Search results for: gene diversity

Authors: Md. Samsuddin Ansari, Ashish Arora

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Every year 10 million people fall ill with one of the oldest diseases known as tuberculosis, caused by Mycobacterium tuberculosis. The success of M. tuberculosis as a pathogen is because of its ability to persist in host tissues. Multidrug resistance (MDR) mycobacteria cases increase every day, which is associated with efflux pumps controlled at the level of transcription. The transcription regulators of MDR transporters in bacteria belong to one of the following four regulatory protein families: AraC, MarR, MerR, and TetR. Phenolic acid decarboxylase repressor (PadR), like a family of transcription regulators, is closely related to the MarR family. Phenolic acid decarboxylase repressor (PadR) was first identified as a transcription factor involved in the regulation of phenolic acid stress response in various microorganisms (including Mycobacterium tuberculosis H37Rv). Recently research has shown that the PadR family transcription factors are global, multifunction transcription regulators. Rv0047c is a PadR subfamily-1 protein. We are exploring the biophysical and structural characterization of Rv0047c. The Rv0047 gene was amplified by PCR using the primers containing EcoRI and HindIII restriction enzyme sites cloned in pET-NH6 vector and overexpressed in DH5α and BL21 (λDE3) cells of E. coli following purification with Ni2+-NTA column and size exclusion chromatography. We did DSC to know the thermal stability; the Tm (transition temperature) of protein is 55.29ºC, and ΔH (enthalpy change) of 6.92 kcal/mol. Circular dichroism to know the secondary structure and conformation and fluorescence spectroscopy for tertiary structure study of protein. To understand the effect of pH on the structure, function, and stability of Rv0047c we employed spectroscopy techniques such as circular dichroism, fluorescence, and absorbance measurements in a wide range of pH (from pH-2.0 to pH-12). At low and high pH, it shows drastic changes in the secondary and tertiary structure of the protein. EMSA studies showed the specific binding of Rv0047c with its own 30-bp promoter region. To determine the effect of complex formation on the secondary structure of Rv0047c, we examined the CD spectra of the complex of Rv0047c with promoter DNA of rv0047. The functional role of Rv0047c was characterized by over-expressing the Rv0047c gene under the control of hsp60 promoter in Mycobacterium tuberculosis H37Rv. We have predicted the three-dimensional structure of Rv0047c using the Swiss Model and Modeller, with validity checked by the Ramachandra plot. We did molecular docking of Rv0047c with dnaA, through PatchDock following refinement through FireDock. Through this, it is possible to easily identify the binding hot-stop of the receptor molecule with that of the ligand, the nature of the interface itself, and the conformational change undergone by the protein pattern. We are using X-crystallography to unravel the structure of Rv0047c. Overall the studies show that Rv0047c may have transcription regulation along with providing an insight into the activity of Rv0047c in the pH range of subcellular environment and helps to understand the protein-protein interaction, a novel target to kill dormant bacteria and potential strategy for tuberculosis control.

Keywords: mycobacterium tuberculosis, phenolic acid decarboxylase repressor, Rv0047c, Circular dichroism, fluorescence spectroscopy, docking, protein-protein interaction

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Authors: Feng Yue, Fei Dai

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With the rapid development of urbanization process in China, its environmental protection pressure is severely tested. So, analyzing and optimizing the landscape pattern is an important measure to ease the pressure on the ecological environment. This paper takes Wuhan Urban Development Zone as the research object, and studies its landscape pattern evolution and quantitative optimization strategy. First, remote sensing image data from 1990 to 2015 were interpreted by using Erdas software. Next, the landscape pattern index of landscape level, class level, and patch level was studied based on Fragstats. Then five indicators of ecological environment based on National Environmental Protection Standard of China were selected to evaluate the impact of landscape pattern evolution on the ecological environment. Besides, the cost distance analysis of ArcGIS was applied to simulate wildlife migration thus indirectly measuring the improvement of ecological environment quality. The result shows that the area of land for construction increased 491%. But the bare land, sparse grassland, forest, farmland, water decreased 82%, 47%, 36%, 25% and 11% respectively. They were mainly converted into construction land. On landscape level, the change of landscape index all showed a downward trend. Number of patches (NP), Landscape shape index (LSI), Connection index (CONNECT), Shannon's diversity index (SHDI), Aggregation index (AI) separately decreased by 2778, 25.7, 0.042, 0.6, 29.2%, all of which indicated that the NP, the degree of aggregation and the landscape connectivity declined. On class level, the construction land and forest, CPLAND, TCA, AI and LSI ascended, but the Distribution Statistics Core Area (CORE_AM) decreased. As for farmland, water, sparse grassland, bare land, CPLAND, TCA and DIVISION, the Patch Density (PD) and LSI descended, yet the patch fragmentation and CORE_AM increased. On patch level, patch area, Patch perimeter, Shape index of water, farmland and bare land continued to decline. The three indexes of forest patches increased overall, sparse grassland decreased as a whole, and construction land increased. It is obvious that the urbanization greatly influenced the landscape evolution. Ecological diversity and landscape heterogeneity of ecological patches clearly dropped. The Habitat Quality Index continuously declined by 14%. Therefore, optimization strategy based on greenway network planning is raised for discussion. This paper contributes to the study of landscape pattern evolution in planning and design and to the research on spatial layout of urbanization.

Keywords: landscape pattern, optimization strategy, ArcGIS, Erdas, landscape metrics, landscape architecture

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Authors: L. Horňák

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The aim of the paper is to analyze a longstanding problem in Slovakia – the effective education of Roma students coming from socially disadvantaged backgrounds. Although it is a relatively small country, there are over 630 communities in the Slovak Republic. The efficiency of the projects was verified by interviews with participants; questionnaires; and direct observations. Evaluation reports which summarized and evaluated the outcomes of the projects only confirmed their success. Slovakia realizes that appropriate social inclusion of marginalized citizens coming from the Roma ethnic group can only be achieved through education based on equality of all students and acceptance of diversity.

Keywords: inclusive education, marginalized communities, Roma pupil, equity in education, socially disadvantaged backgrounds, social inclusion

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Authors: Sandy Elsayed, Aya Mohamed, Noha Nassar

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Introduction: Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by social deficits and repetitive behavior. Multiple studies suggest that oxidative stress and neuroinflammation are key factors in the etiology of ASD and often associated with worsening of ASD-related behaviors. Nuclear factor erythroid 2-related factor 2 (NRF-2) is a transcription factor that promotes expression of antioxidant response element genes in oxidative stress. In ASD subjects, decreased expression of NRF-2 in frontal cortex shifted the redox homeostasis towards oxidative stress, and resulted in inflammation evidenced by elevation of nuclear factor kappa B (NF-κB) transcriptional activity. Dimethyl fumarate (DMF) is a NRF-2 activator that is used in the treatment of psoriasis and multiple sclerosis. It participates in the transcriptional control of inflammatory factors via inhibition of NF-κB and its downstream targets. This study aimed to investigate the role of DMF in alleviating the cognitive impairments and behavior deficits associated with ASD through mitigation of oxidative stress and inflammation in prenatal valproic acid (VPA) rat model of autism. Methods: Pregnant female Wistar rats received a single intraperitoneal injection of VPA (600 mg/kg) to induce autistic-like-behavioral and neurobiological alterations in their offspring. Chronic oral gavage of DMF (150mg/kg/day) started from postnatal day (PND) 24 till PND62 (39 days). Prenatal VPA exposure elicited autistic behaviors including decreased social interaction and stereotyped behavior. Social interaction was evaluated using three-chamber sociability test and calculation of sociability index (SI), while stereotyped repetitive behavior and anxiety associated with ASD were assessed using marble burying test (MBT). Biochemical analyses were done on prefrontal cortex homogenates including NRF-2, and NF-κB expression. Moreover, inducible nitric oxide synthase (iNOS) gene expression and tumor necrosis factor (TNF-) protein expression were evaluated as markers of inflammation. Results: Prenatal VPA elicited decreased social interaction shown by decreased SI compared to control group (p < 0.001) and DMF enhanced SI (p < 0.05). In MBT, prenatal injection of VPA manifested stereotyped behavior and enhanced number of buried marbles compared to control (p < 0.05) and DMF reduced the anxiety-related behavior in rats exhibiting ASD-like behaviors (p < 0.05). In prefrontal cortex, NRF-2 expression was downregulated in prenatal VPA model (p < 0.0001) and DMF reversed this effect (p < 0.0001). The inflammatory transcription factor NF-κB was elevated in prenatal VPA model (p < 0.0001) and reduced (p < 0.0001) upon NRF-2 activation by DMF. Prenatal VPA expressed higher levels of proinflammatory cytokine TNF- compared to control group (p < 0.0001) and DMF reduced it (p < 0.0001). Finally, the gene expression of iNOS was downregulated upon NRF-2 activation by DMF (p < 0.01). Conclusion: This study proposes that DMF is a potential agent that can be used to ameliorate autistic-like-changes through NRF-2 activation along with NF-κB downregulation and therefore, it is a promising novel therapy for ASD.

Keywords: autism spectrum disorders, dimethyl fumarate, neuroinflammation, NRF-2

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Authors: Jing Zhang

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To reduce the carbon footprint of mankind and become more sustainable is one of the major challenges in our era. Internet of Things (IoT) mainly resolves three problems: Things to Things (T2T), Human to Things, H2T), and Human to Human (H2H). Borrowing the classification of IoT, we can find carbon prints of industries also can be divided in these three ways. Therefore, monitoring the routes of generation and circulation of products may help calculate product carbon print. This paper does not consider any technique used by IoT itself, but the ideas of it look at the connection of products. Carbon prints are like a gene or mark of a product from raw materials to the final products, which never leave the products. The contribution of this paper is to combine the characteristics of IoT and the methodology of network science to find a way to calculate the product's carbon footprint. Life cycle assessment, LCA is a traditional and main tool to calculate the carbon print of products. LCA is a traditional but main tool, which includes three kinds.

Keywords: product carbon footprint, Internet of Things, network science, life cycle assessment

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Authors: B. Atika, S. Lehmann, E. Wimmer

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The defense strategy is very common in the insect world. Defensive substances play a wide variety of functions for beetles, such as repellents, toxicants, insecticides, and antimicrobics. Beetles react to predators, invaders, and parasitic microbes with the release of toxic and repellent substances. Defensive substances are directed against a large array of potential target organisms or may function for boiling bombardment or as surfactants. Usually, Coleoptera biosynthesize and store their defensive compounds in a complex secretory organ, known as odoriferous defensive stink glands. The red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae), uses these glands to produce antimicrobial p-benzoquinones and 1-alkenes. In the past, the morphology of stink gland has been studied in detail in tenebrionid beetles; however, very little is known about the genes that are involved in the production of gland secretion. In this study, we studied a subset of genes that are essential for the benzoquinone production in red flour beetle. In the first phase, we selected 74 potential candidate genes from a genome-wide RNA interference (RNAi) knockdown screen named 'iBeetle.' All these 74 candidate genes were functionally characterized by RNAi-mediated gene knockdown. Therefore, they were selected for a subsequent gas chromatography-mass spectrometry (GC-MS) analysis of secretion volatiles in respective RNAi knockdown glands. 33 of them were observed to alter the phenotype of stink gland. In the GC-MS analysis, 7 candidate genes were noted to display a strongly altered gland, in terms of secretion color and chemical composition, upon knockdown, showing their key role in the biosynthesis of gland secretion. Morphologically altered stink glands were found for odorant receptor and protein kinase superfamily. Subsequent GC-MS analysis of secretion volatiles revealed reduced benzoquinone levels in LIM domain, PDZ domain, PBP/GOBP family knockdowns and a complete lack of benzoquinones in the knockdown of sulfatase-modifying factor enzyme 1, sulfate transporter family. Based on stink gland transcriptome data, we analyzed the function of sulfatase-modifying factor enzyme 1 and sulfate transporter family via RNAi-mediated gene knockdowns, GC-MS, in situ hybridization, and enzymatic activity assays. Morphologically altered stink glands were noted in knockdown of both these genes. Furthermore, GC-MS analysis of secretion volatiles showed a complete lack of benzoquinones in the knockdown of these two genes. In situ hybridization showed that these two genes are expressed around the vesicle of certain subgroup of secretory stink gland cells. Enzymatic activity assays on stink gland tissue showed that these genes are involved in p-benzoquinone biosynthesis. These results suggest that sulfatase-modifying factor enzyme 1 and sulfate transporter family play a role specifically in benzoquinone biosynthesis in red flour beetles.

Keywords: Red Flour Beetle, defensive stink gland, benzoquinones, sulfate transporter, sulfatase-modifying factor enzyme 1

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Authors: Khawla Almulla

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It is noticed that many Arab intellectuals have called to the need and the importance of enlightenment and its application in their communities, such as Saudi Arabia. To every Islamic state, the Kingdom of Saudi Arabia represents a strategic cornerstone, since it is considered the cradle of Islam. It is the Land of the Two Holy Mosques: the Holy Mosque in Makkah surrounding the Kaaba, towards which all Muslims around the world turn while performing daily prayers and even travel to if possible in order to perform the Hajj (Pilgrimage). It also has the Prophet'ـ‘s Holy Mosque in Al-Madinah Al-Munawarah, which contains the tomb of Prophet Muhammad (pbuh). Therefore, Saudi Arabia occupies an eminent position among Arab and Islamic countries on a religious level. Saudi Arabia has become the most influential country in the Arab world, since it has one-third of the oil resources outside Central Asia, China and Russia .It is the world’s largest producer and exporter of oil. Discovering oil in Saudi Arabia converted it from an important country for Muslims-only to an important country for the major industrial countries and also the developing countries, as well. For various reasons, the diversity of intellectual currents can play a significant role in each community by way of cultural improvement, the development of civilization and the education of people until they become accustomed to accepting or rejecting opinions or ideas which differ from or oppose their own. In addition, the intellectual pluralism and cultural diversity can play a variety of roles. This helps promote dialogue and understanding between different groups or schools of thought. It can also develop cognitive skills, by exchanging ideas and views between different schools and intellectual currents. However, in Saudi Arabia there is much to oppose this plurality. The situation today shows that having a variety of ideologies and differences of cultures are not considered a reasonable way to develop intellectually as an individual or as a country. Rather the opposite is recommended, such that the ideologies of different groups are enough to bring out intellectual conflict and then to the segregation of society. As a consequence, extremism of thought from the different currents in Saudi Arabia has become apparent. This research is of great importance in its exploration of two significant themes. First, it highlights the Saudi Arabian background, in particular the historical, religious and social contexts, in order to understand the background of each religious or liberal movement and find the core of the intellectual differences between them. In addition, the aim of this research is to show the importance of moderation in Islamic thought in Saudi Arabia by tracing the thoughts and views of Dr Salman Al-Odah, whom he has considered to be the most important moderate thinker in Saudi Arabia.

Keywords: Saudi Arabia, intellectual movements, religious movements, extremism, moderation, Salafism, liberalism, Salman Al-Odah

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Authors: Z. Giagkazoglou, D. Loukovitis, C. Gubili, A. Imsiridou

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Along with the increase in human population, demand for seafood has increased. Despite the strict labeling regulations that exist for most marketed species in the European Union, seafood substitution remains a persistent global issue. Food fraud occurs when food products are traded in a false or misleading way. Mislabeling occurs when one species is substituted and traded under the name of another, and it can be intentional or unintentional. Crustaceans are one of the most regularly consumed seafood in Greece. Shrimps, prawns, lobsters, crayfish, and crabs are considered a delicacy and can be encountered in a variety of market presentations (fresh, frozen, pre-cooked, peeled, etc.). With most of the external traits removed, products as such are susceptible to species substitution. DNA barcoding has proven to be the most accurate method for the detection of fraudulent seafood products. To our best knowledge, the DNA barcoding methodology is used for the first time in Greece, in order to investigate the labeling practices for crustacean products available in the market. A total of 100 tissue samples were collected from various retailers and markets across four Greek cities. In an effort to cover the highest range of products possible, different market presentations were targeted (fresh, frozen and cooked). Genomic DNA was extracted using the DNeasy Blood & Tissue Kit, according to the manufacturer's instructions. The mitochondrial gene selected as the target region of the analysis was the cytochrome c oxidase subunit I (COI). PCR products were purified and sequenced using an ABI 3500 Genetic Analyzer. Sequences were manually checked and edited using BioEdit software and compared against the ones available in GenBank and BOLD databases. Statistical analyses were conducted in R and PAST software. For most samples, COI amplification was successful, and species-level identification was possible. The preliminary results estimate moderate mislabeling rates (25%) in the identified samples. Mislabeling was most commonly detected in fresh products, with 50% of the samples in this category labeled incorrectly. Overall, the mislabeling rates detected by our study probably relate to some degree of unintentional misidentification, and lack of knowledge surrounding the legal designations by both retailers and consumers. For some species of crustaceans (i.e. Squila mantis) the mislabeling appears to be also affected by the local labeling practices. Across Greece, S. mantis is sold in the market under two common names, but only one is recognized by the country's legislation, and therefore any mislabeling is probably not profit-motivated. However, the substitution of the speckled shrimp (Metapenaus monoceros) for the distinct, giant river prawn (Macrobranchium rosenbergii), is a clear example of deliberate fraudulent substitution, aiming for profit. To our best knowledge, no scientific study investigating substitution and mislabeling rates in crustaceans has been conducted in Greece. For a better understanding of Greece's seafood market, similar DNA barcoding studies in other regions with increased touristic importance (e.g., the Greek islands) should be conducted. Regardless, the expansion of the list of species-specific designations for crustaceans in the country is advised.

Keywords: COI gene, food fraud, labelling control, molecular identification

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Authors: Meng-Hui Chen, Chen-Yu Kao, Chia-Yu Hsu, Pei-Chann Chang

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The main idea in this paper is using sequential pattern mining to find the information which is helpful for finding high performance solutions. By combining this information, it is defined as blocks. Using the blocks to generate artificial chromosomes (ACs) could improve the structure of solutions. Estimation of Distribution Algorithms (EDAs) is adapted to solve the combinatorial problems. Nevertheless many of these approaches are advantageous for this application, but only some of them are used to enhance the efficiency of application. Generating ACs uses patterns and EDAs could increase the diversity. According to the experimental result, the algorithm which we proposed has a better performance to solve the permutation flow-shop problems.

Keywords: combinatorial problems, sequential pattern mining, estimationof distribution algorithms, artificial chromosomes

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Authors: Alaa Abdellatif, Gabrièle Breda

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Background. Advanced therapy medicinal products (ATMPs) are innovative therapies that mainly target orphan diseases and high unmet medical needs. ATMP includes gene therapy medicinal products (GTMP), somatic cell therapy medicinal products (CTMP), and tissue-engineered therapies (TEP). Since legislation opened the way in 2007, 25 ATMPs have been approved in the EU, which is about the same amount as the U.S. Food and Drug Administration. However, not all of the ATMPs that have been approved have successfully reached the market and retained their approval. Objectives. We aim to understand all the factors limiting the market access to very promising therapies in a systemic approach, to be able to overcome these problems, in the future, with scientific, regulatory and commercial innovations. Further to recent reviews that focus either on specific countries, products, or dimensions, we will address all the challenges faced by ATMP development today. Methodology. We used mixed methods and a multi-level approach for data collection. First, we performed an updated academic literature review on ATMP development and their scientific and market access challenges (papers published between 2018 and April 2023). Second, we analyzed industry feedback from cell and gene therapy webinars and white papers published by providers and pharmaceutical industries. Finally, we established a comparative analysis of the regulatory guidelines published by EMA and the FDA for ATMP approval. Results: The main challenges in bringing these therapies to market are the high development costs. Developing ATMPs is expensive due to the need for specialized manufacturing processes. Furthermore, the regulatory pathways for ATMPs are often complex and can vary between countries, making it challenging to obtain approval and ensure compliance with different regulations. As a result of the high costs associated with ATMPs, challenges in obtaining reimbursement from healthcare payers lead to limited patient access to these treatments. ATMPs are often developed for orphan diseases, which means that the patient population is limited for clinical trials which can make it challenging to demonstrate their safety and efficacy. In addition, the complex manufacturing processes required for ATMPs can make it challenging to scale up production to meet demand, which can limit their availability and increase costs. Finally, ATMPs face safety and efficacy challenges: dangerous adverse events of these therapies like toxicity related to the use of viral vectors or cell therapy, starting material and donor-related aspects. Conclusion. As a result of our mixed method analysis, we found that ATMPs face a number of challenges in their development, regulatory approval, and commercialization and that addressing these challenges requires collaboration between industry, regulators, healthcare providers, and patient groups. This first analysis will help us to address, for each challenge, proper and innovative solution(s) in order to increase the number of ATMPs approved and reach the patients

Keywords: advanced therapy medicinal products (ATMPs), product development, market access, innovation

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Authors: Berengere Vire, Nicolas Salvetat, Yoann Lannay, Guillaume Marcellin, Siem Van Der Laan, Franck Molina, Dinah Weissmann

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Predicting suicidal behaviors is one of the most complex challenges of daily psychiatric practices. Today, suicide risk prediction using biological tools is not validated and is only based on subjective clinical reports of the at-risk individual. Therefore, there is a great need to identify biomarkers that would allow early identification of individuals at risk of suicide. Alterations of adenosine-to-inosine (A-to-I) RNA editing of neurotransmitter receptors and other proteins have been shown to be involved in etiology of different psychiatric disorders and linked to suicidal behavior. RNA editing is a co- or post-transcriptional process leading to a site-specific alteration in RNA sequences. It plays an important role in the epi transcriptomic regulation of RNA metabolism. On postmortem human brain tissue (prefrontal cortex) of depressed suicide victims, Alcediag found specific alterations of RNA editing activity on the mRNA coding for the serotonin 2C receptor (5-HT2cR). Additionally, an increase in expression levels of ADARs, the RNA editing enzymes, and modifications of RNA editing profiles of prime targets, such as phosphodiesterase 8A (PDE8A) mRNA, have also been observed. Interestingly, the PDE8A gene is located on chromosome 15q25.3, a genomic region that has recurrently been associated with the early-onset major depressive disorder (MDD). In the current study, we examined whether modifications in RNA editing profile of prime targets allow identifying disease-relevant blood biomarkers and evaluating suicide risk in patients. To address this question, we performed a clinical study to identify an RNA editing signature in blood of depressed patients with and without the history of suicide attempts. Patient’s samples were drawn in PAXgene tubes and analyzed on Alcediag’s proprietary RNA editing platform using next generation sequencing technology. In addition, gene expression analysis by quantitative PCR was performed. We generated a multivariate algorithm comprising various selected biomarkers to detect patients with a high risk to attempt suicide. We evaluated the diagnostic performance using the relative proportion of PDE8A mRNA editing at different sites and/or isoforms as well as the expression of PDE8A and the ADARs. The significance of these biomarkers for suicidality was evaluated using the area under the receiver-operating characteristic curve (AUC). The generated algorithm comprising the biomarkers was found to have strong diagnostic performances with high specificity and sensitivity. In conclusion, we developed tools to measure disease-specific biomarkers in blood samples of patients for identifying individuals at the greatest risk for future suicide attempts. This technology not only fosters patient management but is also suitable to predict the risk of drug-induced psychiatric side effects such as iatrogenic increase of suicidal ideas/behaviors.

Keywords: blood biomarker, next-generation-sequencing, RNA editing, suicide

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Authors: Humaira Irfan Khan

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The paper discusses that Pakistan is a multilingual, multicultural, and multiethnic society. The findings from quantitative and qualitative data collected in two public universities look at the importance of English language and the role and status of national and regional languages in the country. The evidence implies that postgraduate students having diverse linguistic, cultural, ethnic, socio-economic, and educational backgrounds display negative attitudes towards the use of English language for academic and interactive functions in universities. It is also discovered that language anxiety of postgraduate students is an outcome of their language learning difficulties. It is suggested that considering the academic needs of students, universities should introduce a language proficiency course to enable them to use English with confidence.

Keywords: Multilingualism, Ethnicity, Cultural Diversity, Importance of English, National language, Regional languages, Language Anxiety

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Authors: Önder M. Özdem

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Producing and circulating films of professional standards have become technically easier with the development and widespread use of digital recording and distribution technologies. Additionally, film festivals on common platforms have rapidly increased in numbers and diversity. On the one hand, no-charge applications result in excessive submissions; thus, it complicates the evaluation and selection process. On the other hand, festival’s high submission fees may make the distribution of films with a limited budget very difficult. Inspired by the author’s engagement with the film industry as both a pre-jury member of an international film festival and an applicant to many festivals, this study discusses the causes and consequences of the increasing commercialization of film festivals. The author’s double identity, both as a jury and an applicant, provides a comparative perspective through which one can unfold the different dimensions and dynamics in the film production and distribution processes.

Keywords: commercialization, film distribution, film festivals, film production

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Authors: Morello Sara, Pederiva Sabina, Bianchi Manila, Martucci Francesca, Marchis Daniela, Decastelli Lucia

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Vegetables represent an integral part of a healthy diet due to their valuable nutritional properties and the growth in consumer demand in recent years is particularly remarkable for a diet rich in vitamins and micronutrients. However, plant-based products are involved in several food outbreaks connected to various sources of contamination and quite often, bacteria responsible for side effects showed high resistance to antibiotics. The abuse of antibiotics can be one of the main mechanisms responsible for increasing antibiotic resistance (AR). Plants grown for food use can be contaminated directly by spraying antibiotics on crops or indirectly by treatments with antibiotics due to the use of manure, which may contain both antibiotics and genes of antibiotic resistance (ARG). Antibiotic residues could represent a potential way of human health risk due to exposure through the consumption of plant-based foods. The presence of antibiotic-resistant bacteria might pose a particular risk to consumers. The present work aims to investigate through a multidisciplinary approach the occurrence of ARG by means of a biomolecular approach (PCR) and the prevalence of antibiotic residues using a multi residues LC-MS/MS method, both in different plant-based products. During the period from July 2020 to October 2021, a total of 74 plant samples (33 lettuces and 41 tomatoes) were collected from 57 farms located throughout the Piedmont area, and18 out of 74 samples (11 lettuces and 7 tomatoes) were selected to LC-MS/MS analyses. DNA extracted (ExtractME, Blirt, Poland) from plants used on crops and isolated bacteria were analyzed with 6 sets of end-point multiplex PCR (Qiagen, Germany) to detect the presence of resistance genes of the main antibiotic families, such as tet genes (tetracyclines), bla (β-lactams) and mcr (colistin). Simultaneous detection of 43 molecules of antibiotics belonging to 10 different classes (tetracyclines, sulphonamides, quinolones, penicillins, amphenicols, macrolides, pleuromotilines, lincosamides, diaminopyrimidines) was performed using Exion LC system AB SCIEX coupled to a triple quadrupole mass spectrometer QTRAP 5500 from AB SCIEX. The PCR assays showed the presence of ARG in 57% (n=42): tetB (4.8%; n=2), tetA (9.5%; n=4), tetE (2.4%; n=1), tetL (12%; n=5), tetM (26%; n=11), blaSHV (21.5%; n=9), blaTEM (4.8%; n =2) and blaCTX-M (19%; n=8). In none of the analyzed samples was the mcr gene responsible for colistin resistance detected. Results obtained from LC-MS/MS analyses showed that none of the tested antibiotics appear to exceed the LOQ (100 ppb). Data obtained confirmed the presence of bacterial populations containing antibiotic resistance determinants such as tet gene (tetracycline) and bla genes (beta-lactams), widely used in human medicine, which can join the food chain and represent a risk for consumers, especially with raw products. The presence of traces of antibiotic residues in vegetables, in concentration below the LOQ of the LC-MS/MS method applied, cannot be excluded. In conclusion, traces of antibiotic residues could be a health risk to the consumer due to potential involvement in the spread of AR. PCR represents a useful and effective approach to characterize and monitor AR carried by bacteria from the entire food chain.

Keywords: plant-based products, ARG, PCR, antibiotic residues

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Authors: Rohita Gupta, G. S. Brah, R. Verma, C. S. Mukhopadhayay

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Although chicken strains show differences in susceptibility to a number of diseases, the underlying immunological basis is yet to be elucidated. In the present study, heterophils were subjected to LPS stimulation and total RNA extraction, further differential gene expression was studied in broiler, layer and indigenous Aseel strain by Real Time RT-PCR at different time periods before and after induction. The expression of the 14 AvBDs and chTLR 1, 2, 3, 4, 5, 7, 15 and 21 was detectable in heterophils. The expression level of most of the AvBDs significantly increased (P<0.05) 3 hours post in vitro lipopolysaccharide challenge. Higher expression level and stronger activation of most AvBDs, NFkB-1 and IRF-3 in heterophils was observed, with the stimulation of LPS in layer compared to broiler, and in Aseel compared to both layer and broiler. This investigation will allow more refined interpretation of immuno-genetic basis of the variable disease resistance/susceptibility in divergent stock of chicken including indigenous breed. Moreover this study will be helpful in formulation of strategy for isolation of antimicrobial peptides from heterophils.

Keywords: differential expression, heterophils, cytokines, defensin, TLR

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Authors: Radek Malinský, Ivan Jelínek

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The current web has become a modern encyclopedia, where people share their thoughts and ideas on various topics around them. Such kind of encyclopedia is very useful for other people who are looking for answers to their questions. However, with the growing popularity of social networking and blogging and ever expanding network services, there has also been a growing diversity of technologies along with different structure of individual websites. It is, therefore, difficult to directly find a relevant answer for a common Internet user. This paper presents a web application for the real-time end-to-end analysis of selected Internet trends; where the trend can be whatever the people post online. The application integrates fully configurable tools for data collection and analysis using selected webometric algorithms, and for its chronological visualization to user. It can be assumed that the application facilitates the users to evaluate the quality of various products that are mentioned online.

Keywords: Trend, visualizer, web analysis, web 2.0.

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Authors: Elena Arquer Cuenca

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The gypsy community has been a mistreated and rejected group since its arrival in the Iberian Peninsula in the 15th century. At present, despite being the largest ethnic minority group in Spain as well as in Europe, the different legal and social initiatives in favour of equality continue to suffer discrimination by the general society. This has fostered a strengthening of the endogroup accompanied by cultural conservatism as a form of self-protection. Despite the current trend of normalization of sexual diversity in modern societies, LGB people continue to suffer discrimination, especially in more traditional environments or communities. This rejection for reasons of sexual orientation within the family or community can hinder the free development of the person and compromise peaceful coexistence. The present work is intended as an approach to the attitudes of the gypsy population towards non-heterosexual sexual orientation. The objective is none other than ‘to know the appreciation that the gypsy population has about homosexual sex-affective relationships, in order to assess whether this has any impact on family and community coexistence’. The following specific objectives are derived from this general objective: ‘To find out whether there is a relationship between the dichotomous Roma gender system and the acceptance/rejection of homosexuality’; ‘to analyse whether sexual orientation has an impact on the coexistence of the Roman family and community’; ‘to analyse whether the historical discrimination suffered by the Roman population favours the maintenance of the patriarchal heterosexual reproductive family’; and lastly ‘to explore whether ICTs have promoted the process of normalisation and/or acceptance of homosexuality within the Roma community’. In order to achieve these objectives, a bibliographical and documentary review has been used, as well as the semi-structured interview technique, in which 4 gypsy people participated (2 women and 2 men of different ages). One of the main findings was the inappropriateness of the use of the homogenising category "Gypsy People" at present, given the great diversity among the Roma communities. Moreover, the difficulty in accepting homosexuality seems to be related to the fact that the heterosexual reproductive family has been the main survival mechanism of Roma communities over centuries. However, it will be concluded that attitudes towards homosexuality will vary depending on the socio-economic and cultural context and factors such as age or professed religion. Three main contributions of this research are: firstly, the inclusion of sexual orientation as a variable to be considered when analysing peaceful coexistence; secondly socio-historical dynamics and structures of inequality have been taken into account when analysing Roma attitudes towards homosexuality; and finally, the processual nature of socio-cultural changes has also been considered.

Keywords: gender, homosexuality, ICTs, peaceful coexistence, Roma community, sexual orientation

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Authors: Hiroshi Yamaguchi, Phillip C. Y. Sheu, Ryo Fujita, Shigeo Tsujii

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In this paper, the secure BioSemantic Scheme is presented to bridge biological/biomedical research problems and computational solutions via semantic computing. Due to the diversity of problems in various research fields, the semantic capability description language (SCDL) plays and important role as a common language and generic form for problem formalization. SCDL is expected the essential for future semantic and logical computing in Biosemantic field. We show several example to Biomedical problems in this paper. Moreover, in the coming age of cloud computing, the security problem is considered to be crucial issue and we presented a practical scheme to cope with this problem.

Keywords: biomedical applications, private information retrieval (PIR), semantic capability description language (SCDL), semantic computing

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Authors: Inas Raafat, Azza El Bassiouny, Waldemar L. Olszewsky, Nagui E. Mikhail, Mona Nossier, Nora E. I. El-Bassiouni, Mona Zoheiry, Houda Abou Taleb, Noha Abd El-Aal, Ali Baioumy, Shimaa Attia

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Background: Orthotopic liver transplantation is an established treatment for patients with severe acute and end-stage chronic liver disease. The shortage of donor organs continues to be the rate-limiting factor for liver transplantation throughout the world. Hepatocyte transplantation is a promising treatment for several liver diseases and can, also, be used as a "bridge" to liver transplantation in cases of liver failure. Aim of the work: This study was designed to develop a highly efficient protocol for isolation and transplantation of hepatocytes in experimental Lewis rat model to provide satisfactory guidelines for future application on humans.Materials and Methods: Hepatocytes were isolated from the liver by double perfusion technique and bone marrow cells were isolated by centrifugation of shafts of tibia and femur of donor Lewis rats. Recipient rats were subjected to sub-lethal dose of irradiation 2 days before transplantation. In a laparotomy operation the spleen was injected by freshly isolated hepatocytes and bone marrow cells were injected intravenously. The animals were sacrificed 45 day latter and splenic sections were prepared and stained with H & E, PAS AFP and Prox1. Results: The data obtained from this study showed that the double perfusion technique is successful in separation of hepatocytes regarding cell number and viability. Also the method used for bone marrow cells separation gave excellent results regarding cell number and viability. Intrasplenic engraftment of hepatocytes and live tissue formation within the splenic tissue were found in 70% of cases. Hematoxylin and eosin stained splenic sections from 7 rats showed sheets and clusters of cells among the splenic tissues. Periodic Acid Schiff stained splenic sections from 7 rats showed clusters of hepatocytes with intensely stained pink cytoplasmic granules denoting the presence of glycogen. Splenic sections from 7 rats stained with anti-α-fetoprotein antibody showed brownish cytoplasmic staining of the hepatocytes denoting positive expression of AFP. Splenic sections from 7 rats stained with anti-Prox1 showed brownish nuclear staining of the hepatocytes denoting positive expression of Prox1 gene on these cells. Also, positive expression of Prox1 gene was detected on lymphocytes aggregations in the spleens. Conclusions: Isolation of liver cells by double perfusion technique using collagenase buffer is a reliable method that has a very satisfactory yield regarding cell number and viability. The intrasplenic route of transplantation of the freshly isolated liver cells in an immunocompromised model was found to give good results regarding cell engraftment and tissue formation. Further studies are needed to assess function of engrafted hepatocytes by measuring prothrombin time, serum albumin and bilirubin levels.

Keywords: Lewis rats, hepatocytes, BMCs, transplantation, AFP, Prox1

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Authors: Beata Lachtara, Renata Szewczyk, Katarzyna Bielinska, Kinga Wieczorek, Jacek Osek

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Listeria monocytogenes is an important human and animal pathogen that causes foodborne outbreaks. The bacteria may be present in different types of food: cheese, raw vegetables, sliced meat products and vacuum-packed sausages, poultry, meat, fish. The most common method, which has been used for the investigation of genetic diversity of L. monocytogenes, is PFGE. This technique is reliable and reproducible and established as gold standard for typing of L. monocytogenes. The aim of the study was characterization by molecular serotyping and PFGE analysis of L. monocytogenes strains isolated from fresh fish and fish products in Poland. A total of 301 samples, including fresh fish (n = 129) and fish products (n = 172) were, collected between January 2014 and March 2016. The bacteria were detected using the ISO 11290-1 standard method. Molecular serotyping was performed with PCR. The isolates were tested with the PFGE method according to the protocol developed by the European Union Reference Laboratory for L. monocytogenes with some modifications. Based on the PFGE profiles, two dendrograms were generated for strains digested separately with two restriction enzymes: AscI and ApaI. Analysis of the fingerprint profiles was performed using Bionumerics software version 6.6 (Applied Maths, Belgium). The 95% of similarity was applied to differentiate the PFGE pulsotypes. The study revealed that 57 of 301 (18.9%) samples were positive for L. monocytogenes. The bacteria were identified in 29 (50.9%) ready-to-eat fish products and in 28 (49.1%) fresh fish. It was found that 40 (70.2%) strains were of serotype 1/2a, 14 (24.6%) 1/2b, two (4.3%) 4b and one (1.8%) 1/2c. Serotypes 1/2a, 1/2b, and 4b were presented with the same frequency in both categories of food, whereas serotype 1/2c was detected only in fresh fish. The PFGE analysis with AscI demonstrated 43 different pulsotypes; among them 33 (76.7%) were represented by only one strain. The remaining 10 profiles contained more than one isolate. Among them 8 pulsotypes comprised of two L. monocytogenes isolates, one profile of three isolates and one restriction type of 5 strains. In case of ApaI typing, the PFGE analysis showed 27 different pulsotypes including 17 (63.0%) types represented by only one strain. Ten (37.0%) clusters contained more than one strain among which four profiles covered two strains; three had three isolates, one with five strains, one with eight strains and one with ten isolates. It was observed that the isolates assigned to the same PFGE type were usually of the same serotype (1/2a or 1/2b). The majority of the clusters had strains of both sources (fresh fish and fish products) isolated at different time. Most of the strains grouped in one cluster of the AscI restriction was assigned to the same groups in ApaI investigation. In conclusion, PFGE used in the study showed a high genetic diversity among L. monocytogenes. The strains were grouped into varied clonal clusters, which may suggest different sources of contamination. The results demonstrated that 1/2a serotype was the most common among isolates from fresh fish and fish products in Poland.

Keywords: Listeria monocytogenes, molecular characteristic, PFGE, serotyping

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Authors: Foad Alzoughool

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Conflicting studies on the association between the rs5219 polymorphism and type 2 diabetes, some studies have confirmed a strong relationship between this variant and type2 diabetes, on the other hand, many studies denied the existence of this association. This study aimed to provide evidence about whether the rs5219 polymorphism has or hasn't a role as a risk factor for diabetes and meta-analysis to investigate the role of the control age group in the association. Genotyping of the rs5219 polymorphism was performed in a cohort of 266 healthy elderly subjects with a mean age (60.2 ± 5.1) with no history of diabetes (HbA1c < 6%) using standard Sanger sequencing methods. Lys/Lys alleles were detected in 20 persons (7.5%), Lys/Glu alleles in 96 persons (36.1%), and Glu/Glu in 150 persons (56.4%). The genotype distribution was consistent with Hardy–Weinberg equilibrium (P =0.7). Meta-analysis notably indicates no association between rs5219 polymorphism and type 2 diabetes in all studies used the younger age of the control group compared to the patient's age. In conclusion, our study sheds light on the importance of age factor among the control group recruited in case-control studies.

Keywords: Type 2 diabetes, rs5219 polymorphism, E23K, KCNJ11 gene

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Authors: Kamaran Fathulla

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Human creativity knows no bounds. More than a millennia ago humans have expressed their knowledge on cave walls and on clay artefacts. Using visuals such as diagrams and paintings have always provided us with a natural and intuitive medium for expressing such creativity. Making sense of human generated visualisation has been influenced by western scientific philosophies which are often reductionist in their nature. Theoretical frameworks such as those delivered by Peirce dominated our views of how to make sense of visualisation where a visual is seen as an emergent property of our thoughts. Others have reduced the richness of human-generated visuals to mere shapes drawn on a piece of paper or on a screen. This paper introduces an alternate framework where the centrality of human functioning is given explicit and richer consideration through the multi aspectual philosophical works of Herman Dooyeweerd. Dooyeweerd's framework of understanding reality was based on fifteen aspects of reality, each having a distinct core meaning. The totality of the aspects formed a ‘rainbow’ like spectrum of meaning. The thesis of this approach is that meaningful human functioning in most cases involves the diversity of all aspects working in synergy and harmony. Illustration of the foundations and applicability of this approach is underpinned in the case of humans use of diagramming for creative purposes, particularly within an educational context. Diagrams play an important role in education. Students and lecturers use diagrams as a powerful tool to aid their thinking. However, research into the role of diagrams used in education continues to reveal difficulties students encounter during both processes of interpretation and construction of diagrams. Their main problems shape up students difficulties with diagrams. The ever-increasing diversity of diagrams' types coupled with the fact that most real-world diagrams often contain a mix of these different types of diagrams such as boxes and lines, bar charts, surfaces, routes, shapes dotted around the drawing area, and so on with each type having its own distinct set of static and dynamic semantics. We argue that the persistence of these problems is grounded in our existing ways of understanding diagrams that are often reductionist in their underpinnings driven by a single perspective or formalism. In this paper, we demonstrate the limitations of these approaches in dealing with the three problems. Consequently, we propose, discuss, and demonstrate the potential of a nonreductionist framework for understanding diagrams based on Symbolic and Spatial Mappings (SySpM) underpinned by Dooyeweerd philosophy. The potential of the framework to account for the meaning of diagrams is demonstrated by applying it to a real-world case study physics diagram.

Keywords: SySpM, drawing style, mapping

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Authors: Chetna Jaswal, Nishi Srivastava, Ahammedul Kabeer AP, Puja Prasad

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We are a culture of mass media consumers and cinema as its integral part has high visibility and potential influence on public attitude towards disability which maintains no sociocultural boundaries but experiences substantial social marginalization. Given the lack of awareness and direct experience with disability, on-screen or film representations can give powerful and memorable definitions for the public that can contribute to framing the perception and attitude change. Social representation refers to common ways of thinking, conceiving about and evaluating social reality. It is a product of collective cognition, common sense and thought system. This study aims at analyzing the representations and narratives of disability in Indian cinema and Hollywood with the help of a conceptual understanding of social representation and its theoretical framework.

Keywords: disability, social representation, mainstream cinema, diversity

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Authors: Mustafa Yorgancılar, Emine Atalay, Necdet Akgün, Ali Topal

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In this study, polymerase chain reaction based Inter-simple sequence repeat (ISSR) from DNA fingerprinting techniques were used to investigate the genetic relationships among barley crossbreed genotypes in Turkey. It is important that selection based on the genetic base in breeding programs via ISSR, in terms of breeding time. 14 ISSR primers generated a total of 97 bands, of which 81 (83.35%) were polymorphic. The highest total resolution power (RP) value was obtained from the F2 (0.53) and M16 (0.51) primers. According to the ISSR result, the genetic similarity index changed between 0.64–095; Lane 3 with Line 6 genotypes were the closest, while Line 36 were the most distant ones. The ISSR markers were found to be promising for assessing genetic diversity in barley crossbreed genotypes.

Keywords: barley, crossbreed, genetic similarity, ISSR

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Authors: Eric Bang

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Interactions between receptors and ligands are crucial for many essential biological processes, including neurotransmission and metabolism. Ligand-receptor analyses that examine cell behavior and interactions often utilize cell type-specific RNA expressions from single-cell RNA sequencing (scRNA-seq) data. Using CellPhoneDB, a public repository consisting of ligands, receptors, and ligand-receptor interactions, the cell-cell interactions were explored in a specific scRNA-seq dataset from kidney tissue and portrayed the results with dot plots and heat maps. Depending on the type of cell, each ligand-receptor pair was aligned with the interacting cell type and calculated the positori probabilities of these associations, with corresponding P values reflecting average expression values between the triads and their significance. Using single-cell data (sample kidney cell references), genes in the dataset were cross-referenced with ones in the existing CellPhoneDB dataset. For example, a gene such as Pleiotrophin (PTN) present in the single-cell data also needed to be present in the CellPhoneDB dataset. Using the single-cell transcriptomics data via slide-seq and reference data, the CellPhoneDB program defines cell types and plots them in different formats, with the two main ones being dot plots and heat map plots. The dot plot displays derived measures of the cell to cell interaction scores and p values. For the dot plot, each row shows a ligand-receptor pair, and each column shows the two interacting cell types. CellPhoneDB defines interactions and interaction levels from the gene expression level, so since the p-value is on a -log10 scale, the larger dots represent more significant interactions. By performing an interaction analysis, a significant interaction was discovered for myeloid and T-cell ligand-receptor pairs, including those between Secreted Phosphoprotein 1 (SPP1) and Fibronectin 1 (FN1), which is consistent with previous findings. It was proposed that an effective protocol would involve a filtration step where cell types would be filtered out, depending on which ligand-receptor pair is activated in that part of the tissue, as well as the incorporation of the CellPhoneDB data in a streamlined workflow pipeline. The filtration step would be in the form of a Python script that expedites the manual process necessary for dataset filtration. Being in Python allows it to be integrated with the CellPhoneDB dataset for future workflow analysis. The manual process involves filtering cell types based on what ligand/receptor pair is activated in kidney cells. One limitation of this would be the fact that some pairings are activated in multiple cells at a time, so the manual manipulation of the data is reflected prior to analysis. Using the filtration script, accurate sorting is incorporated into the CellPhoneDB database rather than waiting until the output is produced and then subsequently applying spatial data. It was envisioned that this would reveal wherein the cell various ligands and receptors are interacting with different cell types, allowing for easier identification of which cells are being impacted and why, for the purpose of disease treatment. The hope is this new computational method utilizing spatially explicit ligand-receptor association data can be used to uncover previously unknown specific interactions within kidney tissue.

Keywords: bioinformatics, Ligands, kidney tissue, receptors, spatial transcriptome

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Authors: Benyapa Thitimapong, Najwa Niyomdecha

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This qualitative descriptive research aimed to explore the self-care experiences during pregnancy of Muslim adolescents. Twenty participants were first-time Muslim mothers who had pregnancy experienceห under 20 years of age in three Southern border provinces of Thailand. Participants were selected by purposive sampling with inclusion criteria. Data were collected from the in-depth interview and analyzed using content analysis. The findings revealed that Muslim pregnant adolescents take care of themselves in the context of combining self-care in an Islamic way and conventional medicine. There are 2 subthemes: 1) antenatal care with Tok Bidan and 2) health promotion during pregnancy. The finding will help to understand self-care during pregnancy of Muslim adolescents among three Southern border provinces and can apply to nurse educators as a guide to educate and manage an appropriate self-care program for Muslim pregnant adolescents based on cultural diversity.

Keywords: adolescents, muslim, pregnancy, selfcare

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Authors: André Pasti

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Historically, the communication sector in Argentina is characterized by intense monopolization and geographical concentration in the city of Buenos Aires. In 2000, the four major media conglomerates in operation – Clarín, Telefónica, America and Hadad – controlled 84% of the national media market. By 2009, new policies were implemented as a result of civil society organizations demands. Legally, a new regulatory framework was approved: the law 26,522 of Audiovisual Communications Services. Supposedly, these policies intend to create new conditions for the development of the audiovisual economy in the territory of Argentina. The regionalization of audiovisual production and the democratization of channels and access to media were among the priorities. This paper analyses the main changes and continuities in the organization of the spatial circuit of the audiovisual industry in Argentina provoked by these new policies. These new policies aim at increasing the diversity of audiovisual producers and promoting regional audiovisual industries. For this purpose, a national program for the development of audiovisual centers within the country was created. This program fostered a federalized production network, based on nine audiovisual regions and 40 nodes. Each node has created technical, financial and organizational conditions to gather different actors in audiovisual production – such as SMEs, social movements and local associations. The expansion of access to technical networks was also a concern of other policies, such as ‘Argentina connected’, whose objective was to expand access to broadband Internet. The Open Digital Television network also received considerable investments. Furthermore, measures have been carried out in order to impose limits on the concentration of ownership as well as to eliminate the oligopolies and to ensure more competition in the sector. These actions intended to force a divide of the media conglomerates into smaller groups. Nevertheless, the corporations that compose these conglomerates resist strongly, making full use of their economic and judiciary power. Indeed, the absence of effective impact of such measures can be testified by the fact that the audiovisual industry remains strongly concentrated in Argentina. Overall, these new policies were designed properly to decentralize audiovisual production and expand the regional diversity of the audiovisual industry. However, the effective transformation of the organization of the audiovisual circuit in the territory faced several resistances. This can be explained firstly and foremost by the ideological and economic power of the media conglomerates. In the second place, there is an inherited inertia from the unequal distribution of the objects needed for the audiovisual production and consumption. Lastly, the resistance also relies on financial needs and in the excessive dependence of the state for the promotion of regional audiovisual production.

Keywords: Argentina, audiovisual industry, communication policies, geographic concentration, regionalization, spatial circuit

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Authors: Baljeet Singh Saharan

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Bioremediation is the demand of the day. Tannery and textile effluents/waste waters have lots of pollution due to presence of hexavalent Chromium. Methodologies used in the present investigations include isolation, cultivation and purification of bacterial strain. Further characterization techniques and 16S rRNA sequencing were performed. Efficient bacterial strain capable of reducing hexavalent chromium was obtained. The strain can be used for bioremediation of industrial effluents containing hexavalent Cr. A gram negative, rod shaped and yellowish pigment producing bacterial strain from tannery effluent was isolated using nutrient agar. The 16S rRNA gene sequence similarity indicated that isolate SA13A is associated with genus Luteimonas (99%). This isolate has been found to reduce 100% of hexavalent chromium Cr (VI) (100 mg L-1) 100% in 16 h. Growth conditions were optimized for Cr (VI) reduction. Maximum reduction was observed at a temperature of 37 °C and pH 8.0. Additionally, Luteimonas aestuarii SA13A showed resistance against various heavy metals like Cr+6, Cr+3, Cu+2, Zn+2, Co+2, Ni+2 and Cd+2 . Hence, Luteimonas aestuarii SA13A could be used as potent Cr (VI) reducing strain as well as significant bioremediator in heavy metal contaminated sites.

Keywords: bioremediation, chromium, eco-friendly, heavy metals

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Authors: H. Sakamoto, Y. Nakagawara, S. Oguri

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Drought stress is a critical environmental factor that adversely affects crop productivity and quality. Because of their immobile nature, plants have evolved mechanisms to sense and respond to drought stress. We identified a novel locus of Arabidopsis, designated DRA1 (drought responsive ankyrin 1), whose disruption leads to increased drought stress tolerance. DRA1 encodes a transmembrane protein with an ankyrin repeat motif that has been implicated in diverse cellular processes such as signal transduction. RT-PCR analysis revealed that there were at least two splicing variants of DRA1 transcripts in wild type plants. In response to drought stress, the levels of DRA1 transcripts retaining second and third introns were increased, whereas these introns were removed under unstressed conditions. These results suggest that DRA1 protein may negatively regulate plant drought tolerance and that the expression of DRA1 is regulated in response to drought stress by alternative splicing.

Keywords: alternative splicing, ankyrin repeat, Arabidopsis, drought tolerance

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Authors: Chung-Young Lee, Se-Hee Ahn, Ilhwan Kim, Du-Min Go, Dae-Yong Kim, Jun-Gu Choi, Youn-Jeong Lee, Jae-Hong Kim, Hyuk-Joon Kwon

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The polymerase of avian influenza A virus (AIV) is a heterotrimer composed of PB2, PB1 and PA. PB2 plays a role in overcoming the host barrier; however, the genetic prerequisites for avian PB2 to acquire mammalian pathogenic mutations have not been well elucidated. Here, we demonstrated that key amino acid mutations (I66M, I109V and I133V, collectively referred to as MVV) of prototypic avian PB2 increase the replication efficiency of recombinant PR8 virus carrying the mutated PB2 in both avian and mammalian hosts. The MVV mutations caused no weight loss in mice, but they did allow replication in infected lungs, and the viruses acquired fatal mammalian pathogenic mutations such as Q591R/K, E627K, or D701N in the infected lungs. The MVV mutations are located at the interfaces of the trimer and are predicted to increase the strength of this structure. Thus, gaining MVV mutations might be the first step for AIV to acquire mammalian pathogenicity. These results provide new insights into the evolution of AIV in birds and mammals.

Keywords: avian influenza A virus, prototypic PB2, polymerase activity, mammalian pathogenicity, first-step mutations

Procedia PDF Downloads 342