocandin B nucleus. The membrane-bound nature of acylase and low levels of expression contributes to the rate-limiting process of enzymatic deacylation, hence low yields of ECB nucleus and anidulafungin. In the present study, this is addressed through novel genetic engineering approaches of overexpression and heterologous expression studies, immobilization of whole cells of Actinoplanes utahensis (NRRL 12052) and Co-cultivation studies. Overexpression of the acylase gene in Actinoplanes utahensis (NRRL 12052) was done by increasing the gene copy number to increase the echinocandin B nucleus production. Echinocandin B acylase gene, under the control of a PermE* promoter, was cloned in pSET152 vector and introduced into Actinoplanes utahensis (NRRL12052) by a ɸC31-directed site-specific recombination method. The resultant recombinant strain (C2-18) showed a 3-fold increase in acylase expression, which was confirmed by HPLC analysis. Pichia pastoris is one of the most effective and versatile host systems for the production of heterologous proteins. The ECB acylase gene was cloned into pPIC9K vector with AOX1 promoter and was transformed into Pichia pastoris (GS115). The acylase expression was confirmed by protein expression and bioconversion studies. The heterologous expression of acylase in Pichia pastoris, is a milestone in the development of antifungals. Actively growing cells of Actinoplanes utahensis (NRRL 12052) were immobilized and tested for bioconversion ability which showed >90% conversion in each cycle. The stability of immobilized cell beads retained the deacylation ability up to 60 days and reusability was confirmed up to 4 cycles. The significant findings from the study have revealed that immobilization of whole cells of Actinoplanes utahensis (NRRL 12052) could be an alternative option for bioconversion of echinocandin B to echinocandin B nucleus, which has not been reported to date. The concept of co-cultivation of Aspergillus nidulans and Actinoplanes utahensis strains for the production of the echinocandin B nucleus was also carried out in order to produce echinocandin B nucleus. The process completely reduced the ECB purification step and, therefore, could be recommended as an ingenious method to improve the yield of the ECB nucleus.Keywords: acylase, anidulafungin, antifungals, Aspergillus nidulans
Procedia PDF Downloads 1071672 The Importance of Knowledge Innovation for External Audit on Anti-Corruption
Authors: Adel M. Qatawneh
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This paper aimed to determine the importance of knowledge innovation for external audit on anti-corruption in the entire Jordanian bank companies are listed in Amman Stock Exchange (ASE). The study importance arises from the need to recognize the Knowledge innovation for external audit and anti-corruption as the development in the world of business, the variables that will be affected by external audit innovation are: reliability of financial data, relevantly of financial data, consistency of the financial data, Full disclosure of financial data and protecting the rights of investors to achieve the objectives of the study a questionnaire was designed and distributed to the society of the Jordanian bank are listed in Amman Stock Exchange. The data analysis found out that the banks in Jordan have a positive importance of Knowledge innovation for external audit on anti-corruption. They agree on the benefit of Knowledge innovation for external audit on anti-corruption. The statistical analysis showed that Knowledge innovation for external audit had a positive impact on the anti-corruption and that external audit has a significantly statistical relationship with anti-corruption, reliability of financial data, consistency of the financial data, a full disclosure of financial data and protecting the rights of investors.Keywords: knowledge innovation, external audit, anti-corruption, Amman Stock Exchange
Procedia PDF Downloads 4641671 Addressing Urban Security Challenges in Nigeria through Neighborhood Renewal: A Reflection of Mokola World Bank Slum Upgrading Pilot Project
Authors: Tabiti S. Tabiti, A. M. Jinadu, Daramola Japheth
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Urban insecurity is among the challenges militating against sustainable urban governance; in the first place it distorts the peace of urban areas making them unsafe. On the other hand it hinders the effective performance of urban functions. Urban security challenges manifest in different forms such as, street violence, theft and robbery, accidents of different types kidnapping, killings etc.. Efforts to address urban security challenges in Nigeria have been concentrated in legislative, law enforcement and the use of community vigilante groups. However in this study, the place of physical planning strategy through effective neighbourhood renewal as practiced in Mokola is presented as an effective complementary approach for addressing urban insecurity. On this backdrop, the paper recommends the need for gradual rehabilitation of urban slum neighborhoods by the state government in collaboration with World Bank and other development financiers. The local governments should be made autonomy in Nigeria so as to make them more responsible to the people. Other recommendations suggested in the paper include creating enabling environment that will promote economic empowerment and public enlightment on personal and community sanitation. It is certain that if these recommendations are adopted the challenge of urban insecurity will reduce significantly in Nigerian cities.Keywords: neighbourhood renewal, pilot project, slum upgrading, urban security
Procedia PDF Downloads 4371670 Demographic Dividend and Creation of Human and Knowledge Capital in Liberal India: An Endogenous Growth Process
Authors: Arjun K., Arumugam Sankaran, Sanjay Kumar, Mousumi Das
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The paper analyses the existence of endogenous growth scenario emanating from the demographic dividend in India during the liberalization period starting from 1980. Demographic dividend creates a fertile ground for the cultivation of human and knowledge capitals contributing to technological progress which can be measured using total factor productivity. The relationship among total factor productivity, human and knowledge capitals are examined in an open endogenous framework for the period 1980-2016. The control variables such as foreign direct investment, trade openness, energy consumption are also employed. The data are sourced from Reserve Bank of India, World Bank, International Energy Agency and The National Science and Technology Management Information System. To understand the dynamic association among variables, ARDL bounds approach to cointegration followed by Toda-Yamamoto causality test are used. The results reveal a short run and long run relationship among the variables supported by the existence of causality. This calls for an integrated policy to build and augment human capital and research and development activities to sustain and pace up growth and development in the nation.Keywords: demographic dividend, young population, open endogenous growth models, human and knowledge capital
Procedia PDF Downloads 1511669 Motif Search-Aided Screening of the Pseudomonas syringae pv. Maculicola Genome for Genes Encoding Tertiary Alcohol Ester Hydrolases
Authors: M. L. Mangena, N. Mokoena, K. Rashamuse, M. G. Tlou
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Tertiary alcohol ester (TAE) hydrolases are a group of esterases (EC 3.1.1.-) that catalyze the kinetic resolution of TAEs and as a result, they are sought-after for the production of optically pure tertiary alcohols (TAs) which are useful as building blocks for number biologically active compounds. What sets these enzymes apart is, the presence of a GGG(A)X-motif in the active site which appears to be the main reason behind their activity towards the sterically demanding TAEs. The genome of Pseudomonas syringae pv. maculicola (Psm) comprises a multitude of genes that encode esterases. We therefore, hypothesize that some of these genes encode TAE hydrolases. In this study, Psm was screened for TAE hydrolase activity using the linalyl acetate (LA) plate assay and a positive reaction was observed. As a result, the genome of Psm was screened for esterases with a GGG(A)X-motif using the motif search tool and two potential TAE hydrolase genes (PsmEST1 and 2, 1100 and 1000bp, respectively) were identified, PsmEST1 was amplified by PCR and the gene sequenced for confirmation. Analysis of the sequence data with the SingnalP 4.1 server revealed that the protein comprises a signal peptide (22 amino acid residues) on the N-terminus. Primers specific for the gene encoding the mature protein (without the signal peptide) were designed such that they contain NdeI and XhoI restriction sites for directional cloning of the PCR products into pET28a. The gene was expressed in E. coli JM109 (DE3) and the clones screened for TAE hydrolase activity using the LA plate assay. A positive clone was selected, overexpressed and the protein purified using nickel affinity chromatography. The activity of the esterase towards LA was confirmed using thin layer chromatography.Keywords: hydrolases, tertiary alcohol esters, tertiary alcohols, screening, Pseudomonas syringae pv., maculicola genome, esterase activity, linalyl acetate
Procedia PDF Downloads 3551668 Genetic Screening of Sahiwal Bulls for Higher Fertility
Authors: Atul C. Mahajan, A. K. Chakravarty, V. Jamuna, C. S. Patil, Neeraj Kashyap, Bharti Deshmukh, Vijay Kumar
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The selection of Sahiwal bulls on the basis of dams best lactation milk yield under breeding programme in herd of the country neglecting fertility traits leads to deterioration in their performances and economy. The goal of this study was to explore polymorphism of CRISP2 gene and their association with semen traits (Post Thaw Motility, Hypo-osmotic Swelling Test, Acrosome Integrity, DNA Fragmentation and capacitation status), scrotal circumference, expected predicted difference (EPD) for milk yield and fertility. Sahiwal bulls included in present study were 60 bulls used in breeding programme as well as 50 young bulls yet to be included in breeding programme. All the Sahiwal bulls were found to be polymorphic for CRISP2 gene (AA, AG and GG) present within exon 7 to the position 589 of CRISP2 mRNA by using PCR-SSCP and Sequencing. Semen analysis were done on 60 breeding bulls frozen semen doses pertaining to four season (winter, summer, rainy and autumn). The scrotal circumference was measured from existing Sahiwal breeding bulls in the herd (n=47). The effect of non-genetic factors on reproduction traits were studied by least-squares technique and the significant difference of means between subclasses of season, period, parity and age group were tested. The data were adjusted for the significant non-genetic factors to remove the differential environmental effects. The adjusted data were used to generate traits like Waiting Period (WP), Pregnancy Rate (PR), Expected Predicted Difference (EPD) of fertility, respectively. Genetic and phenotypic parameters of reproduction traits were estimated. The overall least-squares means of Age at First Calving (AFC), Service Period (SP) and WP were estimated as 36.69 ± 0.18 months, 120.47 ± 8.98 days and 79.78 ± 3.09 days respectively. Season and period of birth had significant effect (p < 0.01) on AFC. AFC was highest during autumn season of birth followed by summer, winter and rainy. Season and period of calving had significant effect (p < 0.01) on SP and WP of sahiwal cows. The WP for Sahiwal cows was standardized based on four developed predicted model for pregnancy rate 42, 63, 84 and 105 days using all lactation records. The WP for Sahiwal cows were standardized as 42 days. A selection criterion was developed for Sahiwal breeding bulls and young Sahiwal bulls on the basis of EPD of fertility. The genotype has significant effect on expected predicted difference of fertility and some semen parameters like post thaw motility and HOST. AA Genotype of CRISP2 gene revealed better EPD for fertility than EPD of milk yield. AA genotype of CRISP2 gene has higher scrotal circumference than other genotype. For young Sahiwal bulls only AA genotypes were present with similar patterns. So on the basis of association of genotype with seminal traits, EPD of milk yield and EPD for fertility status, AA and AG genotype of CRISP2 gene was better for higher fertility in Sahiwal bulls.Keywords: expected predicted difference, fertility, sahiwal, waiting period
Procedia PDF Downloads 5841667 Biomedicine, Suffering, and Sacrifice: Myths and Prototypes in Cell and Gene Therapies
Authors: Edison Bicudo
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Cell and gene therapies (CGTs) result from the intense manipulation of cells or the use of techniques such as gene editing. They have been increasingly used to tackle rare diseases or conditions of genetic origin, such as cancer. One might expect such a complex scientific field to be dominated by scientific findings and evidence-based explanations. However, people engaged in scientific argumentation also mobilize a range of cognitive operations of which they are not fully aware, in addition to drawing on widely available oral traditions. This paper analyses how experts discussing the potentialities and challenges of CGTs have recourse to a particular kind of prototypical myth. This sociology study, conducted at the University of Sussex (UK), involved interviews with scientists, regulators, and entrepreneurs involved in the development or governance of CGTs. It was observed that these professionals, when voicing their views, sometimes have recourse to narratives where CGTs appear as promising tools for alleviating or curing diseases. This is said to involve much personal, scientific, and financial sacrifice. In his study of traditional narratives, Hogan identified three prototypes: the romantic narrative, moved by the ideal of romantic union; the heroic narrative, moved by the desire for political power; and the sacrificial narrative, where the ideal is plenty, well-being, and health. It is argued here that discourses around CGTs often involve some narratives – or myths – that have a sacrificial nature. In this sense, the development of innovative therapies is depicted as a huge sacrificial endeavor involving biomedical scientists, biotech and pharma companies, and decision-makers. These sacrificial accounts draw on oral traditions and benefit from an emotional intensification that can be easily achieved in stories of serious diseases and physical suffering. Furthermore, these accounts draw on metaphorical understandings where diseases and vectors of diseases are considered enemies or invaders while therapies are framed as shields or protections. In this way, this paper aims to unravel the cognitive underpinnings of contemporary science – and, more specifically, biomedicine – revealing how myths, prototypes, and metaphors are highly operative even when complex reasoning is at stake. At the same time, this paper demonstrates how such hidden cognitive operations underpin the construction of powerful ideological discourses aimed at defending certain ways of developing, disseminating, and governing technologies and therapies.Keywords: cell and gene therapies, myths, prototypes, metaphors
Procedia PDF Downloads 171666 The Impact of CYP2C9 Gene Polymorphisms on Warfarin Dosing
Authors: Weaam Aldeeban, Majd Aljamali, Lama A. Youssef
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Background & Objective: Warfarin is considered a problematic drug due to its narrow therapeutic window and wide inter-individual response variations, which are attributed to demographic, environmental, and genetic factors, particularly single nucleotide polymorphism (SNPs) in the genes encoding VKORC1 and CYP2C9 involved in warfarin's mechanism of action and metabolism, respectively. CYP2C9*2rs1799853 and CYP2C9*3rs1057910 alleles are linked to reduced enzyme activity, as carriers of either or both alleles are classified as moderate or slow metabolizers, and therefore exhibit higher sensitivity of warfarin compared with wild type (CYP2C9*1*1). Our study aimed to assess the frequency of *1, *2, and *3 alleles in the CYP2C9 gene in a cohort of Syrian patients receiving a maintenance dose of warfarin for different indications, the impact of genotypes on warfarin dosing, and the frequency of adverse effects (i.e., bleedings). Subjects & Methods: This retrospective cohort study encompassed 94 patients treated with warfarin. Patients’ genotypes were identified by sequencing the polymerase chain reaction (PCR) specific products of the gene encoding CYP2C9, and the effects on warfarin therapeutic outcomes were investigated. Results: Sequencing revealed that 43.6% of the study population has the *2 and/or *3 SNPs. The mean weekly maintenance dose of warfarin was 37.42 ± 15.5 mg for patients with the wild-type allele (CYP2C9*1*1), whereas patients with one or both variants (*2 and/or *3) demanded a significantly lower dose (28.59 ±11.58 mg) of warfarin, (P= 0.015). A higher percentage (40.7%) of patients with allele *2 and/or *3 experienced hemorrhagic accidents compared with only 17.9% of patients with the wild type *1*1, (P = 0.04). Conclusions: Our study proves an association between *2 and *3 genotypes and higher sensitivity to warfarin and a tendency to bleed, which necessitates lowering the dose. These findings emphasize the significance of CYP2C9 genotyping prior to commencing warfarin therapy in order to achieve optimal and faster dose control and to ensure effectiveness and safety.Keywords: warfarin, CYP2C9, polymorphisms, Syrian, hemorrhage
Procedia PDF Downloads 1461665 Effects of Obesity and Family History of Diabetes on the Association of Cholesterol Ester Transfer Protein Gene with High-Density Lipoprotein Cholesterol Levels in Korean Population
Authors: Jae Woong Sull
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Lipid levels are related to the risk of cardiovascular diseases. Cholesterol ester transfer protein (CETP) gene is one of the candidate genes of cardiovascular diseases. A total of 2,304 persons were chosen from a Hospital (N=4,294) in South Korea. Female subjects with the CG/GG genotype had a 2.03 -fold (p=0.0001) higher risk of having abnormal HDL cholesterol levels (<40 mg/dL) than subjects with the CC genotype. Male subjects with the CG/GG genotype had a 1.34 -fold (p=0.0019) higher risk than subjects with the CC genotype. When analyzed by body mass index, the association with CETP was much stronger in male subjects with BMI>=25.69 (OR=1.55, 95% CI: 1.15-2.07, P=0.0037) than in male lean subjects. When analyzed by family history of diabetes, the association with CETP was much stronger in male subjects with positive family history of low physical activity (OR=4.82, 95% CI: 1.86-12.5, P=0.0012) than in male subjects with negative family history of diabetes. This study clearly demonstrates that genetic variants in CETP influence HDL cholesterol levels in Korean adults.Keywords: CETP, diabetes, obesity, polymorphisms
Procedia PDF Downloads 1431664 Recovery of Helicobacter Pylori from Stagnant and Moving Water Biofilms
Authors: Maryam Zafar, Sajida Rasheed, Imran Hashmi
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Water as an environmental reservoir is reported to act as a habitat and transmission route to microaerophilic bacteria such as Heliobacter pylori. It has been studied that in biofilms are the predominant dwellings for the bacteria to grow in water and protective reservoir for numerous pathogens by protecting them against harsh conditions, such as shear stress, low carbon concentration and less than optimal temperature. In this study, influence of these and many other parameters was studied on H. pylori in stagnant and moving water biofilms both in surface and underground aquatic reservoirs. H. pylori were recovered from pipe of different materials such as Polyvinyl Chloride, Polypropylene and Galvanized iron pipe cross sections from an urban water distribution network. Biofilm swabbed from inner cross section was examined by molecular biology methods coupled with gene sequencing and H. pylori 16S rRNA peptide nucleic acid probe showing positive results for H. pylori presence. Studies showed that pipe material affect growth of biofilm which in turn provide additional survival mechanism for pathogens like H. pylori causing public health concerns.Keywords: biofilm, gene sequencing, heliobacter pylori, pipe materials
Procedia PDF Downloads 3591663 Biosynthesis of L-Xylose from Xylitol Using a Dual Enzyme Cascade in Escherichia coli
Authors: Mesfin Angaw Tesfay
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L-xylose is an important intermediate in the pharmaceutical industry, playing a key role in the production of various antiviral and anticancer drugs. Despite its significance, L-xylose is a rare and costly sugar with limited availability in nature. In recent years, enzymatic production methods have garnered considerable attention due to their benefits over conventional chemical synthesis. In this research, a dual enzyme cascade system was developed to synthesize L-xylose from an inexpensive substrate, xylitol. The study involved cloning and co-expressing two key genes: the L-fucose isomerase (L-fucI) gene from Escherichia coli K-12 and the xylitol-4-dehydrogenase (xdh) gene from Pantoea ananatis ATCC 43072 in Escherichia coli. The resulting recombinant cells, engineered with the PET28a-xdh/L-fucI vector, were able to effectively convert xylitol to L-xylose. The system showed optimal performance at 40°C and a pH of 10.0. Moreover, Zn²⁺ (7.5 mM) enhanced the catalytic activity by 1.34 times. This approach yielded 52.2 g/L of L-xylose from an initial 80 g/L xylitol concentration, with a 65% conversion efficiency and a productivity rate of 1.86. The study highlights a practical method for producing L-xylose from xylitol through a co-expression system carrying the L-fucI and xdh genes.Keywords: l-fucose isomerase, xylitol-4-dehydrogenase, l-xylose, xylitol, co-expression
Procedia PDF Downloads 251662 Enhancing Dispute Resolution in Construction: The Potential Contributions of Dispute Boards and the Roadblock to Vaster Adoption
Authors: Zeyad M. Abdelgawad, A. Samer Ezeldin, Waleed El Nemr
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The Egyptian construction industry has evolved significantly over the past decade, driven by enhanced economic sectors and the need for industrial development. This complexity requires diverse and flexible alternative dispute resolution (ADR) techniques. Dispute boards (DB) are globally recognized as effective ADR methods, especially since their introduction to World Bank projects in 1995. Despite their advantages, dispute boards remain underutilized in Egypt aside from the World Bank-financed projects due to several misconceptions. The study reveals the perceptions hindering the wider adoption of dispute boards in the Egyptian construction industry through detailed literature review and interviews with the experts. The perceptions encompassed the lack of awareness and understanding of dispute boards and implementation procedures, misconceptions about the costs associated with implementing dispute boards and the impact on the bid prices, the common orientation of resolving disputes internally and avoid resorting to external parties to preserve the long-term relationship, and lack of trust in the ability of the dispute boards to positively affect the project performance. In response to these identified misconceptions, a proposed alternative draft to the FIDIC 2017 clause twenty-one “Disputes and Arbitration” is provided, offering a way for a practical application of the dispute boards within the Egyptian context.Keywords: alternative dispute resolution, claim management system, dispute boards, Egyptian construction industry, FIDIC
Procedia PDF Downloads 211661 Board Composition and Performance of Listed Deposit Money Banks in Nigeria
Authors: Mary David, Denis Basila
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This study assessed the Impact of Board Composition on the Performance of Listed Deposit Money Banks in Nigeria. A sample of ten (10) deposit money banks formed the sample of this study. Board size, gender diversity, and board independence were used as the independent variables, and firm size as a control variable, whiles the bank performance was proxy with Tobin’s Q (TQ) as the dependent variable. Secondary data was collected from secondary source through the annual report and account of the banks and was analyzed through the support of STATA 14 versions. Descriptive statistics, correlation matrix, and OLS multiple regression model were adopted for the study. Breusch and pagan lagrangian multiplier test for random effect was conducted. The findings of the study reveal that board size has positive and significant impact on Tobin’s Q, gender diversity has positive and significant impact on Tobin’s Q, while board independent had a negative and nonsignificant influence on the Tobin’s Q, Similarly, firm size was found to have a negative and nonsignificant impact on Tobin’s Q of the study banks. This study recommended that policy makers, stakeholders, and corporate managers of deposit money banks of Nigeria and related industries are encouraged to adopt board sizes and gender diversity that impact positively on bank performance.Keywords: board composition, performance, deposit money banks, nigeria
Procedia PDF Downloads 731660 New Technologies in Corporate Finance Management in the Digital Economy: Case of Kyrgyzstan
Authors: Marat Kozhomberdiev
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The research will investigate the modern corporate finance management technologies currently used in the era of digitalization of the global economy and the degree to which financial institutions are utilizing these new technologies in the field of corporate finance management in Kyrgyzstan. The main purpose of the research is to reveal the role of financial management technologies as joint service centers, intercompany banks, specialized payment centers in the third-world country. Particularly, the analysis of the implacability of automated corporate finance management systems such as enterprise resource planning system (ERP) and treasury management system (TMS) will be carried out. Moreover, the research will investigate the role of cloud accounting systems in corporate finance management in Kyrgyz banks and whether it has any impact on the field of improving corporate finance management. The study will utilize a data collection process via surveying 3 banks in Kyrgyzstan, namely Mol-Bulak, RSK, and KICB. The banks were chosen based on their ownerships, such as state banks, private banks with local authorized capital, and private bank with international capital. The regression analysis will be utilized to reveal the correlation between the ownership of the bank and the use of new financial management technologies. The research will provide policy recommendations to both private and state banks on developing strategies for switching and utilizing modern corporate finance management technologies in their daily operations.Keywords: digital economy, corporate finance, digital environment, digital technologies, cloud technologies, financial management
Procedia PDF Downloads 701659 Effects of Exercise on Klotho Expression and Klotho DNA Methylation in Obese Mice
Authors: Yao Huang, Hongjie Yu, Fangrong Xu, Longbiao Cai, Qiqiang He
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The Klotho gene has been found to be involved in cardiovascular health, and epigenetic mechanism has risen as good candidates to understand the role of lifestyle factors in obesity. The aim of this study was to investigate the effect of exercise intervention on the expression and DNA methylation of Klotho gene in high-fat diet induced obese mice. C57BL/6 male mice were fed a normal diet (ND) or a high-fat diet (HFD) for 12 weeks. HFD induced obese mice were divided into secondary group (SED) and exercise group (EX) randomly. The treadmill exercise was performed in EX group for 8 weeks. The expression and DNA methylation of Klotho were evaluated by Western blot, RT-PCR, and Methylation-specific PCR. Results indicated that Klotho protein and mRNA expression were significantly lower in the SED group than those in the ND and EX groups (P<0.01), whereas no significant difference, was found between ND group and EX group (P>0.05). Furthermore, mice in the ND group and SED group showed significantly lower levels of completely methylated Klotho DNA in ND group (0%) and SED group (50%) compared with the EX group (90%), and unmethylated Klotho DNA level in ND group (80%) was significantly higher than those in the SED (0%) and EX (0%) groups. These results suggested that exercise leads to increased Klotho expression and reduced Klotho DNA methylation level in HFD induced obese mice.Keywords: DNA methylation, exercise intervention, klotho, obese mice
Procedia PDF Downloads 3541658 Target and Biomarker Identification Platform to Design New Drugs against Aging and Age-Related Diseases
Authors: Peter Fedichev
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We studied fundamental aspects of aging to develop a mathematical model of gene regulatory network. We show that aging manifests itself as an inherent instability of gene network leading to exponential accumulation of regulatory errors with age. To validate our approach we studied age-dependent omic data such as transcriptomes, metabolomes etc. of different model organisms and humans. We build a computational platform based on our model to identify the targets and biomarkers of aging to design new drugs against aging and age-related diseases. As biomarkers of aging, we choose the rate of aging and the biological age since they completely determine the state of the organism. Since rate of aging rapidly changes in response to an external stress, this kind of biomarker can be useful as a tool for quantitative efficacy assessment of drugs, their combinations, dose optimization, chronic toxicity estimate, personalized therapies selection, clinical endpoints achievement (within clinical research), and death risk assessments. According to our model, we propose a method for targets identification for further interventions against aging and age-related diseases. Being a biotech company, we offer a complete pipeline to develop an anti-aging drug-candidate.Keywords: aging, longevity, biomarkers, senescence
Procedia PDF Downloads 2741657 Development of Transgenic Tomato Immunity to Pepino Mosaic Virus and Tomato Yellow Leaf Curl Virus by Gene Silencing Approach
Authors: D. Leibman, D. Wolf, A. Gal-On
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Viral diseases of tomato crops result in heavy yield losses and may even jeopardize the production of these crops. Classical tomato breeding for disease resistance against Tomato yellow leaf curl virus (TYLCV), leads to partial resistance associated with a number of recessive genes. To author’s best knowledge Pepino mosaic virus (PepMV) genetic resistance is not yet available. The generation of viral resistance by means of genetic engineering was reported and implemented for many crops, including tomato. Transgenic resistance against viruses is based, in most cases, on Post Transcriptional Gene Silencing (PTGS), an endogenous mechanism which destroys the virus genome. In this work, we developed immunity against PepMV and TYLCV in a tomato based on a PTGS mechanism. Tomato plants were transformed with a hairpin-construct-expressed transgene-derived double-strand-RNA (tr-dsRNA). In the case of PepMV, the binary construct harbored three consecutive fragments of the replicase gene from three different PepMV strains (Italian, Spanish and American), to provide resistance against a range of virus strains. In the case of TYLCV, the binary vector included three consecutive fragments of the IR, V2 and C2 viral genes constructed in a hairpin configuration. Selected transgenic lines (T0) showed a high accumulation of transgene siRNA of 21-24 bases, and T1 transgenic lines showed complete immunity to PepMV and TYLCV. Graft inoculation displayed immunity of the transgenic scion against PepMV and TYLCV. The study presents the engineering of resistance in tomato against two serious diseases, which will help in the production of high-quality tomato. However, unfortunately, these resistant plants have not been implemented due to public ignorance and opposition against breeding by genetic engineering.Keywords: PepMV, PTGS, TYLCV, tr-dsRNA
Procedia PDF Downloads 1331656 Relationship of Epidermal Growth Factor Receptor Gene Mutations Andserum Levels of Ligands in Non-Small Cell Lung Carcinoma Patients
Authors: Abdolamir Allameh, Seyyed Mortaza Haghgoo, Adnan Khosravi, Esmaeil Mortaz, Mihan Pourabdollah-Toutkaboni, Sharareh Seifi
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Non-Small Cell Lung Carcinoma (NSCLC) is associated with a number of gene mutations in epidermal growth factor receptor (EGFR). The prognostic significance of mutations in exons 19 and 21, together with serum levels of EGFR, amphiregulin (AR), and Transforming Growth Factor-alpha (TGF-α) are implicated in diagnosis and treatment. The aim of this study was to examine the relationship of EGFR mutations in selected exons with the expression of relevant ligands in sera samples of NSCLC patients. For this, a group of NSCLC patients (n=98) referred to the hospital for lung surgery with a mean age of 59±10.5 were enrolled (M/F: 75/23). Blood specimen was collected from each patient. Besides, formalin fixed paraffin embedded tissues were processed for DNA extraction. Gene mutations in exons 19 and 21 were detected by direct sequencing, following DNA amplification which was done by PCR (Polymerase Chain Reaction). Also, serum levels of EGFR, AR, and TGF-α were measured by ELISA. The results of our study show that EGFR mutations were present in 37% of Iranian NSCLC patients. The most frequently identified mutations were deletions in exon 19 (72.2%) and substitutions in exon 21 (27.8%). The most frequently identified alteration, which is considered as a rare mutation, was the E872K mutation in exon 21, which was found in 90% (9 out of 10) cases. EGFR mutation detected in exon 21 was significantly (P<0.05) correlated with the levels of its ligands, EGFR and TGF-α in serum samples. Furthermore, it was found that increased serum AR (>3pg/ml) and TGF-α (>10.5 pg/ml) were associated with shorter overall survival (P<0.05). The results clearly showed a close relationship between EGFR mutations and serum EGFR and serum TGF-α. Increased serum EGFR was associated with TGF-α and AR and linked to poor prognosis of NSCLC. These findings are implicated in clinical decision-making related to EGFR-Tyrosine kinase inhibitors (TKIs).Keywords: lung cancer, Iranian patients, epidermal growth factor, mutation, prognosis
Procedia PDF Downloads 801655 Cash Management and the Impact of Cashless Policy in a Developing Nation: Nigeria as a Case Study
Authors: Ossai Paulinus Edwin
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Cash Management is a broad area having to do with the collection, concentration, and disbursement of cash including measuring the level of liquidity and managing the cash balance and short-Term Investments. Cash Management involves the efficient collection and disbursement of cash and cash equivalents. It also includes management of marketable securities because, in modern Terminology, money comprises marketable securities and actual cash in hand or in a bank. This cash management is concerned with management of cash inflow and cash outflow of a business especially as it concerns a developing nation like Nigeria. The paper throws light on the impact of cashless policy in Nigeria as it was introduced by the Central Bank of Nigeria (CBN) in December 2011 and was kick-started in Lagos in January 2012. Survey research was adopted with the questionnaires as data collection instrument. Responses show that cashless policy if adopted generally shall increase employment opportunities, reduce cash related robbery thereby reducing risk of carrying cash; it shall also reduce cash related corruption and attract more foreign investors to the country. It is expected that the introduction of cashless policy in Nigeria is a step in the right direction as it shall bring about modernization of Nigeria payment system, reduction in the cost of banking services, reduction in high security and safety risk and also curb banking related corruptions.Keywords: cashless economy, cash management, cashless policy, e-banking, Nigeria
Procedia PDF Downloads 5391654 Molecular Identification and Genotyping of Human Brucella Strains Isolated in Kuwait
Authors: Abu Salim Mustafa
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Brucellosis is a zoonotic disease endemic in Kuwait. Human brucellosis can be caused by several Brucella species with Brucella melitensis causing the most severe and Brucella abortus the least severe disease. Furthermore, relapses are common after successful chemotherapy of patients. The classical biochemical methods of culture and serology for identification of Brucellae provide information about the species and serotypes only. However, to differentiate between relapse and reinfection/epidemiological investigations, the identification of genotypes using molecular methods is essential. In this study, four molecular methods [16S rRNA gene sequencing, real-time PCR, enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus variable-number tandem-repeat analysis (MLVA)-16] were evaluated for the identification and typing of 75 strains of Brucella isolated in Kuwait. The 16S rRNA gene sequencing suggested that all the strains were B. melitensis and real-time PCR confirmed their species identity as B. melitensis. The ERIC-PCR band profiles produced a dendrogram of 75 branches suggesting each strain to be of a unique type. The cluster classification, based on ~ 80% similarity, divided all the ERIC genotypes into two clusters, A and B. Cluster A consisted of 9 ERIC genotypes (A1-A9) corresponding to 9 individual strains. Cluster B comprised of 13 ERIC genotypes (B1-B13) with B5 forming the largest cluster of 51 strains. MLVA-16 identified all isolates as B. melitensis and divided them into 71 MLVA-types. The cluster analysis of MLVA-16-types suggested that most of the strains in Kuwait originated from the East Mediterranean Region, a few from the African group and one new genotype closely matched with the West Mediterranean region. In conclusion, this work demonstrates that B. melitensis, the most pathogenic species of Brucella, is prevalent in Kuwait. Furthermore, MLVA-16 is the best molecular method, which can identify the Brucella species and genotypes as well as determine their origin in the global context. Supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.Keywords: Brucella, ERIC-PCR, MLVA-16, RT-PCR, 16S rRNA gene sequencing
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