Search results for: spatial gene
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3916

Search results for: spatial gene

1516 Upconversion Nanomaterials for Applications in Life Sciences and Medicine

Authors: Yong Zhang

Abstract:

Light has proven to be useful in a wide range of biomedical applications such as fluorescence imaging, photoacoustic imaging, optogenetics, photodynamic therapy, photothermal therapy, and light controlled drug/gene delivery. Taking photodynamic therapy (PDT) as an example, PDT has been proven clinically effective in early lung cancer, bladder cancer, head, and neck cancer and is the primary treatment for skin cancer as well. However, clinical use of PDT is severely constrained by the low penetration depth of visible light through thick tissue, limiting its use to target regions only a few millimeters deep. One way to enhance the range is to use invisible near-infrared (NIR) light within the optical window (700–1100nm) for biological tissues, extending the depth up to 1cm with no observable damage to the intervening tissue. We have demonstrated use of NIR-to-visible upconversion fluorescent nanoparticles (UCNPs), emitting visible fluorescence when excited by a NIR light at 980nm, as a nanotransducer for PDT to convert deep tissue-penetrating NIR light to visible light suitable for activating photosensitizers. The unique optical properties of UCNPs enable the upconversion wavelength to be tuned and matched to the activation absorption wavelength of the photosensitizer. At depths beyond 1cm, however, tissue remains inaccessible to light even within the NIR window, and this critical depth limitation renders existing phototherapy ineffective against most deep-seated cancers. We have demonstrated some new treatment modalities for deep-seated cancers based on UCNP hydrogel implants and miniaturized, wirelessly powered optoelectronic devices for light delivery to deep tissues.

Keywords: upconversion, fluorescent, nanoparticle, bioimaging, photodynamic therapy

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1515 Housing and Urban Refugee: An Introspective Study on Bihari Camp of Mirpur, Dhaka

Authors: Fahmida Nusrat, Sumaia Nasrin, Pinak Sarker

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Biharis as an urban refugee are a significant urban dweller in Dhaka since their forced migration on the partition of 1947. There are many such refugee settlements in Bangladesh, particularly in Dhaka where they often live in dire conditions, facing discrimination from mainstream society. Their camps have become slums. Housing for urban refugee is still not a strategic concern for overall housing policy of Dhaka. The study has been conducted in a significant refugee settlement located in Mirpur-11, Dhaka, to observe their way of living in these camps to understand the socio-cultural aspects that are shaping their settlement morphology, hence to identify the key issues of their built environment to suggest an inclusive and sustainable housing solution for improving their life in urban environment. The methods included first-hand data collection on their household spaces and community spaces accompanied with the overall spatial organization of the settlement pattern which later on followed by a semi-structured interview with randomly selected samples from the camp dwellers to get users’ feedback on the research aspects. The outcome of the study will help initiating housing strategies as well as formulating design issues for this case specific inhabitants of urban Dhaka.

Keywords: Bihari camp, Dhaka, housing strategy, the way of living, urban refugee

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1514 Genetic Diversity Based Population Study of Freshwater Mud Eel (Monopterus cuchia) in Bangladesh

Authors: M. F. Miah, K. M. A. Zinnah, M. J. Raihan, H. Ali, M. N. Naser

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As genetic diversity is most important for existing, breeding and production of any fish; this study was undertaken for investigating genetic diversity of freshwater mud eel, Monopterus cuchia at population level where three ecological populations such as flooded area of Sylhet (P1), open water of Moulvibazar (P2) and open water of Sunamganj (P3) districts of Bangladesh were considered. Four arbitrary RAPD primers (OPB-12, C0-4, B-03 and OPB-08) were screened and RAPD banding patterns were analyzed among the populations considering 15 individuals of each population. In total 174, 138 and 149 bands were detected in the populations of P1, P2 and P3 respectively; however, each primer revealed less number of bands in each population. 100% polymorphic loci were recorded in P2 and P3 whereas only one monomorphic locus was observed in P1, recorded 97.5% polymorphism. Different genetic parameters such as inter-individual pairwise similarity, genetic distance, Nei genetic similarity, linkage distances, cluster analysis and allelic information, etc. were considered for measuring genetic diversity. The average inter-individual pairwise similarity was recorded 2.98, 1.47 and 1.35 in P1, P2 and P3 respectively. Considering genetic distance analysis, the highest distance 1 was recorded in P2 and P3 and the lowest genetic distance 0.444 was found in P2. The average Nei genetic similarity was observed 0.19, 0.16 and 0.13 in P1, P2 and P3, respectively; however, the average linkage distance was recorded 24.92, 17.14 and 15.28 in P1, P3 and P2 respectively. Based on linkage distance, genetic clusters were generated in three populations where 6 clades and 7 clusters were found in P1, 3 clades and 5 clusters were observed in P2 and 4 clades and 7 clusters were detected in P3. In addition, allelic information was observed where the frequency of p and q alleles were observed 0.093 and 0.907 in P1, 0.076 and 0.924 in P2, 0.074 and 0.926 in P3 respectively. The average gene diversity was observed highest in P2 (0.132) followed by P3 (0.131) and P1 (0.121) respectively.

Keywords: genetic diversity, Monopterus cuchia, population, RAPD, Bangladesh

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1513 The Impact of Artificial Intelligence on Sustainable Architecture and Urban Design

Authors: Alfons Aziz Asaad Hozain

Abstract:

The goal of sustainable architecture is to design buildings that have the least negative impact on the environment and provide better conditions for people. What forms of development enhance the area? This question was asked at the Center for the Study of Spatial Development and Building Forms in Cambridge in the late 1960s. This has resulted in many influential articles that have had a profound impact on the practice of urban planning. This article focuses on the sustainability outcomes caused by the climatic conditions of traditional Iranian architecture in hot and dry regions. Since people spend a lot of time at home, it is very important that these homes meet their physical and spiritual needs as well as the cultural and religious aspects of their lifestyle. In a country as large as Iran with different climates, traditional builders have put forward a number of logical solutions to ensure human comfort. With these solutions, the environmental problems of the have long been solved. Taking into account the experiences of traditional architecture in Iran's hot and dry climate, sustainable architecture can be achieved.

Keywords: sustainable development, human rights, the right to development, the human rights-based approach to development, environmental rights, economic development, social sustainability human rights protection, human rights violations, workers’ rights, justice, security

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1512 A Small-Scale Study of Fire Whirls and Investigation of the Effects of Near-Ground Height on the Behavior of Fire Whirls

Authors: M. Arabghahestani, A. Darwish Ahmad, N. K. Akafuah

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In this work, small-scale experiments of fire whirl were conducted to study the spinning fire phenomenon and to gain comprehensive understandings of fire tornadoes and the factors that affect their behavior. High speed imaging was used to track the flames at both temporal and spatial scales. This allowed us to better understand the role of the near-ground height in creating a boundary layer flow profile that, in turn contributes to formation of vortices around the fire, and consequent fire whirls. Based on the results obtained from these observations, we were able to spot the differences in the fuel burning rate of the fire itself as a function of a newly defined specific non-dimensional near-ground height. Based on our observations, there is a cutoff non-dimensional height, beyond which a normal fire can be turned into a fire whirl. Additionally, the results showed that the fire burning rate decreases by moving the fire to a height higher than the ground level. These effects were justified by the interactions between vortices formed by, the back pressure and the boundary layer velocity profile, and the vortices generated by the fire itself.

Keywords: boundary layer profile, fire whirls, near-ground height, vortex interactions

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1511 Seroprevalence and Associated Factors of Hepatitis B and Hepatitis C Viral Infections Among Prisoners in Tigray, Northern Ethiopia

Authors: Belaynesh Tsegay, Teklay Gebrecherkos, Atsebaha Gebrekidan Kahsay, Mahmud Abdulkader

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Background: Hepatitis B and C viruses are important health and socioeconomic problem across the globe, with a remarkable number of diseases and deaths in sub-Saharan African countries. The burden of hepatitis is unknown in the prison settings of Tigray. Therefore, we aimed to describe the seroprevalence and associated factors of hepatitis B and C viruses among prisoners in Tigray, Ethiopia. Methods: A cross-sectional study was carried out from February 2020 to May 2020 at the prison facilities of Tigray. Demographics and associated factors were collected from 315 prisoners prospectively. Five milliliters of blood were collected and tested using rapid tests kits of HBsAg (Zhejiang orient Gene Biotech Co., Ltd., China) and HCV antibodies (Volkan Kozmetik Sanayi Ve Ticaret Ltd. STI, Turkey). Positive samples were confirmed using ELISA (Beijing Wantai Biological Pharmacy Enterprise Co. Ltd). Data were analyzed using the SPSS version 20, and p<0.05 was considered statistically significant. Results: The overall seroprevalence of HBV and HCV were 25 (7.9%) and 1 (0.3%), respectively. The majority of hepatitis B viral infections were identified from the age groups of 18–25 years (10.7%) and unmarried prisoners (11.8%). Prisoners greater than 100 per cell (AOR=3.95, 95% CI=1.15–13.6, p=0.029) and with a history of alcohol consumption (AOR=3.01, 95% CI=1.17–7.74, p=0.022) were significantly associated with HBV infections. Conclusion: The seroprevalence of HBV among prisoners was nearly high or borderline, with a very low HCV prevalence. HBV was most prevalent among young adults, those housed with a large number of prisoners per cell, and those who had a history of alcohol consumption. This study recommends that there should be prison-focused intervention, including regular health education, with the emphasis on the mode of transmission and introducing HBV screening policy for prisoners, especially when they enter the prison.

Keywords: seroprevalence, HBV, HCV, prisoners, tigray

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1510 Survey of the Effect of the Probiotic Bacterium Lactobacillus plantarum and Streptococcus mutans on Casp3, AKT/PTEN, and MAPK Signaling Pathways at Co-Culture with KB Oral Cancer Cell Line and HUVEC Cells

Authors: Negar Zaheddoust, Negin Zaheddoust, Abbas Asoudeh-Fard

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Probiotic bacteria have been employed as a novel and less side-effect strategy for anticancer therapy. Since the oral cavity is a host for probiotic and pathogen bacteria to colonize, more investigation is needed to evaluate the effectiveness of this novel adjunctive treatment for oral cancer. We considered Lactobacillus plantarum as a probiotic and Streptococcus mutans as a pathogen bacterium in our study. The aim of this study is to examine the effect of Lactobacillus plantarum and Streptococcus mutans on Casp3, AKT / PTEN, and MAPK signaling pathway, which is involved in apoptosis or survival of oral cancer KB cells. On the other hand, to study the effects of these bacteria on normal cells, we used HUVEC cells. The KB and HUVEC cell lines were co-cultured with Lactobacillus plantarum and Streptococcus mutans isolated from traditional Iranian dairy and dental plaque, respectively. The growth-inhibitory effects of these two bacteria on KB and HUVEC cells were determined by (3-(4, 5-dimethylthiazolyl-2)-2,5diphenyltetrazolium bromide) MTT assay. MTT results demonstrated that the proliferation of KB cells was affected in a time, dose, and strain-dependent manner. In the following, the examination of induced apoptosis or necrosis in co-cultured KB cells with the best IC50 concentration of the Lactobacillus plantarum and Streptococcus mutans will be analyzed by FACS flow cytometry, and the changes in gene expression of Casp3, AKT / PTEN, MAPK genes will be evaluated using real-time polymerase chain reaction.

Keywords: cancer therapy, induced apoptosis, oral cancer, probiotics

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1509 Biodegradation of Direct Red 23 by Bacterial Consortium Isolated from Dye Contaminated Soil Using Sequential Air-lift Bioreactor

Authors: Lata Kumari Dhanesh Tiwary, Pradeep Kumar Mishra

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The effluent coming from various industries such as textile, carpet, food, pharmaceutical and many other industries is big challenge due to its recalcitrant and xenobiotiocs in nature. Recently, biodegradation of dye wastewater through biological means was widely used due to eco-friendly and cost effective with the higher percentage of removal of dye from wastewater. The present study deals with the biodegradation and decolourization of Direct Red 23 dye using indigenously isolated bacterial consortium. The bacterial consortium was isolated from soil sample from dye contaminated site near a cluster of Carpet industries of Bhadohi, Uttar Pradesh, India. The bacterial strain formed consortia were identified and characterized by morphological, biochemical and 16S rRNA gene sequence analysis. The bacterial strain mainly Staphylococcus saprophyticus strain BHUSS X3 (KJ439576), Microbacterium sp. BHUMSp X4 (KJ740222) and Staphylococcus saprophyticus strain BHUSS X5 (KJ439576) were used as consortia for further studies of dye decolorization. Experimental investigations were made in a Sequencing Air- lift bioreactor using the synthetic solution of Direct Red 23 dye by optimizing various parameters for efficient degradation of dye. The effect of several operating parameters such as flow rate, pH, temperature, initial dye concentration and inoculums size on removal of dye was investigated. The efficiency of isolated bacterial consortia from dye contaminated area in Sequencing Air- lift Bioreactor with different concentration of dye between 100-1200 mg/l at different hydraulic rate (HRTs) 26h and 10h. The maximum percentage of dye decolourization 98% was achieved when operated at HRT of 26h. The percentage of decolourization of dye was confirmed by using UV-Vis spectrophotometer and HPLC.

Keywords: carpet industry, bacterial consortia, sequencing air-lift bioreactor

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1508 A Comparative Study of Environment Risk Assessment Guidelines of Developing and Developed Countries Including Bangladesh

Authors: Syeda Fahria Hoque Mimmi, Aparna Islam

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Genetically engineered (GE) plants are the need of time for increased demand for food. A complete set of regulations need to be followed from the development of a GE plant to its release into the environment. The whole regulation system is categorized into separate stages for maintaining the proper biosafety. Environmental risk assessment (ERA) is one of such crucial stages in the whole process. ERA identifies potential risks and their impacts through science-based evaluation where it is done in a case-by-case study. All the countries which deal with GE plants follow specific guidelines to conduct a successful ERA. In this study, ERA guidelines of 4 developing and 4 developed countries, including Bangladesh, were compared. ERA guidelines of countries such as India, Canada, Australia, the European Union, Argentina, Brazil, and the US were considered as a model to conduct the comparison study with Bangladesh. Initially, ten parameters were detected to compare the required data and information among all the guidelines. Surprisingly, an adequate amount of data and information requirements (e.g., if the intended modification/new traits of interest has been achieved or not, the growth habit of GE plants, consequences of any potential gene flow upon the cultivation of GE plants to sexually compatible plant species, potential adverse effects on the human health, etc.) matched between all the countries. However, a few differences in data requirement (e.g., agronomic conventions of non-transformed plants, applicants should clearly describe experimental procedures followed, etc.) were also observed in the study. Moreover, it was found that only a few countries provide instructions on the quality of the data used for ERA. If these similarities are recognized in a more framed manner, then the approval pathway of GE plants can be shared.

Keywords: GE plants, ERA, harmonization, ERA guidelines, Information and data requirements

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1507 Osteogenesis in Thermo-Sensitive Hydrogel Using Mesenchymal Stem Cell Derived from Human Turbinate

Authors: A. Reum Son, Jin Seon Kwon, Seung Hun Park, Hai Bang Lee, Moon Suk Kim

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These days, stem cell therapy is focused on for promising source of treatment in clinical human disease. As a supporter of stem cells, in situ-forming hydrogels with growth factors and cells appear to be a promising approach in tissue engineering. To examine osteogenic differentiation of hTMSCs which is one of mesenchymal stem cells in vivo in an injectable hydrogel, we use a methoxy polyethylene glycol-polycaprolactone blockcopolymer (MPEG-PCL) solution with osteogenic factors. We synthesized MPEG-PCL hydrogel and measured viscosity to check sol-gel transition. In order to demonstrate osteogenic ability of hTMSCs, we conducted in vitro osteogenesis experiment. Then, to confirm the cell cytotoxicity, we performed WST-1 with hTMSCs and MPEG-PCL. As the result of in vitro experiment, we implanted cell and hydrogel mixture into animal model and checked degree of osteogenesis with histological analysis and amount of expression genes. Through these experimental data, MPEG-PCL hydrogel has sol-gel transition in temperature change and is biocompatible with stem cells. In histological analysis and gene expression, hTMSCs are very good source of osteogenesis with hydrogel and will use it to tissue engineering as important treatment method. hTMSCs could be a good adult stem cell source for usability of isolation and high proliferation. When hTMSCs are used as cell therapy method with in situ-formed hydrogel, they may provide various benefits like a noninvasive alternative for bone tissue engineering applications.

Keywords: injectable hydrogel, stem cell, osteogenic differentiation, tissue engineering

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1506 Design and Implementation of Image Super-Resolution for Myocardial Image

Authors: M. V. Chidananda Murthy, M. Z. Kurian, H. S. Guruprasad

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Super-resolution is the technique of intelligently upscaling images, avoiding artifacts or blurring, and deals with the recovery of a high-resolution image from one or more low-resolution images. Single-image super-resolution is a process of obtaining a high-resolution image from a set of low-resolution observations by signal processing. While super-resolution has been demonstrated to improve image quality in scaled down images in the image domain, its effects on the Fourier-based technique remains unknown. Super-resolution substantially improved the spatial resolution of the patient LGE images by sharpening the edges of the heart and the scar. This paper aims at investigating the effects of single image super-resolution on Fourier-based and image based methods of scale-up. In this paper, first, generate a training phase of the low-resolution image and high-resolution image to obtain dictionary. In the test phase, first, generate a patch and then difference of high-resolution image and interpolation image from the low-resolution image. Next simulation of the image is obtained by applying convolution method to the dictionary creation image and patch extracted the image. Finally, super-resolution image is obtained by combining the fused image and difference of high-resolution and interpolated image. Super-resolution reduces image errors and improves the image quality.

Keywords: image dictionary creation, image super-resolution, LGE images, patch extraction

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1505 Assessment of Lactic Acid Bacteria of Probiotic Potentials in Dairy Produce in Saudi Arabia

Authors: Rashad R. Al-Hindi

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The aim of this study was to isolate and identify lactic acid bacteria and evaluate their therapeutic and food preservation importance. Ninety-three suspected lactic acid bacteria (LAB) were isolated from thirteen different raw and fermented milk of indigenous sources in the Kingdom of Saudi Arabia. The identification of forty-six selected LAB strains and genetic relatedness were performed based on 16S rDNA gene sequence comparison. The LAB counts in certain samples were higher under microaerobic than anaerobic conditions. The identified LAB belonged to genera Enterococcus (16 strains), Lactobacillus (9 strains), Weissella (10 strains), Streptococcus (8 strains) and Lactococcus (3 strains). Phylogenetic tree generated from the full-length (~1.6 kb) sequences confirmed previous findings. Utilization of shorter 16S rDNA sequences (~1.0 kb) also discriminated among strains of which V2 region was the most effective. None of the strains exhibited resistance to clinically relevant antibiotics or undesirable hemolytic activity, while they differed in other probiotic characteristics, e.g., tolerance to acidic pH, resistance to bile salt, and antibacterial activity. In conclusion, the isolates Lactobacillus casei MSJ1, Lactobacillus casei Dwan5, Lactobacillus plantarum EyLan2 and Enterococcus faecium Gail-BawZir8 are likely the best probiotic LAB and we speculate that studying the synergistic effects of bacterial combinations might result in the occurrence of more effective probiotic potential. We argue that the raw and fermented milk of animals hosted in Saudi Arabia, especially stirred yogurt (Laban) made from camel milk, are rich in LAB with promising probiotics potential.

Keywords: fermented foods, lactic acid bacteria, probiotics, Saudi Arabia

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1504 Review of Urban Vitality in China: Exploring the Theoretical Framework, Characteristics, and Assessment Systems

Authors: Dong Wei, Wu Jinxiu

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As China's urban construction enters a new phase of 'stock optimization,' the key point of urban development has shifted to the development and reuse of existing public space. However, cities still face a series of challenges, such as the shortage of space quantity and insufficient space quality, which indirectly affect urban vitality. A review of the vitality of urban public space will significantly contribute to optimizing the quality of the urban built environment. It firstly analyses the research hotspots of urban vitality at home and abroad, based on a semi-systematic literature review. Then this paper summarizes the theoretical definitions of the vitality of urban public space and sorts out the influencing factors from the perspectives of society, environment, and users. Lastly, the paper concludes with the mainstream quantitative and evaluation methods, such as linear evaluation and integrated evaluation. This paper renders a multi-theoretical perspective to understand the characteristics and evaluation system of the vitality of public space, which helps to acknowledge the dynamic relationship between users, urban environment, and vitality. It also looks forward to providing optimal design strategies for constructing a vigorous public space in future cities.

Keywords: public space, quantification of vitality, spatial vitality, urban vitality

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1503 From Vertigo to Verticality: An Example of Phenomenological Design in Architecture

Authors: E. Osorio Schmied

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Architects commonly attempt a depiction of organic forms when their works are inspired by nature, regardless of the building site. Nevertheless it is also possible to try matching structures with natural scenery, by applying a phenomenological approach in terms of spatial operations, regarding perceptions from nature through architectural aspects such as protection, views, and orientation. This method acknowledges a relationship between place and space, where intentions towards tangible facts then become design statements. Although spaces resulting from such a process may present an effective response to the environment, they can also offer further outcomes beyond the realm of form. The hypothesis is that, in addition to recognising a bond between architecture and nature, it is also plausible to associate such perceptions with the inner ambient of buildings, by analysing features such as daylight. The case study of a single-family house in a rainforest near Valdivia, Chilean Patagonia is presented, with the intention of addressing the above notions through a discussion of the actual effects of inhabiting a place by way of a series of insights, including a revision of diagrams and photographs that assist in understanding the implications of this design practice. In addition, figures based on post-occupancy behaviour and daylighting performance relate both architectural and environmental issues to a decision-making process motivated by the observation of nature.

Keywords: architecture, design statements, nature, perception

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1502 ArcGIS as a Tool for Infrastructure Documentation and Asset Management: Establishing a GIS for Computer Network Documentation

Authors: John Segars

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Built out of a real-world need to have better, more detailed, asset and infrastructure documentation, this project will lay out the case for using the database functionality of ArcGIS as a tool to track and maintain infrastructure location, status, maintenance and serviceability. Workflows and processes will be presented and detailed which may be applied to an organizations’ infrastructure needs that might allow them to make use of the robust tools which surround the ArcGIS platform. The end result is a value-added information system framework with a geographic component e.g., the spatial location of various I.T. assets, a detailed set of records which not only documents location but also captures the maintenance history for assets along with photographs and documentation of these various assets as attachments to the numerous feature class items. In addition to the asset location and documentation benefits, the staff will be able to log into the devices and pull SNMP (Simple Network Management Protocol) based query information from within the user interface. The entire collection of information may be displayed in ArcGIS, via a JavaScript based web application or via queries to the back-end database. The project is applicable to all organizations which maintain an IT infrastructure but specifically targets post-secondary educational institutions where access to ESRI resources is generally already available in house.

Keywords: ESRI, GIS, infrastructure, network documentation, PostgreSQL

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1501 The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by miR-375 and Anti-miR-9

Authors: Arefeh Jafarian, Mohammad Taghikani, Saied Abroun, Amir Allahverdi, Masoud Soleimani

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Introduction: The miRNAs have key roles in control of pancreatic islet development and insulin secretion. In this regards, current study investigated the pancreatic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) by up-regulation of miR-375 and down-regulation of miR-9 by lentiviruses containing miR-375 and anti-miR-9. Findings: After 21 days of induction, islet-like clusters containing insulin producing cells (IPCs) were confirmed by dithizone (DTZ) staining. The IPCs and β cell specific related genes and proteins were detected using qRT-PCR and immunofluorescence on days 7, 14 and 21 of differentiation. Glucose challenge test was performed at different concentrations of glucose as well as extracellular and intracellular insulin and C-peptide were assayed using ELISA kit. In derived IPCs by miR-375 alone are capable to express insulin and other endocrine specific transcription factors, the cells lack the machinery to respond to glucose. The differentiated hMSCs by miR-375 and anti-miR-9 lentiviruses could secrete insulin and c-peptide in a glucose-regulated manner. Conclusion: It was found that over-expression of miR-375 led to a reduction in levels of Mtpn protein in derived IPCs, while treatment with anti-miR-9 following miR-375 over-expression had synergistic effects on MSCs differentiation and insulin secretion in a glucose-regulated manner. The researchers reported that silencing of miR-9 increased OC-2 protein in IPCs that may contribute to the observed glucose-regulated insulin secretion. These findings highlight miRNAs functions in stem cells differentiation and suggest that they could be used as therapeutic tools for gene-based therapy in diabetes mellitus.

Keywords: diabetes, differentiation, MSCs, insulin producing cells, miR-375, miR-9

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1500 Positive effect of Cu2+ and Ca2+ on the Thermostability of Bambara Groundnut Peroxidase A6, and its Catalytic Efficiency Toward the Oxidation of 3,3,5,5 -Tetramethyl Benzidine

Authors: Yves Mann Elate Lea Mbassi, Marie Solange Evehe Bebandoue, Wilfred Fon Mbacham

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Improving the catalytic performance of enzymes has been a long-standing theme of analytical biochemistry research. Induction of peroxidase activity by metals is a common reaction in higher plants. We thought that this increase in peroxidase activity may be due, on the one hand, to the stimulation of the gene expression of these enzymes but also to a modification of their chemical reactivity following the binding of some metal ions on their active site. We tested the effect of some metal salts (MgCl₂, MnCl₂, ZnCl₂, CaCl₂ and CuSO₄) on the activity and thermostability of peroxidase A6, a thermostable peroxidase that we discovered and purified in a previous study. The chromogenic substrate used was 3,3′,5,5′-tetramethylbenzidine. Of all the metals tested for their effect on A6, only magnesium and copper had a significant effect on the activity of the enzyme at room temperature. The Mann-Whitney test shows a slight inhibitory effect of activity by the magnesium salt (P = 0.043), while the activity of the enzyme is 5 times higher in the presence of the copper salt (P = 0.002). Moreover, the thermostability of peroxidase A6 is increased when calcium and copper salts are present. The activity in the presence of CaCl₂ is 8 times higher than the residual activity of the enzyme alone after incubation at 80°C for 10 min and 35 times higher in the presence of CuSO4 under the same conditions. In addition, manganese and zinc salts slightly reduce the thermostability of the enzyme. The activity and structural stability of peroxidase A6 can clearly be activated by Cu₂+, which therefore enhance the oxidation of 3,3′,5,5′-tetramethylbenzidine, which was used in this study as a chromogenic substrate. Ca₂+ likely has a more stabilizing function for the catalytic site.

Keywords: peroxidase activity, copper ions, calcium ions, thermostability

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1499 Formulation Development and Characterization of Oligonucleotide Containing Chitosan Nanoparticles

Authors: Gyati Shilakari Asthana, Abhay Asthana

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Purpose: The therapeutic potential of oligonucleotide (ODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study is focused to design low molecular weight chitosan nanoconstructs to meet the requirements of safe and effectual delivery of ODNs. LMW-chitosan is a biodegradable, water soluble, biocompatible polymer and is useful as a non-viral vector for gene delivery due to its better stability in water. Methods: LMW chitosan ODN nanoparticles (CHODN NPs) were formulated by self assembled method using various N/P ratios (moles ratio of amine groups of CH to phosphate moieties of ODNs; 0.5:1, 1:1, 3:1, 5:1 and 7:1) of CH to ODN. The developed CHODN NPs were evaluated with respect to gel retardation assay, particle size, zeta potential and cytotoxicity and transfection efficiency. Results: Complete complexation of CH/ODN was achieved at the charge ratio of 0.5:1 or above and CHODN NPs displayed resistance against DNase I. On increasing the N/P ratio of CH/ODN, particle size of the NPs decreased whereas zeta potential (ZV) value increased. No significant toxicity was observed at all CH concentrations. The transfection efficiency was increased on increasing N/P ratio from 1:1 to 3:1, whereas it was decreased with further increment in N/P ratio upto 7:1. Maximum transfection of CHODN NPs with both the cell lines (Raw 267.4 cells and Hela cells) was achieved at N/P ratio of 3:1. The results suggest that transfection efficiency of CHODN NPs is dependent on N/P ratio. Conclusion: Thus the present study states that LMW chitosan nanoparticulate carriers would be acceptable choice to improve transfection efficiency in vitro as well as in vivo delivery of oligonucleotide.

Keywords: LMW-chitosan, chitosan nanoparticles, biocompatibility, cytotoxicity study, transfection efficiency, oligonucleotide

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1498 Web-GIS Technology: A Tool for Farm-to-Market Road Project Profiling and Proposal Prioritization of the Philippines’ Department of Agriculture

Authors: Elbert S. Moyon, Edsel Matt O. Morales, Jaymer M. Jayoma, Kent C. Espejon, Jayson C. Dollete, Mark Phil B. Pacot

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This research paper focuses on the potential of using Web-GIS technology in prioritizing farm-to-market road projects by the Philippines’ Department of Agriculture (DA). The study aimed to explore the benefits of Web-GIS in addressing the limitations faced by the DA in terms of Farm to Market Road profiling and project prioritization, which include a lack of access to updated data, limited spatial analysis capabilities, and difficulties in sharing information between stakeholders. The research methodology involves a comprehensive literature review and a case study of a Web-GIS application developed for the DA, which was used to profile and prioritize farm-to-market road projects in the Philippines. The results showed that the Web-GIS technology provides the DA with an effective tool for analyzing and visualizing data, which can help in profiling and prioritizing road projects based on various criteria such as economic, social, and environmental impacts. The study also showed that Web-GIS technology could help in reducing the time and effort required for road project prioritization and improve communication between stakeholders.

Keywords: GIS, web application, farm-to-market road, FMR prioritization, Django, GeoServer

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1497 Genome-Wide Expression Profiling of Cicer arietinum Heavy Metal Toxicity

Authors: B. S. Yadav, A. Mani, S. Srivastava

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Chickpea (Cicer arietinum L.) is an annual, self-pollinating, diploid (2n = 2x = 16) pulse crop that ranks second in world legume production after common bean (Phaseolus vulgaris). ICC 4958 flowers approximately 39 days after sowing under peninsular Indian conditions and the crop matures in less than 90 days in rained environments. The estimated collective yield losses due to abiotic stresses (6.4 million t) have been significantly higher than for biotic stresses (4.8 million t). Most legumes are known to be salt sensitive, and therefore, it is becoming increasingly important to produce cultivars tolerant to high-salinity in addition to other abiotic and biotic stresses for sustainable chickpea production. Our aim was to identify the genes that are involved in the defence mechanism against heavy metal toxicity in chickpea and establish the biological network of heavy metal toxicity in chickpea. ICC4958 variety of chick pea was taken and grown in normal condition and 150µM concentration of different heavy metal salt like CdCl₂, K₂Cr2O₇, NaAsO₂. At 15th day leave samples were collected and stored in RNA Later solution microarray was performed for checking out differential gene expression pattern. Our studies revealed that 111 common genes that involved in defense mechanism were up regulated and 41 genes were commonly down regulated during treatment of 150µM concentration of CdCl₂, K₂Cr₂O₇, and NaAsO₂. Biological network study shows that the genes which are differentially expressed are highly connected and having high betweenness and centrality.

Keywords: abiotic stress, biological network, chickpea, microarray

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1496 Illumina MiSeq Sequencing for Bacteria Identification on Audio-Visual Materials

Authors: Tereza Branyšová, Martina Kračmarová, Kateřina Demnerová, Michal Ďurovič, Hana Stiborová

Abstract:

Microbial deterioration threatens all objects of cultural heritage, including audio-visual materials. Fungi are commonly known to be the main factor in audio-visual material deterioration. However, although being neglected, bacteria also play a significant role. In addition to microbial contamination of materials, it is also essential to analyse air as a possible contamination source. This work aims to identify bacterial species in the archives of the Czech Republic that occur on audio-visual materials as well as in the air in the archives. For sampling purposes, the smears from the materials were taken by sterile polyurethane sponges, and the air was collected using a MAS-100 aeroscope. Metagenomic DNA from all collected samples was immediately isolated and stored at -20 °C. DNA library for the 16S rRNA gene was prepared using two-step PCR and specific primers and the concentration step was included due to meagre yields of the DNA. After that, the samples were sent to the University of Fairbanks, Alaska, for Illumina MiSeq sequencing. Subsequently, the analysis of the sequences was conducted in R software. The obtained sequences were assigned to the corresponding bacterial species using the DADA2 package. The impact of air contamination and the impact of different photosensitive layers that audio-visual materials were made of, such as gelatine, albumen, and collodion, were evaluated. As a next step, we will take a deeper focus on air contamination. We will select an appropriate culture-dependent approach along with a culture-independent approach to observe a metabolically active species in the air. Acknowledgment: This project is supported by grant no. DG18P02OVV062 of the Ministry of Culture of the Czech Republic.

Keywords: cultural heritage, Illumina MiSeq, metagenomics, microbial identification

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1495 Sperm Flagellum Center-Line Tracing in 4D Stacks Using an Iterative Minimal Path Method

Authors: Paul Hernandez-Herrera, Fernando Montoya, Juan Manuel Rendon, Alberto Darszon, Gabriel Corkidi

Abstract:

Intracellular calcium ([Ca2+]i) regulates sperm motility. The analysis of [Ca2+]i has been traditionally achieved in two dimensions while the real movement of the cell takes place in three spatial dimensions. Due to optical limitations (high speed cell movement and low light emission) important data concerning the three dimensional movement of these flagellated cells had been neglected. Visualizing [Ca2+]i in 3D is not a simple matter since it requires complex fluorescence microscopy techniques where the resulting images have very low intensity and consequently low SNR (Signal to Noise Ratio). In 4D sequences, this problem is magnified since the flagellum oscillates (for human sperm) at least at an average frequency of 15 Hz. In this paper, a novel approach to extract the flagellum’s center-line in 4D stacks is presented. For this purpose, an iterative algorithm based on the fast-marching method is proposed to extract the flagellum’s center-line. Quantitative and qualitative results are presented in a 4D stack to demonstrate the ability of the proposed algorithm to trace the flagellum’s center-line. The method reached a precision and recall of 0.96 as compared with a semi-manual method.

Keywords: flagellum, minimal path, segmentation, sperm

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1494 A Cognitive Semantic Analysis of the Metaphorical Extensions of Come out and Take Over

Authors: Raquel Rossini, Edelvais Caldeira

Abstract:

The aim of this work is to investigate the motivation for the metaphorical uses of two verb combinations: come out and take over. Drawing from cognitive semantics theories, image schemas and metaphors, it was attempted to demonstrate that: a) the metaphorical senses of both 'come out' and 'take over' extend from both the verbs and the particles central (spatial) senses in such verb combinations; and b) the particles 'out' and 'over' also contribute to the whole meaning of the verb combinations. In order to do so, a random selection of 579 concordance lines for come out and 1,412 for take over was obtained from the Corpus of Contemporary American English – COCA. One of the main procedures adopted in the present work was the establishment of verb and particle central senses. As per the research questions addressed in this study, they are as follows: a) how does the identification of trajector and landmark help reveal patterns that contribute for the identification of the semantic network of these two verb combinations?; b) what is the relationship between the schematic structures attributed to the particles and the metaphorical uses found in empirical data?; and c) what conceptual metaphors underlie the mappings from the source to the target domains? The results demonstrated that not only the lexical verbs come and take, but also the particles out and over play an important whole in the different meanings of come out and take over. Besides, image schemas and conceptual metaphors were found to be helpful in order to establish the motivations for the metaphorical uses of these linguistic structures.

Keywords: cognitive linguistics, English syntax, multi-word verbs, prepositions

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1493 Evaluation of Adaptive Fitness of Indian Teak (Tectona grandis L. F.) Metapopulation through Inter Simple Sequence Repeat Markers

Authors: Vivek Vaishnav, Shamim Akhtar Ansari

Abstract:

Teak (Tectona grandis L.f.) belonging to plant family Lamiaceae and the most commercialized timber species is endemic to South-Asia. The adaptive fitness of the species metapopulation was evaluated through its genetic differentiation and assessing the influence of geo-climatic conditions. 290 genotypes were sampled from 29 locations of its natural distribution and the genetic data was incorporated with geo-climatic parameters. Through Bayesian approach based analysis of 43 highly polymorphic ISSR markers, six homogeneous clusters (0.8% genetic variability) were identified. The six clusters were found with the various regimes of the temperature range, i.e., I - 9.10±1.35⁰C, II -6.35±0.21⁰C, III -12.21±0.43⁰C, IV - 10.8±1.06⁰C, V - 11.67±3.04⁰C, and VI - 12.35±0.21⁰C. The population had a very high percentage of LD (21.48%) among the amplified loci possibly due to experiencing restricted gene flow as well as co-adaptation and association of distant/diverse loci/alleles as a result of the stabilized climatic conditions and countless cycles of historical recombination events on a large geological timescale. The same possibly accounts for the narrow distribution of teak as a climax species in the tropical deciduous forests of the country. The regions of strong LD in teak genome significantly associated with climatic parameters also reflect that the species is tolerant to the wide regimes of the temperature range and may possibly withstand global warming and climate change in the coming millennium.

Keywords: Bayesian analysis, inter simple sequence repeat, linkage disequilibrium, marker-geoclimatic association

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1492 Exploring Re-Configuration of Ordinary Spaces into Recreation and Leisure Space in Compact Unplanned Settlements: Experience from Manzese Informal Settlement-Dar Es Salaam Tanzania

Authors: Edson Ephraim Sanga

Abstract:

This paper stems to explore possible places used for recreation in unplanned settlements in order to avail knowledge on how to create and shape urban spaces essential for recreation and leisure. The context of unplanned settlements is spatially characterized compactness and congestions of buildings developed by residents without professional inputs. These characteristics surpass greenery landscapes such as parks and squares essential for health, happiness and wellbeing. The lack of recreational greenery landscape arises a question on how possible can recreation take places in the settlements? This study used qualitative methods mainly observation and in-depth interview to explore the recreational situation in Manzese informal settlements as an instrumental case and found that ordinary spaces are re-configured into recreational spaces and used as ‘parks’ and ‘squares’ in the settlements. The spaces are diverse and complex as they possess different spatial characteristics based on their physical attributes and the way they are used and interpreted by respective users. This paper argues that the re-configuration processes of ordinary spaces should not be taken for granted because they portray the appropriation of spaces from quotidian dimensions in a particular context.

Keywords: ordinary spaces, recreation, unplanned settlement, urban spaces

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1491 Role of Inflammatory Markers in Arthritic Rats Treated with Ethanolic Bark Extract of Albizia procera

Authors: M. Sangeetha, D. Chamundeeswari, C. Saravanababu, C. Rose, V. Gopal

Abstract:

Rheumatoid arthritis (RA) is a chronic, progressive, systemic inflammatory disorder affecting the synovial joints and typically producing symmetrical arthritis that leads to joint destruction, which is responsible for the deformity and disability. Despite improvements in the treatment of RA over the past decade, there still is a need for new therapeutic agents that are efficacious, less expensive, and free of severe adverse reactions. The present study aimed to investigate role of inflammatory markers in arthritic rats treated with ethanolic bark extract of Albizia procera. The protective effect of ethanolic bark extract of Albizia procera against complete Freund’s adjuvant (CFA) induced arthritis in rats. Arthritis was induced by an intradermal injection of 0.1 ml FCA in the foot pad of left hind limb of rats. ETBE (100 and 200 mg/kg b.wt./p.o) and the reference drug diclofenac (25 mg/kg b.wt./p.o) were administered to arthritic rats. Paw volume was measured for all the animals before inducing arthritis and thereafter once in seven days by using plethysmometer for 42 days. Gene expression of inflammatory markers such as IL-1β and IL-10 were investigated in paw tissues. Up regulation of IL-1β and Down regulation IL-10 were observed in CFA injected rats when compared to normal rats. ETBE attenuated these alterations dose dependently when compared to the vehicle treated rats. These results provide insights into the mechanism of anti-arthritic activity, and unravel potential therapeutic use of Albizia procera in arthritis.

Keywords: CFA-Complete Freund’s adjuvant, ETBE – ethanolic bark extract, IL- interleukins, RA-rheumatoid arthritis

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1490 Understanding the Impact of Spatial Light Distribution on Object Identification in Low Vision: A Pilot Psychophysical Study

Authors: Alexandre Faure, Yoko Mizokami, éRic Dinet

Abstract:

These recent years, the potential of light in assisting visually impaired people in their indoor mobility has been demonstrated by different studies. Implementing smart lighting systems for selective visual enhancement, especially designed for low-vision people, is an approach that breaks with the existing visual aids. The appearance of the surface of an object is significantly influenced by the lighting conditions and the constituent materials of the objects. Appearance of objects may appear to be different from expectation. Therefore, lighting conditions lead to an important part of accurate material recognition. The main objective of this work was to investigate the effect of the spatial distribution of light on object identification in the context of low vision. The purpose was to determine whether and what specific lighting approaches should be preferred for visually impaired people. A psychophysical experiment was designed to study the ability of individuals to identify the smallest cube of a pair under different lighting diffusion conditions. Participants were divided into two distinct groups: a reference group of observers with normal or corrected-to-normal visual acuity and a test group, in which observers were required to wear visual impairment simulation glasses. All participants were presented with pairs of cubes in a "miniature room" and were instructed to estimate the relative size of the two cubes. The miniature room replicates real-life settings, adorned with decorations and separated from external light sources by black curtains. The correlated color temperature was set to 6000 K, and the horizontal illuminance at the object level at approximately 240 lux. The objects presented for comparison consisted of 11 white cubes and 11 black cubes of different sizes manufactured with a 3D printer. Participants were seated 60 cm away from the objects. Two different levels of light diffuseness were implemented. After receiving instructions, participants were asked to judge whether the two presented cubes were the same size or if one was smaller. They provided one of five possible answers: "Left one is smaller," "Left one is smaller but unsure," "Same size," "Right one is smaller," or "Right one is smaller but unsure.". The method of constant stimuli was used, presenting stimulus pairs in a random order to prevent learning and expectation biases. Each pair consisted of a comparison stimulus and a reference cube. A psychometric function was constructed to link stimulus value with the frequency of correct detection, aiming to determine the 50% correct detection threshold. Collected data were analyzed through graphs illustrating participants' responses to stimuli, with accuracy increasing as the size difference between cubes grew. Statistical analyses, including 2-way ANOVA tests, showed that light diffuseness had no significant impact on the difference threshold, whereas object color had a significant influence in low vision scenarios. The first results and trends derived from this pilot experiment clearly and strongly suggest that future investigations could explore extreme diffusion conditions to comprehensively assess the impact of diffusion on object identification. For example, the first findings related to light diffuseness may be attributed to the range of manipulation, emphasizing the need to explore how other lighting-related factors interact with diffuseness.

Keywords: Lighting, Low Vision, Visual Aid, Object Identification, Psychophysical Experiment

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1489 Cytotoxicity and Androgenic Potential of Antifungal Drug Substances on MDA-KB2 Cells

Authors: Benchouala Amira, Bojic Clement, Poupin Pascal, Cossu Leguille-carole

Abstract:

The objective of this study is to evaluate in vitro the cytotoxic and androgenic potential of several antifungal molecules (amphotericin B, econazole, ketoconazole and miconazole) on MDA-Kb2 cell lines. This biological model is an effective tool for the detection of endocrine disruptors because it responds well to the main agonist of the androgen receptor (testosterone) and also to an antagonist: flutamide. The cytotoxicity of each chemical compound tested was measured using an MTT assay (tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) which measures the activity of the reductase function of mitochondrial succinate dehydrogenase enzymes of cultured cells. This complementary cytotoxicity test is essential to ensure that the effects of reduction in luminescence intensity observed during androgenic tests are only attributable to the anti-androgenic action of the compounds tested and not to their possible cytotoxic properties. Tests of the androgenic activity of antifungals show that these compounds do not have the capacity to induce transcription of the luciferase gene. These compounds do not exert an androgenic effect on MDA-Kb2 cells in culture for the environmental concentrations tested. The addition of flutamide for the same tested concentrations of antifungal molecules reduces the luminescence induced by amphotericin B, econazole and miconazole, which is explained by a strong interaction of these molecules with flutamide which may have a greater toxic effect than when tested alone. The cytotoxicity test shows that econazole and ketoconazole can cause cell death at certain concentrations tested. This cell mortality is perhaps induced by a direct or indirect action on deoxyribonucleic acid (DNA), ribonucleic acid (RNA) or proteins necessary for cell division.

Keywords: cytotoxicity, androgenic potential, antifungals, MDA-Kb2

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1488 African Swine Fewer Situation and Diagnostic Methods in Lithuania

Authors: Simona Pileviciene

Abstract:

On 24th January 2014, Lithuania notified two primary cases of African swine fever (ASF) in wild boars. The animals were tested positive for ASF virus (ASFV) genome by real-time PCR at the National Reference Laboratory for ASF in Lithuania (NRL), results were confirmed by the European Union Reference Laboratory for African swine fever (CISA-INIA). Intensive wild and domestic animal monitoring program was started. During the period of 2014-2017 ASF was confirmed in two large commercial pig holding with the highest biosecurity. Pigs were killed and destroyed. Since 2014 ASF outbreak territory from east and south has expanded to the middle of Lithuania. Diagnosis by PCR is one of the highly recommended diagnostic methods by World Organization for Animal Health (OIE) for diagnosis of ASF. The aim of the present study was to compare singleplex real-time PCR assays to a duplex assay allowing the identification of ASF and internal control in a single PCR tube and to compare primers, that target the p72 gene (ASF 250 bp and ASF 75 bp) effectivity. Multiplex real-time PCR assays prove to be less time consuming and cost-efficient and therefore have a high potential to be applied in the routine analysis. It is important to have effective and fast method that allows virus detection at the beginning of disease for wild boar population and in outbreaks for domestic pigs. For experiments, we used reference samples (INIA, Spain), and positive samples from infected animals in Lithuania. Results show 100% sensitivity and specificity.

Keywords: African swine fewer, real-time PCR, wild boar, domestic pig

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1487 Antibacterial Activity of Bacillus thuringiensis Activated Delta-endotoxins

Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

Abstract:

For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

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