Search results for: gene bank

Authors: Yina A. Cifuentes Triana, Andrés M. Pinzón Velásco, Marío E. Velásquez Lozano

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In this work the sequencing and genome characterization of a natural isolate of Saccharomyces cerevisiae yeast (strain 202-3), identified with potential for the production of second generation ethanol from sugarcane bagasse hydrolysates is presented. This strain was selected because its capability to consume xylose during the fermentation of sugarcane bagasse hydrolysates, taking into account that many strains of S. cerevisiae are incapable of processing this sugar. This advantage and other prominent positive aspects during fermentation profiles evaluated in bagasse hydrolysates made the strain 202-3 a candidate strain to improve the production of second-generation ethanol, which was proposed as a first step to study the strain at the genomic level. The molecular characterization was carried out by genome sequencing with the Illumina HiSeq 2000 platform paired end; the assembly was performed with different programs, finally choosing the assembler ABYSS with kmer 89. Gene prediction was developed with the approach of hidden Markov models with Augustus. The genes identified were scored based on similarity with public databases of nucleotide and protein. Records were organized from ontological functions at different hierarchical levels, which identified central metabolic functions and roles of the S. cerevisiae strain 202-3, highlighting the presence of four possible new proteins, two of them probably associated with the positive consumption of xylose.

Keywords: cellulosic ethanol, Saccharomyces cerevisiae, genome sequencing, xylose consumption

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Authors: Azza Gaber Farag, Yasser Atta Shehata, Sara Elsayed Elghazouly, Mustafa Elsayed Elshaib, Nesreen Gamal Elden Elhelbawy

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Background: Androgen receptor (AR) polymorphism in cytosine adenineguanine (CAG) repeat has an effect on the functional capacity of AR in males. However, little researches in this field are available regarding female sexual function. Aim: To investigate the possible link between polymorphism in the CAG repeat of AR gene and female sexual function in a sample of the Egyptian population. Materials and methods: 500 Egyptian married females completed a questionnaire regarding sociodemographic, reproductive, and sexual data. AR CAG repeat length was analyzed for those having female sexual dysfunctions (FSD) using real-time PCR. Results: The most sensitive domain to AR CAG repeat length was the orgasm domain that showed significant positive correlations with short allele (p=0.001), long allele (p=.015), biallellic mean (p=.000), and X weighted biallelic mean (p=.000). The satisfaction domain had significant positive correlations with the biallelic mean (p=.035), and the X weighted biallelic mean (p=. 032). However, the pain domain was of significant negative correlations with AR polymorphism of short allele (p=.002), biallelic mean (p=.013), and X weighted biallelic mean (p = . 011). Conclusions: AR polymorphism could represent a non-negligible aspect in female sexual function. The lower AR CAG repeat polymorphism was of significant impact on FSD, affecting mainly female orgasm followed by pain disorders that finally reflected On her sexual satisfaction.

Keywords: female sexual dysfunction, androgen receptor, CAG repeat polymorphism, androgen

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Authors: Sanxiong Fu, Yanyan Lv, Song Chen, Wei Zhang, Cunkou Qi

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Ethylene response factor proteins are known to play an important role in regulating a variety of stress responses in plants, but their exact functions in submergence stress are not completely understood. In this study, we isolated BnERF from Brassica napus L. to study the function of BnERF in submergence tolerance. The expression of BnERF gene in Brassica napus L. and the expression of antioxidant enzyme genes in transgenic Arabidopsis were analyzed by Quantitative RT-PCR. It was found that expression of BnERF is apparently induced by submergence in Brassica napus L. and overexpression of BnERF in Arabidopsis increases the tolerance level to submergence and oxidative stress. Histochemical method detected lower level of H2O2, O2•− and malondialdehyde (MDA) in the transgenic Arabidopsis. Compared to wild type, transgenic lines also have higher soluble sugar content and higher activity of antioxidant enzymes, which helps protect the plants against the oxidative damage caused by submergence. It was concluded that BnERF can increase the tolerance of plants to submergence stress and BnERF might be involved in regulating soluble sugar content and the antioxidant system in the defense against submergence stress.

Keywords: antioxidant enzyme, Arabidopsis, ethylene response factor, submergence

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Authors: Yen-Ni Teng, Hsing-Yi Chen, Hsien-An Pan, Yung-Ming Lin, Hany A. Omar, Jui-Hsiang Hung

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Leucine-rich repeats and WD repeat domain containing 1 (LRWD1) is highly expressed in the testes of healthy males. On the other hand, LRWD1 is significantly down-regulated in the testicular tissues of patients with severe spermatogenic defects. In our study, the downregulation of LRWD1 expression by shRNA caused a significant reduction of cell growth and mitosis and a noteworthy increase in the cell microtubule atrophy rate. Here, we used EMBOSS CpG plot analysis to explore the promoter region of LRWD1 gene. We found that CpG islands are located between positions -253 to +5 nucleotides upstream from the LRWD1 transcription start site. Luciferase reporter assay revealed that the hypermethylation of the LRWD1 promoter reduced the transcription activity in cells. In addition, quantitative methylation-specific PCR and immunostaining showed that the methylation inhibitor, 5-Aza-2'-deoxycytidine, increased LRWD1 promoter activity, LRWD1 mRNA, protein expression and cell viability. Whereas, the methylation activator, S-adenosylmethionine, caused opposite effects. The overexpression of p53 and Nrf2 in NT2/D1 cells increased LRWD1 promoter activity while 5-fluorodeoxyuridine decreased it. In conclusion, this study highlights evidence that the methylation status of LRWD1 promoter is associated with LRWD1 expression. Since the expression level of LRWD1 plays an important role in spermatogenesis, the methylation status of LRWD1 may serve as a novel molecular diagnostic or therapeutic approach in male's infertility.

Keywords: LRWD1, DNA methylation, p53, Nrf2

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Authors: Luciana C. Ramos, Leandro J. Sousa, Antônio Ferreira da Silva, Valéria Gomes Oliveira Falcão, Suzana T. Cunha Lima

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The commercial production of biodiesel using microalgae demands a high-energy input for harvesting biomass, making production economically unfeasible. Methods currently used involve mechanical, chemical, and biological procedures. In this work, a flocculation system is presented as a cost and energy effective process to increase biomass production of Phaeodactylum tricornutum. This diatom is the only species of the genus that present fast growth and lipid accumulation ability that are of great interest for biofuel production. The algae, selected from the Bank of Microalgae, Institute of Biology, Federal University of Bahia (Brazil), have been bred in tubular reactor with photoperiod of 12 h (clear/dark), providing luminance of about 35 μmol photons m^-2s^-1, and temperature of 22 °C. The medium used for growing cells was the Conway medium, with addition of silica. The seaweed growth curve was accompanied by cell count in Neubauer camera and by optical density in spectrophotometer, at 680 nm. The precipitation occurred at the end of the stationary phase of growth, 21 days after inoculation, using two methods: centrifugation at 5000 rpm for 5 min, and electro-flocculation at 19 EPD and 95 W. After precipitation, cells were frozen at -20 °C and, subsequently, lyophilized. Biomass obtained by electro-flocculation was approximately four times greater than the one achieved by centrifugation. The benefits of this method are that no addition of chemical flocculants is necessary and similar cultivation conditions can be used for the biodiesel production and pharmacological purposes. The results may contribute to improve biodiesel production costs using marine microalgae.

Keywords: biomass, diatom, flocculation, microalgae

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Authors: Damanhur Munardi, Muhammad Hafiz, Dina Nurmalita Sari, Syarifah Ridani Alifa

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Purpose: This research aims to analyze the Benefits, Opportunity, Cost, and Risk aspects of applying the Green Finance concept and to obtain the right Green Finance financing strategy to be implemented within a long-term and short-term strategic framework.Methodology: This research method uses a qualitative-descriptive analysis approach. The analysis technique uses Analytical Network Process (ANP) with a BOCR network structure approach.Findings: The research results show that the most priority long-term strategic alternative based on the long-term BOCR analysis is increasing awareness among the public and industry by 52% and the importance of coordination between related institutions by 50%. Meanwhile, the most priority short-term strategic alternatives are the importance of coordination between related institutions 29%, increasing awareness among the public and industry 28%, the banking industry proactively funding environmentally friendly companies and technology 23%, the existence of Green Finance POS (Standard Operating Procedures) 20%.Implications: This research can be used as a reference for regulators and policymakers in making strategic decisions that can increase green finance financing. The novelty of this research is identifying problems that occur in green finance financing in Aceh province by analyzing opinions from experts in related fields and financial regulators in Aceh to create a strategy that can be implemented to increase green finance financing in Aceh province through BPD in Aceh, namely Bank Aceh.

Keywords: green financing, banking, sharia, islamic

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Authors: Abhishikta Ghosh Roy

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Breast cancer is the most common malignancy among women globally, including India. Human breast cancer results from the genetic and environmental interaction. The present study attempts to understand the molecular heterogeneity of BRCA1 and BRCA2 genes, as well as to understand the association of various lifestyle and reproductive variables for the Breast Cancer risk. The study was conducted amongst 110 patients and 128 controls with total DNA sequencing of flanking and coding regions of BRCA1 BRCA2 genes that revealed ten Single Nucleotide Polymorphisms (SNPs) (6 novels). The controls selected for the study were age, sex and ethnic group matched. After written and informed consent biological samples were collected from the subjects. After detailed molecular analysis, significant (p < 0.005) molecular heterogeneity is revealed in terms of SNPs in BRCA1 (4 Exonic & 1 Intronic) and BRCA2 (2exonic and 3 Intronic) genes. The augmentation study investigated significant (p < 0.05) association with positive family history, early age at menarche, irregular menstrual periods, menopause, prolong contraceptive use, nulliparity, history of abortions, consumption of alcohol and smoking for breast cancer risk. To the best of authors knowledge, this study is the first of its kind, envisaged that the identification of the SNPs and modification of the lifestyle factors might aid to minimize the risk among the Bengalee Hindu females.

Keywords: breast cancer, BRCA, lifestyle, India

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Authors: Zhou Shangcheng, Yi Sicen

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Objective: To understand the mortality patterns of Zhangwan District in 2010 and provide the basis for the development of scientific and rational health policy. Methods: Data are collected from the Sixth Census of Zhangwan District and disease surveillance system. The statistical analysis include death difference between age, gender, region and time and the related factors. Methods developed for the Global Burden of Disease (GBD) Study by the World Bank and World Health Organization (WHO) were adapted and applied to Zhangwan District population health data. DALY rate per 1,000 was calculated for varied causes of death. SPSS 16 is used by statistic analysis. Results: From the data of death population of Zhangwan District we know the crude mortality rate was 6.03 ‰. There are significant differences of mortality rate in male and female population which was respectively 7.37 ‰ and 4.68 ‰. 0 age group population life expectancy in Zhangwan District in 2010 was 78.40 years old(Male 75.93, Female 81.03). The five leading causes of YLL in descending order were: cardiovascular diseases(42.63DALY/1000), malignant neoplasm (23.73DALY/1000), unintentional injuries (5.84DALY/1000), Respiratory diseases(5.43 DALY/1000), Respiratory infections (2.44DALY/1000). In addition, there are strong relation between the marital status , educational level and mortality in some to a certain extend. Conclusion Zhangwan District, as city level, is at lower mortality levels. The mortality of the total population of Zhangwan District has a downward trend and life expectancy is rising.

Keywords: sixth census, Zhangwan district, death level differences, influencing factors, cause of death

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Authors: Prasanthi Malapati, R. Reshma, Vijay Soni, Perumal Yogeeswari, Dharmarajan Sriram

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In the present study, we attempted to develop Mycobacterium tuberculosis (Mtb) inhibitors by exploring the pharmaceutically underexploited enzyme targets which are majorly involved in cell wall biosynthesis of mycobacteria. For this purpose, glutamate racemase (coded by MurI gene) was selected. This enzyme racemize L-glutamate to D-glutamate required for the construction of peptidoglycan in the bacterial cell wall synthesis process. Furthermore this enzyme is neither expressed nor its product, D-glutamate is normally found in mammals, and hence designing inhibitors against this enzyme will not affect the host system as well act as potential antitubercular drugs. A library of BITS in house compounds were screened against Mtb MurI enzyme. Based on docking score, interactions and synthetic feasibility one hit lead was identified. Further optimization of lead was attempted and its derivatives were synthesized. Forty eight derivatives of 2-phenylbenzo[d]oxazole and 2-phenylbenzo[d]thiazole were synthesized and evaluated for Mtb MurI inhibition study, in vitro activities against Mtb, cytotoxicity against RAW 264.7 cell line. Chemical derivatization of the lead resulted in compounds NR-1213 AND NR-1124 as the potent M. tuberculosis glutamate racemase inhibitors with IC50 of 4-5µM which are remarkable and were found to be non-cytotoxic. Molecular dynamics, dormant models and cardiotoxicity studies of the most active molecules are in process.

Keywords: cell wall biosynthesis, dormancy, glutamate racemase, tuberculosis

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Authors: Muhammad Izzam Bin Mohd Khazar, Ruqayyah Binti Mohamad Ali, Nurul Atiqah Binti Yusri

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Value-based intermediation (VBI) has been introduced by Bank Negara Malaysia (BNM) as the next strategic direction and growth driver for Islamic banking institutions. The aim of VBI is to deliver the intended outcome of Shariah through practices, conducts, and offerings that generate positive and sustainable impact to the economy, community and environment which is aligned to Maqasid Shariah in preserving the common interest of society by preventing harm and maximizing benefit. Hence, upon its implementation, VBI will experiment the current Shariah compliance treatment and revolutionise new policies and systems that can meritoriously entrench and convey the objectives of Shariah. However, discussion revolving VBI in the light of Maqasid analysis is still scarce hence further research needs to be undertaken. The idea of implementation of VBI vision into quantifiable Maqasid Shariah measurement is yet to be explored due to the nature of Maqasid that is variable. The contemporary scholars also have different views on the implementation of VBI. This paper aims to discuss on the importance of Maqasid Shariah in the current Islamic finance transactions by providing Shariah index measurement in the application of VBI. This study also intends to explore basic Shariah guidelines and parameters based on the objectives of Shariah; preservation of the five pillars (religion, life, progeny, intellect and wealth) with further elaboration on preservation of wealth under five headings: rawaj (circulation and marketability); wuduh (transparency); hifz (preservation); thabat (durability and tranquillity); and ‘adl (equity and justice). In alignment with these headings, Islamic finance can be innovated for VBI implementation, particularly in Maybank Islamic being a significant leader in the IFI market.

Keywords: Islamic Financial Institutions, Maqasid Index, Maqasid Shariah, sustainability, value-based intermediation

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Authors: Nounamo Nguedie Yann Harold

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The factors behind the debt have been the subject of several studies in the literature. Pioneering studies based on the 'double deficit' approach linked indebtedness to the imbalance between savings and investment, the budget deficit and the current account deficit. Most studies on identifying factors that may stimulate or reduce the level of external public debt agree that the following variables are important explanatory variables in leveraging debt: the budget deficit, trade opening, current account and exchange rate, import, export, interest rate, term variation exchange rate, economic growth rate and debt service, capital flight, and over-indebtedness. Few studies addressed the impact of political factors on the level of external debt. In general, however, the IMF's stabilization programs in developing countries following the debt crisis have resulted in economic recession and the advent of political crises that have resulted in changes in governments. In this sense, political institutions are recognised as factors of accumulation of external debt in most developing countries. This paper assesses the role of political factors on the external debt level of African countries in the Franc Zone over the period 1985-2016. Data used come from World Bank and ICRG. Using a logit in panel, the results show that the more a country is politically stable, the lower the external debt compared to the gross domestic product. Political stability multiplies 1.18% the chances of being in the sustainable debt zone. For example, countries with good political institutions experience less severe external debt burdens than countries with bad political institutions.

Keywords: African countries, external debt, Franc Zone, political factors

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Authors: Francisco Dos Santos, Diogo Fonseca-Pereira, Sílvia Arroz-Madeira, Henrique Veiga-Fernandes

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Haematopoiesis is a developmental process that generates all blood cell lineages in health and disease. This relies on quiescent haematopoietic stem cells (HSCs) that are able to differentiate, self renew and expand upon physiological demand. HSCs have great interest in regenerative medicine, including haematological malignancies, immunodeficiencies and metabolic disorders. However, the limited yield from existing HSC sources drives the global need for reliable techniques to expand harvested HSCs at high quality and sufficient quantities. With the extensive use of cord blood progenitors for clinical applications, there is a demand for a safe and efficient expansion protocol that is able to overcome the limitations of the cord blood as a source of HSC. StemCell2MAXTM developed a technology that enhances the survival, proliferation and transplantation efficiency of HSC, leading the way to a more widespread use of HSC for research and clinical purposes. StemCell2MAXTM MIX is a solution that improves HSC expansion up to 20x, while preserving stemness, when compared to state-of-the-art. In a recent study by a leading cord blood bank, StemCell2MAX MIX was shown to support a selective 100-fold expansion of CD34+ Hematopoietic Stem and Progenitor Cells (when compared to a 10-fold expansion of Total Nucleated Cells), while maintaining their multipotent differentiative potential as assessed by CFU assays. The technology developed by StemCell2MAXTM opens new horizons for the usage of expanded hematopoietic progenitors for both research purposes (including quality and functional assays in Cord Blood Banks) and clinical applications.

Keywords: cord blood, expansion, hematopoietic stem cell, transplantation

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Authors: Musarat Ishaq, Tara Karnezis, Ramin Shayan

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Lipedema, a poorly understood chronic disease of adipose hyper-deposition, is often mistaken for obesity and causes significant impairment to mobility and quality-of-life. To identify molecular mechanisms underpinning lipedema, we employed comprehensive omics-based comparative analyses of whole tissue, adipocyte precursors (adipose-derived stem cells (ADSCs)), and adipocytes from patients with or without lipedema. Transcriptional profiling revealed significant differences in lipedema tissue, adipocytes, and ADSCs, with altered levels of mRNAs involved inproliferation and cell adhesion. One highly up-regulated gene in lipedema adipose tissue, adipocytes and ADSCs, ZIC4, encodes Zinc Finger Protein ZIC 4, a class of transcription factor which may be involved in regulating metabolism and adipogenesis. ZIC4 inhibition impaired the adipogenesis of ADSCs into mature adipocytes. Epigenetic regulation study revealed overexpression of ZIC4 is involved in decreased promoter DNA methylation and subsequent decrease in adipogenesis. These epigenetic modifications can alter adipocytes microenvironment and adipocytes differentiation. Our study show that epigenetic events regulate the ability of ADSCs to commit and differentiate into mature adipocytes by modulating ZIC4.

Keywords: lipedema, adipose-derived stem cells, adipose tisue, adipocytes, zinc finger protein, epigenetic

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Authors: Mohamed El-Khateeb

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Encompassing inventory, warehousing, and transportation management, logistics is a crucial predictor of firm performance. This has been extensively proven by extant literature in business and operations management. Logistics is also a fundamental determinant of a country's ability to access international markets. Available studies in international and transport economics have shown that limited transport infrastructure and underperforming transport services can severely affect international competitiveness. However, the evidence lacks the overall impact of logistics performance-encompassing all inventory, warehousing, and transport components- on global trade. In order to fill this knowledge gap, the paper uses a gravitational trade model with 155 countries from all geographical regions between 2007 and 2018. Data on logistics performance is obtained from the World Bank's Logistics Performance Index (LPI). First, the relationship between logistics performance and a country’s total trade is estimated, followed by a breakdown by the economic sector. Then, the analysis is disaggregated according to the level of technological intensity of traded goods. Finally, after evaluating the intensive margin of trade, the relevance of logistics infrastructure and services for the extensive trade margin is assessed. Results suggest that: (i) improvements in both logistics infrastructure and services are associated with export growth; (ii) manufactured goods can significantly benefit from these improvements, especially when both exporting and importing countries increase their logistics performance; (iii) the quality of logistics infrastructure and services becomes more important as traded goods are technology-intensive; and (iv) improving the exporting country's logistics performance is essential in the intensive margin of trade while enhancing the importing country's logistics performance is more relevant in the extensive margin.

Keywords: low-cost, recycling, reuse, solid waste, wastewater treatment

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Authors: Syeda Kiran Shahzadi, Nasir Mahmood, Muhammad Abdul Qadir

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In the current research, three recombinant human interferon alpha-2b proteins (two modified and one normal form) were produced and Pegylated with an aim to produce more effective drugs against viral infections and cancers. The modified recombinant human interferon alpha-2b proteins were produced by site-directed modifications of interferon alpha 2b gene, targeting the amino acids at positions ‘R23’ and ‘H34’. The resulting chemically modified and unmodified forms of human interferon alpha 2b were conjugated with methoxy-polyethylene glycol propanealdehyde (400 KDa) and methoxy-polyethylene glycol succinimidyl succinate (400 KDa). Pegylation of normal and modified forms of Interferon alpha-2b prolong their release time and enhance their efficacy. The conjugation of PEG with modified and unmodified human interferon alpha 2b protein drugs was also characterized with 1H-NMR, HPLC, and SDS-PAGE. Antiproliferative assays of modified and unmodified forms of drugs were performed in cell based bioassays using MDBK cell lines. The results indicated that experimentally produced recombinant human interferon alpha-2b proteins were biologically active and resulted in significant inhibition of cell growth.

Keywords: protein refolding, antiproliferative activities, biomedical applications, human interferon alpha-2b, pegylation, mPEG-propionaldehyde, site directed mutagenesis, E. coli expression

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Authors: Liu Xiaohan

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Epstein–Barr virus (EBV) is a human herpesvirus and is closely related to many malignancies of lymphocyte and epithelial origins, such as gastric cancer, Burkitt’s lymphoma, and nasopharyngeal carcinoma (NPC). NPC is a malignant epithelial tumor which is 100% associated with EBV latent infection. Most of the NPC cases are densely populated in southern China, especially in Guangdong and Hong Kong. To our knowledge, overexpression of pro-inflammatory cytokines may result in a loss of balance of the immune system and cause damage to human bodies. Interleukin-6 (IL6) is a pro-inflammatory cytokine which plays an important role in tumor progression. In addition, gene expression is regulated by both transcriptional and post-transcriptional pathways, while post-transcriptional regulation is an important mechanism to modulate the mature mRNA level in mammalian cells. AU-rich element binding factor 1 (AUF1)/heterogeneous nuclear RNP D (hnRNP D) is known for its function in destabilizing mRNAs, including cytokines and cell cycle regulators. Previous studies have found that overexpression of hnRNP D would lead to tumorigenesis. In this project, our aim is to determine the role played by hnRNP D in EBV-infected cells and how our anti-EBV agents can affect the function of hnRNP D. The results of this study will provide a new insight into how the pro-inflammatory cytokine expression can be regulated by EBV.

Keywords: interleukin 6 (IL6), epstein-barr virus (EBV), nasopharyngeal carcinoma (NPC, epstein-barr nuclear antigen-1 (EBNA1)

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Authors: Isaac Makundi, Kazuo Nishigaki

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The Tsushima leopard cat (TLC) Prionailurus bengalensis euptilurus, designated a National Natural Monument of Japan, inhabits Tsushima Island, Nagasaki Prefecture, Japan. TLC is considered a subspecies of P. bengalensis, and lives only on Tsushima Island. TLCs are threatened by various infectious diseases. Feline leukemia virus (FeLV) causes a serious infectious disease with a poor prognosis in cats. Therefore, the transmission of FeLV from Tsushima domestic cats (TDCs) to TLCs may threaten the TLC population. We investigated the FeLV infection status of both TDCs and TLCs on Tsushima Island by screening blood samples for FeLV p27 antigen and using PCR to amplify the full-length FeLV env gene. The prevalence of FeLV was 6.4% in TDCs and 0% in TLCs. We also demonstrated that the virus can replicate in the cells of TLCs, suggesting its potential cross-species transmission. The viruses in TDCs were classified as genotype I/clade 3, which is prevalent on a nearby island, based on previous studies of FeLV genotypes and FeLV epidemiology. The FeLV viruses identified on Tsushima Island can be further divided into two lineages within genotype I/clade 3, which are geographically separated in Kamijima and Shimojima, indicating that FeLV may have been transmitted to Tsushima Island at least twice. Monitoring FeLV infection in the TDC and TLC populations is highly recommended as part of the TLC surveillance and management strategy.

Keywords: epidemiology, Feline leukemia virus, Tsushima Island, wildlife management

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Authors: Jae-Hyung Jin, Kyung-Tae Lee, Yee-Seul So, Eunji Jeong, Yeonseon Lee, Dongpil Lee, Dong-Gi Lee, Yong-Sun Bahn

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Cryptococcus neoformans causes cryptococcal meningoencephalitis mainly in immunocompromised patients as well as immunocompetent people. But therapeutic options are limited to treat cryptococcosis. Some signaling pathways including cyclic AMP pathway, MAPK pathway, and calcineurin pathway play a central role in the regulation of the growth, differentiation, and virulence of C. neoformans. To understand signaling networks regulating the virulence of C. neoformans, we selected the 114 putative phosphatase genes, one of the major components of signaling networks, in the genome of C. neoformans. We identified putative phosphatases based on annotation in C. neoformans var. grubii genome database provided by the Broad Institute and National Center for Biotechnology Information (NCBI) and performed a BLAST search of phosphatases of Saccharomyces cerevisiae, Aspergillus nidulans, Candida albicans and Fusarium graminearum to Cryptococcus neoformans. We classified putative phosphatases into 14 groups based on InterPro phosphatase domain annotation. Here, we constructed 170 signature-tagged gene-deletion strains through homologous recombination methods for 91 putative phosphatases. We examined their phenotypic traits under 30 different in vitro conditions, including growth, differentiation, stress response, antifungal resistance and virulence-factor production.

Keywords: human fungal pathogen, phosphatase, deletion library, functional genomics

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Authors: A. Qasmaoui, F. Ohmani, Z. Zaine, I. El Akrad, J. Hamamouchi, K. Halout, B. Belkadi, R. Charof

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The evolution of antibiotic resistance is a crucial aspect of the problem related to the intensive use of these substances in medicine for humans and animals. The production of ESBL extended spectrum β-lactamase enzymes is the main mechanism of resistance to β-lactam antibiotics in Escherichia coli. The objective of our work is to determine the resistance genes in E. coli strains.ESBL coli stored at the epidemic diseases laboratory of the National Institute of Hygiene. The strains were identified according to the classic bacteriological criteria. An antibiogram was performed on the strains isolated by the Mueller Hinton agar disc diffusion method. The production of ESBL in the strains was detected by the synergy assay technique and confirmed for the presence of the blaCTX-M1, blaCTX-M2, blaTEM, blaSHV, blaOXA-48 genes by gene amplification . Of the 27 observed strains of E.coli, 17 isolated strains present the phenotype of extended-spectrum Beta-lactamase with a percentage of 63%.. All 18 cefotaxime-resistant strains were analyzed for an ESBL phenotype. All strains were positive in the double-disc synergy assay. The fight against the emergence and spread of these multi-resistant antibiotic-resistant strains requires the reasonable use of antibiotics.

Keywords: E coli, BLSE, CTX, TEM, SHV, OXA, résistance aux antibiotique

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Authors: Z. Mmango, U. Nwodo, L. V. Mabinya, A. I. Okoh

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Cellulose and hemicellulose account for a large portion of the world‘s plant biomass. In nature, these polysaccharides are intertwined forming complex materials that requires multiple and expensive treatment processes to free up the raw materials trapped in the matrix. Enzymatic degradation remains as the preferred technique as it is inexpensive and eco-friendly. However, the insufficiencies of enzyme battery systems in the degradation of lignocellulosic complex motivate the search for effective degrading enzymes from bacterial isolates from uncommon environment. The study aimed at the evaluation of actinomycetes isolated from saw dust samples collected from wood factory under bed. Cellulase and xylanase production was screened through organism culture on carboxyl methyl cellulose agar and Birchwood xylan. Halo zone indicating lignocellose utilization was shown by an isolate identified through 16S rRNA gene as Micrococcus luteus. The optimum condition for the production of cellulase and xylanase were incubation temperature of 25 °C, fermentation medium pH 5 and 10, agitation speed of 50 and 200 (rpm) and fermentation incubation time of 96 and 84 (h) respectively. The high cellulose and xylanase activity obtained from this isolate portends industrial relevance.

Keywords: carboxyl methyl cellulose, birchwood xylan, optimization, cellulase, xylanase, micrococcus, DNS method

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Authors: Harish Rajak, Preeti Patel

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HDAC inhibitors can reactivate gene expression and inhibit the growth and survival of cancer cells. The 3D-QSAR and Pharmacophore modeling studies were performed to identify important pharmacophoric features and correlate 3D-chemical structure with biological activity. The pharmacophore hypotheses were developed using e-pharmacophore script and phase module. Pharmacophore hypothesis represents the 3D arrangement of molecular features necessary for activity. A series of 55 compounds with well-assigned HDAC inhibitory activity was used for 3D-QSAR model development. Best 3D-QSAR model, which is a five PLS factor model with good statistics and predictive ability, acquired Q2 (0.7293), R2 (0.9811) and standard deviation (0.0952). Molecular docking were performed using Histone Deacetylase protein (PDB ID: 1t69) and prepared series of hydroxamic acid based HDAC inhibitors. Docking study of compound 43 show significant binding interactions Ser 276 and oxygen atom of dioxine cap region, Gly 151 and amino group and Asp 267 with carboxyl group of CONHOH, which are essential for anticancer activity. On docking, most of the compounds exhibited better glide score values between -8 to -10.5. We have established structure activity correlation using docking, energetic based pharmacophore modelling, pharmacophore and atom based 3D QSAR model. The results of these studies were further used for the design and testing of new HDAC analogs.

Keywords: Docking, e-pharmacophore, HDACIs, QSAR, Suberoylanilidehydroxamic acid.

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Authors: Benhammadi Hocine, Djamel Boubaya, Chaffai Hicham

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Meskiana Area is characterized by a semi-arid climate where the water supply for irrigation and industry is not sufficient as the priority goes for domestic use. To meet the increasing population growth and development, the authorities have considered building a new water retaining structure on some major temporary water streams. For this purpose Chebabta site on Oued Meskiana was chosen as the future dam site. It is large enough to store the desired volume of water. This study comes to investigate the conditions of the site and the adequacy of the ground as a foundation for the projected dam. The conditions of the site include the geological structure and mainly the presence of discontinuities in the formation on which the dam will be built, the nature of the lithologies under the foundation and the future lake, and the presence of any hazard. This site characterization is usually carried out using different methods in order to highlight any underground buried problematic structure. In this context, the different geophysical technics remain the most used ones. Three geophysical methods were used in the case of the Chebabta dam site, namely, electric survey, seismic refraction, and tomography. The choice of the technics and the location of the scan line was made on the basis of the available geological data. In this sense, profiles have been established on both banks of Oued Meskiana. The obtained results have allowed a better characterization of the geological structure, defining the limit between the surface cover and the bedrock, which is, in other words, the limit between the weathered zone and the bedrock. Their respective thicknesses were also determined by seismic refraction and electrical resistivity sounding. However, the tomography imaging technic has succeeded in positioning a fault structure passing through the right bank of the wadi.

Keywords: dam site, fault, geophysic, investigation, Meskiana

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Authors: Liang Jiansheng, Zhu Wei

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Light and brassinosteroids are essential external and internal cues for plant survival. Although the coordination of light with phytohormone signals is crucial for plant growth and development, the molecular connection between light and brassinosteroid signaling during root soil penetration remains elusive. Here, we reveal that light-stabilized RACK1 couples a brassinosteroid signaling cascade to drive cell division in root meristems. RACK1 family scaffold proteins positively regulate light-induced the promotion of root elongation during soil penetration. Under the light condition, RACK1A interacts with both phyB and SPA1, then reinforces the phyB-SPA1 association to accumulate its abundance in roots. In response to brassinosteroid signals, RACK1A competes with BKI1 to attenuate the BRI1-BKI1 interaction, thereby leading to activating BRI1 actions in root development. Furthermore, RACK1A binds to BES1 to repress its DNA binding activity toward the target gene CYCD3;1. This ultimately allows to release the inhibition of CYCD3;1 transcription, and promotes cell division during root growth. Our study illustrates a new mechanistic model of how plants engage scaffold proteins in transducing light information to facilitate brassinosteroid signaling for root growth in the soil.

Keywords: root growth, cell division, light signaling, brassinosteroid signaling, soil penetration, scaffold protein, RACK1

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Authors: Yusuf Ali Abdulle, Azhar Uddin Keerio

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Background: Protein elicitors play a key role in signaling or displaying plant defense mechanisms and emerging as vital tools for bio-control of insects. This study was aimed at the characterization of the novel protein elicitor isolated from entomopathogenic fungi Lecanicillium lecanii (V3) strain and its activity against Whitefly, Bemisia tabaci in cotton. The sequence of purified elicitor protein showed 100% similarity with hypothetical protein LEL_00878 [Cordyceps confragosa RCEF 1005], GenBank no (OAA81333.1). This novel protein elicitor has 253 amino acid residues and 762bp with a molecular mass of 29 kDa. The protein recombinant was expressed in Escherichia coli using pET‐28a (+) plasmid. Effects of purified novel protein elicitor on Bemisia tabaci were determined at three concentrations of protein (i.e., 58.32, 41.22, 35.41 μg mL⁻¹) on cotton plants and were exposed to newly molted adult B.tabaci. Bioassay results showed a significant effect of the exogenous application of novel protein elicitor on B. tabaci in cotton. In addition, the gene expression analysis found a significant up-regulation of the major genes associated with salicylic acid (SA) and jasmonic acid (JA) linked plant defense pathways in elicitor protein-treated plants. Our results suggested the potential application of a novel protein elicitor derived from Lecanicillium lecanii as a future bio-intensive controlling approach against the whitefly, Bemisia tabaci.

Keywords: resistance, Lecanicillium lecanii, secondary metabolites, whitefly

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Authors: S. Hallal, Z. Chami, A. Hadji-Lehtihet, S. Sokhal-Boudella, A. Berhoune, L. Yargui

Abstract:

Gaucher disease is the most common lysosomal storage in our population, it is due to a deficiency of β –glucosidase acid. The enzyme deficiency causes a pathological accumulation of undegraded substrate in lysosomes. This metabolic overload is responsible for a multisystemic disease with hepatosplenomegaly, anemia, thrombocytopenia, and bone involvement. Neurological involvement is rare. The laboratory diagnosis of Gaucher disease consists of phenotypic diagnosis by determining the enzymatic activity of β - glucosidase by fluorimetric method, a study by genotypic diagnosis in the GBA gene, limiting the search recurrent mutations (N370S, L444P, 84 GG); PCR followed by an enzymatic digestion. Abnormal profiles were verified by sequencing. Monitoring of treated patients is provided by the determination of chitotriosidase. Our experience spaning a period of 6 years (2007-2014) has enabled us to diagnose 78 patients out of a total of 328 requests from the various departments of pediatrics, internal medicine, neurology. Genotypic diagnosis focused on the entire family of 9 children treated at pediatric CHU Mustapha, which help define the clinical form; or 5 of them had type III disease, carrying the L444P mutation in the homozygous state. Three others were composite (N370/L444P) (N370S/other unintended mutation in our study), and only in one family no recurrent mutation has been found. This molecular study permits screening of heterozygous essential for genetic counseling.

Keywords: Gaucher disease, mutations, N370S, L444P

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Authors: Jafar Ahmadi, Alireza Pour-Aboughadareh

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Wheat is the first and the most important grain of the world and its bakery property is due to glutenin and gliadin qualities. Wheat seed proteins were divided into four groups according to solubility. Two groups are albumin and globulin dissolving in water and salt solutions possessing metabolic activities. Two other groups are inactive and non-dissolvable and contain glutelins or glutenins and prolamins or gliadins. Gliadins are major components of the storage proteins in wheat endosperm. Gliadin proteins are separated into three groups based on electrophoretic mobility: α/β-gliadin, γ-gliadin, and ω-gliadin. It seems that little information is available about gliadin genes in Iranian wild relatives of wheat. Thus, the aim of this study was the evaluation of the wheat wild relatives collected from different origins of Zagros Mountains in Iran, involving coding gliadin genes using specific primers. For this, forty accessions of Triticum boeoticum and Triticum urartu were selected. For each accession, genomic DNA was extracted and PCRs were performed in total volumes of 15 μl. The amplification products were separated on 1.5% agarose gels. In results, for Gli-2A locus, three allelic variants were detected by Gli-2As primer pairs. The sizes of PCR products for these alleles were 210, 490 and 700 bp. Only five (13%) and two accessions (5%) produced 700 and 490 bp fragments when their DNA was amplified with the Gli.As.2 primer pairs. However, 37 of the 40 accessions (93%) carried 210 bp allele, and three accessions (8%) did not yield any product for this marker. Therefore, these germplasm could be used as rich gene pool to broaden the genetic base of bread wheat.

Keywords: diploied wheat, gliadin, Triticum boeoticum, Triticum urartu

Procedia PDF Downloads 251

Authors: Parmjit S. Panesar, Rupinder Kaur, Ram S. Singh

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Enzymes are the biocatalysts which catalyze the biochemical processes and thus have a wide variety of applications in the industrial sector. β-Galactosidase (E.C. 3.2.1.23) also known as lactase, is one of the prime enzymes, which has significant potential in the dairy and food processing industries. It has the capability to catalyze both the hydrolytic reaction for the production of lactose hydrolyzed milk and transgalactosylation reaction for the synthesis of prebiotics such as lactulose and galactooligosaccharides. These prebiotics have various nutritional and technological benefits. Although, the enzyme is naturally present in almonds, peaches, apricots and other variety of fruits and animals, the extraction of enzyme from these sources increases the cost of enzyme. Therefore, focus has been shifted towards the production of low cost enzyme from the microorganisms such as bacteria, yeast and fungi. As compared to yeast and bacteria, fungal β-galactosidase is generally preferred as being extracellular and thermostable in nature. Keeping the above in view, the present study was carried out for the isolation of the β-galactosidase producing fungal strain from the food as well as the agricultural wastes. A total of more than 100 fungal cultures were examined for their potential in enzyme production. All the fungal strains were screened using X-gal and IPTG as inducers in the modified Czapek Dox Agar medium. Among the various isolated fungal strains, the strain exhibiting the highest enzyme activity was chosen for further phenotypic and genotypic characterization. The strain was identified as Rhizomucor pusillus on the basis of 5.8s RNA gene sequencing data.

Keywords: beta-galactosidase, enzyme, fungal, isolation

Procedia PDF Downloads 253

Authors: M. Zafar, S. Rasheed, Imran Hashmi

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Very little is concerned about the bacterial contamination in drinking water biofilm which provide a potential source for bacteria to grow and increase rapidly. So as to understand the microbial density in DWDs, a three-month study was carried out. The aim of this study was to examine biofilm in three different pipe materials including PVC, PPR and GI. A set of all these pipe materials was installed in DWDs at nine different locations and assessed on monthly basis. Drinking water quality was evaluated by different parameters and characterization of biofilm. Among various parameters are Temperature, pH, turbidity, TDS, electrical conductivity, BOD, COD, total phosphates, total nitrates, total organic carbon (TOC) free chlorine and total chlorine, coliforms and spread plate counts (SPC) according to standard methods. Predominant species were Bacillus thuringiensis, Pseudomonas fluorescens , Staphylococcus haemolyticus, Bacillus safensis and significant increase in bacterial population was observed in PVC pipes while least in cement pipes. The quantity of DWDs bacteria was directly depended on biofilm bacteria and its increase was correlated with growth and detachment of bacteria from biofilms. Pipe material also affected the microbial community in drinking water distribution network biofilm while Similarity in bacterial species was observed between systems due to same disinfectant dose, time period and plumbing pipes.

Keywords: biofilm, DWDs, pipe material, bacterial population

Procedia PDF Downloads 347

Authors: Zhang Zhenzhen

Abstract:

Valproate (VPA) is commonly used in the treatment of bipolar disorder and epilepsy. The mechanism is complicated, including its ability to inhibit histone deacetylases (HDACs). Here, we show that VPA attenuated VEGF gene expression and the morphological changes in choroidal neovascularization (CNV) induced by photocoagulation in retina. C57BL/6 mice were injected subcutaneously at 300mg/kg twice daily with VPA before insult. Vascular endothelial growth factor (VEGF)-A and VEGF-B were examined in the eyes of VPA-treated mice and in human retinal pigment epithelial cell lines (ARPE-19) exposed to VPA. In addition, CNV was induced by photocoagulation in mice injected with VPA, and the volume of CNV was compared by fluorescence-labeled choroidal flat mount. Morphological changes were analyzed on stained histological sections. Western blot analysis was used to determine protein levels of VEGF-A and VEGF-B, and acetylation of histone H3 in each group. VPA injected intraperitoneally attenuated the VEGF-A and VEGF-B expression in the retina, accompanied by the hyperacetylation of retina tissue, indicating that VPA acts directly on retina tissues through acetylation to reduce the expression of VEGF. VPA also attenuated the VEGF-A mRNA expression in the retinal pigment epithelium showed by immunohistochemistry. Moreover, the administration of VPA significantly attenuated photocoagulation-induced CNV in mice. These results demonstrate that VPA attenuated VEGF production in retina associated with choroidal neovascularization possibly via the HDAC inhibition.

Keywords: retina, acetylation, chorodial neovascularization, vascular endothelial growth factor

Procedia PDF Downloads 204

Authors: Bin Mu, Site Li, Shijin Yuan

Abstract:

Under the circumstance of environment deterioration, people are increasingly concerned about the quality of the environment, especially air quality. As a result, it is of great value to give accurate and timely forecast of AQI (air quality index). In order to simplify influencing factors of air quality in a city, and forecast the city’s AQI tomorrow, this study used MATLAB software and adopted the method of constructing a mathematic model of PCA-GABP to provide a solution. To be specific, this study firstly made principal component analysis (PCA) of influencing factors of AQI tomorrow including aspects of weather, industry waste gas and IAQI data today. Then, we used the back propagation neural network model (BP), which is optimized by genetic algorithm (GA), to give forecast of AQI tomorrow. In order to verify validity and accuracy of PCA-GABP model’s forecast capability. The study uses two statistical indices to evaluate AQI forecast results (normalized mean square error and fractional bias). Eventually, this study reduces mean square error by optimizing individual gene structure in genetic algorithm and adjusting the parameters of back propagation model. To conclude, the performance of the model to forecast AQI is comparatively convincing and the model is expected to take positive effect in AQI forecast in the future.

Keywords: AQI forecast, principal component analysis, genetic algorithm, back propagation neural network model

Procedia PDF Downloads 229