Search results for: nucleotide diversity

Authors: Hamidreza Saberkari, Mousa Shamsi, Hossein Ahmadi, Saeed Vaali, , MohammadHossein Sedaaghi

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In the recent years, using signal processing tools for accurate identification of the protein coding regions has become a challenge in bioinformatics. Most of the genomic signal processing methods is based on the period-3 characteristics of the nucleoids in DNA strands and consequently, spectral analysis is applied to the numerical sequences of DNA to find the location of periodical components. In this paper, a novel ensemble algorithm for gene selection in DNA sequences has been presented which is based on the combination of Goertzel algorithm and anti-notch filter (ANF). The proposed algorithm has many advantages when compared to other conventional methods. Firstly, it leads to identify the coding protein regions more accurate due to using the Goertzel algorithm which is tuned at the desired frequency. Secondly, faster detection time is achieved. The proposed algorithm is applied on several genes, including genes available in databases BG570 and HMR195 and their results are compared to other methods based on the nucleotide level evaluation criteria. Implementation results show the excellent performance of the proposed algorithm in identifying protein coding regions, specifically in identification of small-scale gene areas.

Keywords: protein coding regions, period-3, anti-notch filter, Goertzel algorithm

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Authors: Nebiyou Masebo

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The rapid conversion of an old aged agroforestry (AF) based agricultural system to monocropping farming system in southern Ethiopia is increasing. The consequence of this, combined with climate change, has been impaired biodiversity, soil microbial biomass carbon (MBC), and soil organic carbon (SOC). The AF system could curb such problems due it is an ecologically and economically sustainable strategies. This study was aimed to investigate different agroforestry practices (AFPs) on MBC and SOC in southern Ethiopia. Soil samples were collected from homegarden based agroforestry practice (HAFP), crop land based agroforestry practice (ClAFP), woodlot based agroforestry practice (WlAFP), and trees on soil and water conservation based agroforestry practice (TSWAFP) using two depth layer (0-30 & 30-60 cm) by systematic sampling. Moreover, woody species inventorywas also collected. The chloroform fumigation extraction method was employed to determine MBC from different AFP types. In this study, the value of MBC and SOC decreased significantly with soil depth (p< 0.05). Besides, AFP type, soil depth, woody species diversity, and key soil properties also strongly influenced MBC and SOC (p< 0.05). In this study, the MBC was the highest (786 mg kg⁻¹ soil) in HAFP, followed by WlAFP (592 mg kg⁻¹ soil), TSWAFP (421 mg kg⁻¹ soil), and ClAFP (357 mg kg⁻¹ soil). The highest mean value of SOC (43.5Mg C ha⁻¹) was recorded in HAFP, followed by WlAFP (35.1Mg C ha⁻¹), TSWAFP (22.3 Mg C ha⁻¹), while the lowest (21.8 Mg C ha⁻¹) was recorded in ClAFP. The HAFP had high woody species diversity, and the lowest was recorded in ClAFP. The finding indicated that SOC and MBC were significantly affected by land management practices, and HAFP has the potential to improve MBC and SOC through good management practices of AFP.

Keywords: agroforestry practices, microbial biomass carbon, soil carbon, rapid conversion

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Authors: Jean Pierre Kabue, Emma Meader, Afsatou Ndama Traore, Paul R. Hunter, Natasha Potgieter

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Norovirus is now considered the most common cause of outbreaks of nonbacterial gastroenteritis. Limited data are available for Norovirus strains in Africa, especially in rural and peri-urban areas. Despite the excessive burden of diarrhea disease in developing countries, Norovirus infections have been to date mostly reported in developed countries. There is a need to investigate intensively the role of viral agents associated with diarrhea in different settings in Africa continent. To determine the prevalence and genetic diversity of Norovirus strains circulating in the rural communities in the Limpopo Province, South Africa and investigate the genetic relationship between Norovirus strains, a cross-sectional study was performed on human stools collected from rural communities. Between July 2014 and April 2015, outpatient children under 5 years of age from rural communities of Vhembe District, South Africa, were recorded for the study. A total of 303 stool specimens were collected from those with diarrhea (n=253) and without (n=50) diarrhea. NoVs were identified using real-time one-step RT-PCR. Partial Sequence analyses were performed to genotype the strains. Phylogenetic analyses were performed to compare identified NoVs genotypes to the worldwide circulating strains. Norovirus detection rate was 41.1% (104/253) in children with diarrhea. There was no significant difference (OR=1.24; 95% CI 0.66-2.33) in Norovirus detection between symptomatic and asymptomatic children. Comparison of the median CT values for NoV in children with diarrhea and without diarrhea revealed significant statistical difference of estimated GII viral load from both groups, with a much higher viral burden in children with diarrhea. To our knowledge, this is the first study reporting on the differences in estimated viral load of GII and GI NoV positive cases and controls. GII.Pe (n=9) were the predominant genotypes followed by GII.Pe/GII.4 Sydney 2012 (n=8) suspected recombinant and GII.4 Sydney 2012 variants(n=7). Two unassigned GII.4 variants and an unusual RdRp genotype GII.P15 were found. With note, the rare GIIP15 identified in this study has a common ancestor with GIIP15 strain from Japan previously reported as GII/untypeable recombinant strain implicated in a gastroenteritis outbreak. To our knowledge, this is the first report of this unusual genotype in the African continent. Though not confirmed predictive of diarrhea disease in this study, the high detection rate of NoV is an indication of subsequent exposure of children from rural communities to enteric pathogens due to poor sanitation and hygiene practices. The results reveal that the difference between asymptomatic and symptomatic children with NoV may possibly be related to the NoV genogroups involved. The findings emphasize NoV genetic diversity and predominance of GII.Pe/GII.4 Sydney 2012, indicative of increased NoV activity. An uncommon GII.P15 and two unassigned GII.4 variants were also identified from rural settings of the Vhembe District/South Africa. NoV surveillance is required to help to inform investigations into NoV evolution, and to support vaccine development programmes in Africa.

Keywords: asymptomatic, common, outpatients, norovirus genetic diversity, sporadic gastroenteritis, South African rural communities, symptomatic

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Authors: Abraha Hatsey, Nesibu Yahya, Abeje Eshete

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Ethiopian Orthodox Tewahido Church has a long tradition of conserving the Church vegetation and is an area treated as a refugee camp for many endangered indigenous tree species in Northern Ethiopia. Though around 36,000 churches exist in Ethiopia, only a few churches have been studied so far. Thus, this study assessed the land use land cover change of 3km buffer (1986-2018) and the woody species diversity and regeneration status of Tsimur St. Gebriel monastery in Tigray region, Northern Ethiopia. For vegetation study, systematic sampling was used with 100m spacing between plots and between transects. Plot size was 20m*20m for the main plot and 2 subplots (5m*5m each) for the regeneration study. Tree height, diameter at breast height(DBH) and crown area were measured in the main plot for all trees with DBH ≥ 5cm. In the subplots, all seedlings and saplings were counted with DBH < 5cm. The data was analyzed on excel and Pass biodiversity software for diversity and evenness analysis. The major land cover classes identified include bare land, farmland, forest, shrubland and wetland. The extents of forest and shrubland were declined considerably due to bare land and agricultural land expansions within the 3km buffer, indicating an increasing pressure on the church forest. Regarding the vegetation status, A total of 19 species belonging to 13 families were recorded in the monastery. The diversity (H’) and evenness recorded were 2.4 and 0.5, respectively. The tree density (DBH ≥ 5cm) was 336/ha and a crown cover of 65%. Olea europaea was the dominant (6.4m2/ha out of 10.5m2 total basal area) and a frequent species (100%) with good regeneration in the monastery. The rest of the species are less frequent and are mostly confined to water sources with good site conditions. Juniperus procera (overharvested) and the other indigenous species were with few trees left and with no/very poor regeneration status. The species having poor density, frequency and regeneration (Junperus procera, Nuxia congesta Fersen and Jasminium abyssinica) need prior conservation and enrichment planting. The indigenous species could also serve as a potential seed source for the reproduction and restoration of nearby degraded landscapes. The buffer study also demonstrated expansion of agriculture and bare land, which could be a threat to the forest of the isolated monastery. Hence, restoring the buffer zone is the only guarantee for the healthy existence of the church forest.

Keywords: church forests, regeneration, land use change, vegetation status

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Authors: Mona T. Kashef, Omneya M. Helmy

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Multidrug resistant (MDR) bacteria have become a significant public health threat. The prevalence rates, of Gram negative MDR bacteria, are in continuous increase. However, few data are available about these resistant strains. Since, third generation cephalosporins are one of the most commonly used antimicrobials, we set out to investigate the prevalence, different mechanisms and clonal relatedness of multidrug resistance among third generation resistant Gram negative clinical isolates. A total of 114 Gram negative clinical isolates, previously characterized as being resistant to at least one of 3rd generation cephalosporins, were included in this study. Each isolate was tested, using Kirby Bauer disk diffusion method, against its assigned categories of antimicrobials. The role of efflux pump in resistance development was tested by the efflux pump inhibitor-based microplate assay using chloropromazine as an inhibitor. Detecting different aminoglycosides, β-lactams and quinolones resistance genes was done using polymerase chain reaction. The genetic diversity of MDR isolates was investigated using Random Amplification of Polymorphic DNA technique. MDR phenotype was detected in 101 isolates (89%). Efflux pump mediated resistance was detected in 49/101 isolates. Aminoglycosides resistance genes; armA and aac(6)-Ib were detected in one and 53 isolates, respectively. The aac(6)-Ib-cr allele, that also confers resistance to floroquinolones, was detected in 28/53 isolates. β-lactam resistance genes; blaTEM, blaSHV, blaCTX-M group 1 and group 9 were detected in 52, 29, 61 and 35 isolates, respectively. Quinolone resistance genes; qnrA, qnrB and qnrS were detectable in 2, 14, 8 isolates respectively, while qepA was not detectable at all. High diversity was observed among tested MDR isolates. MDR is common among 3rd generation cephalosporins resistant Gram negative bacteria, in Egypt. In most cases, resistance was caused by different mechanisms. Therefore, new treatment strategies should be implemented.

Keywords: gram negative, multidrug resistance, RAPD typing, resistance genes

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Authors: Azusa Katsumata

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Several universities in Japan have introduced activities aimed at the mutual enlightenment of a diversity of people in career education. However, several programs emphasize on delivering results, and on practicing the prepared materials as planned. Few programs focus on unexpected failures and setbacks. This way of learning is important in career education so that classmates can help each other, overcome difficulties, draw out each other’s strengths, and learn from them. Seijo University in Tokyo offered excursion focusing Various Problems of Society, as second year career education course, Students will learn about contraception, infertility, homeless people, LGBT, and they will discuss based on the excursion. This paper aims to study the ‘learning platform’ created by a series of processes such as the excursion, the discussion, and the presentation. In this course, students looked back on their lives and imagined the future in concrete terms, performing tasks in groups. The students came across a range of values through lectures and conversations, thereby developing feelings of self-efficacy. We conducted a questionnaire to measure the development of career in class. From the results of the questionnaire, we can see, in the example of this class, that students respected diversity and understood the importance of uncertainty and discontinuity. Whereas the students developed career awareness, they actually did not come across that scene and would do so only in the future when it became necessary. In this class, students consciously considered social problems, but did not develop the practical skills necessary to deal with these. This is appropriate for one of project, but we need to consider how this can be incorporated into future courses. University constitutes only a single period in life-long career formation. Thus, further research may be indicated to determine whether the positive effects of career education at university continue to contribute to individual careers going forward.

Keywords: career education of university, excursion, learning platform, problems of society

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Authors: Mark Osborn

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Clustered, regularly interspaced short palindromic repeat (CRISPR/Cas) proteins can be designed to bind specified DNA and RNA sequences and hold great promise for the accurate detection of nucleic acids for diagnostics. Commercially available reagents were integrated into a CRISPR/Cas9-based lateral flow assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequences with single-base specificity. This approach requires minimal equipment and represents a simplified platform for field-based deployment. A rapid, multiplex fluorescence CRISPR/Cas9 nuclease cleavage assay capable of detecting and differentiating SARS-CoV-2, influenza A and B, and respiratory syncytial virus in a single reaction was also developed. These findings provide proof of principle for CRISPR/Cas9 point-of-care diagnosis that can detect specific SARS-CoV-2 strain(s). Further, Cas9 cleavage allows for a scalable fluorescent platform for identifying respiratory viral pathogens with overlapping symptomology. Collectively, this approach is a facile platform for diagnostics with broad application to user-defined sequence interrogation and detection.

Keywords: CRISPR/Cas9, lateral flow assay, SARS-Co-V2, single-nucleotide resolution

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

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

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

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Authors: Byeong-Uk Lim, Young-Ran Song, Sang-Ho Baik

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This study aimed to develop yeast starters for scientific alcohol production and systematic quality control of whisky. A total of 389 yeast strains were isolated from traditional Korean fermentation starter (nuruk) and rice wine (makgeolli), and ten strains were finally selected for their high alcohol productivities, in which their alcohol productions were above 17.3% (v/v) during 10 days under two steps of glucose feeding condition (30% and then 15%, w/v). By 18s rDNA sequence analysis, all strains were identified as Saccharomyces cerevisiae (SC), and they can grow well under 50% (w/v) glucose and 10% (v/v) ethanol conditions. Furthermore, the capability of ten different SC strains to ferment rice wine for whisky was studied. Rice wine was fermented with only steamed rice, water, and two types of enzymes (glucoamylase and α-amylase) during 14 days at 25 °C, and then their oenological properties have been determined. As the results, the fermented rice wines indicated the final pH range of 4.24-4.38 and acidity range of 0.12-0.18. The highest ethanol production of 20.2% (v/v) was found in the fermentation with a SC-156 strain, whereas SC-92 (16.8%) and SC-119 (16.4%) showed significantly lowest ethanol productions. In addition, the residual sugar contents showed negative correlation with alcohol contents. Moreover, this study focused on nucleotide polymorphisms in the MSN2 and MSN4 genes to investigate the cause of the defective stress responses in yeast. Consequently, it was also confirmed that the deletion of the N termini of Msn4p from identified point mutations in SC-63, SC-95, SC-156, SC-158, and SC-160 strains.

Keywords: yeast, high-alcohol, whisky, rice wine

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Authors: Tsegay Teklebrhan Gebremariam, Zhiliang, Arjan Jonker

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The study investigated that using corn gluten (CG) instead of cornmeal (CM) increased dietary sulfur shifted H₂ metabolism from methanogenesis to alternative sink and modulated microbiome in the rumen as well as hindgut segments of goats. Ruminal fermentation, CH₄ emissions and microbial abundance in goats (n = 24). The experiment was performed using a randomized block design with two dietary treatments (CM and CG with 400 g/kg DM each). Goats in CG increased sulfur, NDF and CP intake and decreased starch intake as compared with those in CM. Goats that received CG diet had decreased dissolved hydrogen (dH₂) (P = 0.01) and dissolved methane yield and emission (dCH₄) (P = 0.001), while increased dH₂S both in the rumen and hindgut segments than those fed CM. Goats fed CG had higher (p < 0.01) gene copies of microbiota and cellulolytic bacteria, whereas starch utilizing bacterial species were less in the rumen and hindgut than those fed CM. Higher (P < 0.05) methanogenic diversity and abundances of Methanimicrococcus and Methanomicrobium were observed in goats that consumed CG, whilst containing lower Methanobrevibacter populations than those receiving CM. The study suggested that goats fed corn gluten improved the gene copies of microbiota and fibrolytic bacterial species while reducing starch utilizing species in the rumen and hindgut segments as compared with that fed cornmeal. Goats consuming corn gluten had a more enriched methanogenic diversity and reduced Methanobrevibacter, a contributor to CH₄ emissions, as compared with goats fed CM. Corn gluten could be used as an alternative feed to decrease the enteric CH₄ emission in ruminant production.

Keywords: dissolved gasses, methanogenesis, microbial community, metagenomics

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Authors: Masoud Sheidai, Fahimeh Koohdar, Hashem Sharifi

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Juglans regia (L.) commonly known as Persian walnut of the genus Juglans L. (Juglandaceae) is one of the most important cultivated plant species due to its high-quality wood and edible nuts. The genetic diversity analysis is essential for conservation and management of tree species. Persian walnut is native from South-Eastern Europe to North-Western China through Tibet, Nepal, Northern India, Pakistan, and Iran. The species like Persian walnut, which has a wide range of geographical distribution, should harbor extensive genetic variability to adapt to environmental fluctuations they face. We aimed to study the population genetic structure of seven Persian walnut populations including three wild and four cultivated populations by using ISSR (Inter simple sequence repeats) and SRAP (Sequence related amplified polymorphism) molecular markers. We also aimed to compare the genetic variability revealed by ISSR neutral multilocus marker and rDNA ITS sequences. The studied populations differed in morphological features as the samples in each population were clustered together and were separate from the other populations. Three wild populations studied were placed close to each other. The mantel test after 5000 times permutation performed between geographical distance and morphological distance in Persian walnut populations produced significant correlation (r = 0.48, P = 0.002). Therefore, as the populations become farther apart, they become more divergent in morphological features. ISSR analysis produced 47 bands/ loci, while we obtained 15 SRAP bands. Gst and other differentiation statistics determined for these loci revealed that most of the ISSR and SRAP loci have very good discrimination power and can differentiate the studied populations. AMOVA performed for these loci produced a significant difference (< 0.05) supporting the above-said result. AMOVA produced significant genetic difference based on ISSR data among the studied populations (PhiPT = 0.52, P = 0.001). AMOVA revealed that 53% of the total variability is due to among population genetic difference, while 47% is due to within population genetic variability. The results showed that both multilocus molecular markers and ITS sequences can differentiate Persian walnut populations. The studied populations differed genetically and showed isolation by distance (IBD). ITS sequence based MP and Bayesian phylogenetic trees revealed that Iranian walnut cultivars form a distinct clade separated from the cultivars studied from elsewhere. Almost all clades obtained have high bootstrap value. The results indicated that a combination of multilpcus and sequencing molecular markers can be used in genetic differentiation of Persian walnut.

Keywords: genetic diversity, population, molecular markers, genetic difference

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Authors: So Youn Park, Yi Sle Lee, Ki Whan Hong, Chi Dae Kim

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The role of metabolism in the pathogenesis of osteoarthritis is an emerging field. Metabolic alterations may be a role in osteoarthritis (OA) pathogenesis, and these changes influence joint destruction via several cytokine. Especially, in OA patients, levels of IL-1β are elevated in the synovial fluid, synovial membrane, subchondral bone, and cartilage. The IL-1β is activated by NLRP3 inflammasomes, and NLRP3 inflammasomes are cytosolic complexes that drive the production of other inflammatory cytokines, including IL-1β. In this study, we examined that SIRT1 suppresses IL-1β through inhibiting NLRP3 inflammasomes and SIRT1 ameliorates osteoarthritis. OA fibroblasts were isolated from synovium of OA patients. IL-1β and NLRP3 were detected in synovium of OA patients by immunohistochemistry. Lipopolysaccharides (LPS) stimulated the expression of active IL-1β mRNA in OA fibroblasts and combination of LPS, and adenosine triphosphate increased more the expression of active IL-1β in OA fibroblasts. The level of IL-1β was measured by western blot and ELISA assay. NLRP3 inflammasomes complex were measured by western blot. SIRT1 did not inhibit expression of NLRP3 inflammasome. So caspase-1, apoptotic speck-like protein containing a caspase recruitment domain (ASC) and NLRP3 protein were expressed in OA fibroblasts. But SIRT1 suppressed activation of IL-1β by inhibiting activity of caspase-1 via NLRP3 inflammasome in OA fibroblasts under LPS plus ATP stimulation. These results suggest that SIRT1 is a modulator of NLRP3 inflammasomes in OA fibroblasts and ameliorate IL-1β, so expression of SIRT1 in OA fibroblast may be a potential strategy for OA inflammation treatment.

Keywords: osteoarthritis, inflammasome, SIRT1, IL-1beta

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Authors: Ikpesu, Thomas Ohwofasa, Babtunde Ilesanmi

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The effects of anthropogenic activities on fish and fisheries products in Niger Delta water bodies were investigated. The rivers were selected based on their close proximity to contaminants and dredging activities. Three stations were chosen per river. The stations chosen to depicting downstream and upstream stations were visited and samples collected on monthly basis. The down streams stations are the polluted and heavily dredged sites, where the upstream station is far, without any evidence of pollution or human activities. During these periods, the fishes of the same species were collected and analyzed for morphological and physiological changes, after which they were returned back to the rivers. The physico-chemicals parameters of these stations were also taken. Morphological changes such as skin ulcerations and other lesions, as well as fungi infections were observed in the down streams fishes. The fish in up streams look healthier and bigger (though the age could not be affirmed) than the downstream fishes. The physico-chemical parameters between the up streams and down streams stations vary significantly (p < 0.01). These anthropogenic effects must have interfere with the normal migration pattern of these fishes, because there were changes in the composition of population and species diversity in the samples sites, with the upstream having true species diversity. The release of pollutants into the water in the Niger Delta areas may triggers off naturally occurring bio toxicity cycles and other fish poisoning. There is risk of biomagnifications of these poisons along the tropic level. This makes the normally valuable food resource dangerous for human consumption and thereby instances of human death caused by such poisoning.

Keywords: anthropogenic, dredging, fisheries, niger delta, pollution, rivers

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Authors: Abhishikta Ghosh Roy

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

Keywords: breast cancer, BRCA, lifestyle, India

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Authors: I. Boroňová, J. Bernasovská, J. Kmec, E. Petrejčíková

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Dilated cardiomyopathy (DCM) is a severe cardiovascular disorder characterized by progressive systolic dysfunction due to cardiac chamber dilatation and inefficient myocardial contractility often leading to chronic heart failure. Recently, a genome-wide association studies (GWASs) on DCM indicate that the ZBTB17 gene rs10927875 single nucleotide polymorphism is associated with DCM. The aim of the study was to identify the distribution of ZBTB17 gene rs10927875 polymorphism in 50 Slovak patients with DCM and 80 healthy control subjects using the Custom Taqman®SNP Genotyping assays. Risk factors detected at baseline in each group included age, sex, body mass index, smoking status, diabetes and blood pressure. The mean age of patients with DCM was 52.9±6.3 years; the mean age of individuals in control group was 50.3±8.9 years. The distribution of investigated genotypes of rs10927875 polymorphism within ZBTB17 gene in the cohort of Slovak patients with DCM was as follows: CC (38.8%), CT (55.1%), TT (6.1%), in controls: CC (43.8%), CT (51.2%), TT (5.0%). The risk allele T was more common among the patients with dilated cardiomyopathy than in normal controls (33.7% versus 30.6%). The differences in genotype or allele frequencies of ZBTB17 gene rs10927875 polymorphism were not statistically significant (p=0.6908; p=0.6098). The results of this study suggest that ZBTB17 gene rs10927875 polymorphism may be a risk factor for susceptibility to DCM in Slovak patients with DCM. Studies of numerous files and additional functional investigations are needed to fully understand the roles of genetic associations.

Keywords: ZBTB17 gene, rs10927875 polymorphism, dilated cardiomyopathy, cardiovascular disorder

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Authors: Priyanka Sharma, Ranjita Das, Mohan C. Kalita, Debajit Thakur

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Background: Actinomycetes are the richest source of novel bioactive secondary metabolites such as antibiotics, enzymes and other therapeutically useful metabolites with diverse biological activities. The present study aims at the antimicrobial potential and genetic diversity of culturable Actinomycetes isolated from protected forest ecosystems of Assam which includes Kaziranga National Park (26°30˝-26°45˝N and 93°08˝-93°36˝E), Pobitora Wildlife Sanctuary (26º12˝-26º16˝N and 91º58˝-92º05˝E) and Gibbon Wildlife Sanctuary (26˚40˝-26˚45˝N and 94˚20˝-94˚25˝E) which are located in the North-eastern part of India. Northeast India is a part of the Indo-Burma mega biodiversity hotspot and most of the protected forests of this region are still unexplored for the isolation of effective antibiotic-producing Actinomycetes. Thus, there is tremendous possibility that these virgin forests could be a potential storehouse of novel microorganisms, particularly Actinomycetes, exhibiting diverse biological properties. Methodology: Soil samples were collected from different ecological niches of the protected forest ecosystems of Assam and Actinomycetes were isolated by serial dilution spread plate technique using five selective isolation media. Preliminary screening of Actinomycetes for an antimicrobial activity was done by spot inoculation method and the secondary screening by disc diffusion method against several test pathogens, including multidrug resistant Staphylococcus aureus (MRSA). The strains were further screened for the presence of antibiotic synthetic genes such as type I polyketide synthases (PKS-I), type II polyketide synthases (PKS-II) and non-ribosomal peptide synthetases (NRPS) genes. Genetic diversity of the Actinomycetes producing antimicrobial metabolites was analyzed through 16S rDNA-RFLP using Hinf1 restriction endonuclease. Results: Based on the phenotypic characterization, a total of 172 morphologically distinct Actinomycetes were isolated and screened for antimicrobial activity by spot inoculation method on agar medium. Among the strains tested, 102 (59.3%) strains showed activity against Gram-positive bacteria, 98 (56.97%) against Gram-negative bacteria, 92 (53.48%) against Candida albicans MTCC 227 and 130 (75.58%) strains showed activity against at least one of the test pathogens. Twelve Actinomycetes exhibited broad spectrum antimicrobial activity in the secondary screening. The taxonomic identification of these twelve strains by 16S rDNA sequencing revealed that Streptomyces was found to be the predominant genus. The PKS-I, PKS-II and NRPS genes detection indicated diverse bioactive products of these twelve Actinomycetes. Genetic diversity by 16S rDNA-RFLP indicated that Streptomyces was the dominant genus amongst the antimicrobial metabolite producing Actinomycetes. Conclusion: These findings imply that Actinomycetes from the protected forest ecosystems of Assam, India, are a potential source of bioactive secondary metabolites. These areas are as yet poorly studied and represent diverse and largely unscreened ecosystem for the isolation of potent Actinomycetes producing antimicrobial secondary metabolites. Detailed characterization of the bioactive Actinomycetes as well as purification and structure elucidation of the bioactive compounds from the potent Actinomycetes is the subject of ongoing investigation. Thus, to exploit Actinomycetes from such unexplored forest ecosystems is a way to develop bioactive products.

Keywords: Actinomycetes, antimicrobial activity, forest ecosystems, RFLP

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Authors: Fayad Ghawbar

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The significance of the Multiple Input Multiple Output (MIMO) system in the 5G wireless communication system is essential to enhance channel capacity and provide a high data rate resulting in a need for dual-polarization in vertical and horizontal. Furthermore, size reduction is critical in a MIMO system to deploy more antenna elements requiring a compact, low-profile design. A compact dual-band 4-MIMO antenna system has been presented in this paper with pattern and polarization diversity. The proposed single antenna structure has been designed using two antenna layers with a C shape in the front layer and a partial slot with a U-shaped cut in the ground to enhance isolation. The single antenna is printed on an FR4 dielectric substrate with an overall size of 18 mm×18 mm×1.6 mm. The 4-MIMO antenna elements were printed orthogonally on an FR4 substrate with a size dimension of 36 × 36 × 1.6 mm3 with zero edge-to-edge separation distance. The proposed compact 4-MIMO antenna elements resonate at 3.4-3.6 GHz and 4.8-5 GHz. The s-parameters measurement and simulation results agree, especially in the lower band with a slight frequency shift of the measurement results at the upper band due to fabrication imperfection. The proposed design shows isolation above -15 dB and -22 dB across the 4-MIMO elements. The MIMO diversity performance has been evaluated in terms of efficiency, ECC, DG, TARC, and CCL. The total and radiation efficiency were above 50 % across all parameters in both frequency bands. The ECC values were lower than 0.10, and the DG results were about 9.95 dB in all antenna elements. TARC results exhibited values lower than 0 dB with values lower than -25 dB in all MIMO elements at the dual-bands. Moreover, the channel capacity losses in the MIMO system were depicted using CCL with values lower than 0.4 Bits/s/Hz.

Keywords: compact antennas, MIMO antenna system, 5G communication, dual band, ECC, DG, TARC

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Authors: M. Chukhryaeva, R. Skhalyakho, J. Kagazegeva, E. Pocheshkhova, L. Yepiskopossyan, O. Balanovsky, E. Balanovska

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The Middle East is crossroad of different populations at different times. The Kurds are of particular interest in this region. Historical sources suggested that the origin of the Kurds is associated with Medes. Therefore, it was especially interesting to compare gene pool of Kurds with other supposed descendants of Medes-Tats. Yezidis are ethno confessional group of Kurds. Yezidism as a confessional teaching was formed in the XI-XIII centuries in Iraq. Yezidism has caused reproductively isolation of Yezidis from neighboring populations for centuries. Also, isolation helps to retain Yezidian caste system. It is unknown how the history of Yezidis affected its genу pool because it has never been the object of researching. We have examined the Y-chromosome variation in Yezidis and Kurdish males to understand their gene pool. We collected DNA samples from 90 Yezidi males and 24 Kurdish males together with their pedigrees. We performed Y-STR analysis of 17 loci in the samples collected (Yfiler system from Applied Biosystems) and analysis of 42 Y-SNPs by real-time PCR. We compared our data with published data from other Kurdish groups and from European, Caucasian, and West Asian populations. We found that gene pool of Yezidis contains haplogroups common in the Middle East (J-M172(xM67,M12)- 24%, E-M35(xM78)- 9%) and in South Western Asia (R-M124- 8%) and variant with wide distribution area - R-M198(xM458- 9%). The gene pool of Kurdish has higher genetic diversity than Yezidis. Their dominants haplogroups are R-M198- 20,3 %, E-M35- 9%, J-M172- 9%. Multidimensional scaling also shows that the Kurds and Yezidis are part of the same frontier Asian cluster, which, in addition, included Armenians, Iranians, Turks, and Greeks. At the same time, the peoples of the Caucasus and Europe form isolated clusters that do not overlap with the Asian clusters. It is noteworthy that Kurds from our study gravitate towards Tats, which indicates that most likely these two populations are descendants of ancient Medes population. Multidimensional scaling also reveals similarity between gene pool of Yezidis, Kurds with Armenians and Iranians. The analysis of Yezidis pedigrees and their STR variability did not reveal a reliable connection between genetic diversity and caste system. This indicates that the Yezidis caste system is a social division and not a biological one. Thus, we showed that, despite many years of isolation, the gene pool of Yezidis retained a common layer with the gene pool of Kurds, these populations have common spectrum of haplogroups, but Yezidis have lower genetic diversity than Kurds. This study received primary support from the RSF grant No. 16-36-00122 to MC and grant No. 16-06-00364 to EP.

Keywords: gene pool, haplogroup, Kurds, SNP and STR markers, Yezidis

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Authors: Fanny Soulard

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Today work environment is more multicultural than ever: spatial limits have been blown out, encouraging people and ideas mobility all around the globe. Indeed, opportunities to design with culturally diverse team workers, clients, or end-users, have become within everyone's reach. We enjoy traveling to discover other civilizations, but when it comes to business, we often take for granted that our own work methodology will be generic enough to federate each party and cover the project needs. This paper aims to explore why, by skipping cultural awareness, we often create misunderstandings, frustration, and even counterproductive design. Tree lighting projects successively developed by a French lighting studio, a Vietnamese lighting studio, and an Australian Engineering company will be assessed from their concept stage to completion. All these study cases are based in Vietnam, where the construction market is equally led by local and international consultants. Core criteria such as lighting standard reference, service scope, communication tools, internal team organization, delivery package content, key priorities, and client relationship will help to spot and list when and how cultural diversity has impacted the design output and effectiveness. On the second hand, we will demonstrate through the same selected projects how intercultural intelligence tools and mindset can not only respond positively to previous situations and avoid major clashes but also turn cultural differences into a key added value to generate significant benefits for individuals, teams, and companies. By understanding the major importance of including a cultural factor within any design, intercultural intelligence will quickly turn out as a “must have” skill to be developed and acquired by any designer.

Keywords: intercultural intelligence, lighting design, work methodology, multicultural diversity

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Authors: Stephanie Khater, Ali Chehade, Lamis Chalak

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The progressive replacement of the Lebanese autochthonous grapevine cultivars during the last decade by the imported foreign varieties almost resulted in the genetic erosion of the local germplasm and the confusion with cultivars' names. Hence there is a need to characterize these local cultivars and to assess the possible existing variability at the cultivar level. This work was conducted in an attempt to evaluate the intra-varietal diversity within Lebanese traditional cultivars 'Aswad', 'Maghdoushe', 'Maryame', 'Merweh', 'Meksese' and 'Obeide'. A total of 50 accessions distributed over five main geographical areas in Lebanon were collected and submitted to both ampelographic description and ISSR DNA analysis. A set of 35 ampelographic descriptors previously established by the International Office of Vine and Wine and related to leaf, bunch, berry, and phenological stages, were examined. Variability was observed between accessions within cultivars for blade shape, density of prostrate and erect hairs, teeth shape, berry shape, size and color, cluster shape and size, and flesh juiciness. At the molecular level, nine ISSR (inter-simple sequence repeat) primers, previously developed for grapevine, were used in this study. These primers generated a total of 35 bands, of which 30 (85.7%) were polymorphic. Totally, 29 genetic profiles were differentiated, of which 9 revealed within 'Obeide', 6 for 'Maghdoushe', 5 for 'Merweh', 4 within 'Maryame', 3 for 'Aswad' and 2 within 'Meksese'. Findings of this study indicate the existence of several genotypes that form the basis of the main indigenous cultivars grown in Lebanon and which should be further considered in the establishment of new vineyards and selection programs.

Keywords: ampelography, autochthonous cultivars, ISSR markers, Lebanon, Vitis vinifera L.

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Authors: Samuel Ntsanwisi

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This study investigates the transformative role of Artificial Intelligence (AI) and Virtual Reality (VR) in the preservation of endangered African languages. The study is contextualized within the film and television industry, highlighting disparities in screen representation for certain languages in South Africa, underscoring the need for increased visibility and preservation efforts; with globalization and cultural shifts posing significant threats to linguistic diversity, this research explores approaches to language preservation. By leveraging AI technologies, such as speech recognition, translation, and adaptive learning applications, and integrating VR for immersive and interactive experiences, the study aims to create a framework for teaching and passing on endangered African languages. Through digital documentation, interactive language learning applications, storytelling, and community engagement, the research demonstrates how these technologies can empower communities to revitalize their linguistic heritage. This study employs a dual-method approach, combining a rigorous literature review to analyse existing research on the convergence of AI, VR, and language preservation with primary data collection through interviews and surveys with ten filmmakers. The literature review establishes a solid foundation for understanding the current landscape, while interviews with filmmakers provide crucial real-world insights, enriching the study's depth. This balanced methodology ensures a comprehensive exploration of the intersection between AI, VR, and language preservation, offering both theoretical insights and practical perspectives from industry professionals.

Keywords: language preservation, endangered languages, artificial intelligence, virtual reality, interactive learning

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Authors: Sharon Diaz Ruiz

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This qualitative study addresses college English instructors’ perceptions of classroom diversity and culturally diverse teaching strategies within the adult English language learning classroom environment. Every year, English college instructors face numerous challenges as the adult Latinx population keeps rising. To better understand the Latinx adult learners and the language classroom dynamics, research should focus on the experiences, pedagogical methods, and teaching insights of full-time and adjunct minority professors at degree-granting postsecondary institutions. Culturally responsive teaching is used as the framework to understand and explore the perceptions of English instructors on the realities and needs of Latinx adult emergent bilinguals enrolled in developmental English courses. Snowball sampling allows the researcher to locate members who meet these specific criteria: adjunct and part-time English instructors of adult Latinx language learners. Participants answered a demographic questionnaire and then contributed to 45-minute in-depth interviews to explore their perceptions of culturally responsive practices in the Latinx adult emergent bilinguals’ basic and intermediate developmental English courses. The interviews shed light on topics such as teaching biases, educators’ cultural experiences, and resources and strategies faculty recommend for effective culturally responsive teaching strategies. The result of this investigation will shed light on the gap in the literature documenting the application of culturally responsive pedagogy to Latino adult language learners.

Keywords: Latinx, English language learners, English faculty, adult learners, critical theory, culturally responsive theory

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Authors: J. Dharmaraj, C. Gunasekaran

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Invertebrates are the reliable ecological indicators of disturbance of the forest ecosystems and they respond to environment changes more quickly than other fauna. Among these the terrestrial invertebrates are vital to functioning ecosystems, contributing to processes such as decomposition, nutrient cycling and soil fertility. The natural ecosystems of the forests have been subject to various types of disturbances, which lead to decline of flora and fauna. The comparative diversity of micro-arthropods in natural forest, wattle plantation and eucalyptus plantations were studied in Nilgiris. The study area was divided in to five major sites (Emerald (Site-I), Thalaikundha (Site-II), Kodapmund (Site-III), Aravankad (Site-IV), Kattabettu (Site-V). The research was conducted during period from March 2014 to August 2014. The leaf and soil samples were collected and isolated by using Berlese funnel extraction methods. Specimens were isolated and identified according to their morphology (Balogh 1972). In the present study results clearly showed the variation in soil pH, NPK (Major Nutrients) and organic carbon among the study sites. The chemical components of the leaf litters of the plantation decreased the diversity of micro-arthropods and decomposition rate leads to low amount of carbon and other nutrients present in the soil. Moreover eucalyptus and wattle plantations decreases the availability of the ground water source to other plantations and micro-arthropods and hences affects the soil fertility. Hence, the present study suggests to minimize the growth of wattle and eucalyptus tree plantations in the natural areas which may help to reduce the decline of forests.

Keywords: micro-arthropods, assemblage, berlese funnel, morphology, NPK, nilgiris

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Authors: Luai Z. Hasoun, Steven W. Bailey, Kitti K. Outlaw, June E. Ayling

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Human skin color is thought to have evolved to balance sufficient photochemical synthesis of vitamin D versus the need to protect not only DNA but also folate from degradation by ultraviolet light (UV). Although the risk of DNA damage and subsequent skin cancer is related to light skin color, the effect of UV on skin folate of any species is unknown. Here we show that UVA irradiation at 13 mW/cm2 for a total exposure of 187 J/cm2 (similar to a maximal daily equatorial dose) induced a significant loss of total folate in epidermis of ex vivo white skin. No loss was observed in black skin samples, or in the dermis of either color. Interestingly, while the concentration of 5 methyltetrahydrofolate (5-MTHF) fell in white epidermis, a concomitant increase of tetrahydrofolic acid was found, though not enough to maintain the total pool. These results demonstrate that UVA indeed not only decreases folate in skin, but also rearranges the pool components. This could be due in part to the reported increase of NADPH oxidase activity upon UV irradiation, which in turn depletes the NADPH needed for 5-MTHF biosynthesis by 5,10-methylenetetrahydrofolate reductase. The increased tetrahydrofolic acid might further support production of the nucleotide bases needed for DNA repair. However, total folate was lost at a rate that could, with strong or continuous enough exposure to ultraviolet radiation, substantially deplete light colored skin locally, and also put pressure on total body stores for individuals with low intake of folate.

Keywords: depletion, folate, human skin, ultraviolet

Procedia PDF Downloads 370

Authors: Margaret K. Spoo-Chupka

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The Metropolitan Water District of Southern California (MWDSC) manages surface water reservoirs that are a source of drinking water for more than 19 million people in Southern California. These reservoirs experience periodic planktonic cyanobacteria blooms that can impact water quality. MWDSC imports water from two sources – the Colorado River (CR) and the State Water Project (SWP). The SWP brings supplies from the Sacramento-San Joaquin Delta that are characterized as having higher nutrients than CR water. Above average precipitation in 2017 after five years of drought allowed the majority of the reservoirs to fill. Phytoplankton was analyzed during the drought and after the drought at three reservoirs: Diamond Valley Lake (DVL), which receives SWP water exclusively, Lake Skinner, which can receive a blend of SWP and CR water, and Lake Mathews, which generally receives only CR water. DVL experienced a significant increase in water elevation in 2017 due to large SWP inflows, and there were no significant changes to total phytoplankton biomass, Shannon-Wiener diversity of the phytoplankton, or cyanobacteria biomass in 2017 compared to previous drought years despite the higher nutrient loads. The biomass of cyanobacteria that could potentially impact DVL water quality (Microcystis spp., Aphanizomenon flos-aquae, Dolichospermum spp., and Limnoraphis birgei) did not differ significantly between the heavy precipitation year and drought years. Compared to the other reservoirs, DVL generally has the highest concentration of cyanobacteria due to the water supply having greater nutrients. Lake Mathews’ water levels were similar in drought and wet years due to a reliable supply of CR water and there were no significant changes in the total phytoplankton biomass, phytoplankton diversity, or cyanobacteria biomass in 2017 compared to previous drought years. The biomass of cyanobacteria that could potentially impact water quality at Lake Mathews (L. birgei and Microcystis spp.) did not differ significantly between 2017 and previous drought years. Lake Mathews generally had the lowest cyanobacteria biomass due to the water supply having lower nutrients. The CR supplied most of the water to Lake Skinner during drought years, while the SWP was the primary source during 2017. This change in water source resulted in a significant increase in phytoplankton biomass in 2017, no significant change in diversity, and a significant increase in cyanobacteria biomass. Cyanobacteria that could potentially impact water quality at Skinner included: Microcystis spp., Dolichospermum spp., and A.flos-aquae. There was no significant difference in Microcystis spp. biomass in 2017 compared to previous drought years, but biomass of Dolichospermum spp. and A.flos-aquae were significantly greater in 2017 compared to previous drought years. Dolichospermum sp. and A. flos-aquae are two cyanobacteria that are more sensitive to nutrients than Microcystis spp., which are more sensitive to temperature. Patterns in problem cyanobacteria abundance among Southern California reservoirs as a result of above-average precipitation after more than five years of drought were most closely related to nutrient loading.

Keywords: drought, reservoirs, cyanobacteria, and phytoplankton ecology

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Authors: Henry Kuppen

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In the last decade, tree managers have faced many threats of pests and diseases and the effects of climate change. Managers will recognize that they have to put more energy and more money into tree management. By recognizing the cause behind this, the opportunity will arise to build sustainable tree populations for the future. More and more, unwanted larvae are sprayed, ash dieback infected trees are pruned or felled, and emerald ash borer is knocking at the door of West Europe. A lot of specific knowledge is needed to produce management plans and best practices. If pest and disease have a large impact, society loses complete tree species and need to start all over again building urban forest. But looking at the cause behind it, landscape design, and tree species selection, the sustainable solution does not present itself in managing these threats. Every pest or disease needs two important basic ingredients to be successful: climate and food. The changing climate is helping several invasive pathogens to survive. Food is often designed by the landscapers and managers of the urban forest. Monocultures promote the success of pathogens. By looking more closely at the basics, tree managers will realise very soon that the solution will not be the management of pathogens. The long-term solution for sustainable tree populations is a different design of our urban landscape. The use of tree species diversity can help to reduce the impact of climate change and pathogens. Therefore landscapers need to be supported. They are the specialists in designing the landscape using design values like canopy volume, ecosystem services, and seasonal experience. It’s up to the species specialist to show what the opportunities are for different species that meet the desired interpretation of the landscape. Based on landscapers' criteria, selections can be made, including tree species related requirements. Through this collaboration and formation of integral teams, sustainable plant design will be possible.

Keywords: climate change, landscape design, resilient landscape, tree species selection

Procedia PDF Downloads 101

Authors: Fatemeh Noori

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The fashion industry is a dynamic and ever-evolving realm that continuously shapes societal perceptions of beauty and style. Within this industry, fashion modeling plays a crucial role, acting as the visual representation of brands and designers. However, behind the glamorous façade lies a complex web of legal considerations that govern the rights, responsibilities, and ethical practices within the field. This paper aims to explore the legal landscape surrounding fashion modeling, shedding light on key issues such as contract law, intellectual property, labor rights, and the increasing importance of ethical considerations in the industry. Fashion modeling involves the collaboration of various stakeholders, including models, designers, agencies, and photographers. To ensure a fair and transparent working environment, it is imperative to establish a comprehensive legal framework that addresses the rights and obligations of each party involved. One of the primary legal considerations in fashion modeling is the contractual relationship between models and agencies. Contracts define the terms of engagement, including payment, working conditions, and the scope of services. This section will delve into the essential elements of modeling contracts, the negotiation process, and the importance of clarity to avoid disputes. Models are not just individuals showcasing clothing; they are integral to the creation and dissemination of artistic and commercial content. Intellectual property rights, including image rights and the use of a model's likeness, are critical aspects of the legal landscape. This section will explore the protection of models' image rights, the use of their likeness in advertising, and the potential for unauthorized use. Models, like any other professionals, are entitled to fair and ethical treatment. This section will address issues such as working conditions, hours, and the responsibility of agencies and designers to prioritize the well-being of models. Additionally, it will explore the global movement toward inclusivity, diversity, and the promotion of positive body image within the industry. The fashion industry has faced scrutiny for perpetuating harmful standards of beauty and fostering a culture of exploitation. This section will discuss the ethical responsibilities of all stakeholders, including the promotion of diversity, the prevention of exploitation, and the role of models as influencers for positive change. In conclusion, the legal considerations in fashion modeling are multifaceted, requiring a comprehensive approach to protect the rights of models and ensure ethical practices within the industry. By understanding and addressing these legal aspects, the fashion industry can create a more transparent, fair, and inclusive environment for all stakeholders involved in the art of modeling.

Keywords: fashion modeling contracts, image rights in modeling, labor rights for models, ethical practices in fashion, diversity and inclusivity in modeling

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Authors: Caroline F. Moeser

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The skin-gut axis defines the relationship between the intestinal microbiota and the development of pathological skin diseases. Low diversity within the gut can predispose to the development of allergic skin conditions, and a greater diversity of the gastrointestinal microflora has been associated with a reduction of skin flares in people with atopic dermatitis. Manipulation of the gut microflora has been used as a treatment option for several conditions in people, but there is limited data available on the use of faecal transplantation as a preventative measure in either people or dogs. Six, 4-month-old pups from a litter of ten were presented for diarrhea and/or signs of skin disease (chronic scratching, otitis externa). Of these pups, two were given probiotics with a resultant resolution of diarrhea. The other four pups were given faecal transplantation, either as a sole treatment or in combination with other treatments. Follow-up on the litter of ten pups was performed at 18 months of age. At this stage, the four pups that had received faecal transplantation had resolved all clinical signs and had no recurrence of either skin or gastrointestinal symptoms. Of the remaining six pups from the litter, all had developed at least one episode of Malassezia otitis externa within the period of 5 months to 18 months of age. Two pups had developed two Malassezia otitis infections, and one had developed three Malassezia otitis infections during this period. Favrot’s criteria for the diagnosis of canine atopic dermatitis include chronic or recurrent Malassezia infections by the age of three years. Early results from this litter predict a reduction in the development of canine atopic disease in dogs given faecal microbial transplantation. Follow-up studies at three years of age and within a larger population of dogs can enhance understanding of the impact of early faecal transplantation in the prevention of canine atopic dermatitis.

Keywords: canine atopic dermatitis, faecal microbial transplant, skin-gut axis, otitis

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Authors: Rueyhung Weng, Chia-En Huang, Jung-Chi-Liao

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The transition zone (TZ) serves as a diffusion barrier to regulate the ins and outs of the proteins recruited to the primary cilia. TCTN2 is one of the TZ proteins and its mutation causes Joubert syndrome, a serious multi-organ disease. Despite its important medical relevance, the functions of TCTN2 remain elusive. Here we created a TCTN2 gene deleted retinal pigment epithelial cells (RPE1) using CRISPR/Cas9-based genome editing technique and used this knockout line to reveal roles of TCTN2. TCTN2 knockout RPE1 cells displayed a significantly reduced ciliogenesis or a shortened primary cilium length in the cilium-remaining population. Intraflagellar transport protein IFT88 aberrantly accumulated at the tip of TCTN2 deficient cells. Guanine nucleotide exchange factor Arl13B was mostly absent from the ciliary compartment, with a small population localizing at the ciliary tip. The deficient TZ was corroborated with the mislocalization of two other TZ proteins TMEM67 and MKS1. In addition, TCTN2 deficiency induced TZ impairment led to the suppression of Sonic hedgehog signaling in response to Smoothened (Smo) agonist. Together, depletion of TCTN2 destabilizes other TZ proteins and considerably alters the localization of key transport and signaling-associated proteins, including IFT88, Arl13B, and Smo.

Keywords: CRISPR/Cas9, primary cilia, Sonic hedgehog signaling, transition zone

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Authors: Nishal Parbhoo, John B. Dewar, Samantha Gildenhuys

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Rotavirus is the major cause of severe gastroenteritis worldwide in children aged 5 and younger. Death rates are high particularly in developing countries. The mature rotavirus is a non-enveloped triple-layered nucleocapsid containing 11 double-stranded RNA segments. Here a global view on the sequence and structure of the three main capsid proteins, VP7, VP6, and VP2 is taken by generating a consensus sequence for each of these rotavirus proteins, for each species obtained from published data of representative rotavirus genotypes from across the world and across species. The degree of conservation between species was represented on homology models for each of the proteins. VP7 shows the highest level of variation with 14 - 45 amino acids showing conservation of less than 60%. These changes are localized to the outer surface which is exposed to antibodies alluding to a possible mechanism in evading the immune system. The middle layer, VP6 shows lower variability with only 14-32 sites having lower than 70% conservation. The inner structural layer made up of VP2 showed the lowest variability with only 1-16 sites having less than 70% conservation across species. The results correlate with proteins’ multiple structural roles. Although the nucleotide sequences vary due to an error-prone replication and lack of proofreading, the corresponding amino acid sequence of VP2, 6 and 7 remains conserved. Sequence conservation maintained for the virus results in stable protein structures, fit for function. This can be exploited in drug design, molecular studies and biotechnological applications.

Keywords: amino acid sequence conservation, capsid protein, protein structure, vaccine candidate

Procedia PDF Downloads 279