Search results for: gene pool
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1819

Search results for: gene pool

829 MicroRNA Expression Distinguishes Neutrophil Subtypes

Authors: R. I. You, C. L. Ho, M. S. Dai, H. M. Hung, S. F. Yen, C. S. Chen, T. Y. Chao

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Neutrophils are the most abundant innate immune cells to against invading microorganisms. Numerous data shown neutrophils have plasticity in response to physiological and pathological conditions. Tumor-associated neutrophils (TAN) exist in distinct types of tumor and play an important role in cancer biology. Different transcriptomic profiles of neutrophils in tumor and non-tumor samples have been identified. Several miRNAs have been recognized as regulators of gene expression in neutrophil, which may have key roles in neutrophil activation. However, the miRNAs expression patterns in TAN are not well known. To address this question, magnetic bead isolated neutrophils from tumor-bearing mice were used in this study. We analyzed production of reactive oxygen species (ROS) by luminol-dependent chemiluminescence assay. The expression of miRNAs targeting NADPH oxidase, ROS generation and autophagy was explored using quantitative real-time polymerase chain reaction. Our data suggest that tumor environment influence neutrophil develop to differential states of activation via miRNAs regulation.

Keywords: tumor-associated neutrophil, miRNAs, neutrophil, ROS

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828 Architectural Experience of the Everyday in Phuket Old Town

Authors: Thirayu Jumsai na Ayudhya

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Initial attempts to understand about what architecture means to people as they go about their everyday life through my previous research revealed that knowledge such as environmental psychology, environmental perception, environmental aesthetics, did not adequately address a perceived need for the contextualized and holistic theoretical framework. In my previous research, it is found that people’s making senses of their everyday architecture can be described in terms of four super‐ordinate themes; (1) building in urban (text), (2) building in (text), (3) building in human (text), (4) and building in time (text). For more comprehensively understanding of how people make sense of their everyday architectural experience, in this ongoing research Phuket Old town was selected as the focal urban context where the distinguish character of Chino-Portuguese is remarkable. It is expected that in a unique urban context like Phuket old town unprecedented super-ordinate themes will be unveiled through the reflection of people’s everyday experiences. The ongoing research of people’s architectural experience conducted in Phuket Island, Thailand, will be presented succinctly. The research will address the question of how do people make sense of their everyday architecture/buildings especially in a unique urban context, Phuket Old town, and identify ways in which people make sense of their everyday architecture. Participant-Produced-Photograph (PPP) and Interpretative Phenomenological Analysis (IPA) are adopted as main methodologies. PPP allows people to express experiences of their everyday urban context freely without any interference or forced-data generating by researchers. With IPA methodology a small pool of participants is considered desirable given the detailed level of analysis required and its potential to produce a meaningful outcome.

Keywords: architectural experience, the everyday architecture, Phuket, Thailand

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827 History and Its Significance in Modern Visual Graphic: Its Niche with Respect to India

Authors: Hemang Madhusudan Anglay, Akash Gaur

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Value of visual perception in today’s context is vulnerable. Visual Graphic broadly and conveniently expresses culture, language and science of art that satisfactorily is a mould to cast various expressions. It is one of the essential parts of communication design which relatively can be used to approach the above areas of expressions. In between the receptors and interpreters, there is an expanse of comprehension and cliché in relation to the use of Visual Graphics. There are pedagogies, commodification and honest reflections where Visual Graphic is a common area of interest. The traditional receptors amidst the dilemma of this very situation find themselves in the pool of media, medium and interactions. Followed by a very vague interpretation the entire circle of communication becomes a question of comprehension vs cliché. Residing in the same ‘eco-system’ these communities who make pedagogies and multiply its reflections sometimes with honesty and sometimes on commercial values tend to function differently. With the advent of technology, which is a virtual space allows the user to access various forms of content. This diminishes the core characteristics and creates a vacuum even though it satisfies the user. The symbolic interpretation of visual form and structure is transmitted in a culture by the means of contemporary media. Starting from a very individualistic approach, today it is beyond Print & Electronic media. The expected outcome will be a study of Ahmedabad City, situated in the Gujarat State of India. It is identity with respect to socio-cultural as well as economic changes. The methodology will include process to understand the evolution and narratives behind it that will encompass diverse community, its reflection and it will sum up the salient features of communication through combination of visual and graphic that is relevant in Indian context trading its values to global scenario.

Keywords: communication, culture, graphic, visual

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826 Prediction and Identification of a Permissive Epitope Insertion Site for St Toxoid in cfaB from Enterotoxigenic Escherichia coli

Authors: N. Zeinalzadeh, Mahdi Sadeghi

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Enterotoxigenic Escherichia coli (ETEC) is the most common cause of non-inflammatory diarrhea in the developing countries, resulting in approximately 20% of all diarrheal episodes in children in these areas. ST is one of the most important virulence factors and CFA/I is one of the frequent colonization factors that help to process of ETEC infection. ST and CfaB (CFA/I subunit) are among vaccine candidates against ETEC. So, ST because of its small size is not a good immunogenic in the natural form. However to increase its immunogenic potential, here we explored candidate positions for ST insertion in CfaB sequence. After bioinformatics analysis, one of the candidate positions was selected and the chimeric gene (cfaB*st) sequence was synthesized and expressed in E. coli BL21 (DE3). The chimeric recombinant protein was purified with Ni-NTA columns and characterized with western blot analysis. The residue 74-75 of CfaB sequence could be a good candidate position for ST and other epitopes insertion.

Keywords: bioinformatics, CFA/I, enterotoxigenic E. coli, ST toxoid

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825 Identifying Promoters and Their Types Based on a Two-Layer Approach

Authors: Bin Liu

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Prokaryotic promoter, consisted of two short DNA sequences located at in -35 and -10 positions, is responsible for controlling the initiation and expression of gene expression. Different types of promoters have different functions, and their consensus sequences are similar. In addition, their consensus sequences may be different for the same type of promoter, which poses difficulties for promoter identification. Unfortunately, all existing computational methods treat promoter identification as a binary classification task and can only identify whether a query sequence belongs to a specific promoter type. It is desired to develop computational methods for effectively identifying promoters and their types. Here, a two-layer predictor is proposed to try to deal with the problem. The first layer is designed to predict whether a given sequence is a promoter and the second layer predicts the type of promoter that is judged as a promoter. Meanwhile, we also analyze the importance of feature and sequence conversation in two aspects: promoter identification and promoter type identification. To the best knowledge of ours, it is the first computational predictor to detect promoters and their types.

Keywords: promoter, promoter type, random forest, sequence information

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824 Major Histocompatibility Complex (MHC) Polymorphism and Disease Resistance

Authors: Oya Bulut, Oguzhan Avci, Zafer Bulut, Atilla Simsek

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Livestock breeders have focused on the improvement of production traits with little or no attention for improvement of disease resistance traits. In order to determine the association between the genetic structure of the individual gene loci with possibility of the occurrence and the development of diseases, MHC (major histocompatibility complex) are frequently used. Because of their importance in the immune system, MHC locus is considered as candidate genes for resistance/susceptibility against to different diseases. Major histocompatibility complex (MHC) molecules play a critical role in both innate and adaptive immunity and have been considered candidate molecular markers of an association between polymorphisms and resistance/susceptibility to diseases. The purpose of this study is to give some information about MHC genes become an important area of study in recent years in terms of animal husbandry and determine the relation between MHC genes and resistance/susceptibility to disease.

Keywords: MHC, polymorphism, disease, resistance

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823 Malaria Vector Situation in Tanjung Subdistrict, West Lombok Regency, West Nusa Tenggara Province, Indonesia

Authors: Subagyo Yotopranoto, Sri Wijayanti Sulistyawati, Sukmawati Basuki, Budi Armika, Yoes Prijatna Dachlan

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Malaria is a parasitic infectious disease that still remains a health problem in the world, including Indonesia. There is an outbreak happen at West Nusa Tenggara in 2007. A tourist spot in West Nusa Tenggara called West Lombok is mesoendemic area for malaria. Tanjung is the highest malaria morbidity subdistrict in West Lombok. Thus, the research conducted for the presence of a new species of malaria vectors, that are suspected of one factors which caused high morbidity of malaria in this region. The study was conducted in coastal and highland areas. We collected and identified Anopheles larvae from their breeding places. We also collected and identified Anopheles adult mosquitoes with outdoor cow net, indoor and outdoor human bait. In coastal area (Tembobor village), we found Anopheles vagus larvae from rivers as its breeding places. In highland area (Dasan Tengah village), we found An. subpictus from pool, lagoon, and river as its breeding places. In coastal area, with outdoor human bait, we collected An. vagus and An. subpictus adult mosquitoes. With indoor human bait, we collected An. subpictus adult mosquitoes. Whereas with outdoor cow net, we collected An. subpictus and An. maculatus, the first was more dominant. Furthermore, An subpictus strong suspected as malaria vector in coastal area. Anopheles subpictus was an anthropozoophylic mosquitoes, because it was found at indoor and outdoor places.

Keywords: malaria, vector, Tanjung, West Nusa Tenggara

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822 Computational Fluid Dynamics Simulation on Heat Transfer of Hot Air Bubble Injection into Water Column

Authors: Jae-Yeong Choi, Gyu-Mok Jeon, Jong-Chun Park, Yong-Jin Cho, Seok-Tae Yoon

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When air flow is injected into water, bubbles are formed in various types inside the water pool along with the air flow rate. The bubbles are floated in equilibrium with forces such as buoyancy, surface tension and shear force. Single bubble generated at low flow rate maintains shape, but bubbles with high flow rate break up to make mixing and turbulence. In addition to this phenomenon, as the hot air bubbles are injected into the water, heat affects the interface of phases. Therefore, the main scope of the present work reveals how to proceed heat transfer between water and hot air bubbles injected into water. In the present study, a series of CFD simulation for the heat transfer of hot bubbles injected through a nozzle near the bottom in a cylindrical water column are performed using a commercial CFD software, STAR-CCM+. The governing equations for incompressible and viscous flow are the continuous and the RaNS (Reynolds- averaged Navier-Stokes) equations and discretized by the FVM (Finite Volume Method) manner. For solving multi-phase flow, the Eulerian multiphase model is employed and the interface is defined by VOF (Volume-of-Fluid) technique. As a turbulence model, the SST k-w model considering the buoyancy effects is introduced. For spatial differencing the 3th-order MUSCL scheme is adopted and the 2nd-order implicit scheme for time integration. As the results, the dynamic behavior of the rising hot bubbles with the flow rate injected and regarding heat transfer mechanism are discussed based on the simulation results.

Keywords: heat transfer, hot bubble injection, eulerian multiphase model, flow rate, CFD (Computational Fluid Dynamics)

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821 The Role of Fluid Catalytic Cracking in Process Optimisation for Petroleum Refineries

Authors: Chinwendu R. Nnabalu, Gioia Falcone, Imma Bortone

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Petroleum refining is a chemical process in which the raw material (crude oil) is converted to finished commercial products for end users. The fluid catalytic cracking (FCC) unit is a key asset in refineries, requiring optimised processes in the context of engineering design. Following the first stage of separation of crude oil in a distillation tower, an additional 40 per cent quantity is attainable in the gasoline pool with further conversion of the downgraded product of crude oil (residue from the distillation tower) using a catalyst in the FCC process. Effective removal of sulphur oxides, nitrogen oxides, carbon and heavy metals from FCC gasoline requires greater separation efficiency and involves an enormous environmental significance. The FCC unit is primarily a reactor and regeneration system which employs cyclone systems for separation.  Catalyst losses in FCC cyclones lead to high particulate matter emission on the regenerator side and fines carryover into the product on the reactor side. This paper aims at demonstrating the importance of FCC unit design criteria in terms of technical performance and compliance with environmental legislation. A systematic review of state-of-the-art FCC technology was carried out, identifying its key technical challenges and sources of emissions.  Case studies of petroleum refineries in Nigeria were assessed against selected global case studies. The review highlights the need for further modelling investigations to help improve FCC design to more effectively meet product specification requirements while complying with stricter environmental legislation.

Keywords: design, emission, fluid catalytic cracking, petroleum refineries

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820 Gender and Geographical Disparity in Editorial Boards of Lithuanian Scientific Journals: An Overview of Different Science Disciplines

Authors: Andrius Suminas

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Editors-in-chief and members of editorial boards of scientific journals play an extremely important role in the development of science and assure research integrity, as scientific publications are the major results of research. While gender parity in tenure-track hiring decisions and promotion rates has improved, female academics remain underrepresented in senior career phases, including editors-in-chief and members of editorial boards positions of scientific journals. Journal editors and members of editorial boards exert considerable power over what is published and in certain cases the direction of an academic discipline and the career advancement of authors. For this reason it is important to minimize biases extrinsic to the merit of the work impacting publication decisions. One way to achieve this is to ensure a diverse pool of editors and members of editorial boards, ensuring the widest possible coverage of different competencies. This is in line with a diversity model of editorial appointment where editorial boards are structured to dismantle wider conditions of inequality. Another possible option, a distributive model would seek an editorial board reflective of existing proportions in the field at large. Paper presents comprehensive results of Lithuanian scientific journals study. During the research process were reviewed publicly available information from all scientific journals published in Lithuania to infer the proportions of members of editorial boards by gender and country of affiliation. The results of the study revealed differences the proportions of male and female members of editorial boards in different disciplines of science, as well as clear geographical disparity in Lithianian scientific journals editorial boards.

Keywords: scientific journals, editorial boards of scientific journals, gender disparity, geographical disparity, scientific communication

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819 Mesoporous Tussah Silk Fibroin Microspheres for Drug Delivery

Authors: Weitao Zhou, Qing Wang, Jianxin He, Shizhong Cui

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Mesoporous Tussah silk fibroin (TSF) spheres were fabricated via the self-assembly of TSF molecules in aqueous solutions. The results showed that TSF particles were approximately three-dimensional spheres with the diameter ranging from 500nm to 6μm without adherence. More importantly, the surface morphology is mesoporous structure with nano-pores of 20nm - 200nm in size. Fourier transform infrared (FT-IR) and X-ray diffraction (XRD) studies demonstrated that mesoporous TSF spheres mainly contained beta-sheet conformation (44.1 %) as well as slight amounts of random coil (13.2 %). Drug release test was performed with 5-fluorouracil (5-Fu) as a model drug and the result indicated the mesoporous TSF microspheres had a good capacity of sustained drug release. It is expected that these stable and high-crystallinity mesoporous TSF sphere produced without organic solvents, which have significantly improved drug release properties, is a very promising material for controlled gene medicines delivery.

Keywords: Tussah silk fibroin, porous materials, microsphere, drug release

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818 The Effect of Dopamine D2 Receptor TAQ A1 Allele on Sprinter and Endurance Athlete

Authors: Öznur Özge Özcan, Canan Sercan, Hamza Kulaksız, Mesut Karahan, Korkut Ulucan

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Genetic structure is very important to understand the brain dopamine system which is related to athletic performance. Hopefully, there will be enough studies about athletics performance in the terms of addiction-related genetic markers in the future. In the present study, we intended to investigate the Receptor-2 Gene (DRD2) rs1800497, which is related to brain dopaminergic system. 10 sprinter and 10 endurance athletes were enrolled in the study. Real-Time Polymerase Chain Reaction method was used for genotyping. According to results, A1A1, A1A2 and A2A2 genotypes in athletes were 0 (%0), 3 (%15) and 17 (%85). A1A1 genotype was not found and A2 allele was counted as the dominating allele in our cohort. These findings show that dopaminergic mechanism effects on sport genetic may be explained by the polygenic and multifactorial view.

Keywords: addiction, athletic performance, genotype, sport genetics

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817 Mindfulness and Mental Resilience Training for Pilots: Enhancing Cognitive Performance and Stress Management

Authors: Nargiza Nuralieva

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The study delves into assessing the influence of mindfulness and mental resilience training on the cognitive performance and stress management of pilots. Employing a meticulous literature search across databases such as Medline and Google Scholar, the study used specific keywords to target a wide array of studies. Inclusion criteria were stringent, focusing on peer-reviewed studies in English that utilized designs like randomized controlled trials, with a specific interest in interventions related to mindfulness or mental resilience training for pilots and measured outcomes pertaining to cognitive performance and stress management. The initial literature search identified a pool of 123 articles, with subsequent screening resulting in the exclusion of 77 based on title and abstract. The remaining 54 articles underwent a more rigorous full-text screening, leading to the exclusion of 41. Additionally, five studies were selected from the World Health Organization's clinical trials database. A total of 11 articles from meta-analyses were retained for examination, underscoring the study's dedication to a meticulous and robust inclusion process. The interventions varied widely, incorporating mixed approaches, Cognitive behavioral Therapy (CBT)-based, and mindfulness-based techniques. The analysis uncovered positive effects across these interventions. Specifically, mixed interventions demonstrated a Standardized Mean Difference (SMD) of 0.54, CBT-based interventions showed an SMD of 0.29, and mindfulness-based interventions exhibited an SMD of 0.43. Long-term effects at a 6-month follow-up suggested sustained impacts for both mindfulness-based (SMD: 0.63) and CBT-based interventions (SMD: 0.73), albeit with notable heterogeneity.

Keywords: mindfulness, mental resilience, pilots, cognitive performance, stress management

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816 Intended Use of Genetically Modified Organisms, Advantages and Disadvantages

Authors: Pakize Ozlem Kurt Polat

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GMO (genetically modified organism) is the result of a laboratory process where genes from the DNA of one species are extracted and artificially forced into the genes of an unrelated plant or animal. This technology includes; nucleic acid hybridization, recombinant DNA, RNA, PCR, cell culture and gene cloning techniques. The studies are divided into three groups of properties transferred to the transgenic plant. Up to 59% herbicide resistance characteristic of the transfer, 28% resistance to insects and the virus seems to be related to quality characteristics of 13%. Transgenic crops are not included in the commercial production of each product; mostly commercial plant is soybean, maize, canola, and cotton. Day by day increasing GMO interest can be listed as follows; Use in the health area (Organ transplantation, gene therapy, vaccines and drug), Use in the industrial area (vitamins, monoclonal antibodies, vaccines, anti-cancer compounds, anti -oxidants, plastics, fibers, polyethers, human blood proteins, and are used to produce carotenoids, emulsifiers, sweeteners, enzymes , food preservatives structure is used as a flavor enhancer or color changer),Use in agriculture (Herbicide resistance, Resistance to insects, Viruses, bacteria, fungi resistance to disease, Extend shelf life, Improving quality, Drought , salinity, resistance to extreme conditions such as frost, Improve the nutritional value and quality), we explain all this methods step by step in this research. GMO has advantages and disadvantages, which we explain all of them clearly in full text, because of this topic, worldwide researchers have divided into two. Some researchers thought that the GMO has lots of disadvantages and not to be in use, some of the researchers has opposite thought. If we look the countries law about GMO, we should know Biosafety law for each country and union. For this Biosecurity reasons, the problems caused by the transgenic plants, including Turkey, to minimize 130 countries on 24 May 2000, ‘the United Nations Biosafety Protocol’ signed nudes. This protocol has been prepared in addition to Cartagena Biosafety Protocol entered into force on September 11, 2003. This protocol GMOs in general use by addressing the risks to human health, biodiversity and sustainable transboundary movement of all GMOs that may affect the prevention, transit covers were dealt and used. Under this protocol we have to know the, ‘US Regulations GMO’, ‘European Union Regulations GMO’, ‘Turkey Regulations GMO’. These three different protocols have different applications and rules. World population increasing day by day and agricultural fields getting smaller for this reason feeding human and animal we should improve agricultural product yield and quality. Scientists trying to solve this problem and one solution way is molecular biotechnology which is including the methods of GMO too. Before decide to support or against the GMO, should know the GMO protocols and it effects.

Keywords: biotechnology, GMO (genetically modified organism), molecular marker

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815 The Research On The Necessity Of Launching Environmental Programs For Studies In Universities As Well As Training Specialists In This Sphere.

Authors: Anastasia V. Lazareva

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Nowadays in the light of the evolving multifocal challenges in the sphere of environmental and social difficulties and despite the strong opposition of globalist and anti-globalist movements, we are facing the urgent need of the creation of a vast pool of educated environmentalists through the implementation of relevant university faculties and programs. Considering the threats humanity has tackled these years portrayed in every tiny detail in AGENDA – 2030 –namely, poverty, biodiversity loss, marine and terrestrial pollution, lack of sanitation, and equal rights for all, we must admit that professionals are required to address them all. With this purpose, we have conducted research based on the questionnaires of students, faculty members, and companies’ chief executives and human resources managers on what particular disciplines should be incorporated into the programs in universities and higher institutions to meet the millennium goals and tests. The research is based on the Linkert scale and covers various age groups of students. The topicality of this issue is predetermined by modern reality. The subject of the research is a questionnaire database filled in by 97 students, 17 faculty members of MGIMO University, and 14 companies’ representatives concerning their attitudes towards the implementation of environmental programs of studies in universities and the choice of disciplines required. The study has a limitation -it is based only on one university students' and faculty members’ questionnaires. The methods applied are a questionnaire, content analysis, sampling, and categorization. The findings of this survey imply that all three groups of respondents admit the necessity of implementing environmental programs for studies in higher education. Nevertheless, different groups favor various programs and disciplines to be incorporated into the curriculum.

Keywords: ecology, university studies, environmentalists, education, global challenges

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814 An Experimental Approach to the Influence of Tipping Points and Scientific Uncertainties in the Success of International Fisheries Management

Authors: Jules Selles

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The Atlantic and Mediterranean bluefin tuna fishery have been considered as the archetype of an overfished and mismanaged fishery. This crisis has demonstrated the role of public awareness and the importance of the interactions between science and management about scientific uncertainties. This work aims at investigating the policy making process associated with a regional fisheries management organization. We propose a contextualized computer-based experimental approach, in order to explore the effects of key factors on the cooperation process in a complex straddling stock management setting. Namely, we analyze the effects of the introduction of a socio-economic tipping point and the uncertainty surrounding the estimation of the resource level. Our approach is based on a Gordon-Schaefer bio-economic model which explicitly represents the decision making process. Each participant plays the role of a stakeholder of ICCAT and represents a coalition of fishing nations involved in the fishery and decide unilaterally a harvest policy for the coming year. The context of the experiment induces the incentives for exploitation and collaboration to achieve common sustainable harvest plans at the Atlantic bluefin tuna stock scale. Our rigorous framework allows testing how stakeholders who plan the exploitation of a fish stock (a common pool resource) respond to two kinds of effects: i) the inclusion of a drastic shift in the management constraints (beyond a socio-economic tipping point) and ii) an increasing uncertainty in the scientific estimation of the resource level.

Keywords: economic experiment, fisheries management, game theory, policy making, Atlantic Bluefin tuna

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813 The Effect of Vitamin D Supplementation on Prostate Cancer: A Systematic Review and Meta-Analysis of Clinical Trials

Authors: Simin Shahvazi, Sepideh Soltani, Seyed Mehdi Ahmadi, Russell J. De Souza, Amin Salehi-Abargouei

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Background and Objectives: Vitamin D has received attention for its potential to disrupt cancer processes such as attenuating cell proliferation and exacerbating differentiation and apoptosis. However, whether there exists a role for vitamin D in the treatment of prostate cancer specifically remains controversial. We systematically review the literature to assess whether supplementation with vitamin D influences PSA response and overall survival in patients with prostate cancer. Methods: We searched PubMed, Scopus, ISI Web of Science and Google scholar from inception through up to 10 September 2017 for both before-and-after and randomized trials that evaluated the effect of vitamin D supplementation on the prostate specific antigen (PSA) response rate in participants with prostate cancer. The DerSimonian and Laird, inverse-weighted random-effects model was used to pool effect estimates from the studies. Heterogeneity and potential publication bias were evaluated. Subgroup analyses were also performed. Results: Twenty-two studies (16 before-after and 6 randomized controlled trials) were found and included in meta-analysis. The analysis on controlled clinical trials revealed that PSA change from baseline [weighted mean difference (WMD) = -1.66 ng/ml, 95%CI: -0.69, 0.36, P= 0.543)], PSA response (RR=1.18, 95%CI: 0.97, 1.45, P=0.104) and mortality rate (risk ratio (RR) = 1.05, 95% CI: 0.81-1.36; P=0.713) was not significantly different between vitamin D supplementation and placebo groups. Single arm trials revealed that vitamin D supplementation had had a modest effect on PSA response rate: 19% of those enrolled had at least a 50% reduction in PSA by the end of treatment (95% CI: 7% to 31%; p=0.002). Conclusion: We found that vitamin D modestly increases the PSA response rate in single arm studies. No effect on serum PSA levels, PSA response and mortality was seen in randomized controlled clinical trials. It does not seem patients with prostate cancer benefit from vitamin D supplementation.

Keywords: mortality, prostatic neoplasms, PSA response, vitamin D

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812 Resilience and Mindfulness as Individual Resources Building Communication Skills for Physicians

Authors: Malgorzata Sekulowicz, Krystyna Boron-Krupinska, Paulina Morga, Blazej Cieslik

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Burnout is highly prevalent in health care employees, especially in physicians. It significantly reduces the efficiency of these employees, which can have negative consequences for both physicians and patients. Resilience and mindfulness enhancing positive emotions, leading to sustainable development and personal commitment, can have a significant impact on burnout. Therefore, the aim of this study was to determine the relationship between burnout symptoms and mindfulness and resilience among physicians. The authors conducted a cross-sectional survey study among seventy-four polish physicians. Participants filled out the following psychometric tools: the Maslach Burnout Inventory - Human Services Survey (MBI-HSS), Five Facet Mindfulness Questionnaire (FFMQ), Areas of Work-Life Survey (AWS), International Personality Item Pool (IPIP), the Resilience Assessment Scale (SPP-25) and the Mini-COPE Inventory. The relationship between burnout and resilience and mindfulness was assessed with path analysis. Analyzing the relationship between MBI-HSS components and mindfulness, a significant negative correlation of the FFMQ score with emotional exhaustion (-0.50, p < 0.05) and depersonalization (-0.43, p < 0.05) and a positive correlation with personal accomplishment (0.50, p < 0.05) was demonstrated. Analyzing resilience, a statistically significant relationship of SPP-25 with all tested components of MBI-HSS was demonstrated: emotional exhaustion (-0.54, p < 0.05), depersonalization (-0.31, p < 0.05) and personal accomplishment (0.35, p < 0.05). In the group of medical doctors, the higher the level of mindfulness and resilience, the lower the risk of burnout. Furthermore, the more frequently used active coping strategies (planning, acceptance), the lower the risk of burnout, while the use of passive, evasive strategies increases the risk of burnout. It may be worth considering implementing mindfulness intervention to effectively manage burnout symptoms in this group.

Keywords: burnout, medical doctors, mindfulness, physicians, resilience

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811 Artificial Cells Capable of Communication by Using Polymer Hydrogel

Authors: Qi Liu, Jiqin Yao, Xiaohu Zhou, Bo Zheng

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The first artificial cell was produced by Thomas Chang in the 1950s when he was trying to make a mimic of red blood cells. Since then, many different types of artificial cells have been constructed from one of the two approaches: a so-called bottom-up approach, which aims to create a cell from scratch, and a top-down approach, in which genes are sequentially knocked out from organisms until only the minimal genome required for sustaining life remains. In this project, bottom-up approach was used to build a new cell-free expression system which mimics artificial cell that capable of protein expression and communicate with each other. The artificial cells constructed from the bottom-up approach are usually lipid vesicles, polymersomes, hydrogels or aqueous droplets containing the nucleic acids and transcription-translation machinery. However, lipid vesicles based artificial cells capable of communication present several issues in the cell communication research: (1) The lipid vesicles normally lose the important functions such as protein expression within a few hours. (2) The lipid membrane allows the permeation of only small molecules and limits the types of molecules that can be sensed and released to the surrounding environment for chemical communication; (3) The lipid vesicles are prone to rupture due to the imbalance of the osmotic pressure. To address these issues, the hydrogel-based artificial cells were constructed in this work. To construct the artificial cell, polyacrylamide hydrogel was functionalized with Acrylate PEG Succinimidyl Carboxymethyl Ester (ACLT-PEG2000-SCM) moiety on the polymer backbone. The proteinaceous factors can then be immobilized on the polymer backbone by the reaction between primary amines of proteins and N-hydroxysuccinimide esters (NHS esters) of ACLT-PEG2000-SCM, the plasmid template and ribosome were encapsulated inside the hydrogel particles. Because the artificial cell could continuously express protein with the supply of nutrients and energy, the artificial cell-artificial cell communication and artificial cell-natural cell communication could be achieved by combining the artificial cell vector with designed plasmids. The plasmids were designed referring to the quorum sensing (QS) system of bacteria, which largely relied on cognate acyl-homoserine lactone (AHL) / transcription pairs. In one communication pair, “sender” is the artificial cell or natural cell that can produce AHL signal molecule by synthesizing the corresponding signal synthase that catalyzed the conversion of S-adenosyl-L-methionine (SAM) into AHL, while the “receiver” is the artificial cell or natural cell that can sense the quorum sensing signaling molecule form “sender” and in turn express the gene of interest. In the experiment, GFP was first immobilized inside the hydrogel particle to prove that the functionalized hydrogel particles could be used for protein binding. After that, the successful communication between artificial cell-artificial cell and artificial cell-natural cell was demonstrated, the successful signal between artificial cell-artificial cell or artificial cell-natural cell could be observed by recording the fluorescence signal increase. The hydrogel-based artificial cell designed in this work can help to study the complex communication system in bacteria, it can also be further developed for therapeutic applications.

Keywords: artificial cell, cell-free system, gene circuit, synthetic biology

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810 Study on the Expression of Drought Tolerant Genes in Water-Stressed Basella Alba and Basella Rubra

Authors: T. O. Ajewole, K. S. Olorunmiaye, D. A. Animasaun, M. Okpeku

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Drought impact on the production of food crops for the benefit of mankind cannot be overemphasized. This study shows the different kind of genes expressed at various level of drought regimes on Basella alba and rubra using a real-time PCR machine. The planting was done in the screen house while the gene expression study was carried out in the laboratory. Sandy-loamy soil was collected and four levels of drought regime was used as treatment and a control experiment was set up for the two vegetables. Drought interval of 5, 10, 15 and 20 days were used as treatments while a control experiment which was not starved of water at any point was also set up, five replicates were set up for each treatment. Stress was introduced at 12 Weeks after planting (WAP). From the result of this study, Basella alba shows the highest amplicon size of 34.6 and 52.32 for GmPCS5 and HVA1 respectively which by implication means these genes were expressed the more as the stress period interval increases.

Keywords: water stress, basella alba, basella rubra, HVA1

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809 Water Quality Calculation and Management System

Authors: H. M. B. N Jayasinghe

Abstract:

The water is found almost everywhere on Earth. Water resources contain a lot of pollution. Some diseases can be spread through the water to the living beings. So to be clean water it should undergo a number of treatments necessary to make it drinkable. So it is must to have purification technology for the wastewater. So the waste water treatment plants act a major role in these issues. When considering the procedures taken after the water treatment process was always based on manual calculations and recordings. Water purification plants may interact with lots of manual processes. It means the process taking much time consuming. So the final evaluation and chemical, biological treatment process get delayed. So to prevent those types of drawbacks there are some computerized programmable calculation and analytical techniques going to be introduced to the laboratory staff. To solve this problem automated system will be a solution in which guarantees the rational selection. A decision support system is a way to model data and make quality decisions based upon it. It is widely used in the world for the various kind of process automation. Decision support systems that just collect data and organize it effectively are usually called passive models where they do not suggest a specific decision but only reveal information. This web base system is based on global positioning data adding facility with map location. Most worth feature is SMS and E-mail alert service to inform the appropriate person on a critical issue. The technological influence to the system is HTML, MySQL, PHP, and some other web developing technologies. Current issues in the computerized water chemistry analysis are not much deep in progress. For an example the swimming pool water quality calculator. The validity of the system has been verified by test running and comparison with an existing plant data. Automated system will make the life easier in productively and qualitatively.

Keywords: automated system, wastewater, purification technology, map location

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808 Long Non-Coding RNAs Mediated Regulation of Diabetes in Humanized Mouse

Authors: Md. M. Hossain, Regan Roat, Jenica Christopherson, Colette Free, Zhiguang Guo

Abstract:

Long noncoding RNA (lncRNA) mediated post-transcriptional gene regulation, and their epigenetic landscapes have been shown to be involved in many human diseases. However, their regulation in diabetes through governing islet’s β-cell function and survival needs to be elucidated. Due to the technical and ethical constraints, it is difficult to study their role in β-cell function and survival in human under in vivo condition. In this study, humanized mice have been developed through transplanting human pancreatic islet under the kidney capsule of NOD.SCID mice and induced β-cell death leading to diabetes condition to study lncRNA mediated regulation. For this, human islets from 3 donors (3000 IEQ, purity > 80%) were transplanted under the kidney capsule of STZ induced diabetic NOD.scid mice. After at least 2 weeks of normoglycecemia, lymphocytes from diabetic NOD mice were adoptively transferred and islet grafts were collected once blood glucose reached > 200 mg/dl. RNA from human donor islets, islet grafts from humanized mice with either adoptive lymphocyte transfer (ALT) or PBS control (CTL) were ribodepleted; barcoded fragment libraries were constructed and sequenced on the Ion Proton sequencer. lncRNA expression in isolated human islets, islet grafts from humanized mice with and without induced β-cell death and their regulation in human islets function in vitro under glucose challenge, cytokine mediated inflammation and induced apoptotic condition were investigated. Out of 3155 detected lncRNAs, 299 that highly expressed in islets were found to be significantly downregulated and 224 upregulated in ALT compared to CTL. Most of these are found to be collocated within 5 kb upstream and 1 kb downstream of 788 up- and 624 down-regulated mRNAs. Genomic Regions Enrichment of Annotations Analysis revealed deregulated and collocated genes are related to pancreas endocrine development; insulin synthesis, processing, and secretion; pancreatitis and diabetes. Many of them, that found to be located within enhancer domains for islet specific gene activity, are associated to the deregulation of known islet/βcell specific transcription factors and genes that are important for β-cell differentiation, identity, and function. RNA sequencing analysis revealed aberrant lncRNA expression which is associated to the deregulated mRNAs in β-cell function as well as in molecular pathways related to diabetes. A distinct set of candidate lncRNA isoforms were identified as highly enriched and specific to human islets, which are deregulated in human islets from donors with different BMIs and with type 2 diabetes. These RNAs show an interesting regulation in cultured human islets under glucose stimulation and with induced β-cell death by cytokines. Aberrant expression of these lncRNAs was detected in the exosomes from the media of islets cultured with cytokines. Results of this study suggest that the islet specific lncRNAs are deregulated in human islet with β-cell death, hence important in diabetes. These lncRNAs might be important for human β-cell function and survival thus could be used as biomarkers and novel therapeutic targets for diabetes.

Keywords: β-cell, humanized mouse, pancreatic islet, LncRNAs

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807 People's Perspective on Water Commons in Trans-Boundary Water Governance: A Case Study from Nepal

Authors: Sristi Silwal

Abstract:

South Asian rivers support ecosystems and sustain well-being of thousands of riparian communities. Rivers however are also sources of conflict between countries and one of the contested issues between governments of the region. Governments have signed treaties to harness some of the rivers but their provisions have not been successful in improving the quality of life of those who depend on water as common property resources. This paper will present a case of the study of the status of the water commons along the lower command areas of Koshi, Gandka and Mahakali rivers. Nepal and India have signed treaties for development and management of these rivers in 1928, 1954 and 1966. The study investigated perceptions of the local community on climate-induced disasters, provision of the treaties such as water for irrigation, participation in decision-making and specific impact of women. It looked at how the local community coped with adversities. The study showed that the common pool resources are gradually getting degraded, flood events increasing while community blame ‘other state’ and state administration for exacerbating these ills. The level of awareness about provisions of existing treatise is poor. Ongoing approach to trans-boundary water management has taken inadequate cognizance of these realities as the dominant narrative perpetuates cooperation between the governments. The paper argues that on-going discourses on trans-boundary water development and management need to use a new metrics of taking cognizance of the condition of the commons and that of the people depended on them for sustenance. In absence of such narratives, the scale of degradation would increase making those already marginalized more vulnerable to impacts of global climate change.

Keywords: climate change vulnerability, conflict, cooperation, water commons

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806 Evaluation of the Incidence of Mycobacterium Tuberculosis Complex Associated with Soil, Hayfeed and Water in Three Agricultural Facilities in Amathole District Municipality in the Eastern Cape Province

Authors: Athini Ntloko

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Mycobacterium bovis and other species of Mycobacterium tuberculosis complex (MTBC) can result to a zoonotic infection known as Bovine tuberculosis (bTB). MTBC has members that may contaminate an extensive range of hosts, including wildlife. Diverse wild species are known to cause disease in domestic livestock and are acknowledged as TB reservoirs. It has been a main study worldwide to deliberate on bTB risk factors as a result and some studies focused on particular parts of risk factors such as wildlife and herd management. The significance of the study was to determine the incidence of Mycobacterium tuberculosis complex that is associated with soil, hayfeed and water. Questionnaires were administered to thirty (30) smallholding farm owners in the two villages (kwaMasele and Qungqwala) and three (3) three commercial farms (Fort Hare dairy farm, Middledrift dairy farm and Seven star dairy farm). Detection of M. tuberculosis complex was achieved by Polymerase Chain Reaction using primers for IS6110; whereas a genotypic drug resistance mutation was detected using Genotype MTBDRplus assays. Nine percent (9%) of respondents had more than 40 cows in their herd, while 60% reported between 10 and 20 cows in their herd. Relationship between farm size and vaccination for TB differed from forty one percent (41%) being the highest to the least five percent (5%). The highest number of respondents who knew about relationship between TB cases and cattle location was ninety one percent (91%). Approximately fifty one percent (51%) of respondents had knowledge about wild life access to the farms. Relationship between import of cattle and farm size ranged from nine percent (9%) to thirty five percent (35%). Cattle sickness in relation to farm size differed from forty three (43%) being the highest to the least three percent (3%); while thirty three percent (33%) of respondents had knowledge about health management. Respondents with knowledge about the occurrence of TB infections in farms were forty-eight percent (48%). The frequency of DNA isolation from samples ranged from the highest forty-five percent (45%) from water to the least twenty two percent (22%) from soil. Fort Hare dairy farm had the highest number of positive samples, forty four percent (44%) from water samples; whereas Middledrift dairy farm had the lowest positive from water, seventeen percent (17%). Twelve (22%) out of 55 isolates showed resistance to INH and RIF that is, multi-drug resistance (MDR) and nine percent (9%) were sensitive to either INH or RIF. The mutations at rpoB gene differed from 58% being the highest to the least (23%). Fifty seven percent (57%) of samples showed a S315T1 mutation while only 14% possessed a S531L in the katG gene. The highest inhA mutations were detected in T8A (80 %) and the least was observed in A16G (17%). The results of this study reveal that risk factors for bTB in cattle and dairy farm workers are a serious issue abound in the Eastern Cape of South Africa; with the possibility of widespread dissemination of multidrug resistant determinants in MTBC from the environment.

Keywords: hayfeed, isoniazid, multi-drug resistance, mycobacterium tuberculosis complex, polymerase chain reaction, rifampicin, soil, water

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805 Gut Microbial Dynamics in a Mouse Model of Inflammation-Linked Carcinogenesis as a Result of Diet Supplementation with Specific Mushroom Extracts

Authors: Alvarez M., Chapela M. J., Balboa E., Rubianes D., Sinde E., Fernandez de Ana C., Rodríguez-Blanco A.

Abstract:

The gut microbiota plays an important role as gut inflammation could contribute to colorectal cancer development; however, this role is still not fully understood, and tools able to prevent this progression are yet to be developed. The main objective of this study was to monitor the effects of a mushroom extracts formulation in gut microbial community composition of an Azoxymethane (AOM)/Dextran sodium sulfate (DSS) mice model of inflammation-linked carcinogenesis. For the in vivo study, 41 adult male mice of the C57BL / 6 strain were obtained. 36 of them have been induced in a state of colon carcinogenesis by a single intraperitoneal administration of AOM at a dose of 12.5 mg/kg; the control group animals received instead of the same volume of 0.9% saline. DSS is an extremely toxic polysaccharide sulfate that causes chronic inflammation of the colon mucosa, favoring the appearance of severe colitis and the production of tumors induced by AOM. Induction by AOM/DSS is an interesting platform for chemopreventive intervention studies. This time the model was used to monitor gut microbiota changes as a result of supplementation with a specific mushroom extracts formulation previously shown to have prebiotic activity. The animals have been divided into three groups: (i) Cancer + mushroom extracts formulation experimental group: to which the MicoDigest2.0 mushroom extracts formulation developed by Hifas da Terra S.L has been administered dissolved in drinking water at an estimated concentration of 100 mg / ml. (ii) Control group of animals with Cancer: to which normal water has been administered without any type of treatment. (iii) Control group of healthy animals: these are the animals that have not been induced cancer or have not received any treatment in drinking water. This treatment has been maintained for a period of 3 months, after which the animals were sacrificed to obtain tissues that were subsequently analyzed to verify the effects of the mushroom extract formulation. A microbiological analysis has been carried out to compare the microbial communities present in the intestines of the mice belonging to each of the study groups. For this, the methodology of massive sequencing by molecular analysis of the 16S gene has been used (Ion Torrent technology). Initially, DNA extraction and metagenomics libraries were prepared using the 16S Metagenomics kit, always following the manufacturer's instructions. This kit amplifies 7 of the 9 hypervariable regions of the 16S gene that will then be sequenced. Finally, the data obtained will be compared with a database that makes it possible to determine the degree of similarity of the sequences obtained with a wide range of bacterial genomes. Results obtained showed that, similarly to certain natural compounds preventing colorectal tumorigenesis, a mushroom formulation enriched the Firmicutes and Proteobacteria phyla and depleted Bacteroidetes. Therefore, it was demonstrated that the consumption of the mushroom extracts’ formulation developed could promote the recovery of the microbial balance that is disrupted in the mice model of carcinogenesis. More preclinical and clinical studies are needed to validate this promising approach.

Keywords: carcinogenesis, microbiota, mushroom extracts, inflammation

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804 Oral Lichen Planus a Manifestation of Grinspan's Syndrome or a Lichenoid Reaction to Medication

Authors: Sahar Iqrar, Malik Adeel Anwar, Zain Akram, Maria Noor

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Introduction: Oral lichen planus is a chronic inflammatory condition of unknown etiology. Oral lichen planus may be related with several other diseases. Grinspan's Syndrome is characterized by a triad of oral lichen planus, hypertension, and diabetes mellitus. Other associations reported in the literature are with chronic liver disease and, with dyslipidemia. The nature of these associations is still not fully understood. Material and methods: Study was conducted in Department of Oral Medicine, Fatima Memorial Hospital College of Medicine and Dentistry, Lahore, Pakistan. A total of n=89 clinically diagnosed patients of oral lichen planus of both gender and all age groups were recruited and detailed history were recorded in the designed performs. Results: A total of n=89 patients were taken with male to female ratio of 3:8 in which 24 were male and 65 females. Mean age was 48.8 ± 13.8 years. Age range of 10-74 years was seen. Among these patients suffering from oral lichen planus, 41.6% (n=37) had a positive history for hypertension with 59.5% (n=22) of these patients were taking different medication for their condition. Whereas Diabetes Mellitus was found in 24.7% (n=22) patients with 72.7% (n=16) of these patients using the hypoglycemic drug (oral or injectable) to control their blood glucose levels. Out of these n=89 lichen planus patients 21.3% had both hypertension and diabetes mellitus (fulfilling the criteria for Grinspan's Syndrome). Out of this Grinspan's Syndrome pool 94.7% (n=19) were taking drug atleast for one of the two conditions. Conclusion: As noticed form the medical history of the patients, most of them were using hypoglycemic drugs for diabetes mellitus and beta blockers, diuretics and calcium channel blockers for hypertension. These drugs are known for lichenoid reaction. Therefore, it should be ruled out at histopathological/ immunological and molecular level whether these patients are suffering from lichen planus or lichenoid drug reaction to truly declare them as patients with Grinspan’s Syndrome.

Keywords: diabetes mellitus, grinspan's syndrome, lichenoid drug reaction, oral lichen planus

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803 Right Atrial Tissue Morphology in Acquired Heart Diseases

Authors: Edite Kulmane, Mara Pilmane, Romans Lacis

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Introduction: Acquired heart diseases remain one of the leading health care problems in the world. Changes in myocardium of the diseased hearts are complex and pathogenesis is still not fully clear. The aim of this study was to identify appearance and distribution of apoptosis, homeostasis regulating factors, and innervation and ischemia markers in right atrial tissue in different acquired heart diseases. Methods: During elective open heart surgery were taken right atrial tissue fragments from 12 patients. All patients were operated because of acquired heart diseases- aortic valve stenosis (5 patients), coronary heart disease (5 patients), coronary heart disease and secondary mitral insufficiency (1 patient) and mitral disease (1 patient). The mean age was (mean±SD) 70,2±7,0 years (range 58-83 years). The tissues were stained with haematoxylin and eosin methods for routine light-microscopical examination and for immunohistochemical detection of protein gene peptide 9.5 (PGP 9.5), human atrial natriuretic peptide (hANUP), vascular endothelial growth factor (VEGF), chromogranin A and endothelin. Apoptosis was detected by TUNEL method. Results: All specimens showed degeneration of cardiomyocytes with lysis of myofibrils, diffuse vacuolization especially in perinuclear region, different size of cells and their nuclei. The severe invasion of connective tissue was observed in main part of all fragments. The apoptotic index ranged from 24 to 91%. One specimen showed region of newly performed microvessels with cube shaped endotheliocytes that were positive for PGP 9.5, endothelin, chromogranin A and VEGF. From all fragments, taken from patients with coronary heart disease, there were observed numerous PGP 9.5-containing nerve fibres, except in patient with secondary mitral insufficiency, who showed just few PGP 9.5 positive nerves. In majority of specimens there were regions observed with cube shaped mixed -VEGF immunoreactive endocardial and epicardial cells. Only VEGF positive endothelial cells were observed just in few specimens. There was no significant difference of hANUP secreting cells among all specimens. All patients operated due to the coronary heart disease moderate to numerous number of chromogranin A positive cells were seen while in patients with aortic valve stenosis tissue demonstrated just few factor positive cells. Conclusions: Complex detection of different factors may indicate selectively disordered morphopathogenetical event of heart disease: decrease of PGP 9.5 nerves suggests the decreased innervation of organ; increased apoptosis indicates the cell death without ingrowth of connective tissue; persistent presence of hANUP proves the unchanged homeostasis of cardiomyocytes probably supported by expression of chromogranins. Finally, decrease of VEGF detects the regions of affected blood vessels in heart affected by acquired heart disease.

Keywords: heart, apoptosis, protein-gene peptide 9.5, atrial natriuretic peptide, vascular endothelial growth factor, chromogranin A, endothelin

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802 An Invertebrate-Type Lysozyme from Chinese Mitten Crab Eriocheir Sinensis: Cloning and Characterization

Authors: Fengmei Li, Li Xu, Guoliang Xia

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Lysozyme is a catalytic enzyme that performs bacterial cell lysis by cleaving the β-1,4-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine of peptidoglycan in cell walls. In the present study, an invertebrate-type (i-type) lysozyme gene was cloned from Chinese mitten crab Eriocheir sinensis (designated as EsLysozyme) based on PCR-based rapid amplification of cDNA ends (RACE) technology. The full-length cDNA of EsLysozyme was of 831 bp. SMART and SIGNALP 3.0 program analysis revealed that EsLysozyme contained a signal peptide and a destabilase domain. The five amino acid residues (Tyr63, Trp64, Tyr91, His110, Pro114) and the conserved motif GSLSCG(P/Y)FQI and CL(E/L/R/H)C(I/M)C in i-type lysozymes were also found in EsLysozyme. The high similarity of EsLysozyme with L. vannamei lysozymes and phylogenetic analysis suggested that EsLysozyme should be a new member of i-type lysozyme family.

Keywords: i-type lysozyme, Eriocheir sinensis, cloning, characterization

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801 Characterization of Calcium-Signalling Mediated by Human GPR55 Expressed in HEK293 Cells

Authors: Yousuf M. Al Suleimani, Robin Hiley

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The endogenous phospholipid lysophosphatidylinositol (LPI) was recently identified as a novel ligand for the G protein-coupled receptor 55 (GPR55) and an inducer of intracellular Ca2+ [Ca2+]i release. This study attempts to characterize Ca2+ signals provoked by LPI in HEK293 cells engineered to stably express human GPR55 and to test cannabinoid ligand activity at GPR55. The study shows that treatment with LPI stimulates a sustained, oscillatory Ca2+ release. The response is characterized by an initial rapid rise, which is mediated by the Gαq-PLC-IP3 pathway, and this is followed by prolonged oscillations that require RhoA activation. Ca2+ oscillations are initiated by intracellular mechanisms and extracellular Ca2+ is only required to replenish Ca2+ lost from the cytoplasm. Analysis of cannabinoid ligand activity at GPR55 revealed no clear effect of the endocannabinoid anandamide, however, rimonabant and the CB1 receptor antagonist AM251 evoked GPR55-mediated [Ca2+]i. Thus, LPI is likely to be a key plasma membrane mediator of signaling events and changes in gene expression through GPR55 activation.

Keywords: lysophosphatidylinositol, calcium, GPR55, cannabinoid

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800 Analysis of Taxonomic Compositions, Metabolic Pathways and Antibiotic Resistance Genes in Fish Gut Microbiome by Shotgun Metagenomics

Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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