Search results for: late cornified envelope gene 1C

Authors: Leonard Lipovich, Pattaraporn Thepsuwan, Anton-Scott Goustin, Juan Cai, Donghong Ju, James B. Brown

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Two-thirds of human genes do not encode any known proteins. Aside from long non-coding RNA (lncRNA) genes with recently-discovered functions, the ~40,000 non-protein-coding human genes remain poorly understood, and a role for their transcripts as de-facto unconventional messenger RNAs has not been formally excluded. Ribosome profiling (Riboseq) predicts translational potential, but without independent evidence of proteins from lncRNA open reading frames (ORFs), ribosome binding of lncRNAs does not prove translation. Previously, we mass-spectrometrically documented translation of specific lncRNAs in human K562 and GM12878 cells. We now examined lncRNA translation in human MCF7 cells, integrating strand-specific Illumina RNAseq, Riboseq, and deep mass spectrometry in biological quadruplicates performed at two core facilities (BGI, China; City of Hope, USA). We excluded known-protein matches. UCSC Genome Browser-assisted manual annotation of imperfect (tryptic-digest-peptides)-to-(lncRNA-three-frame-translations) alignments revealed three peptides hypothetically explicable by 'stop-to-nonstop' in-frame replacement of stop codons by amino acids in two ORFs of the lncRNA MMP24-AS1. To search for this phenomenon genomewide, we designed and implemented a novel pipeline, matching tryptic-digest spectra to wildcard-instead-of-stop versions of repeat-masked, six-frame, whole-genome translations. Along with singleton putative stop-to-nonstop events affecting four other lncRNAs, we identified 24 additional peptides with stop-to-nonstop in-frame substitutions from multiple positive-strand MMP24-AS1 ORFs. Only UAG and UGA, never UAA, stop codons were impacted. All MMP24-AS1-matching spectra met the same significance thresholds as high-confidence known-protein signatures. Targeted resequencing of MMP24-AS1 genomic DNA and cDNA from the same samples did not reveal any mutations, polymorphisms, or sequencing-detectable RNA editing. This unprecedented apparent gene-specific violation of the genetic code highlights the importance of matching peptides to whole-genome, not known-genes-only, ORFs in mass-spectrometry workflows, and suggests a new mechanism enhancing the combinatorial complexity of the proteome. Funding: NIH Director’s New Innovator Award 1DP2-CA196375 to LL.

Keywords: genetic code, lncRNA, long non-coding RNA, mass spectrometry, proteogenomics, ribo-seq, ribosome, RNAseq

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Authors: Purva Nemavarkar, Badri Narain Pandey, Jitendra Kumar

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Introduction: The objective was to understand the effect of various radioprotectors on DNA damage repair enzyme and survival in gamma-irradiated wild and cdc mutants of S. pombe (fission yeast) cultured under permissive and restrictive conditions. DNA repair process, as influenced by radioprotectors, was measured by activity of DNA polymerase in the cells. The use of single cell gel electrophoresis assay (SCGE) or Comet Assay to follow gamma-irradiation induced DNA damage and effect of radioprotectors was employed. In addition, studying the effect of caffeine at different concentrations on S-phase of cell cycle was also delineated. Materials and Methods: S. pombe cells grown at permissive temperature (250C) and/or restrictive temperature (360C) were followed by gamma-radiation. Percentage survival and activity of DNA Polymerase (yPol II) were determined after post-irradiation incubation (5 h) with radioprotectors such as Caffeine, Curcumin, Disulphiram, and Ellagic acid (the dose depending on individual D 37 values). The gamma-irradiated yeast cells (with and without the radioprotectors) were spheroplasted by enzyme glusulase and subjected to electrophoresis. Radio-resistant cells were obtained by arresting cells in S-phase using transient treatment of hydroxyurea (HU) and studying the effect of caffeine at different concentrations on S-phase of cell cycle. Results: The mutants of S. pombe showed insignificant difference in survival when grown under permissive conditions. However, growth of these cells under restrictive temperature leads to arrest in specific phases of cell cycle in different cdc mutants (cdc10: G1 arrest, cdc22: early S arrest, cdc17: late S arrest, cdc25: G2 arrest). All the cdc mutants showed decrease in survival after gamma radiation when grown at permissive and restrictive temperatures. Inclusion of the radioprotectors at respective concentrations during post irradiation incubation showed increase in survival of cells. Activity of DNA polymerase enzyme (yPol II) was increased significantly in cdc mutant cells exposed to gamma-radiation. Following SCGE, a linear relationship was observed between doses of irradiation and the tail moments of comets. The radioprotection of the fission yeast by radioprotectors can be seen by the reduced tail moments of the yeast comets. Caffeine also exhibited its radio-protective ability in radio-resistant S-phase cells obtained after HU treatment. Conclusions: The radioprotectors offered notable radioprotection in cdc mutants when added during irradiation. The present study showed activation of DNA damage repair enzyme (yPol II) and an increase in survival after treatment of radioprotectors in gamma irradiated wild type and cdc mutants of S. pombe cells. Results presented here showed feasibility of applying SCGE in fission yeast to follow DNA damage and radioprotection at high doses, which are not feasible with other eukaryotes. Inclusion of caffeine at 1mM concentration to S phase cells offered protection and did not decrease the cell viability. It can be proved that at minimal concentration, caffeine offered marked radioprotection.

Keywords: radiation protection, cell cycle, fission yeast, comet assay, s-phase, DNA repair, radioprotectors, caffeine, curcumin, SCGE

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Authors: Nima Samie, Sekaran Muniandy, Zahurin Mohamed, M. S. Kanthimathi

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Although number of indole containing compounds have been reported to have anticancer properties in vitro but only a few of them show potential as anticancer compounds in vivo. The current study was to evaluate the mechanism of cytotoxicity of selected indolic compound in vivo and in vitro. In this context, we determined the potency of the compound in the induction of apoptosis, cell cycle arrest, and cytoskeleton rearrangement. HT-29, WiDr, CCD-18Co, human monocyte/macrophage CRL-9855, and B lymphocyte CCL-156 cell lines were used to determine the IC50 of the compound using the MTT assay. Analysis of apoptosis was carried out using immunofluorescence, acridine orange/ propidium iodide double staining, Annexin-V-FITC assay, evaluation of the translocation of NF-kB, oxygen radical antioxidant capacity, quenching of reactive oxygen species content, measurement of LDH release, caspase-3/-7, -8 and -9 assays and western blotting. The cell cycle arrest was examined using flowcytometry and gene expression was assessed using qPCR array. Results displayed a potent suppressive effect on HT-29 and WiDr after 24 h of treatment with IC50 value of 2.52±0.34 µg/ml and 2.13±0.65 µg/ml respectively. This cytotoxic effect on normal, monocyte/macrophage and B-cells was insignificant. Dipping in the mitochondrial membrane potential and increased release of cytochrome c from the mitochondria indicated induction of the intrinsic apoptosis pathway by the compound. Activation of this pathway was further evidenced by significant activation of caspase-9 and 3/7. The compound was also shown to activate the extrinsic pathways of apoptosis via activation of caspase-8 which is linked to the suppression of NF-kB translocation to the nucleus. Cell cycle arrest in the G1 phase and up-regulation of glutathione reductase, based on excessive ROS production were also observed. These findings were further investigated for inhibitory efficiency of the compound on colonic aberrant crypt foci in male rats. Rats were divided in to 5 groups: vehicle, cancer control, positive control groups and the groups treated with 25 and 50 mg/kg of compounds for 10 weeks. Administration of compound suppressed total colonic ACF formation up to 73.4%. The results also showed that treatment with the compound significantly reduced the level of malondialdehyde while increasing superoxide dismutase and catalase activities. Furthermore, the down-regulation of PCNA and Bcl2 and the up-regulation of Bax was confirmed by immunohistochemical staining. The outcome of this study suggest sthat the indolic compound is a potent anti-cancer agent against colon cancer and can be further evaluated by animal trial.

Keywords: indolic compound, chemoprevention, crypt, azoxymethane, colon cancer

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Authors: Evgenia Papathanasiou, Georgios Kokkotis, Georgios Axiaris, Theodoros Argyropoulos, Nikos Viazis, Olga Giouleme, Konstantinos Gkoumas, Αnthia Gatopoulou, Αggelos Theodoulou, Georgios Theocharis, Αngeliki Theodoropoulou, Μaria Κalogirou, Pantelis Karatzas, Κonstantinos Κatsanos, Theodora Kafetzi, Κonstantinos Κarmiris, Αnastasia Κourikou, Ιoannis E Κoutroubakis, Christos Liatsos, Gerassimos J. Mantzaris, Νicoletta Μathou, Georgia Bellou, George Michalopoulos Αikaterini Μantaka, Penelope Nikolaou, Μichael Oikonomou, Dimitrios Polymeros, George Papatheodoridis, Εvdoxia Stergiou, Κonstantinos Soufleris, Εpameinondas Skouloudis, Μaria Tzouvala, Georgia Tsiolakidou, Εftychia Tsironi, Styliani Tsafaraki, Kalliopi Foteinogiannopoulou, Konstantina Chalakatevaki, Αngeliki Christidou, Dimitrios K. Christodoulou, Giorgos Bamias, Spyridon Michopoulos, Εvanthia Zampeli

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Background: Inflammatory bowel disease (IBD) commonly affects female patients of reproductive age, making the interaction between fertility, pregnancy and IBD an important issue in disease management. The effect of disease activity on the outcome of pregnancy and its impact on neonatal growth is a field of intense research. Close follow-up of pregnant IBD patients by a multidisciplinary team improves maternal and neonatal outcomes. Aim – Methods: Α national retrospective study of pregnancies in women with IBD between 2010-2020 was carried out in 22 IBD reference centers in Greece. Patient characteristics such as disease profile, type of treatment, and disease activity during gestation were analyzed in correlation to the method of delivery, pregnancy outcomes, as well as breastfeeding and offspring health. Results: Two-hundred and twenty-three pregnancies in 175 IBD patients were registered in the study. 122 with Crohn’s disease (CD). Median age during diagnosis was 25.6 years (12-44), with median disease duration of 7.4 years (0-23). One-hundred and twenty-nine patients (58%) were recorded during their first pregnancy. Early pregnancy termination was reported by 48 patients (22%). Pregnancy as a result of in vitro fertilization (IVF) occurred in 15 cases (6.7%). At the beginning of gestation, 165 patients (74%) were under treatment: 48 with anti-TNF agents (29%), 43 with azathioprine (26%), 101 with 5-aminosalicylic acid formulations (61%) and 12 with steroids (7%). We recorded 49 cases of IBD flares (22%) during pregnancy. Two-thirds of them (n=30) were in remission at the onset of the pregnancy. Almost half of them (n=22) required corticosteroid treatment. Patients with ulcerative colitis (UC) were in greater risk of disease flare during pregnancy (p<0.001). All but 3 pregnancies (99.1%) resulted in uncomplicated delivery. In 147 cases (67.1%), cesarean delivery was performed. Two late fetal deaths (0.9%) were reported, both in patients with continuously active disease since the beginning of pregnancy. After delivery, 75 patients (34%) presented with a disease flare, which was associated with active disease at the beginning of pregnancy (p <0.001). Conclusion: The majority of female, Greek IBD patients, had a favorable pregnancy outcome. Active inflammation during gestation and UC diagnosis were associated with a negative impact on pregnancy outcomes. The results of this study are in favor of the continuation of IBD treatment during pregnancy.

Keywords: pregnancy, ulcerative colitis, Crohn disease, flare

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Authors: A. K. Tursunova, O. V. Chebonenko, A. Zh. Amirkulova, A. O. Abaildayev, O. A. Sapko, Y. M. Dyo, A. Sh. Utarbaeva

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Fusarium solani and its variants cause root and stem rot of plants. Dry rot is the most common disease of potato tubers during storage. The causative agents of fusariosis in contact with plants behave as antagonists, growth stimulants or parasites. The diversity of host-parasite relationships is explained by the parasite’s ability to produce a wide spectrum of biologically active compounds including toxins, enzymes, oligosaccharides, antibiotic substances, enniatins and gibberellins. Many of these metabolites contribute to the creation of compatible relations; others behave as elicitors, inducing various protective responses in plants. An important part of the strategy for developing plant resistance against pathogens is the activation of protein synthesis to produce protective ‘pathogenesis-related’ proteins. The family of PR-proteins known to confer the most protective response is chitinases (EC 3.2.1.14, Cht) and β-1,3-glucanases (EC 3.2.1.39, Glu). PR-proteins also include a large multigene family of peroxidases (EC 1.11.1.7, Pod), and increased activity of Pod and expression of the Pod genes leads to the development of resistance to a broad class of pathogens. Despite intensive research on the role of PR-proteins, the question of their participation in the mechanisms of formation of the F.solani–S.tuberosum pathosуstem is not sufficiently studied. Our aim was to investigate the effect of different classes of F. solani metabolites on the activity of chitinase, β-1,3-glucanases and peroxidases in tubers of Solanum tuberosum. Metabolite culture filtrate (CF) and cytoplasmic components were fractionated by extraction of the mycelium with organic solvents, salting out techniques, dialysis, column chromatography and ultrafiltration. Protein, lipid, carbohydrate and polyphenolic fractions of fungal metabolites were derived. Using enzymatic hydrolysis we obtained oligo glycans from fungal cell walls with different molecular weights. The activity of the metabolites was tested using potato tuber discs (d = 16mm, h = 5mm). The activity of PR-proteins of tubers was analyzed in a time course of 2–24 hours. The involvement of the analysed metabolites in the modulation of both early non-specific and late related to pathogenesis reactions was demonstrated. The most effective inducer was isolated from the CF (fraction of total phenolic compounds including naphtazarins). Induction of PR-activity by this fraction was: chitinase - 340-360%, glucanase - 435-450%, soluble forms of peroxidase - 400-560%, related forms of peroxidase - 215-237%. High-inducing activity was observed by the chloroform and acetonitrile extracts of the mycelium (induction of chitinase and glucanase activity was 176-240%, of soluble and bound forms of peroxidase - 190-400%). The fraction of oligo glycans mycelium cell walls of 1.2 kDa induced chitinase and β-1,3-glucanase to 239-320%; soluble forms and related peroxidase to 198-426%. Oligo glycans cell walls of 5-10 kDa had a weak suppressor effect - chitinase (21-25%) and glucanase (25-28%) activity; had no effect on soluble forms of peroxidase, but induced to 250-270% activity related forms. The CF polysaccharides of 8.5 kDa and 3.1 kDa inhibited synchronously the glucanase and chitinase specific response in step (after 24 hours at 42-50%) and the step response induced nonspecific peroxidase activity: soluble forms 4.8 -5.2 times, associated forms 1.4-1.6 times.

Keywords: fusarium solani, PR-proteins, peroxidase, solanum tuberosum

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Authors: Serhat Tüzün, Tufan Demirel

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Decision Support Systems (DSS) have been investigated by researchers and technologists for more than 35 years. This paper analyses the developments in the architecture and software of these systems, provides a systematic analysis for different Web-based DSS approaches and Intelligent Decision-making Technologies (IDT), with the suggestion for future studies. Decision Support Systems literature begins with building model-oriented DSS in the late 1960s, theory developments in the 1970s, and the implementation of financial planning systems and Group DSS in the early and mid-80s. Then it documents the origins of Executive Information Systems, online analytic processing (OLAP) and Business Intelligence. The implementation of Web-based DSS occurred in the mid-1990s. With the beginning of the new millennia, intelligence is the main focus on DSS studies. Web-based technologies are having a major impact on design, development and implementation processes for all types of DSS. Web technologies are being utilized for the development of DSS tools by leading developers of decision support technologies. Major companies are encouraging its customers to port their DSS applications, such as data mining, customer relationship management (CRM) and OLAP systems, to a web-based environment. Similarly, real-time data fed from manufacturing plants are now helping floor managers make decisions regarding production adjustment to ensure that high-quality products are produced and delivered. Web-based DSS are being employed by organizations as decision aids for employees as well as customers. A common usage of Web-based DSS has been to assist customers configure product and service according to their needs. These systems allow individual customers to design their own products by choosing from a menu of attributes, components, prices and delivery options. The Intelligent Decision-making Technologies (IDT) domain is a fast growing area of research that integrates various aspects of computer science and information systems. This includes intelligent systems, intelligent technology, intelligent agents, artificial intelligence, fuzzy logic, neural networks, machine learning, knowledge discovery, computational intelligence, data science, big data analytics, inference engines, recommender systems or engines, and a variety of related disciplines. Innovative applications that emerge using IDT often have a significant impact on decision-making processes in government, industry, business, and academia in general. This is particularly pronounced in finance, accounting, healthcare, computer networks, real-time safety monitoring and crisis response systems. Similarly, IDT is commonly used in military decision-making systems, security, marketing, stock market prediction, and robotics. Even though lots of research studies have been conducted on Decision Support Systems, a systematic analysis on the subject is still missing. Because of this necessity, this paper has been prepared to search recent articles about the DSS. The literature has been deeply reviewed and by classifying previous studies according to their preferences, taxonomy for DSS has been prepared. With the aid of the taxonomic review and the recent developments over the subject, this study aims to analyze the future trends in decision support systems.

Keywords: decision support systems, intelligent decision-making, systematic analysis, taxonomic review

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Authors: Christine P. Gibson, Ramona Alex, Michael Farney, Johana Vallejo-Elias, Mitra Esfandiarei

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Aortic aneurysm is the leading cause of death in Marfan syndrome (MFS), a connective tissue disorder caused by mutations in fibrillin-1 gene (FBN1). MFS aneurysm is characterized by weakening of the aortic wall due to elastin fibers fragmentation and disorganization. The above-average height and distinct physical features make young adults with MFS desirable candidates for competitive sports; but little is known about the exercise limit at which they will be at risk for aortic rupture. On the other hand, aerobic cardiovascular exercise has been shown to have protective effects on the heart and aorta. We have previously reported that mild aerobic exercise can delay the formation of aortic aneurysm in a mouse model of MFS. In this study, we aimed to investigate the effects of various levels of exercise intensity on the progression of aortic aneurysm in the mouse model. Starting at 4 weeks of age, we subjected control and MFS mice to different levels of exercise intensity (8m/min, 10m/min, 15m/min, and 20m/min, corresponding to 55%, 65%, 75%, and 85% of VO2 max, respectively) on a treadmill for 30 minutes per day, five days a week for the duration of the study. At 24 weeks of age, aortic tissue were isolated and subjected to structural and functional studies using histology and wire myography in order to evaluate the effects of different exercise routines on elastin fragmentation and organization and aortic wall elasticity/stiffness. Our data shows that exercise training at the intensity levels between 55%-75% significantly reduces elastin fragmentation and disorganization, with less recovery observed in 85% MFS group. The reversibility of elasticity was also significantly restored in MFS mice subjected to 55%-75% intensity; however, the recovery was less pronounced in MFS mice subjected to 85% intensity. Furthermore, our data shows that smooth muscle cells (SMCs) contractilion in response to vasoconstrictor agent phenylephrine (100nM) is significantly reduced in MFS aorta (54.84 ± 1.63 mN/mm2) as compared to control (95.85 ± 3.04 mN/mm2). At 55% of intensity, exercise did not rescue SMCs contraction (63.45 ± 1.70 mN/mm2), while at higher intensity levels, SMCs contraction in response to phenylephrine was restored to levels similar to control aorta [65% (81.88 ± 4.57 mN/mm2), 75% (86.22 ± 3.84 mN/mm2), and 85% (83.91 ± 5.42 mN/mm2)]. This study provides the first time evidence that high intensity exercise (e.g. 85%) may not provide the most beneficial effects on aortic function (vasoconstriction) and structure (elastin fragmentation, aortic wall elasticity) during the progression of aortic aneurysm in MFS mice. On the other hand, based on our observations, medium intensity exercise (e.g. 65%) seems to provide the utmost protective effects on aortic structure and function in MFS mice. These findings provide new insights into the potential capacity, in which MFS patients could participate in various aerobic exercise routines, especially in young adults affected by cardiovascular complications particularly aortic aneurysm. This work was funded by Midwestern University Research Fund.

Keywords: aerobic exercise, aortic aneurysm, aortic wall elasticity, elastin fragmentation, Marfan syndrome

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Authors: Ojochenemi J. Onubi, Debbi Marais, Lorna Aucott, Friday Okonofua, Amudha Poobalan

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Background: Obesity is a worldwide epidemic with major health and economic consequences. Pregnancy is a trigger point for the development of obesity, and maternal obesity is associated with significant adverse effects in the mother and child. Nigeria is experiencing a double burden of under- and over-nutrition with rising levels of obesity particularly in women. However, there is scarcity of data on maternal obesity in Nigeria and other African countries. Aims and Objectives: This project aimed at identifying crucial components of potential interventions for maternal obesity in Nigeria. The objectives were to assess the prevalence, effects, and distribution of maternal obesity; knowledge, attitude and practice (KAP) of pregnant women and maternal healthcare providers; and identify existing interventions for maternal obesity in Nigeria. Methodology: A systematic review and meta-analysis were initially conducted to appraise the existing literature on maternal obesity in Africa. Following this, a quantitative questionnaire survey of the KAP of pregnant women and a qualitative interview study of the KAP of Health Care Workers (HCW) were conducted in seven secondary and tertiary hospitals across Nigeria. Quantitative data was analysed using SPSS statistical software, while thematic analysis was conducted for qualitative data. Results: Twenty-nine studies included in the systematic review showed significant prevalence, socio-demographic associations, and adverse effects of maternal obesity on labour, maternal, and child outcomes in Africa. The questionnaire survey of 435 mothers revealed a maternal obesity prevalence of 17.9% among mothers who registered for antenatal care in the first trimester. The mothers received nutrition information from different sources and had insufficient knowledge of their own weight category or recommended Gestational Weight Gain (GWG), causes, complications, and safe ways to manage maternal obesity. However, majority of the mothers were of the opinion that excess GWG is avoided in pregnancy and some practiced weight management (diet and exercise) during pregnancy. For the qualitative study, four main themes were identified: ‘Concerns about obesity in pregnancy’, ‘Barriers to care for obese pregnant women’, ‘Practice of care for obese pregnant women’, and ‘Improving care for obese pregnant women’. HCW expressed concerns about rising levels of maternal obesity, lack of guidelines for the management of obese pregnant women and worries about unintended consequences of antenatal interventions. ‘Barriers’ included lack of contact with obese women before pregnancy, late registration for antenatal care, and perceived maternal barriers such as socio-cultural beliefs of mothers and poverty. ‘Practice’ included anticipatory care and screening for possible complications, general nutrition education during antenatal care and interdisciplinary care for mothers with complications. HCW offered suggestions on improving care for obese women including timing, type, and settings of interventions; and the need for involvement of other stake holders in caring for obese pregnant women. Conclusions: Culturally adaptable/sensitive interventions should be developed for the management of obese pregnant women in Africa. Education and training of mothers and health care workers, and provision of guidelines are some of the components of potential interventions in Nigeria.

Keywords: Africa, maternal, obesity, pregnancy

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Authors: Anantha S. Babbili

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This paper analyzes the current media practices dominant in global journalistic practices within the framework of world press theories of Libertarian, Authoritarian, Communist, and Social Responsibility to evaluate their efficacy in addressing their role in the coverage of the coronavirus, also known as COVID-19. The global media flows, determinants of news coverage, and international awareness and the Western view of the world will be critically analyzed within the context of the prevalent news values that underpin free press and media coverage of the world. While evaluating the global discourse paramount to a sustained and dispassionate understanding of world events, this paper proposes an ethical framework that brings clarity devoid of sensationalism, partisanship, right-wing and left-wing interpretations to a breaking and dangerous development of a pandemic. As the world struggles to contain the coronavirus pandemic with death climbing close to 6,000 from late January to mid-March, 2020, the populations of the developed as well as the developing nations are beset with news media renditions of the crisis that are contradictory, confusing and evoking anxiety, fear and hysteria. How are we to understand differing news standards and news values? What lessons do we as journalism and mass media educators, researchers, and academics learn in order to construct a better news model and structure of media practice that addresses science, health, and media literacy among media practitioners, journalists, and news consumers? As traditional media struggles to cover the pandemic to its audience and consumers, social media from which an increasing number of consumers get their news have exerted their influence both in a positive way and in a negative manner. Even as the world struggles to grasp the full significance of the pandemic, the World Health Organization (WHO) has been feverishly battling an additional challenge related to the pandemic in what it termed an 'infodemic'—'an overabundance of information, some accurate and some not, that makes it hard for people to find trustworthy sources and reliable guidance when they need it.' There is, indeed, a need for journalism and news coverage in times of pandemics that reflect social responsibility and ethos of public service journalism. Social media and high-tech information corporations, collectively termed GAMAF—Google, Apple, Microsoft, Amazon, and Facebook – can team up with reliable traditional media—newspapers, magazines, book publishers, radio and television corporates—to ease public emotions and be helpful in times of a pandemic outbreak. GAMAF can, conceivably, weed out sensational and non-credible sources of coronavirus information, exotic cures offered for sale on a quick fix, and demonetize videos that exploit peoples’ vulnerabilities at the lowest ebb. Credible news of utility delivered in a sustained, calm, and reliable manner serves people in a meaningful and helpful way. The world’s consumers of news and information, indeed, deserve a healthy and trustworthy news media – at least in the time of pandemic COVID-19. Towards this end, the paper will propose a practical model for news media and journalistic coverage during times of a pandemic.

Keywords: COVID-19, international news flow, social media, social responsibility

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Authors: Kimberley Adamek

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According to the Council on Tall Buildings, there has been a rapid increase in the construction of tall or “megatall” buildings over the past two decades. Simultaneously, the New England Journal of Medicine has reported that there has been a steady increase in climate related natural disasters since the 1970s; the eastern expansion of the USA's infamous Tornado Alley being just one of many current issues. In the future, this could mean that tall buildings, which already guide high speed winds down to pedestrian levels would have to withstand stronger forces and protect pedestrians in more extreme ways. Although many projects are required to be verified within wind tunnels and a handful of cities such as San Francisco have included wind testing within building code standards, there are still many examples where wind is only considered for basic loading. This typically results in and an increase of structural expense and unwanted mitigation strategies that are proposed late within a project. When building cities, architects rarely consider how each building alters the invisible patterns of wind and how these alterations effect other areas in different ways later on. It is not until these forces move, overpower and even destroy cities that people take notice. For example, towers have caused winds to blow objects into people (Walkie-Talkie Tower, Leeds, England), cause building parts to vibrate and produce loud humming noises (Beetham Tower, Manchester), caused wind tunnels in streets as well as many other issues. Alternatively, there exist towers which have used their form to naturally draw in air and ventilate entire facilities in order to eliminate the needs for costly HVAC systems (The Met, Thailand) and used their form to increase wind speeds to generate electricity (Bahrain Tower, Dubai). Wind and weather exist and effect all parts of the world in ways such as: Science, health, war, infrastructure, catastrophes, tourism, shopping, media and materials. Working in partnership with a leading wind engineering company RWDI, a series of tests, images and animations documenting discovered interactions of different building forms with wind will be collected to emphasize the possibilities for wind use to architects. A site within San Francisco (due to its increasing tower development, consistently wind conditions and existing strict wind comfort criteria) will host a final design. Iterations of this design will be tested within the wind tunnel and computational fluid dynamic systems which will expose, utilize and manipulate wind flows to create new forms, technologies and experiences. Ultimately, this thesis aims to question the amount which the environment is allowed to permeate building enclosures, uncover new programmatic possibilities for wind in buildings, and push the boundaries of working with the wind to ensure the development and safety of future cities. This investigation will improve and expand upon the traditional understanding of wind in order to give architects, wind engineers as well as the general public the ability to broaden their scope in order to productively utilize this living phenomenon that everyone constantly feels but cannot see.

Keywords: wind engineering, climate, visualization, architectural aerodynamics

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Authors: Raha Bahreini

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This paper discusses the roll-back of women’s sexual and reproductive rights in the Islamic Republic of Iran, which has come in the wake of a striking shift in the country’s official population policies. Since the late 1980s, Iran has won worldwide praise for its sexual and reproductive health and services, which have contributed to a steady decline in the country’s fertility rate–from 7.0 births per women in 1980 to 5.5 in 1988, 2.8 in 1996 and 1.85 in 2014. This is owed to a significant increase in the voluntary use of modern contraception in both rural and urban areas. In 1976, only 37 per cent of women were using at least one method of contraception; by 2014 this figure had reportedly risen to a high of nearly 79 per cent for married girls and women living in urban areas and 73.78 per cent for those living in rural areas. Such progress may soon be halted. In July 2012, Iran’s Supreme Leader Ayatollah Sayed Ali Khamenei denounced Iran’s family planning policies as an imitation of Western lifestyle. He exhorted the authorities to increase Iran’s population to 150 to 200 million (from around 78.5 million), including by cutting subsidies for contraceptive methods and dismantling the state’s Family and Population Planning Programme. Shortly thereafter, Iran’s Minister of Health and Medical Education announced the scrapping of the budget for the state-funded Family and Population Planning Programme. Iran’s Parliament subsequently introduced two bills; the Comprehensive Population and Exaltation of Family Bill (Bill 315), and the Bill to Increase Fertility Rates and Prevent Population Decline (Bill 446). Bill 446 outlaws voluntary tubectomies, which are believed to be the second most common method of modern contraception in Iran, and blocks access to information about contraception, denying women the opportunity to make informed decisions about the number and spacing of their children. Coupled with the elimination of state funding for Iran’s Family and Population Programme, the move would undoubtedly result in greater numbers of unwanted pregnancies, forcing more women to seek illegal and unsafe abortions. Bill 315 proposes various discriminatory measures in the areas of employment, divorce, and protection from domestic violence in order to promote a culture wherein wifedom and child-bearing is seen as women’s primary duty. The Bill, for example, instructs private and public entities to prioritize, in sequence, men with children, married men without children and married women with children when hiring for certain jobs. It also bans the recruitment of single individuals as family law lawyers, public and private school teachers and members of the academic boards of universities and higher education institutes. The paper discusses the consequences of these initiatives which would, if continued, set the human rights of women and girls in Iran back by decades, leaving them with a future shaped by increased inequality, discrimination, poor health, limited choices and restricted freedoms, in breach of Iran’s international human rights obligations.

Keywords: family planning and reproductive health, gender equality and empowerment of women, human rights, population growth

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Authors: Eva Filová, Jana Daňková, Věra Sovková, Matej Daniel

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Titanium alloys are biocompatible metals that are widely used in clinical practice as load bearing implants. The chemical modification may influence cell adhesion, proliferation, and differentiation as well as stiffness of the material. The aim of the study was to evaluate the adhesion, growth and differentiation of pig mesenchymal stem cells on the novel beta titanium alloy Ti36Nb6Ta compared to standard medical titanium alloy Ti6Al4V. Discs of Ti36Nb6Ta and Ti6Al4V alloy were sterilized by ethanol, put in 48-well plates, and seeded by pig mesenchymal stem cells at the density of 60×103/cm2 and cultured in Minimum essential medium (Sigma) supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cell viability was evaluated using MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay;Promega), cell proliferation using Quant-iT™ ds DNA Assay Kit (Life Technologies). Cells were stained immunohistochemically using monoclonal antibody beta-actin, and secondary antibody conjugated with AlexaFluor®488 and subsequently the spread area of cells was measured. Cell differentiation was evaluated by alkaline phosphatase assay using p-nitrophenyl phosphate (pNPP) as a substrate; the reaction was stopped by NaOH, and the absorbance was measured at 405 nm. Osteocalcin, specific bone marker was stained immunohistochemically and subsequently visualized using confocal microscopy; the fluorescence intensity was analyzed and quantified. Moreover, gene expression of osteogenic markers osteocalcin and type I collagen was evaluated by real-time reverse transcription-PCR (qRT-PCR). For statistical evaluation, One-way ANOVA followed by Student-Newman-Keuls Method was used. For qRT-PCR, the nonparametric Kruskal-Wallis Test and Dunn's Multiple Comparison Test were used. The absorbance in MTS assay was significantly higher on titanium alloy Ti6Al4V compared to beta titanium alloy Ti36Nb6Ta on days 7 and 14. Mesenchymal stem cells were well spread on both alloys, but no difference in spread area was found. No differences in alkaline phosphatase assay, fluorescence intensity of osteocalcin as well as the expression of type I collagen, and osteocalcin genes were observed. Higher expression of type I collagen compared to osteocalcin was observed for cells on both alloys. Both beta titanium alloy Ti36Nb6Ta and titanium alloy Ti6Al4V Ti36Nb6Ta supported mesenchymal stem cellsˈ adhesion, proliferation and osteogenic differentiation. Novel beta titanium alloys Ti36Nb6Ta is a promising material for bone implantation. The project was supported by the Czech Science Foundation: grant No. 16-14758S, the Grant Agency of the Charles University, grant No. 1246314 and by the Ministry of Education, Youth and Sports NPU I: LO1309.

Keywords: beta titanium, cell growth, mesenchymal stem cells, titanium alloy, implant

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Authors: Chang-Hui Shen, Paulina Konarzewska

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Cell homeostasis is regulated by vacuolar activity and it has been shown that lipid composition of the vacuole plays an important role in vacuolar function. The major phosphoinositide species present in the vacuolar membrane include phosphatidylinositol 3,5-biphosphate (PI(3,5)P₂) which is generated from PI(3)P controlled by Fab1p. Deletion of FAB1 gene reduce the synthesis of PI(3,5)P₂ and thus result in enlarged or fragmented vacuoles, with neutral vacuolar pH due to reduced vacuolar H⁺-ATPase activity. These mutants also exhibited poor growth at high extracellular pH and in the presence of CaCl₂. Conversely, VPS34 regulates the synthesis of PI(3)P from phosphatidylinositol (PI), and the lack of Vps34p results in the reduction of vacuolar activity. Although the cellular observations are clear, it is still unknown about the molecular mechanism between the phospholipid biosynthesis pathway and vacuolar activity. Since both VPS34 and FAB1 are important in vacuolar activity, we hypothesize that the molecular mechanism of vacuolar function might be regulated by the transcriptional regulators of phospholipid biosynthesis. In this study, we study the role of the major phospholipid biosynthesis transcription factor, INO2, in the regulation of vacuolar activity. We first performed qRT-PCR to examine the effect of Ino2p on the expression of VPS34 and FAB1. Our results showed that VPS34 was upregulated in the presence of inositol for both WT and ino2Δ cells. However, FAB1 was only upregulated significantly in ino2Δ cells. This indicated that Ino2p might be the negative regulator for FAB1 expression. Next, growth sensitivity experiment showed that WT, vma3Δ, and ino2Δ grew well in growth medium buffered to pH 5.5 containing 10 mM CaCl₂. As cells were switched to growth medium buffered to pH 7 containing CaCl₂ WT, ino2Δ and opi1Δ showed growth reduction, whereas vma3Δ was completely nonviable. As the concentration of CaCl₂ was increased to 60 mM, ino2Δ cells showed moderate growth reduction compared to WT. This result suggests that ino2Δ cells have better vacuolar activity. Microscopic analysis and vacuolar acidification were employed to further elucidate the importance of INO2 in vacuolar homeostasis. Analysis of vacuolar morphology indicated that WT and vma3Δ cells displayed vacuoles that occupied a small area of the cell when grown in media buffered to pH 5.5. Whereas, ino2Δ displayed fragmented vacuoles. On the other hand, all strains grown in media buffered to pH 7, exhibited enlarged vacuoles that occupied most of the cell’s surface. This indicated that the presence of INO2 may play negative effect in vacuolar morphology when cells are grown in media buffered to pH 5.5. Furthermore, vacuolar acidification assay showed that only vma3Δ cells displayed notably less acidic vacuoles as cells were grown in media buffered to pH 5.5 and pH 7. Whereas, ino2Δ cells displayed more acidic pH compared to WT at pH7. Taken together, our results demonstrated the molecular mechanism of the vacuolar activity regulated by the phospholipid biosynthesis transcription factors Ino2p. Ino2p negatively regulates vacuolar activity through the expression of FAB1.

Keywords: vacuole, phospholipid, homeostasis, Ino2p, FAB1

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Authors: Jim O'brien

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Contemporary global representation and consumption of La Liga across a plethora of media platform outlets has resulted in significant implications for the historical, political and cultural developments which shaped the development of Spanish club football. This has established and reinforced a hierarchy of a small number of teams belonging to or aspiring to belong to a cluster of global elite clubs seeking to imitate the blueprint of the English Premier League in respect of corporate branding and marketing in order to secure a global fan base through success and exposure in La Liga itself and through the Champions League. The synthesis between globalization, global sport and the status of high profile clubs has created radical change within the folkloric iconography of Spanish football. The main focus of this paper is to critically evaluate the consequences of globalization on the rich tapestry at the core of the game’s distinctive history in Spain. The seminal debate underpinning the study considers whether the divergent aspects of globalization have acted as a malevolent force, eroding tradition, causing financial meltdown and reducing much of the fabric of club football to the status of by standers, or have promoted a renaissance of these traditions, securing their legacies through new fans and audiences. The study draws on extensive sources on the history, politics and culture of Spanish football, in both English and Spanish. It also uses primary and archive material derived from interviews and fieldwork undertaken with scholars, media professionals and club representatives in Spain. The paper has four main themes. Firstly, it contextualizes the key historical, political and cultural forces which shaped the landscape of Spanish football from the late nineteenth century. The seminal notions of region, locality and cultural divergence are pivotal to this discourse. The study then considers the relationship between football, ethnicity and identity as a barometer of continuity and change, suggesting that tradition is being reinvented and re-framed to reflect the shifting demographic and societal patterns within the Spanish state. Following on from this, consideration is given to the paradoxical function of ‘El Clasico’ and the dominant duopoly of the FC Barcelona – Real Madrid axis in both eroding tradition in the global nexus of football’s commodification and in protecting historic political rivalries. To most global consumers of La Liga, the mega- spectacle and hyperbole of ‘El Clasico’ is the essence of Spanish football, with cultural misrepresentation and distortion catapulting the event to the global media audience. Finally, the paper examines La Liga as a sporting phenomenon in which elite clubs, cult managers and galacticos serve as commodities on the altar of mass consumption in football’s global entertainment matrix. These processes accentuate a homogenous mosaic of cultural conformity which obscures local, regional and national identities and paradoxically fuses the global with the local to maintain the distinctive hue of La Liga, as witnessed by the extraordinary successes of Athletico Madrid and FC Eibar in recent seasons.

Keywords: Spanish football, globalization, cultural identity, tradition, folklore

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Authors: Luminiţa Duţică, Gheorghe Duţică

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One of the most powerful language innovation in the twentieth century music was the heterophony–hypostasis of the vertical syntax entered into the sphere of interest of many composers, such as George Enescu, Pierre Boulez, Mauricio Kagel, György Ligeti and others. The heterophonic syntax has a history of its growth, which means a succession of different concepts and writing techniques. The trajectory of settling this phenomenon does not necessarily take into account the chronology: there are highly complex primary stages and advanced stages of returning to the simple forms of writing. In folklore, the plurimelodic simultaneities are free or random and originate from the (unintentional) differences/‘deviations’ from the state of unison, through a variety of ornaments, melismas, imitations, elongations and abbreviations, all in a flexible rhythmic and non-periodic/immeasurable framework, proper to the parlando-rubato rhythmics. Within the general framework of the multivocal organization, the heterophonic syntax in elaborate (academic) version has imposed itself relatively late compared with polyphony and homophony. Of course, the explanation is simple, if we consider the causal relationship between the sound vocabulary elements – in this case, the modalism – and the typologies of vertical organization appropriate for it. Therefore, adding up the ‘classic’ pathway of the writing typologies (monody – polyphony – homophony), heterophony - applied equally to the structures of modal, serial or synthesis vocabulary – reclaims necessarily an own macrotemporal form, in the sense of the analogies enshrined by the evolution of the musical styles and languages: polyphony→fugue, homophony→sonata. Concerned about the prospect of edifying a new musical ontology, the composer Ştefan Niculescu experienced – along with the mathematical organization of heterophony according to his own original methods – the possibility of extrapolation of this phenomenon in macrostructural plan, reaching this way to the unique form of ‘synchrony’. Founded on coincidentia oppositorum principle (involving the ‘one-multiple’ binom), the sound architecture imagined by Ştefan Niculescu consists in one (temporal) model / algorithm of articulation of two sound states: 1. monovocality state (principle of identity) and 2. multivocality state (principle of difference). In this context, the heterophony becomes an (auto)generative mechanism, with macrotemporal amplitude, strategy that will be grown by the composer, practically throughout his creation (see the works: Ison I, Ison II, Unisonos I, Unisonos II, Duplum, Triplum, Psalmus, Héterophonies pour Montreux (Homages to Enescu and Bartók etc.). For the present demonstration, we selected one of the most edifying works of Ştefan Niculescu – Simphony II, Opus dacicum – where the form of (heterophony-)synchrony acquires monumental-symphonic features, representing an emblematic case for the complexity level achieved by this type of vertical syntax in the twentieth century music.

Keywords: heterophony, modalism, serialism, synchrony, syntax

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Authors: John G. Skelly, Peter K. Dearden, Thomas W. R. Harrop, Sarah N. Inwood, Joseph Guhlin

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The Argentine stem weevil (ASW; L. bonariensis) is an economically important pasture pest in New Zealand, which causes about $200 million of damage per annum. Microctonus hyperodae (Mh), a parasite of the ASW in its natural range in South America, was introduced into New Zealand to curb the pasture damage caused by the ASW. Mh is an endoparasitic wasp that lays its eggs in the ASW halting its reproduction. Mh was initially successful at preventing ASW proliferation and reducing pasture damage. The effectiveness of Mh has since declined due to decreased parasitism rates and has resulted in increased pasture damage. Although the mechanism through which ASW has developed resistance to Mh has not been discovered, it has been proposed to be due to the different reproductive modes used by Mh and the ASW in New Zealand. The ASW reproduces sexually, whereas Mh reproduces asexually, which has been hypothesised to have allowed the ASW to ‘out evolve’ Mh. Other species within the Microctonus genus reproduce both sexually and asexually. Strains of Microctonus aethiopoides (Ma), a species closely related to Mh, reproduce either by sexual or asexual reproduction. Comparing the genomes of sexual and asexual Microctonus may allow for the identification of the mechanism of asexual reproduction and other characteristics that may improve Mh as a biocontrol agent. The genomes of Mh and three strains of Ma, two of which reproduce sexually and one reproduces asexually, have been sequenced and annotated. The French (MaFR) and Moroccan (MaMO) reproduce sexually, whereas the Irish strain (MaIR) reproduces asexually. Like Mh, The Ma strains are also used as biocontrol agents, but for different weevil species. The genomes of Mh and MaIR were subsequently upgraded using Hi-C, resulting in a set of high quality, highly contiguous genomes. A subset of the genes involved in mitosis and meiosis, which have been identified though the use of Hidden Markov Models generated from genes involved in these processes in other Hymenoptera, have been catalogued in Mh and the strains of Ma. Meiosis and mitosis genes were broadly conserved in both sexual and asexual Microctonus species. This implies that either the asexual species have retained a subset of the molecular components required for sexual reproduction or that the molecular mechanisms of mitosis and meiosis are different or differently regulated in Microctonus to other insect species in which these mechanisms are more broadly characterised. Bioinformatic analysis of the chemoreceptor compliment in Microctonus has revealed some variation in the number of olfactory receptors, which may be related to host preference. Phylogenetic analysis of olfactory receptors highlights variation, which may be able to explain different host range preferences in the Microctonus. Hi-C clustering implies that Mh has 12 chromosomes, and MaIR has 8. Hence there may be variation in gene regulation between species. Genome alignment of Mh and MaIR implies that there may be large scale genome structural variation. Greater insight into the genetics of these agriculturally important group of parasitic wasps may be beneficial in restoring or maintaining their biocontrol efficacy.

Keywords: argentine stem weevil, asexual, genomics, Microctonus hyperodae

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Authors: Alex Konrad

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In 2017, about 4.5 million Muslims live in Germany. They can practice their faith openly, mostly in well-equipped community centers. At the same time, right-wing politicians and media allege that all Muslims tend to be radical and undemocratic. Both perspectives are rooted in an interacting development since the 1970s. German authorities closed the 'King Fahd Academy' international school in Bonn in summer 2017 because they accused the school administration of attracting Islamists. Only 30 years ago, German authorities and labor unions directed their requests for pastoral care of the Muslim communities in Germany to the Turkish and Saudi administrations. This study shows the leading and misleading tracks of Muslim life and its perception in Germany from a historical point of view. Most of the Muslims came as so-called 'Gastarbeiter' (migrant workers) from Turkey and Morocco to West Germany in the 1960s and 1970s. Until the late 1970s, German society recognized them as workforce solely and ignored their religious needs broadly. The Iranian Revolution of 1979 caused widespread hysteria about Islamic radicalization. Likewise, it shifted the German perception of migrant workers in Germany. For the first time, the majority society saw them as religious people. Media and self-proclaimed 'experts' on Islam suspected Muslims in Germany of subversive and undemocratic belief. On the upside, they obtained the opportunity to be heard by German society and authorities. In the ensuing decades, Muslims and Islamophiles fought a discursive struggle against right-wing politicians, 'experts' and media with monolithic views. In the 1990s, Muslims achieved to establish a solid infrastructure of Islamic community center throughout Germany. Their religious life became present and contributed to diversifying the common monolithic images of Muslims as insane fundamentalists in Germany. However, the media and many 'experts' promoted the fundamentalist narrative, which gained more and more acceptance in German society at the same time. This study uses archival sources from German authorities, Islamic communities, together with local and national media to get a close approach to the contemporary historical debates. In addition, contributions by Muslims and Islamophiles in Germany, for example in magazines, event reports, and internal communication, revealing their quotidian struggle for more acceptance are being used as sources. The inclusion of widely publicized books, documentaries and newspaper articles about Islam as a menace to Europe conduces to a balanced analysis of the contemporary debates and views. Theoretically, the study applies the Third Space approach. Muslims in Germany fight the othering by the German majority society. It was their chief purpose not to be marginalized in both spatial meanings, discursively and physically. Therefore, they established realities of life as hybrids in Germany. This study reconstructs the development of the perception of Muslims in Germany. It claims that self-proclaimed experts and politicians with monolithic views maintained the hegemonic discursive positions and coined the German images of Muslims. Nevertheless, Muslims in Germany accomplished that Muslim presence in Germany’s everyday life became an integral part of society and the public sphere. This is how Muslims hybridized religious spaces in Germany.

Keywords: experts, fundamentalism, Germany, hybridization, Islamophobia, migrant workers

Procedia PDF Downloads 200

Authors: Mohamed Emran, Giovanni Pardini, Maria Gispert, Mohamed Rashad

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Land use and abandonment in semi-arid degraded ecosystems may cause regressive dynamics in vegetation cover affecting organic matter contents, soil nutrients and structural stability, thus reducing soil resistance to erosion. Mediterranean areas are generally subjected to climatic fluctuations, which modify soil conditions and hydrological processes, such as runoff and water infiltration within the upper soil horizons. Low erosion rates occur in very fragile and shallow soils with minor clay content progressively decrease organic carbon C and nitrogen N pools in the upper soil horizons. Seven soils were selected representing variant context of land use and abandonment at the Cap de Creus Peninsula, Catalonia, NE Spain, from recent cultivated vines and olive groves, mid abandoned forests standing under cork and pine trees, pasture to late abandoned Cistus and Erica scrubs. The aim of this work was to study the effect of changes in land use and abandonment on the depletion of soil organic carbon and nitrogen transported by runoff water in shallow soils after natural rainfall events during two years with different rainfall patterns (1st year with low rainfall and 2nd year with high rainfall) by i) monitoring the most significant soil erosion parameters at recorded rainfall events, ii) studying the most relevant soil physical and chemical characteristics on seasonal basis and iii) analysing the seasonal trends of depleted carbon and nitrogen and their interaction with soil surface compaction parameters. Significant seasonal variability was observed in the relevant soil physical and chemical parameters and soil erosion parameters in all soils to establish their evolution under land use and abandonment during two years of different rainfall patterns (214 and 487 mm per year), giving important indications on soil response to rainfall impacts. Erosion rates decreased significantly with the increasing of soil C and N under low and high rainfall. In cultivated soils, C and N depletion increased by 144% and 115%, respectively by 13% increase in erosion rates during the 1st year with respect to the 2nd year. Depleted C and N were proportionally higher in soils under vines and olive with vulnerable soil structure and low soil resilience leading to degradation, altering nutrients cycles and causing adverse impact on environmental quality. Statistical analysis underlined that, during the 1st year, soil surface was less effective in preserving stocks of organic resources leading to higher susceptibility to erosion with consequent C and N depletion. During the 2nd year, higher organic reserve and water storage occurred despite the increasing of C and N loss with an effective contribution from soil surface compaction parameters. The overall estimation during the two years indicated clear differences among soils under vines, olive, cork and pines, suggesting on the one hand, that current cultivation practices are inappropriate and that reforestation with pines may delay the achievement of better soil conditions. On the other hand, the natural succession of vegetation under Cistus, pasture and Erica suggests the recovery of good soil conditions.

Keywords: land abandonment, land use, nutrient's depletion, soil erosion

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Authors: Danny Barash

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Riboswitches are RNA genetic control elements that were originally discovered in bacteria and provide a unique mechanism of gene regulation. They work without the participation of proteins and are believed to represent ancient regulatory systems in the evolutionary timescale. One of the biggest challenges in riboswitch research is that many are found in prokaryotes but only a small percentage of known riboswitches have been found in certain eukaryotic organisms. The few examples of eukaryotic riboswitches were identified using sequence-based bioinformatics search methods that include some slight structural considerations. These pattern-matching methods were the first ones to be applied for the purpose of riboswitch detection and they can also be programmed very efficiently using a data structure called affix arrays, making them suitable for genome-wide searches of riboswitch patterns. However, they are limited by their ability to detect harder to find riboswitches that deviate from the known patterns. Several methods have been developed since then to tackle this problem. The most commonly used by practitioners is Infernal that relies on Hidden Markov Models (HMMs) and Covariance Models (CMs). Profile Hidden Markov Models were also carried out in the pHMM Riboswitch Scanner web application, independently from Infernal. Other computational approaches that have been developed include RMDetect by the use of 3D structural modules and RNAbor that utilizes Boltzmann probability of structural neighbors. We have tried to incorporate more sophisticated secondary structure considerations based on RNA folding prediction using several strategies. The first idea was to utilize window-based methods in conjunction with folding predictions by energy minimization. The moving window approach is heavily geared towards secondary structure consideration relative to sequence that is treated as a constraint. However, the method cannot be used genome-wide due to its high cost because each folding prediction by energy minimization in the moving window is computationally expensive, enabling to scan only at the vicinity of genes of interest. The second idea was to remedy the inefficiency of the previous approach by constructing a pipeline that consists of inverse RNA folding considering RNA secondary structure, followed by a BLAST search that is sequence-based and highly efficient. This approach, which relies on inverse RNA folding in general and our own in-house fragment-based inverse RNA folding program called RNAfbinv in particular, shows capability to find attractive candidates that are missed by Infernal and other standard methods being used for riboswitch detection. We demonstrate attractive candidates found by both the moving-window approach and the inverse RNA folding approach performed together with BLAST. We conclude that structure-based methods like the two strategies outlined above hold considerable promise in detecting riboswitches and other conserved RNAs of functional importance in a variety of organisms.

Keywords: riboswitches, RNA folding prediction, RNA structure, structure-based methods

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Authors: Joanna Marie A. Paulino-Morente, Vaneza Valentina L. Penolio, Grace Sabado

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Ovarian cancer is extremely hard to diagnose in its early stages, and those afflicted at the time of diagnosis are typically asymptomatic and in the late stages of the disease, with metastasis to other organs. Ovarian cancers often occur sporadically, with only 5% associated with hereditary mutations. Mutations in the BRCA1 and BRCA2 tumor suppressor genes have been found to be responsible for the majority of hereditary ovarian cancers. One type of ovarian tumor is Malignant Mixed Mullerian Tumor (MMMT), which is a very rare and aggressive type, accounting for only 1% of all ovarian cancers. Reported is a case of a 43-year-old G3P3 (3003), who came into our institution due to a 2-month history of difficulty of breathing. Family history reveals that her eldest and younger sisters both died of ovarian malignancy, with her younger sister having a histopathology report of endometrioid ovarian carcinoma, left ovary stage IIIb. She still has 2 asymptomatic sisters. Physical examination pointed to pleural effusion of right lung, and presence of bilateral ovarian new growth, which had a Sassone score of 13. Admitting Diagnosis was G3P3 (3003), Ovarian New Growth, bilateral, Malignant; Pleural effusion secondary to malignancy. BRCA was requested to establish a hereditary mutation; however, the patient had no funds. Once the patient was stabilized, TAHBSO with surgical staging was performed. Intraoperatively, the pelvic cavity was occupied by firm, irregularly shaped ovaries, with a colorectal metastasis. Microscopic sections from both ovaries and the colorectal metastasis had pleomorphic tumor cells lined by cuboidal to columnar epithelium exhibiting glandular complexity, displaying nuclear atypia and increased nuclear-cytoplasmic ratio, which are infiltrating the stroma, consistent with the features of Malignant Mixed Mullerian Tumor, since MMMT is composed histologically of malignant epithelial and sarcomatous elements. In conclusion, discussed is the clinic-pathological feature of a patient with primary ovarian Malignant Mixed Mullerian Tumor, a rare malignancy comprising only 1% of all ovarian neoplasms. Also, by understanding the hereditary ovarian cancer syndromes and its relation to this patient, it cannot be overemphasized that a comprehensive family history is really fundamental for early diagnosis. The familial association of the disease, given that the patient has two sisters who were diagnosed with an advanced stage of ovarian cancer and succumbed to the disease at a much earlier age than what is reported in the general population, points to a possible hereditary syndrome which occurs in only 5% of ovarian neoplasms. In a low-resource setting, being in a third world country, the following will be recommended for monitoring and/or screening women who are at high risk for developing ovarian cancer, such as the remaining sisters of the patient: 1) Physical examination focusing on the breast, abdomen, and rectal area every 6 months. 2) Transvaginal sonography every 6 months. 3) Mammography annually. 4) CA125 for postmenopausal women. 5) Genetic testing for BRCA1 and BRCA2 will be reserved for those who are financially capable.

Keywords: BRCA, hereditary breast-ovarian cancer syndrome, malignant mixed mullerian tumor, ovarian cancer

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Authors: Isabel N. Sierra-Garcia, Maria J. Ferreira, Sandro Figuereido, Newton Gomes, Helena Silva, Angela Cunha

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Plant-associated microbial communities are considered crucial in the adaptation of halophytes to coastal environments. The plant microbiota can be horizontally acquired from the environment or vertically transmitted from generation to generation via seeds. Recruiting of the microbial communities by the plant is affected by geographical location, soil source, host genotype, and cultivation practice. There is limited knowledge reported on the microbial communities in halophytes the influence of biotic and abiotic factors. In this work, the microbiota associated with the halophyte Salicornia ramosissima was investigated to determine whether the structure of bacterial communities is influenced by host genotype or soil source. For this purpose, two contrasting sites where S. ramosissima is established in the estuarine system of the Ria de Aveiro were investigated. One site corresponds to a natural salt marsh where S. ramosissima plants are present (wild plants), and the other site is a former salt pan that nowadays are subjected to intensive crop production of S. ramosissima (crop plants). Bacterial communities from the rhizosphere, seeds and root endosphere of S. ramossisima from both sites were investigated by sequencing bacterial 16S rRNA gene using the Illumina MiSeq platform. The analysis of the sequences showed that the three plant-associated compartments, rhizosphere, root endosphere, and seed endosphere, harbor distinct microbiomes. However, bacterial richness and diversity were higher in seeds of wild plants, followed by rhizosphere in both sites, while seeds in the crop site had the lowest diversity. Beta diversity measures indicated that bacterial communities in root endosphere and seeds were more similar in both wild and crop plants in contrast to rhizospheres that differed by local, indicating that the recruitment of the similar bacterial communities by the plant genotype is active in regard to the site. Moreover, bacterial communities from the root endosphere and rhizosphere were phylogenetically more similar in both sites, but the phylogenetic composition of seeds in wild and crop sites was distinct. These results indicate that cultivation practices affect the seed microbiome. However, minimal vertical transmission of bacteria from seeds to adult plants is expected. Seeds from the crop site showed higher abundances of Kushneria and Zunongwangia genera. Bacterial members of the classes Alphaprotebacteria and Bacteroidia were the most ubiquitous across sites and compartments and might encompass members of the core microbiome. These findings indicate that bacterial communities associated with S. ramosissima are more influenced by host genotype rather than local abiotic factors or cultivation practices. This study provides a better understanding of the composition of the plant microbiota in S. ramosissima , which is essential to predict the interactions between plant and associated microbial communities and their effects on plant health. This knowledge is useful to the manipulations of these microbial communities to enhance the health and productivity of this commercially important plant.

Keywords: halophytes, plant microbiome, Salicornia ramosissima, agriculture

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Authors: Milu T. Cherian, Sergio C. Chai, Morgan A. Casal, Taosheng Chen

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This study aims to use CINPA1, a recently discovered small-molecule inhibitor of the xenobiotic receptor CAR (constitutive androstane receptor) for understanding the binding modes of CAR and to guide CAR-mediated gene expression profiling studies in human primary hepatocytes. CAR and PXR are xenobiotic sensors that respond to drugs and endobiotics by modulating the expression of metabolic genes that enhance detoxification and elimination. Elevated levels of drug metabolizing enzymes and efflux transporters resulting from CAR activation promote the elimination of chemotherapeutic agents leading to reduced therapeutic effectiveness. Multidrug resistance in tumors after chemotherapy could be associated with errant CAR activity, as shown in the case of neuroblastoma. CAR inhibitors used in combination with existing chemotherapeutics could be utilized to attenuate multidrug resistance and resensitize chemo-resistant cancer cells. CAR and PXR have many overlapping modulating ligands as well as many overlapping target genes which confounded attempts to understand and regulate receptor-specific activity. Through a directed screening approach we previously identified a new CAR inhibitor, CINPA1, which is novel in its ability to inhibit CAR function without activating PXR. The cellular mechanisms by which CINPA1 inhibits CAR function were also extensively examined along with its pharmacokinetic properties. CINPA1 binding was shown to change CAR-coregulator interactions as well as modify CAR recruitment at DNA response elements of regulated genes. CINPA1 was shown to be broken down in the liver to form two, mostly inactive, metabolites. The structure-activity differences of CINPA1 and its metabolites were used to guide computational modeling using the CAR-LBD structure. To rationalize how ligand binding may lead to different CAR pharmacology, an analysis of the docked poses of human CAR bound to CITCO (a CAR activator) vs. CINPA1 or the metabolites was conducted. From our modeling, strong hydrogen bonding of CINPA1 with N165 and H203 in the CAR-LBD was predicted. These residues were validated to be important for CINPA1 binding using single amino-acid CAR mutants in a CAR-mediated functional reporter assay. Also predicted were residues making key hydrophobic interactions with CINPA1 but not the inactive metabolites. Some of these hydrophobic amino acids were also identified and additionally, the differential coregulator interactions of these mutants were determined in mammalian two-hybrid systems. CINPA1 represents an excellent starting point for future optimization into highly relevant probe molecules to study the function of the CAR receptor in normal- and pathophysiology, and possible development of therapeutics (for e.g. use for resensitizing chemoresistant neuroblastoma cells).

Keywords: antagonist, chemoresistance, constitutive androstane receptor (CAR), multi-drug resistance, structure activity relationship (SAR), xenobiotic resistance

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Authors: Ayoti Banerjee, Meenakshi Banerjee

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The study of metamorphic reaction textures is important in contributing to our understanding of the evolution of metamorphic terranes. Where preserved, they provide information on changes in the P-T conditions during the metamorphic history of the rock, and thus allow us to speculate on the P-T-t evolution of the terrane. Mafic dykes have attracted the attention of petrologists because they act as window to mantle. This rock represents a mafic dyke of doleritic composition. It is fine to medium grained in which clinopyroxene are enclosed by the lath shaped plagioclase grains to form spectacular ophitic texture. At places, sub ophitic texture was also observed. Grains of pyroxene and plagioclase show very less deformation typically plagioclase showing deformed lamella along with plagioclase-clinopyroxene-phyric granoblastic fabric within a groundmass of feldspar microphenocrysts and Fe–Ti oxides. Both normal and reverse zoning were noted in the plagioclase laths. The clinopyroxene grains contain exsolved phases such as orthopyroxene, plagioclase, magnetite, ilmenite along the cleavage traces and the orthopyroxene lamella form granules in the periphery of the clinopyroxene grains. Garnet corona also develops preferentially around plagioclase at the contact of clinopyroxene, ilmenite or magnetite. Tiny quartz and K-fs grains showed symplectic intergrowth with garnet at a few places. The product quartz formed along with garnet rims the coronal garnet and the reacting clinopyroxene. Thin amphibole corona formed along the periphery of deformed plagioclase and clinopyroxene occur as patches over the magmatic minerals. The amphibole coronas cannot be assigned to a late magmatic stage and are interpreted as reactive being restricted to the contact between clinopyroxene and plagioclase, thus postdating the crystallization of both. The amphibole and garnet do not share grain boundary in the entire rock and is thus pointing towards simultaneous crystallization. Olivine is absent. Spectacular myrmekitic growth of orthoclase and quartz rimming the plagioclase is consistent with the potash metasomatic effects that is also found in other rocks of this region. These textural features are consistent with a phase of fluid induced metamorphism (retrogression). But the appearance of coronal garnet and amphibole exclusive of each other reflects the participation if Ti as the prime reason. Presence of Ti as a reactant phase is a must for amphibole forming reactions whereas it is not so in case of garnet forming reactions although the reactants are the same plagioclase and clinopyroxene in both cases. These findings are well validated by petrographical and textural analysis. In order to obtain balanced chemical reactions that explain formation of amphibole and garnet in the mafic dyke rocks a matrix operation technique called Singular Value Decomposition (SVD) was adopted utilizing the measured chemical compositions of the minerals. The computer program C-Space was used for this purpose and the required compositional matrix. Data fed to C-Space was after doing cation-calculation of the oxide percentages obtained from EPMA analysis. The Garnet-Clinopyroxene geothermometer yielded a temperature of 650 degrees Celsius. The Garnet-Clinopyroxene-Plagioclase geobarometer and Al-in amphibole yielded roughly 7.5 kbar pressure.

Keywords: corona, dolerite, geothermometer, metasomatism, metamorphic reaction texture, retrogression

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Authors: Waleed Al-Nuaimy, Luke Anastassiou, Manjinder Kainth

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As the process of teaching has evolved with the advent of new technologies over the ages, so has the process of learning. Educators have perpetually found themselves on the lookout for new technology-enhanced methods of teaching in order to increase learning efficiency and decrease ever expanding workloads. Shortly after the invention of the internet, web-based learning started to pick up in the late 1990s and educators quickly found that the process of providing learning material and marking assignments could change thanks to the connectivity offered by the internet. With the creation of early web-based virtual learning environments (VLEs) such as SPIDER and Blackboard, it soon became apparent that VLEs resulted in higher reported computer self-efficacy among students, but at the cost of students being less satisfied with the learning process . It may be argued that the impersonal nature of VLEs, and their limited functionality may have been the leading factors contributing to this reported dissatisfaction. To this day, often faced with the prospects of assigning colossal engineering cohorts their homework and assessments, educators may frequently choose optimally curated assessment formats, such as multiple-choice quizzes and numerical answer input boxes, so that automated grading software embedded in the VLEs can save time and mark student submissions instantaneously. A crucial skill that is meant to be learnt during most science and engineering undergraduate degrees is gaining the confidence in using, solving and deriving mathematical equations. Equations underpin a significant portion of the topics taught in many STEM subjects, and it is in homework assignments and assessments that this understanding is tested. It is not hard to see that this can become challenging if the majority of assignment formats students are engaging with are multiple-choice questions, and educators end up with a reduced perspective of their students’ ability to manipulate equations. Artificial intelligence (AI) has in recent times been shown to be an important consideration for many technologies. In our paper, we explore the use of new AI based software designed to work in conjunction with current VLEs. Using our experience with the software, we discuss its potential to solve a selection of problems ranging from impersonality to the reduction of educator workloads by speeding up the marking process. We examine the software’s potential to increase learning efficiency through its features which claim to allow more customized and higher-quality feedback. We investigate the usability of features allowing students to input equation derivations in a range of different forms, and discuss relevant observations associated with these input methods. Furthermore, we make ethical considerations and discuss potential drawbacks to the software, including the extent to which optical character recognition (OCR) could play a part in the perpetuation of errors and create disagreements between student intent and their submitted assignment answers. It is the intention of the authors that this study will be useful as an example of the implementation of AI in a practical assessment scenario insofar as serving as a springboard for further considerations and studies that utilise AI in the setting and marking of science and engineering assignments.

Keywords: engineering education, assessment, artificial intelligence, optical character recognition (OCR)

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Authors: Yana Posmitnaya, Galina Rudnitskaya, Tatyana Lukashenko, Anton Bukatin, Anatoly Evstrapov

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An important area of biological and medical research is the study of genetic mutations and polymorphisms that can alter gene function and cause inherited diseases and other diseases. The following methods to analyse DNA fragments are used: capillary electrophoresis and electrophoresis on microfluidic chip (MFC), mass spectrometry with electrophoresis on MFC, hybridization assay on microarray. Electrophoresis on MFC allows to analyse small volumes of samples with high speed and throughput. A soft lithography in polydimethylsiloxane (PDMS) was chosen for operative fabrication of MFCs. A master-form from silicon and photoresist SU-8 2025 (MicroChem Corp.) was created for the formation of micro-sized structures in PDMS. A universal topology which combines T-injector and simple cross was selected for the electrophoretic separation of the sample. Glass K8 and PDMS Sylgard® 184 (Dow Corning Corp.) were used for fabrication of MFCs. Electroosmotic flow (EOF) plays an important role in the electrophoretic separation of the sample. Therefore, the estimate of the quantity of EOF and the ways of its regulation are of interest for the development of the new methods of the electrophoretic separation of biomolecules. The following methods of surface modification were chosen to change EOF: high-frequency (13.56 MHz) plasma treatment in oxygen and argon at low pressure (1 mbar); 1% aqueous solution of polyvinyl alcohol; 3% aqueous solution of Kolliphor® P 188 (Sigma-Aldrich Corp.). The electroosmotic mobility was evaluated by the method of Huang X. et al., wherein the borate buffer was used. The influence of physical and chemical methods of treatment on the wetting properties of the PDMS surface was controlled by the sessile drop method. The most effective way of surface modification of MFCs, from the standpoint of obtaining the smallest value of the contact angle and the smallest value of the EOF, was the processing with aqueous solution of Kolliphor® P 188. This method of modification has been selected for the treatment of channels of MFCs, which are used for the separation of mixture of oligonucleotides fluorescently labeled with the length of chain with 10, 20, 30, 40 and 50 nucleotides. Electrophoresis was performed on the device MFAS-01 (IAI RAS, Russia) at the separation voltage of 1500 V. 6% solution of polydimethylacrylamide with the addition of 7M carbamide was used as the separation medium. The separation time of components of the mixture was determined from electropherograms. The time for untreated MFC was ~275 s, and for the ones treated with solution of Kolliphor® P 188 – ~ 220 s. Research of physical-chemical methods of surface modification of MFCs allowed to choose the most effective way for reducing EOF – the modification with aqueous solution of Kolliphor® P 188. In this case, the separation time of the mixture of oligonucleotides decreased about 20%. The further optimization of method of modification of channels of MFCs will allow decreasing the separation time of sample and increasing the throughput of analysis.

Keywords: electrophoresis, microfluidic chip, modification, nucleic acid, polydimethylsiloxane, soft lithography

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Authors: Juergen Junk, Franz Ronellenfitsch, Michael Eickermann

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An appropriate weed management in autumn is a prerequisite for an economically successful harvest in the following year. In Luxembourg oilseed rape, wheat and barley is sown from August until October, accompanied by a chemical weed control with soil active herbicides, depending on the state of the weeds and the meteorological conditions. Based on regular ground and surface water-analysis, high levels of contamination by transformation products of respective herbicide compounds have been found in Luxembourg. The most ideal conditions for incorporating soil active herbicides are single rain events. Weed control may be reduced if application is made when weeds are under drought stress or if repeated light rain events followed by dry spells, because the herbicides tend to bind tightly to the soil particles. These effects have been frequently reported for Luxembourg throughout the last years. In the framework of a multisite long-term field experiment (EFFO) weed monitoring, plants observations and corresponding meteorological measurements were conducted. Long-term time series (1947-2016) from the SYNOP station Findel-Airport (WMO ID = 06590) showed a decrease in the number of days with precipitation. As the total precipitation amount has not significantly changed, this indicates a trend towards rain events with higher intensity. All analyses are based on decades (10-day periods) for September and October of each individual year. To assess the future meteorological conditions for Luxembourg, two different approaches were applied. First, multi-model ensembles from the CORDEX experiments (spatial resolution ~12.5 km; transient projections until 2100) were analysed for two different Representative Concentration Pathways (RCP8.5 and RCP4.5), covering the time span from 2005 until 2100. The multi-model ensemble approach allows for the quantification of the uncertainties and also to assess the differences between the two emission scenarios. Second, to assess smaller scale differences within the country a high resolution model projection using the COSMO-LM model was used (spatial resolution 1.3 km). To account for the higher computational demands, caused by the increased spatial resolution, only 10-year time slices have been simulated (reference period 1991-2000; near future 2041-2050 and far future 2091-2100). Statistically significant trends towards higher air temperatures, +1.6 K for September (+5.3 K far future) and +1.3 K for October (+4.3 K), were predicted for the near future compared to the reference period. Precipitation simultaneously decreased by 9.4 mm (September) and 5.0 mm (October) for the near future and -49 mm (September) and -10 mm (October) in the far future. Beside the monthly values also decades were analyzed for the two future time periods of the CLM model. For all decades of September and October the number of days with precipitation decreased for the projected near and far future. Changes in meteorological variables such as air temperature and precipitation did already induce transformations in weed societies (composition, late-emerging etc.) of arable ecosystems in Europe. Therefore, adaptations of agronomic practices as well as effective weed control strategies must be developed to maintain crop yield.

Keywords: CORDEX projections, dry spells, ensembles, weed management

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Authors: Zhihao Peng, Qiong Zhang, Xiyao Li, Yongzhen Peng

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Nowadays, conventional nitrogen removal process (nitrification and denitrification) was adopted in most wastewater treatment plants, but many problems have occurred, such as: high aeration energy consumption, extra carbon sources dosage and high sludge treatment costs. The emergence of anammox has bring about the great revolution to the nitrogen removal technology, and only the ammonia and nitrite were required to remove nitrogen autotrophically, no demand for aeration and sludge treatment. However, there existed many challenges in anammox applications: difficulty of biomass retention, insufficiency of nitrite substrate, damage from complex organic etc. Much effort was put into the research in overcoming the above challenges, and the payment was rewarded. It was also imperative to establish an innovative process that can settle the above problems synchronously, after all any obstacle above mentioned can cause the collapse of anammox system. Therefore, in this study, a two-stage process was established that the sequencing batch reactor (SBR) and upflow anaerobic sludge blanket (UASB) were used in the pre-stage and post-stage, respectively. The domestic wastewater entered into the SBR first and went through anaerobic/aerobic/anoxic (An/O/A) mode, and the draining at the aerobic end of SBR was mixed with domestic wastewater, the mixture then entering to the UASB. In the long term, organic and nitrogen removal performance was evaluated. All along the operation, most COD was removed in pre-stage (COD removal efficiency > 64.1%), including some macromolecular organic matter, like: tryptophan, tyrosinase and fulvic acid, which could weaken the damage of organic matter to anammox. And the An/O/A operating mode of SBR was beneficial to the achievement and maintenance of partial nitrification (PN). Hence, sufficient and steady nitrite supply was another favorable condition to anammox enhancement. Besides, the flexible mixing ratio helped to gain a substrate ratio appropriate to anammox (1.32-1.46), which further enhance the anammox. Further, the UASB was used and gas recirculation strategy was adopted in the post-stage, aiming to achieve granulation by the selection pressure. As expected, the granules formed rapidly during 38 days, which increased from 153.3 to 354.3 μm. Based on bioactivity and gene measurement, the anammox metabolism and abundance level rose evidently, by 2.35 mgN/gVss·h and 5.3 x109. The anammox bacteria mainly distributed in the large granules (>1000 μm), while the biomass in the flocs (<200 μm) and microgranules (200-500 μm) barely displayed anammox bioactivity. Enhanced anammox promoted the advanced autotrophic nitrogen removal, which increased from 71.9% to 93.4%, even when the temperature was only 12.9 ℃. Therefore, it was feasible to enhance anammox in the multiple favorable conditions created, and the strategy extended the application of anammox to the full-scale mainstream, enhanced the understanding of anammox in the aspects of culturing conditions.

Keywords: anammox, granules, nitrite accumulation, nitrogen removal efficiency

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Authors: Dilnee D. Suraweera, Tim Groom, Brian Chung, Brendan Bond, Andrew Schipp, Marc E. Nicolas

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Pyrethrum (Tanacetum cinerariifolium) is a perennial plant belongs to family Asteraceae. This is cultivated commercially for extraction of natural insecticide pyrethrins, which accumulates in their flower head achenes. Approximately 94% of the pyrethrins are produced within secretory ducts and trichomes of achenes of the mature pyrethrum flower. This is the most widely used botanical insecticide in the world and Australia is the current largest pyrethrum producer in the world. Rainfall in pyrethrum growing regions in Australia during pyrethrum flowering period, in late spring and early summer is significantly less. Due to lack of adequate soil moisture and under elevated temperature conditions during post-anthesis period, resulting in yield reductions. Therefore, understanding of yield responses of pyrethrum to irrigation is important for Pyrethrum as a commercial crop. Irrigation management has been identified as a key area of pyrethrum crop management strategies that could be manipulated to increase yield. Pyrethrum is a comparatively drought tolerant plant and it has some ability to survive in dry conditions due to deep rooting. But in dry areas and in dry seasons, the crop cannot reach to its full yield potential without adequate soil moisture. Therefore, irrigation is essential during the flowering period prevent crop water stress and maximise yield. Irrigation during the water deficit period results in an overall increased rate of water uptake and growth by the plant which is essential to achieve the maximum yield benefits from commercial crops. The effects of irrigation treatments applied at post-anthesis period on pyrethrum yield responses were studied in two irrigation methods. This was conducted in a first harvest commercial pyrethrum field in Waubra, Victoria, during 2012/2013 season. Drip irrigation and overhead sprinkler irrigation treatments applied during whole flowering period were compared with ‘rainfed’ treatment in relation to flower yield and pyrethrin yield responses. The results of this experiment showed that the application of 180mm of irrigation throughout the post-anthesis period, from early flowering stages to physiological maturity under drip irrigation treatment increased pyrethrin concentration by 32%, which combined with the 95 % increase in the flower yield to give a total pyrethrin yield increase of 157%, compared to the ‘rainfed’ treatment. In contrast to that overhead sprinkler irrigation treatment increased pyrethrin concentration by 19%, which combined with the 60 % increase in the flower yield to give a total pyrethrin yield increase of 91%, compared to the ‘rainfed’ treatment. Irrigation treatments applied throughout the post-anthesis period significantly increased flower yield as a result of enhancement of number of flowers and flower size. Irrigation provides adequate soil moisture for flower development in pyrethrum which slows the rate of flower development and increases the length of the flowering period, resulting in a delayed crop harvest (11 days) compared to the ‘rainfed’ treatment. Overall, irrigation has a major impact on pyrethrin accumulation which increases the rate and duration of pyrethrin accumulation resulting in higher pyrethrin yield per flower at physiological maturity. The findings of this study will be important for future yield predictions and to develop advanced agronomic strategies to maximise pyrethrin yield in pyrethrum.

Keywords: achene, drip irrigation, overhead irrigation, pyrethrin

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Authors: Yuriko Takayama, Norihiro Kato

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Quorum sensing (QS) is a cell-to-cell communication system in bacteria to regulate expression of target genes. In gram-negative bacteria, activation on QS is controlled by a concentration increase of N-acylhomoserine lactone (AHL), which can diffuse in and out of the cell. Effective control of QS is expected to avoid virulence factor production in infectious pathogens, biofilm formation, and antibiotic production because various cell functions in gram-negative bacteria are controlled by AHL-mediated QS. In this research, we applied cyclodextrins (CDs) as artificial hosts for the AHL signal to reduce the AHL concentration in the culture broth below its threshold for QS activation. The AHL-receptor complex induced under the high AHL concentration activates transcription of the QS-target gene. Accordingly, artificial reduction of the AHL concentration is one of the effective strategies to inhibit the QS. A hydrophobic cavity of the CD can interact with the acyl-chain of the AHL due to hydrophobic interaction in aqueous media. We studied N-hexanoylhomoserine lactone (C6HSL)-mediated QS in Serratia marcescens; accumulation of C6HSL is responsible for regulation of the expression of pig cluster. Inhibitory effects of added CDs on QS were demonstrated by determination of prodigiosin amount inside cells after reaching stationary phase, because production of prodigiosin depends on the C6HSL-mediated QS. By adding approximately 6 wt% hydroxypropyl-β-CD (HP-β-CD) in Luria-Bertani (LB) medium prior to inoculation of S. maecescens AS-1, the intracellularly accumulated prodigiosin was drastically reduced to 7-10%, which was determined after the extraction of prodigiosin in acidified ethanol. The AHL retention ability of HP-β-CD was also demonstrated by Chromobacterium violacuem CV026 bioassay. The CV026 strain is an AHL-synthase defective mutant that activates QS solely by adding AHLs from outside of cells. A purple pigment violacein is induced by activation of the AHL-mediated QS. We demonstrated that the violacein production was effectively suppressed when the C6HSL standard solution was spotted on a LB agar plate dispersing CV026 cells and HP-β-CD. Physico-chemical analysis was performed to study the affinity between the immobilized CD and added C6HSL using a quartz crystal microbalance (QCM) sensor. The COOH-terminated self-assembled monolayer was prepared on a gold electrode of 27-MHz AT-cut quartz crystal. Mono(6-deoxy-6-N, N-diethylamino)-β-CD was immobilized on the electrode using water-soluble carbodiimide. The C6HSL interaction with the β-CD cavity was studied by injecting the C6HSL solution to a cup-type sensor cell filled with buffer solution. A decrement of resonant frequency (ΔFs) clearly showed the effective C6HSL complexation with immobilized β-CD and its stability constant for MBP-SpnR-C6HSL complex was on the order of 102 M-1. The CD has high potential for engineered control of QS because it is safe for human use.

Keywords: acylhomoserine lactone, cyclodextrin, intracellular signaling, quorum sensing

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Authors: Almudena Espin Perez, Tyler Risom, Carl Pelz, Isabel English, Robert M. Angelo, Rosalie Sears, Andrew J. Gentles

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Pancreatic ductal adenocarcinoma (PDAC) is one of the most deadly malignancies. The role of the tumor microenvironment (TME) is gaining significant attention in cancer research. Despite ongoing efforts, the nature of the interactions between tumors, immune cells, and stromal cells remains poorly understood. The cell-intrinsic properties that govern cell lineage plasticity in PDAC and extrinsic influences of immune populations require technically challenging approaches due to the inherently heterogeneous nature of PDAC. Understanding the cell lineage plasticity of PDAC will improve the development of novel strategies that could be translated to the clinic. Members of the team have demonstrated that the acquisition of ductal to neuroendocrine lineage plasticity in PDAC confers therapeutic resistance and is a biomarker of poor outcomes in patients. Our approach combines computational methods for deconvolving bulk transcriptomic cancer data using CIBERSORTx and high-throughput single-cell imaging using Multiplexed Ion Beam Imaging (MIBI) to study lineage plasticity in PDAC and its relationship to the infiltrating immune system. The CIBERSORTx algorithm uses signature matrices from immune cells and stroma from sorted and single-cell data in order to 1) infer the fractions of different immune cell types and stromal cells in bulked gene expression data and 2) impute a representative transcriptome profile for each cell type. We studied a unique set of 300 genomically well-characterized primary PDAC samples with rich clinical annotation. We deconvolved the PDAC transcriptome profiles using CIBERSORTx, leveraging publicly available single-cell RNA-seq data from normal pancreatic tissue and PDAC to estimate cell type proportions in PDAC, and digitally reconstruct cell-specific transcriptional profiles from our study dataset. We built signature matrices and optimized by simulations and comparison to ground truth data. We identified cell-type-specific transcriptional programs that contribute to cancer cell lineage plasticity, especially in the ductal compartment. We also studied cell differentiation hierarchies using CytoTRACE and predict cell lineage trajectories for acinar and ductal cells that we believe are pinpointing relevant information on PDAC progression. Collaborators (Angelo lab, Stanford University) has led the development of the Multiplexed Ion Beam Imaging (MIBI) platform for spatial proteomics. We will use in the very near future MIBI from tissue microarray of 40 PDAC samples to understand the spatial relationship between cancer cell lineage plasticity and stromal cells focused on infiltrating immune cells, using the relevant markers of PDAC plasticity identified from the RNA-seq analysis.

Keywords: deconvolution, imaging, microenvironment, PDAC

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