Search results for: gene diversity

Authors: Salman Akrokayan

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Synthetic cDNA (ScDNA) of granulocyte colony-stimulating factor (G-CSF) was constructed using a DNA synthesizer with the aim to increase its expression level. 5' end of the ScDNA of G-CSF coding region was modified by decreasing the GC content without altering the predicted amino acids sequence. The identity of the resulting protein from ScDNA was confirmed by the highly specific enzyme-linked immunosorbent assay. In conclusion, a synthetic G-CSF cDNA in combination with the recombinant DNA protocol offers a rapid and reliable strategy for synthesizing the target protein. However, the commercial utilization of this methodology requires rigorous validation and quality control.

Keywords: synthetic cDNA, recombinant G-CSF, cloning, gene expression

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Authors: Otuwose E. Agyeno, Adeniyi A. Jayeola, Bashir A. Ajala

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P. esculentus is indigenous to Nigeria yet no wild relation has been encountered or reported. This has made it difficult to establish proper lineages between the varieties and landraces under cultivation. The present work is the first to determine the apormophy of 135 morphoanatomical characters in organs of 46 accessions drawn from 23 populations of this species based on dicta. The character states were coded in accession x character-state matrices and only 83 were informative and utilised for neighbour joining clustering based on euclidean values, and heuristic search in parsimony analysis using PAST ver. 3.15 software. Compatibility and evolutionary trends between accessions were then explored from values and diagrams produced. The low consistency indices (CI) recorded support monophyly and low homoplasy in this taxon. Agglomerative schedules based on character type and source data sets divided the accessions into mainly 3 clades, each of complexes of accessions. Solenostemon rotundifolius (Poir) J.K Morton was the outgroup (OG) used, and it occurred within the largest clades except when the characters were combined in a data set. The OG showed better compatibility with accessions of populations of landrace Isci, and varieties Riyum and Long’at. Otherwise, its aerial parts are more consistent with those of accessions of variety Bebot. The highly polytomous clades produced due to anatomical data set may be an indication of how stable such characters are in this species. Strict consensus trees with more than 60 nodes outputted showed that the basal nodes were strongly supported by 3 to 17 characters across the data sets, suggesting that populations of this species are more alike. The OG was clearly the first diverging lineage and closely related to accessions of landrace Gwe and variety Bebot morphologically, but different from them anatomically. It was also distantly related to landrace Fina and variety Long’at in terms of root, stem and leaf structural attributes. There were at least 5 other clades with each comprising of complexes of accessions from different localities and terrains within the study area. Spherical stem in cross section, size of vascular bundles at the stem corners as well as the alternate and whorl phyllotaxy are attributes which may have facilitated each other’s evolution in all accessions of the landrace Gwe, and they may be innovative since such states are not characteristic of the larger Lamiaceae, and Plectranthus L’Her in particular. In conclusion, this study has provided valuable information about infraspecific diversity in this taxon. It supports recognition of the varietal statuses accorded to populations of P. esculentus, as well as the hypothesis that the wild gene might have been distributed on the Jos Plateau. However, molecular characterisation of accessions of populations of this species would resolve this problem better.

Keywords: clustering, lineage, morphoanatomical characters, Nigeria, phylogenetics, Plectranthus esculentus, population

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Authors: Sepehr Moradi

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This study investigates the contribution of individual Australian Superfine Merino Wool (ASFW) and Inner Mongolia Cashmere (IMC) fibres strength behaviour to the breaking force variation (CVBF) and minimum fibre diameter (CVₘFD) induced by actual single fibre lengths and the combination of length and diameter groups. Mid-side samples were selected for the ASFW (n = 919) and IMC (n = 691) since it is assumed to represent the average of the whole fleece. The average (LₘFD) varied for ASFW and IMC by 36.6 % and 33.3 % from shortest to longest actual single fibre length and -21.2 % and -21.7 % between longest-coarsest and shortest-finest groups, respectively. The tensile properties of single animal fibres were characterised using Single Fibre Analyser (SIFAN 4). After normalising for diversity in fibre diameter at the position of breakage, the parameters, which explain the strength behaviour within actual fibre lengths and combination of length-diameter groups, were the Intrinsic Fibre Strength (IFS) (MPa), Min IFS (MPa), Max IFS (MPa) and Breaking force (BF) (cN). The average strength of single fibres varied extensively within actual length groups and within a combination of length-diameter groups. IFS ranged for ASFW and IMC from 419 to 355 MPa (-15.2 % range) and 353 to 319 (-9.6 % range) and BF from 2.2 to 3.6 (63.6 % range) and 3.2 to 5.3 cN (65.6 % range) from shortest to longest groups, respectively. Single fibre properties showed no differences within actual length groups and within a combination of length-diameter groups, or was there a strong interaction between the strength of single fibre (P > 0.05) within remaining and removing length-diameter groups. Longer-coarser fibre fractionation had a significant effect on BF and IFS and all of the length groups showed a considerable variance in single fibre strength that is accounted for by diversity in the diameter variation along the fibre. There are many concepts for the improvement of the stress-strain properties of animal fibres as a means of raising a single fibre strength by simultaneous changes in fibre length and diameter. Fibre fractionation over a given length directly for single fibre strength or using the variation traits of fibre diameter is an important process used to increase the strength of the single fibre.

Keywords: single animal fibre fractionation, actual length groups, strength variation, length-diameter groups, diameter variation along fibre

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Authors: Shakila Singh

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Dominant discourses constitute heterosexuality as natural, normal and the only legitimate sexuality, and diverse sexual subjectivities as abnormal, unnatural and socially taboo. Similarly, the cisgender subject is reified. There are ongoing debates about the inclusion and suitability of sexuality education in the school curriculum and research show that teachers are not adequately prepared to teach about such issues in the classroom. Not surprising then, that many young people enter these institutions having had limited previous exposure to, or education about, sex, gender and sexuality diversity. This paper discusses the presence of heterosexism and cissexism at multiple layers in higher education institutions, impacting students and staff. Increasing knowledge and awareness of sex, gender and sexuality diversities is also crucial to challenging existing perceptions of sex, gender and sexuality diversities that marginalise and subordinate a large proportion of students and staff. There is a persistent disjuncture between dominant discourses that generally position higher education institutions as socially progressive, open environments and the discourses that legitimate the ascendency of heterosexual and cisgender identities. This paper argues that such disjuncture must be addressed by providing inclusive physical and emotional spaces if universities are to affirm every individual and produce graduates across all disciplines with the cultural capability to engage with increasingly diverse communities. Given the key role of language in shaping cultural and social attitudes, using gender-inclusive language is a powerful way to promote gender equality and eradicate gender bias. This means speaking and writing in a way that does not discriminate against a particular sex, gender or sexual identity and does not perpetuate gender stereotypes. Individuals must be allowed to present themselves and identify in ways they choose and be addressed by their chosen pronouns.

Keywords: heteronormativity, inclusivity, gender, universities

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Authors: Anil Kumar, Diwakar Aggarwal, M. Sudhakara Reddy

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The wood of Eucalyptus, Populus and bamboos are some important species used as raw material for the manufacture of pulp. However, higher levels of lignin pose a problem during Kraft pulping and yield of pulp is also lower. In order to increase the yield of pulp per unit wood and reduce the use of chemicals during kraft pulping it is important to reduce the lignin content and/or increase cellulose content in wood. Cellulose biosynthesis in wood takes place by the coordinated action of many enzymes. The two important enzymes are KORRIGAN and SUCROSE SYNTHASE. KORRIGAN (Endo-1,4--glucanase) is implicated in the process of editing growing cellulose chains and improvement of the crystallinity of produced cellulose, whereas SUCROSE SYNTHASE is involved in providing substrate (UDP-glucose) for growing cellulose chains. The present study was aimed at the cloning, characterization and overexpression of these genes in Eucalyptus and Populus. An efficient shoot organogenesis protocol from leaf explants taken from micro shoots of the species has been developed. Agrobacterium mediated genetic transformation using Agrobacterium tumefaciens strain EHA105 and LBA4404 harboring binary vector pBI121 was achieved. Both the genes were cloned from cDNA library of Populus deltoides. These were subsequently characterized using various bioinformatics tools. The cloned genes were then inserted into pBI121 under the CaMV35S promotors replacing GUS gene. The constructs were then mobilized into above strains of Agrobacterium and used for the transformation work. Subsequently, genetic transformation of these clones with target genes following already developed protocol is in progress. Four transgenic lines of Eucalyptus tereticornis overexpressing Korrigan gene under the strong constitutive promoters CaMV35S have been developed, which are being further evaluated. Work on development of more transgenic lines overexpressing these genes in Populus and Eucalyptus is also in progress. This presentation will focus on important developments in this direction.

Keywords: Eucalyptus tereticornis, genetic transformation, Kraft pulping Populus deltoides

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Authors: Lena Lapidot, Amir Amnon, Rita Nosenko, Veitsman Ella, Cohen-Ezra Oranit, Davidov Yana, Segev Shlomo, Koren Omry, Safran Michal, Ben-Ari Ziv

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Introduction: Hepatocellular carcinoma (HCC) is one of the leading causes of cancer related mortality worldwide. Liver Cirrhosis is the main predisposing risk factor for the development of HCC. The factor(s) influencing disease progression from Cirrhosis to HCC remain unknown. Gut microbiota has recently emerged as a major player in different liver diseases, however its association with HCC is still a mystery. Moreover, there might be an important association between the gut microbiota, nutrition, life style and the progression of Cirrhosis and HCC. The aim of our study was to characterize the gut microbial signature in association with life style and nutrition of patients with Cirrhosis, HCC-Cirrhosis and healthy controls. Design: Stool samples were collected from 95 individuals (30 patients with HCC, 38 patients with Cirrhosis and 27 age, gender and BMI-matched healthy volunteers). All participants answered lifestyle and Food Frequency Questionnaires. 16S rRNA sequencing of fecal DNA was performed (MiSeq Illumina). Results: There was a significant decrease in alpha diversity in patients with Cirrhosis (qvalue=0.033) and in patients with HCC-Cirrhosis (qvalue=0.032) compared to healthy controls. The microbiota of patients with HCC-cirrhosis compared to patients with Cirrhosis, was characterized by a significant overrepresentation of Clostridium (pvalue=0.024) and CF231 (pvalue=0.010) and lower expression of Alphaproteobacteria (pvalue=0.039) and Verrucomicrobia (pvalue=0.036) in several taxonomic levels: Verrucomicrobiae, Verrucomicrobiales, Verrucomicrobiaceae and the genus Akkermansia (pvalue=0.039). Furthermore, we performed an analysis of predicted metabolic pathways (Kegg level 2) that resulted in a significant decrease in the diversity of metabolic pathways in patients with HCC-Cirrhosis (qvalue=0.015) compared to controls, one of which was amino acid metabolism. Furthermore, investigating the life style and nutrition habits of patients with HCC-Cirrhosis, we found significant correlations between intake of artificial sweeteners and Verrucomicrobia (qvalue=0.12), High sugar intake and Synergistetes (qvalue=0.021) and High BMI and the pathogen Campylobacter (qvalue=0.066). Furthermore, overweight in patients with HCC-Cirrhosis modified bacterial diversity (qvalue=0.023) and composition (qvalue=0.033). Conclusions: To the best of the our knowledge, we present the first report of the gut microbial composition in patients with HCC-Cirrhosis, compared with Cirrhotic patients and healthy controls. We have demonstrated in our study that there are significant differences in the gut microbiome of patients with HCC-cirrhosis compared to Cirrhotic patients and healthy controls. Our findings are even more pronounced because the significantly increased bacteria Clostridium and CF231 in HCC-Cirrhosis weren't influenced by diet and lifestyle, implying this change is due to the development of HCC. Further studies are needed to confirm these findings and assess causality.

Keywords: Cirrhosis, Hepatocellular carcinoma, life style, liver disease, microbiome, nutrition

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Authors: F. O. Arimoro, F. N. Okonkwo, R. B. Ikomi

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The environmental determinants structuring species richness has been generating interest recently but we still lack an understanding of these patterns in various regions (e.g. Afrotropical), and how seasons help to structure these patterns. Our aim was to assessed the environmental drivers importance in regulating species richness and community structure of fish species. The lchthyofauna assemblage of Warri River, Niger Delta area of Nigeria was studied between August 2013 and July 2014. A total of 1152 individuals representing 43 species in 23 families and 30 genera were caught. Of the 43 species recorded, 67.4%, 53.5% and 67.4% of the species occurred in Stations 1, 2 and 3 respectively. Eight taxa representing 18.6% of the total abundance were ubiquitous. The claroteid, Chrysichthys walkeri and the cichlid, Chromidotilapia guentheri were the most dominant species accounting for 19.2% and 6.0% respectively of the total catch. The species richness and general diversity were relatively higher in station 1 although Jaccard similarity index revealed that stations 1 and 3 were significantly similar while station 2 showed complete dissimilarity with stations 1 and 3. Canonical correspondence analysis indicated that dissolved oxygen, electrical conductivity, total nitrogen, Biochemical Oxygen demand and temperature were important variables structuring the overall fish assemblages. The presence of appreciable number of juveniles in this water body suggests that the Warri River is a breeding and nursery ground for fish species particularly those of brackish origin. These findings indicate that the water body is still useful as a good fishing ground for the rural communities and every effort should be put in place to ensure its protection and conservation for the production of healthy fish.

Keywords: Chrysichthys walkeri, fish communities, mangrove ecosystem, physicochemical parameters, Warri River

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Authors: Hossein Rassi, Marjan Moradi Fard, Masoud Houshmand

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Background: Cervical cancer is multistep disease that is thought to result from an interaction between genetic background and environmental factors. Human papillomavirus (HPV) infection is the leading risk factor for cervical intraepithelial neoplasia (CIN) and cervical cancer. In other hand, some of p53 and miRNA polymorphism may plays an important role in carcinogenesis. This study attempts to clarify the relation of p53 genotypes and miR-146a rs2910164 polymorphism in cervical lesions. Method: Forty two archival samples with cervical lesion retired from Khatam hospital and 40 sample from healthy persons used as control group. A simple and rapid method was used to detect the simultaneous amplification of the HPV consensus L1 region and HPV-16,-18, -11, -31, 33 and -35 along with the b-globin gene as an internal control. We use Multiplex PCR for detection of P53 and miR-146a rs2910164 genotypes in our lab. Finally, data analysis was performed using the 7 version of the Epi Info(TM) 2012 software and test chi-square(x2) for trend. Results: Cervix lesions were collected from 42 patients with Squamous metaplasia, cervical intraepithelial neoplasia, and cervical carcinoma. Successful DNA extraction was assessed by PCR amplification of b-actin gene (99bp). According to the results, p53 GG genotype and miR-146a rs2910164 CC genotype was significantly associated with increased risk of cervical lesions in the study population. In this study, we detected 13 HPV 18 from 42 cervical cancer. Conclusion: The connection between several SNP polymorphism and human virus papilloma in rare researches were seen. The reason of these differences in researches' findings can result in different kinds of races and geographic situations and also differences in life grooves in every region. The present study provided preliminary evidence that a p53 GG genotype and miR-146a rs2910164 CC genotype may effect cervical cancer risk in the study population, interacting synergistically with HPV 18 genotype. Our results demonstrate that the testing of p53 codon 72 polymorphism genotypes and miR-146a rs2910164 polymorphism genotypes in combination with HPV18 can serve as major risk factors in the early identification of cervical cancers. Furthermore, the results indicate the possibility of primary prevention of cervical cancer by vaccination against HPV18 in Iran.

Keywords: cervical cancer, HPV18, p53 codon 72 polymorphism, miR-146a rs2910164 polymorphism

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Authors: Lidimma Benjamin, Nimmyel Gwakzing, Wuyep Nanyi

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Technical and Vocational Education and Training (TVET) has many stakeholders, each with their own values and interests. This study will focus on the diversity of the values and interests within and across groups of stakeholders by quantifying the value that stakeholders attached to several quality attributes of TVET, and also find out to what extent TVET stakeholders differ in their values. The quality of TVET therefore, depends on how well it aligns with the values and interests of these stakeholders. The five stakeholders are parents, students, teachers, policy makers, and work place training supervisors. The 9 attributes are employer appreciation of students, graduation rate, obtained computer skills of students, mentoring hours in workplace learning/Students Industrial Work Experience Scheme (SIWES), challenge, structure, students’ appreciation of teachers, schooling hours, and attention to civic education. 346 respondents (comprising Parents, Students, Teachers, Policy Makers, and Workplace Training Supervisors) were repeatedly asked to rank a set of 4 programs, each with a specific value on the nine quality indicators. Conjoint analysis was used to obtain the values that the stakeholders assigned to the 9 attributes when evaluating the quality of TVET programs. Rank-ordered logistic regression was the statistical/tool used for ranking the respondents values assign to the attributes. The similarities and diversity in values and interests of the different stakeholders will be of use by both Nigerian government and TVET colleges, to improve the overall quality of education and the match between vocational programs and their stakeholders simultaneous evaluation and combination of information in product attributes. Such approach models the decision environment by confronting a respondent with choices that are close to real-life choices. Therefore, it is more realistically than traditional survey methods.

Keywords: TVET, vignette study, conjoint analysis, quality perception, educational stakeholders

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Authors: Khin Aye Myint, Mohd Rafii Yusop, Mohd Yusoff Abd Samad, Shairul Izan Ramlee, Mohd Din Amiruddin, Zulkifli Yaakub

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The narrow base of genetic is the main obstacle of breeding and genetic improvement in oil palm industry. In order to broaden the genetic bases, the Malaysian Palm Oil Board has been extensively collected wild germplasm from its original area of 11 African countries which are Nigeria, Senegal, Gambia, Guinea, Sierra Leone, Ghana, Cameroon, Zaire, Angola, Madagascar, and Tanzania. The germplasm collections were established and maintained as a field gene bank in Malaysian Palm Oil Board (MPOB) Research Station in Kluang, Johor, Malaysia to conserve a wide range of oil palm genetic resources for genetic improvement of Malaysian oil palm industry. Therefore, assessing the performance and genetic diversity of the wild materials is very important for understanding the genetic structure of natural oil palm population and to explore genetic resources. Principal component analysis (PCA) and Cluster analysis are very efficient multivariate tools in the evaluation of genetic variation of germplasm and have been applied in many crops. In this study, eight populations of MPOB-Senegal oil palm germplasm were studied to explore the genetic variation pattern using PCA and cluster analysis. A total of 20 yield and yield component traits were used to analyze PCA and Ward’s clustering using SAS 9.4 version software. The first four principal components which have eigenvalue >1 accounted for 93% of total variation with the value of 44%, 19%, 18% and 12% respectively for each principal component. PC1 showed highest positive correlation with fresh fruit bunch (0.315), bunch number (0.321), oil yield (0.317), kernel yield (0.326), total economic product (0.324), and total oil (0.324) while PC 2 has the largest positive association with oil to wet mesocarp (0.397) and oil to fruit (0.458). The oil palm population were grouped into four distinct clusters based on 20 evaluated traits, this imply that high genetic variation existed in among the germplasm. Cluster 1 contains two populations which are SEN 12 and SEN 10, while cluster 2 has only one population of SEN 3. Cluster 3 consists of three populations which are SEN 4, SEN 6, and SEN 7 while SEN 2 and SEN 5 were grouped in cluster 4. Cluster 4 showed the highest mean value of fresh fruit bunch, bunch number, oil yield, kernel yield, total economic product, and total oil and Cluster 1 was characterized by high oil to wet mesocarp, and oil to fruit. The desired traits that have the largest positive correlation on extracted PCs could be utilized for the improvement of oil palm breeding program. The populations from different clusters with the highest cluster means could be used for hybridization. The information from this study can be utilized for effective conservation and selection of the MPOB-Senegal oil palm germplasm for the future breeding program.

Keywords: cluster analysis, genetic variability, germplasm, oil palm, principal component analysis

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Authors: Olatunde Sunday Eludoyin, Charles Obiechina Olisa

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The study examined the spatial interactions between earthworm abundance (EA) and tree growth characteristics in ecological belts of Western Niger Delta, Nigeria. Eight 20m x 20m quadrat were delimited in the natural vegetation in each of the rainforest (RF), mangrove (M), fresh water swamp (FWS), and guinea savanna (GS) ecological belts to gather data about the tree species (TS) characteristics which included individual number of tree species (IN), diversity (Di), density (De) and richness (Ri). Three quadrats of 1m x 1m were delineated in each of the 20m x 20m quadrats to collect earthworm species the topsoil (0-15cm), and subsoil (15-30cm) and were taken to laboratory for further analysis. Descriptive statistics and inferential statistics were used for data analysis. Findings showed that a total of 19 earthworm species was found, with 58.5% individual species recorded in the topsoil and 41.5% recorded in the subsoil. The total population ofEudriliuseugeniae was predominantly highest in both topsoil (38.4%) and subsoil (27.1%). The total population of individual species of earthworm was least in GS in the topsoil (11.9%) and subsoil (8.4%). A total of 40 different species of TS was recorded, of which 55.5% were recorded in FWS, while RF was significantly highest in the species diversity(0.5971). Regression analysis revealed that Ri, IN, DBH, Di, and De of trees explained 65.9% of the variability of EA in the topsoil, while 46.9 % of the variability of earthworm abundance was explained by the floristic parameters in the subsoil.Similarly, correlation statistics revealed that in the topsoil, EA is positively and significantly correlated with Ri (r=0.35; p<0.05), IN (r=0.523; p<0.05) and De (r=0.469; p<0.05) while DBH was negatively and significantly correlated with earthworm abundance (r=-0.437; p<0.05). In the subsoil, only Ri and DBH correlated significantly with EA. The study concluded that EA in the study locations was highly influenced by tree growth species especially Ri, IN, DBH, Di, and De. The study recommended that the TSabundance should be improved in the study locations to ensure the survival of earthworms for ecosystem functions.

Keywords: interactions, earthworm abundance, tree growth, ecological zones, western niger delta

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Authors: Jin-Feng Hu

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Over the past decades, the global biodiversity has continued to decline. The threats to the terrestrial plant species have increased under anthropogenic activities and other massive ecological change impacts. The situation is much more serious in China, the third richest countries regarding plant biodiversity in the world. It was not until 1992 that the first volume of the China Plant Red Data Book was published. Nowadays, a significant number of Chinese endemic plants have been threatened (The IUCN Red List). Nevertheless, plant-originated natural products (NPs) have continued to play a crucial role in the drug discovery and development process. The opportunity for identifying new chemical entities for emerging and malignant diseases depends on a diversity of drug-producing species. Several statistical surveys unveiled that the rare and endangered plants (REPs) have proven to be better sources for drug discovery than other botanic sources. The identification of bioactive NPs from REPs reveals the importance of conservation efforts in preventing species diversity loss and addressing human diseases at the same time. Thus, there is an urgent need to investigate these fragile REPs. Since 2013, our group has initially launched a special program to systematically identify bioactive/novel NPs from REPs native to China. The selected plant species were generally collected from the remote Mountain areas, and have never been chemically or pharmacologically investigated. Due to the difficult collection of the mass-limited samples of REPs, studies on the secondary metabolites of REPs-associated endophytes would provide a promising alternative potential solution. This presentation details the achievements that related to a series of “Phytochemical and biological studies on rare and endangered plants endemic to China”.

Keywords: bioactive naturally-occrring compounds, rare and endengered plants (REPs), plant endophytes, drug discovery

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Authors: Dong-An Wang

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All kinds of microspheres have been extensively employed as carriers for drug, gene and therapeutic cell delivery. Most therapeutic cell delivery microspheres rely on a two-step methodology: fabrication of microspheres and subsequent seeding of cells onto them. In this study, we have developed a novel one-step cell encapsulation technique using a convenient and instant water-in-oil single emulsion approach to form cell-encapsulated gelatin microspheres. This technology is adopted for hyaline cartilage tissue engineering, in which autologous chondrocytes are used as therapeutic cells. Cell viability was maintained throughout and after the microsphere formation (75-100 µm diameters) process that avoids involvement of any covalent bonding reactions or exposure to any further chemicals. Further encapsulation of cell-laden microspheres in alginate gels were performed under 4°C via a prompt process. Upon the formation of alginate constructs, they were immediately relocated into CO2 incubator where the temperature was maintained at 37°C; under this temperature, the cell-laden gelatin microspheres dissolved within hours to yield similarly sized cavities and the chondrocytes were therefore suspended within the cavities inside the alginate gel bulk. Hence, the gelatin cell-laden microspheres served two roles: as cell delivery vehicles which can be removable through temperature curing, and as porogens within an alginate hydrogel construct to provide living space for cell growth and tissue development as well as better permeability for mutual diffusions. These cell-laden microspheres, namely “temperature-cured dissolvable gelatin microsphere based cell carriers” (tDGMCs), were further encapsulated in a chondrocyte-laden alginate scaffold system and analyzed by WST-1, gene expression analyses, biochemical assays, histology and immunochemistry stains. The positive results consistently demonstrated the promise of tDGMC technology in delivering these non-anchorage dependent cells (chondrocytes). It can be further conveniently translated into delivery of other non-anchorage dependent cell species, including stem cells, progenitors or iPS cells, for regeneration of tissues in internal organs, such as engineered hepatogenesis or pancreatic regeneration.

Keywords: biomaterials, tissue engineering, microsphere, hydrogel, porogen, anchorage dependence

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Authors: Tsitsi Nyukurayi Muparari, Walter Timo De Vries, Jaap Zevenbergen

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The nature of Land administration accommodates diversity in terms of both spatial data handling activities and the expertise involved, which supposedly aims to satisfy the unpredictable demands of land data and the diverse demands of the customers arising from the land. However, it is known that strategic decisions of restructuring are in most cases repelled in favour of complex structures that strive to accommodate professional diversity and diverse roles in the field of Land administration. Yet despite of this widely accepted knowledge, there is scanty theoretical knowledge concerning the psychological methodologies that can extract the deeper perceptions from the diverse spatial expertise in order to explain the invisible control arm of the polarised reception of the ideas of change. This paper evaluates Q methodology in the context of a cadastre and land registry merger (under one agency) using the Swedish cadastral system as a case study. Precisely, the aim of this paper is to evaluate the effectiveness of Q methodology towards modelling the diverse psychological perceptions of spatial professionals who are in a widely contested decision of merging the cadastre and land registry components of Land administration using the Swedish cadastral system as a case study. An empirical approach that is prescribed by Q methodology starts with the concourse development, followed by the design of statements and q sort instrument, selection of the participants, the q-sorting exercise, factor extraction by PQMethod and finally narrative development by logic of abduction. The paper uses 36 statements developed from a dominant competing value theory that stands out on its reliability and validity, purposively selects 19 participants to do the Qsorting exercise, proceeds with factor extraction from the diversity using varimax rotation and judgemental rotation provided by PQMethod and effect the narrative construction using the logic abduction. The findings from the diverse perceptions from cadastral professionals in the merger decision of land registry and cadastre components in Sweden’s mapping agency (Lantmäteriet) shows that focus is rather inclined on the perfection of the relationship between the legal expertise and technical spatial expertise. There is much emphasis on tradition, loyalty and communication attributes which concern the organisation’s internal environment rather than innovation and market attributes that reveals customer behavior and needs arising from the changing humankind-land needs. It can be concluded that Q methodology offers effective tools that pursues a psychological approach for the evaluation and gradations of the decisions of strategic change through extracting the local perceptions of spatial expertise.

Keywords: cadastre, factor extraction, land administration merger, land registry, q-methodology, rotation

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Authors: Maffo Maffo Nicole Liliane, Mounmeni Kpoumie Hubert, Mbaire Matindje Karl Marx, Zapfack Louis

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Sacred forests play a valuable role in conserving local biodiversity and provide numerous ecosystem services in Cameroon. The study was carried out in the sacred forests of Bandrefam and Batoufam (western Cameroon). The aim was to estimate the diversity of woody species, carbon stocks and degradation factors in these sacred forests. The floristic inventory was carried out in plots measuring 25m × 25m for trees with diameters greater than 10 cm and 5m × 5m for trees with diameters less than 10 cm. Carbon stocks were estimated using the non-destructive method and the allometric equations. Data on degradation factors were collected using semi-structured surveys in the Bandrefam and Batoufam neighborhoods. The floristic inventory identified 65 species divided into 57 genera and 30 families in the Bandrefam Sacred Forest and 45 species divided into 42 genera and 27 families in the Batoufam Sacres Forest. The families common to both sacred forests are as follows: Phyllanthaceae, Fabaceae, Moraceae, Lamiaceae, Malvaceae, Rubiaceae, Meliaceae, Anacardiaceae, and Sapindaceae. Three genera are present in both sites. These are: Albizia, Macaranga, Trichillia. In addition, there are 27 species in common between the two sites. The total carbon stock is 469.26 tC/ha at Batoufam and 291.41 tC/ha at Bandrefam. The economic value varies between 15 823 877.05 fcfa at Batoufam and 9 825 530.528 fcfa at Bandrefam. The study shows that despite the sacred nature of these forests, they are subject to degradation factors such as bushfires (35.42 %), the creation of plantations (23.96 %), illegal timber exploitation (21.88 %), young people's lack of interest in the notion of conservation (9.38 %), climate change (7.29 %) and growing urbanization (2.08 %). These factors threaten biodiversity and reduce carbon storage in these forests.

Keywords: sacred forests, degradation factors, carbon stocks, semi-structured surveys

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Authors: Sandra Pietri, Helene Dubout, Sabrina Ena, Candice Hoste, Enrico Bastianelli

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Bone Therapeutics is a bone cell therapy company addressing high unmet medical needs in the field of bone fracture repair, more specifically in non-union and delayed-union fractures where the bone repair process is impaired. The company has developed a unique allogeneic osteoblastic cell product (ALLOB®) derived from bone marrow which is currently tested in humans in the indication of delayed-union fractures. The purpose of our study was to directly compare ALLOB® vs. non-differentiated mesenchymal stem cells (MSC) for their in vitro osteogenic characteristics and their in vivo osteogenic potential in order to determine which cellular type would be the most adapted for bone fracture repair. Methods: Healthy volunteers’ bone marrow aspirates (n=6) were expended (i) into BM-MSCs using a complete MSC culture medium or (ii) into ALLOB® cells according to its manufacturing process. Cells were characterized in vitro by morphology, immunophenotype, gene expression and differentiation potential. Additionally, their osteogenic potential was assessed in vivo in the subperiosteal calvaria bone formation model in nude mice. Results: The in vitro side-by-side comparison studies showed that although ALLOB® and BM-MSC shared some common general characteristics such as the 3 minimal MSC criteria, ALLOB® expressed significantly higher levels of chondro/osteoblastic genes such as BMP2 (fold change (FC) > 100), ALPL (FC > 12), CBFA1 (FC > 3) and differentiated significantly earlier than BM-MSC toward the osteogenic lineage. Moreover the bone formation model in nude mice demonstrated that used at the same cellular concentration, ALLOB® was able to induce significantly more (160% vs.107% for control animals) bone formation than BM-MSC (118% vs. 107 % for control animals) two weeks after administration. Conclusion: Our side-by-side comparison studies demonstrated that in vitro and in vivo, ALLOB® displays superior osteogenic capacity to BM-MScs and is therefore a better candidate for the treatment of bone fractures.

Keywords: gene expression, histomorphometry, mesenchymal stem cells, osteogenic differentiation potential, preclinical

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Authors: Yu-Mei Hsueh, Wei-Jen Chen, Yung-Kai Huang, Cheng-Shiuan Tsai, Kuo-Cheng Yeh

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Prostate cancer occurs in men over the age of 50, and rank sixth of the top ten cancers in Taiwan, and the incidence increased gradually over the past decade in Taiwan. Arsenic is confirmed as a carcinogen by International Agency for Research on (IARC). Arsenic induces oxidative stress may be a risk factor for prostate cancer, but the mechanism is not clear. Selenium is an important antioxidant element. Whether the association between plasma selenium levels and risk of prostate cancer are modified by different genotype of selenoprotein is still unknown. Glutathione peroxidase, selenoprotein P (SEPP1) and 15 kDa selenoprotein (SEP 15) are selenoprotein and regulates selenium transport and the oxidation and reduction reaction. However, the association between gene polymorphisms of selenoprotein and prostate cancer is not yet clear. The aim of this study is to determine the relationship between plasma selenium, polymorphism of selenoprotein, urinary total arsenic concentration and prostate cancer. This study is a hospital-based case-control study. Three hundred twenty-two cases of prostate cancer and age (±5 years) 1:1 matched 322 control group were recruited from National Taiwan University Hospital, Taipei Medical University Hospital, and Wan Fang Hospital. Well-trained personnel carried out standardized personal interviews based on a structured questionnaire. Information collected included demographic and socioeconomic characteristics, lifestyle and disease history. Blood and urine samples were also collected at the same time. The Research Ethics Committee of National Taiwan University Hospital, Taipei, Taiwan, approved the study. All patients provided informed consent forms before sample and data collection. Buffy coat was to extract DNA, and the polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) was used to measure the genotypes of SEPP1 rs3797310, SEP15 rs5859, GPX1 rs1050450, GPX2 rs4902346, GPX3 rs4958872, and GPX4 rs2075710. Plasma concentrations of selenium were determined by inductively coupled plasma mass spectrometry (ICP-MS).Urinary arsenic species concentrations were measured by high-performance liquid chromatography links hydride generator and atomic absorption spectrometer (HPLC-HG-AAS). Subject with high education level compared to those with low educational level had a lower prostate cancer odds ratio (OR) Mainland Chinese and aboriginal people had a lower OR of prostate cancer compared to Fukien Taiwanese. After adjustment for age, educational level, subjects with GPX1 rs1050450 CT and TT genotype compared to the CC genotype have lower, OR of prostate cancer, the OR and 95% confidence interval (Cl) was 0.53 (0.31-0.90). SEPP1 rs3797310 CT+TT genotype compared to those with CC genotype had a marginally significantly lower OR of PC. The low levels of plasma selenium and the high urinary total arsenic concentrations had the high OR of prostate cancer in a significant dose-response manner, and SEPP1 rs3797310 genotype modified this joint association.

Keywords: prostate cancer, plasma selenium concentration, urinary total arsenic concentrations, glutathione peroxidase, selenoprotein P, selenoprotein 15, gene polymorphism

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Authors: Endah Tri Lestari, Mustafa Mahmud Amin, Elmeida Effendy

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Background: Vitamin D levels with schizophrenia is lower than the control due to lifestyle and physical health factors such as smoking, increases of body mass index, inactivity, and social withdrawal including decreases of sunlight exposure. Asia has the lowest average of vitamin D serum levels and Europe with lighter colored skin has higher serum levels of vitamin D. Indonesia is a tropical region in the area of the equator, and has only two seasons: the rainy season and summer. The Indonesian people are known as a nation that has a diversity of ethnic groups that exist in many areas. Each tribe has differences in living habits. Ethnic diversity, culture, religion, customs, geographical location, this is reflected in our daily lives that will affect the levels of vitamin D in patients with schizophrenic. Individuals with darker skin tend to be deficient in vitamin D and increases the chance of schizophrenia than the general population. Aims: To determine the difference of vitamin D serum by ethnicity between Bataknese and Malayan schizophrenic patient. Methods: This study was an analytical study to recruited 60 subjects of schizophrenic male patient (30 Bataknese and 30 Malayan), aged between 15 to 55 years old, period at May - November 2016, the acute phase with no agitation. Statistical analysis was using T- independent test. Blood sample for vitamin D serum was using ELFA method. Results: The vitamin D serum levels were lower in Bataknese ethnic group schizophrenic patients in highland than Malayan ethnic group in lowland, reaching statistically (22.9±3.33 ng/ml) vs (27.9±4.19 ng/ml) p < 0,001. Conclusion: There are significant differences of vitamin D serum Status by Ethnicity between Bataknese in the Highland and Malayan in the Lowland on Schizophrenic Patient in North Sumatera.

Keywords: schizophrenia, serum vitamin D, ethnicity

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Authors: Umara Nissar Rafiqi, Irum Gul, Nazima Nasrullah, Monica Saifi, Malik Z. Abdin

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Background: Stevia rebaudiana, a member of Asteraceae family is an important medicinal plant and produces a commercially used non-caloric natural sweetener, which is also an alternate herbal cure for diabetes. Steviol glycosides are the main sweetening compounds present in these plants. Secondary metabolites are crucial to the adaption of plants to the environment and its overcoming stress conditions. In agricultural procedures, the abiotic stresses like salinity, high metal toxicity and drought, in particular, are responsible for the majority of the reduction that differentiates yield potential from harvestable yield. Salt stress and heavy metal toxicity lead to increased production of reactive oxygen species (ROS). To avoid oxidative damage due to ROS and osmotic stress, plants have a system of anti-oxidant enzymes along with several stress induced enzymes. This helps in scavenging the ROS and relieve the osmotic stress in different cell compartments. However, whether stress induced toxicity modulates the activity of these enzymes in Stevia rebaudiana is poorly understood. Aim: The present study focussed on the effect of salinity, heavy metal toxicity (lead and mercury) on physiological traits and transcriptional profiling of Stevia rebaudiana. Method: Stevia rebaudiana plants were collected from the Central Institute of Medicinal and Aromatic plants (CIMAP), Patnagar, India and maintained under controlled conditions in a greenhouse at Hamdard University, Delhi, India. The plants were subjected to different concentrations of salt (0, 25, 50 and 75 mM respectively) and heavy metals, lead and mercury (0, 100, 200 and 300 µM respectively). The physiological traits such as shoot length, root numbers, leaf growth were evaluated. The samples were collected at different developmental stages and analysed for transcription profiling by RT-PCR. Transcriptional studies in stevia rebaudiana involves important antioxidant enzymes: catalase (CAT), superoxide dismutase (SOD), cytochrome P450 monooxygenase (CYP) and stress induced aquaporin (AQU), auxin repressed protein (ARP-1), Ndhc gene. The data was analysed using GraphPad Prism and expressed as mean ± SD. Result: Low salinity and lower metal toxicity did not affect the fresh weight of the plant. However, this was substantially decreased by 55% at high salinity and heavy metal treatment. With increasing salinity and heavy metal toxicity, the values of all studied physiological traits were significantly decreased. Chlorosis in treated plants was also observed which could be due to changes in Fe:Zn ratio. At low concentrations (upto 25 mM) of NaCl and heavy metals, we did not observe any significant difference in the gene expressions of treated plants compared to control plants. Interestingly, at high salt concentration and high metal toxicity, a significant increase in the expression profile of stress induced genes was observed in treated plants compared to control (p < 0.005). Conclusion: Stevia rebaudiana is tolerant to lower salt and heavy metal concentration. This study also suggests that with the increase in concentrations of salt and heavy metals, harvest yield of S. rebaudiana was hampered.

Keywords: Stevia rebaudiana, natural sweetener, salinity, heavy metal toxicity

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Authors: Evans V. Mauda, Stefan H. Foord, Thinandavha C. Munyai

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The greatest threat to biodiversity is a loss of habitat through landscape fragmentation and attrition. Land use changes are therefore among the most immediate drivers of species diversity. Urbanization and agriculture are the main drivers of habitat loss and transformation in the Savanna biomes of South Africa. Agricultural expansion and the intensification in particular, take place at the expense of biodiversity and will probably be the primary driver of biodiversity loss in this century. Arthropods show measurable behavioural responses to changing land mosaics at the smallest scale and heterogeneous environments are therefore predicted to support more complex and diverse biological assemblages. Ants are premier soil turners, channelers of energy and dominate insect fauna, while spiders are a mega-diverse group that can regulate other invertebrate populations. This study aims to quantify the response of these two taxa in a rural-urban mosaic of a rapidly developing communal area. The study took place in and around two villages in the north-eastern corner of South Africa. Two replicates for each of the dominant land use categories, viz. urban settlements, dryland cultivation and cattle rangelands, were set out in each of the villages and sampled during the dry and wet seasons for a total of 2 villages × 3 land use categories × 2 seasons = 24 assemblages. Local scale variables measured included vertical and horizontal habitat structure as well as structural and chemical composition of the soil. Ant richness was not affected by land use but local scale variables such as vertical vegetation structure (+) and leaf litter cover (+), although vegetation complexity at lower levels was negatively associated with ant richness. However, ant richness was largely shaped by regional and temporal processes invoking the importance of dispersal and historical processes. Spider species richness was mostly affected by land use and local conditions highlighting their landscape elements. Spider richness did not vary much between villages and across seasons and seems to be less dependent on context or history. There was a considerable amount of variation in spider richness that was not explained and this could be related to factors which were not measured in this study such as temperature and competition. For both ant and spider assemblages the constrained ordination explained 18 % of variation in these taxa. Three environmental variables (leaf litter cover, active carbon and rock cover) were important in explaining ant assemblage structure, while two (sand and leaf litter cover) were important for spider assemblage structure. This study highlights the importance of disturbance (land use activities) and leaf litter with the associated effects on ant and spider assemblages across the study area.

Keywords: ants, assemblages, biosphere, diversity, land use, spiders, urbanization

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Authors: Surya Prasad Sharma, Suyash Katdare, Syed Ainul Hussain

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Human-induced factors contributed to the population decline of crocodylians in India which became evident by the mid-20th century when authorities forewarned the extinction risk for the crocodile and proposed regulation in the crocodile trade. The proposed action led to the enactment of national and international wildlife regulations to prohibit the trade-in of crocodile skins and parts. Subsequently, conservation translocation programs were initiated to restore the species in the wild through a 'head-start' approach. In India, the crocodile conservation program, which began in the early 1970s, has been one of India's longest-running conservation initiatives. The gharial (Gavialis gangeticus) population has benefitted, and the gharial number increased rapidly owing to these efforts. The immediate risk of extinction was averted as the gharial has recovered due to decades-long cumulative conservation efforts, the consideration of the genetic for monitoring the recovery of the recovered populations is still lacking. Hence, we assessed the genetic diversity of the Girwa gharial population in India using six polymorphic nuclear microsatellites loci and mitochondrial control region. The number of alleles per loci ranged between 2 to 5, and the allelic richness (Ar) was 2.67 ± 0.49, and the observed (Ho) and expected (He) heterozygosities were 0.42 ± 0.08 and 0.42 ± 0.09, respectively. The M-ratio yielded a value of (0.41 ± 0.16) lower than critical M, suggesting a genetic bottleneck in the Girwa population. We observed more mitochondrial control region haplotypes in the Girwa population than previously reported in the largest gharial population in the Chambal River. Overall, our study indicates that genetic diversity remains low despite the recovery in the Girwa population. Hence, we recommend a range-wide genetic assessment of gharial populations using high-throughput techniques to identify the source population and plan future translocation programs.

Keywords: conservation translocation, recovery, crocodile, bottleneck

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Authors: Irena Balaban Cakirpaloglu, Karla Hrbackova

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This paper focuses on the issue of intercultural competencies of university students who are preparing to work in assisting professions. In recent years, the Czech Republic has become a major destination for many people from different cultural environments, and there is a growing need for workers in assisting professions to be able to respond flexibly and adequately to the changing living conditions of multicultural coexistence. The main objective of this study is to analyse the preparedness of students in assisting professions in relation to intercultural competencies. Intercultural competences include several essential skills for working successfully with diversity. Taking into account the main objective of this research, a pilot study was conducted among students of Social Pedagogy at the Faculty of Humanities at Tomas Bata University in Zlin in the academic year 2017/2018. The research sample consisted of 116 students. To obtain the data, we used the Cross-Cultural Adaptability Inventory (CCAI) by Kelley and Meyers. The inventory maps strengths and weaknesses in 4 skill areas: Emotional Resilience, Flexibility/Openness, Perceptual Acuity and Personal Autonomy. This inventory also examines individual ability to succeed in intercultural interaction and communication. The results obtained from the survey were statistically processed and analysed using the relevant statistical methods. The results of the survey point to the fact that students of social pedagogy achieve average to below average results in individual skill areas. At the same time, significant differences have been detected among the students with work experience in multicultural environment and those with no experience.

Keywords: cross–cultural adaptability inventory, diversity, intercultural competences, students of social pedagogy

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Authors: Seyedeh Banafsheh Bagheri Marzouni, Sona Rostampour Yasouri

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Salmonella is one of the most important common pathogens between humans and animals worldwide. Globally, the prevalence of the disease in humans is due to the consumption of food contaminated with animal-derived Salmonella. These foods include eggs, red meat, chicken, and milk. Contamination of chicken and its products with Salmonella may occur at any stage of the chicken processing chain. Salmonella infection is usually not fatal. However, its occurrence is considered dangerous in some individuals, such as infants, children, the elderly, pregnant women, or individuals with weakened immune systems. If Salmonella infection enters the bloodstream, the possibility of contamination of tissues throughout the body will arise. Therefore, determining the potential risk of Salmonella at various stages is essential from the perspective of consumers and public health. The aim of this study is to isolate and identify Salmonella from chicken samples distributed in the Tehran market using the Gold standard culture method and PCR techniques based on specific genes, invA and ent. During the years 2022-2023, sampling was performed using swabs from the liver and intestinal contents of distributed chickens in the Tehran province, with a total of 120 samples taken under aseptic conditions. The samples were initially enriched in buffered peptone water (BPW) for pre-enrichment overnight. Then, the samples were incubated in selective enrichment media, including TT broth and RVS medium, at temperatures of 37°C and 42°C, respectively, for 18 to 24 hours. Organisms that grew in the liquid medium and produced turbidity were transferred to selective media (XLD and BGA) and incubated overnight at 37°C for isolation. Suspicious Salmonella colonies were selected for DNA extraction, and PCR technique was performed using specific primers that targeted the invA and ent genes in Salmonella. The results indicated that 94 samples were Salmonella using the PCR technique. Of these, 71 samples were positive based on the invA gene, and 23 samples were positive based on the ent gene. Although the culture technique is the Gold standard, PCR is a faster and more accurate method. Rapid detection through PCR can enable the identification of Salmonella contamination in food items and the implementation of necessary measures for disease control and prevention.

Keywords: culture, PCR, salmonella spp, salmonella enteritidis

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Authors: Aleksandra Bylinska, Samantha Slight-Webb, Kevin Thomas, Miles Smith, Susan Macwana, Nicolas Dominguez, Eliza Chakravarty, Joan T. Merrill, Judith A. James, Joel M. Guthridge

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Background: Systemic Lupus Erythematosus (SLE) is an interferon-related autoimmune disease characterized by B cell dysfunction. One of the main hallmarks is a loss of tolerance to self-antigens leading to increased levels of autoantibodies against nuclear components (ANAs). However, up to 20% of healthy ANA+ individuals will not develop clinical illness. SLE is more prevalent among women and minority populations (African, Asian American and Hispanics). Moreover, African Americans have a stronger interferon (IFN) signature and develop more severe symptoms. The exact mechanisms involved in ethnicity-dependent B cell dysregulation and the progression of autoimmune disease from ANA+ healthy individuals to clinical disease remains unclear. Methods: Peripheral blood mononuclear cells (PBMCs) from African (AA) and European American (EA) ANA- (n=12), ANA+ (n=12) and SLE (n=12) individuals were assessed by multimodal scRNA-Seq/CITE-Seq methods to examine differential gene signatures in specific B cell subsets. Library preparation was done with a 10X Genomics Chromium according to established protocols and sequenced on Illumina NextSeq. The data were further analyzed for distinct cluster identification and differential gene signatures in the Seurat package in R and pathways analysis was performed using Ingenuity Pathways Analysis (IPA). Results: Comparing all subjects, 14 distinct B cell clusters were identified using a community detection algorithm and visualized with Uniform Manifold Approximation Projection (UMAP). The proportion of each of those clusters varied by disease status and ethnicity. Transitional B cells trended higher in ANA+ healthy individuals, especially in AA. Ribonucleoprotein high population (HNRNPH1 elevated, heterogeneous nuclear ribonucleoprotein, RNP-Hi) of proliferating Naïve B cells were more prevalent in SLE patients, specifically in EA. Interferon-induced protein high population (IFIT-Hi) of Naive B cells are increased in EA ANA- individuals. The proportion of memory B cells and plasma cells clusters tend to be expanded in SLE patients. As anticipated, we observed a higher signature of cytokine-related pathways, especially interferon, in SLE individuals. Pathway analysis among AA individuals revealed an NRF2-mediated Oxidative Stress response signature in the transitional B cell cluster, not seen in EA individuals. TNFR1/2 and Sirtuin Signaling pathway genes were higher in AA IFIT-Hi Naive B cells, whereas they were not detected in EA individuals. Interferon signaling was observed in B cells in both ethnicities. Oxidative phosphorylation was found in age-related B cells (ABCs) for both ethnicities, whereas Death Receptor Signaling was found only in EA patients in these cells. Interferon-related transcription factors were elevated in ABCs and IFIT-Hi Naive B cells in SLE subjects of both ethnicities. Conclusions: ANA+ healthy individuals have altered gene expression pathways in B cells that might drive apoptosis and subsequent clinical autoimmune pathogenesis. Increases in certain regulatory pathways may delay progression to SLE. Further, AA individuals have more elevated activation pathways that may make them more susceptible to SLE.

Keywords:

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Authors: Mei Chen, Yingping Hou, Michelle Hao, Soheil Aghamohammadzadeh, Esteban Terzo, Roger Clark, Vijay Modur

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Background: Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a genetic skin condition characterized by skin tearing and unremitting blistering upon minimal trauma. Repeated blistering, fibrosis, and scarring lead to aggressive squamous cell carcinoma later in life. RDEB is caused by mutations in the COL7A1 gene encoding collagen type VII (C7), the major component of anchoring fibrils mediating epidermis-dermis adherence. Nonsense mutations in the COL7A1 gene of a subset of RDEB patients leads to premature termination codons (PTC). Similarly, most Junctional Epidermolysis Bullosa (JEB) cases are caused by nonsense mutations in the LAMB3 gene encoding the β3 subunit of laminin 332. Currently, there is an unmet need for the treatment of RDEB and JEB. Zikani Therapeutics has discovered an array of macrocyclic compounds with ring structures similar to macrolide antibiotics that can facilitate readthrough activity of nonsense mutations in the COL7A1 and LAMB3 genes by acting as Ribosome Modulating Agents (RMAs). The medicinal chemistry synthetic advancements of these macrocyclic compounds have allowed targeting the human ribosome while preserving the structural elements responsible for the safety and pharmacokinetic profile of clinically used macrolide antibiotics. Methods: C7 expression was used as a measure of readthrough activity by immunoblot assays in two primary human fibroblasts from RDEB patients (R578X/R578X and R163X/R1683X-COL7A1). Similarly, immunoblot assays in C325X/c.629-12T > A-LAMB3 keratinocytes were used to measure readthrough activity for JEB. The relative readthrough activity of each compound was measured relative to Gentamicin. An imaging-based fibroblast migration assay was used as an assessment of C7 functionality in RDEB-fibroblasts over 16-20 hrs. The incubation period for the above experiments was 48 hrs for RDEB fibroblasts and 72 hours for JEB keratinocytes. Results: 9 RMAs demonstrated increased protein expression in both patient RDEB fibroblasts. The highest readthrough activity at tested concentrations without cytotoxicities increased protein expression up to 179% of Gentamicin (400 µg/ml), with favored readthrough activity in R163X/R1683X-COL7A1 fibroblasts. Concurrent with protein expression, fibroblast hypermotility phenotype observed in RDEB was rescued by reducing motility by ~35% to WT levels (the same level as 690 µM Gentamicin treated cells). Laminin β3 expression was also shown to be increased by 6 RMAs in keratinocytes to 33-83% of (400 µg/ml) Gentamicin. Conclusions: To date, 9 RMAs have been identified that enhance the expression of functional C7 in a mutation-dependent manner in two different RDEB patient fibroblast backgrounds (R578X/R578X and R163X/R1683X-COL7A1). A further 6 RMAs have been identified that enhance the readthrough of C325X-LAMB3 in JEB patient keratinocytes. Based on the clinical trial conducted by us with topical gentamycin in 2017, Zikani’s RMAs achieve clinically significant levels of read-through for the treatment of recessive dystrophic and Junctional Epidermolysis Bullosa.

Keywords: epidermolysis bullosa, nonsense mutation, readthrough, ribosome modulation

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Authors: Natalia Moreno-Castellanos, Amaia Rodriguez, Gema Frühbeck

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Introduction: In adipocytes, the cytoskeleton undergoes important expression and distribution in adipocytes rearrangements during adipogenesis and in obesity. Indeed, a role for these proteins in the regulation of adipocyte differentiation and response to insulin has been demonstrated. Recently, septins have been considered as new components of the cytoskeletal network that interact with other cytoskeletal elements (actin and tubulin) profoundly modifying their dynamics. However, these proteins have not been characterized as yet in adipose tissue. In this work, were examined the cellular, molecular and functional features of a member of this family, septin 11 (SEPT11), in adipocytes and evaluated the impact of obesity on the expression of this protein in human adipose tissue. Methods: Adipose gene and protein expression levels of SEPT11 were analysed in human samples. SEPT11 distribution was evaluated by immunocytochemistry, electronic microscopy, and subcellular fractionation techniques. GST-pull down, immunoprecipitation and a Yeast-Two Hybrid (Y2H) screening were used to identify the SEPT11 interactome. Gene silencing was employed to assess the role of SEPT11 in the regulation of insulin signaling and lipid metabolism in adipocytes. Results: SEPT11 is expressed in human adipocytes, and its levels increased in both omental and subcutaneous adipose tissue in obesity, with SEPT11 mRNA content positively correlating with parameters of insulin resistance in subcutaneous fat. In non-stimulated adipocytes, SEPT11 immunoreactivity showed a ring-like distribution at the cell surface and associated to caveolae. Biochemical analyses showed that SEPT11 interacted with the main component of caveolae, caveolin-1 (CAV1) as well as with the fatty acid-binding protein, FABP5. Notably, the three proteins redistributed and co-localized at the surface of lipid droplets upon exposure of adipocytes to oleate. In this line, SEPT11 silencing in 3T3-L1 adipocytes impaired insulin signaling and decreased insulin-induced lipogenesis. Conclusions: Those findings demonstrate that SEPT11 is a novel component of the adipocyte cytoskeleton that plays an important role in the regulation of lipid traffic, metabolism and can thus represent a potential biomarker of insulin resistance in obesity in adipocytes through its interaction with both CAV1 and FABP5.

Keywords: caveolae, lipid metabolism, obesity, septins

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Authors: David Ahiamadia, Ramilan Thiagarajah, Peter Tozer

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In Sub Saharan Africa, farm typologies have been used as a practical way to address heterogeneity among farming systems which is mostly done by grouping farms into subsets with similar characteristics. Due to the complexity in farming systems among farm households, it is not possible to formulate policy recommendations for individual farmers. As a result, this study employs a multivariate statistical approach using Principal Component Analysis (PCA) coupled with cluster analysis to reduce heterogeneity in a 615-household data set from the Africa Rising Baseline Evaluation Survey for 25 farming communities in Northern Ghana. Variables selected for the study were mostly socio-economic, production potential, production intensity, production orientation, crop diversity, food security, resource endowments, and climate risk variables. To avoid making some individuals in the subpopulation worse off when aclimate risk intervention is broadly implemented, the findings of the study also account for diversity in climate risk perception among the different farm types identified and their response strategies towards climate risk. The climate risk variables used in this study involve the most severeclimate shock types perceived by the household, household response to climate shock type, and reason for crop failure (i.e., maize, rice, and groundnut). Eventually, four farm types, each with an adequate level of homogeneity in climate risk perception and response strategies, were identified. Farm type 1 and 3 were wealthy with a lower degree of climate risk perception compared to farm type 2 and 4. Also, relatively wealthy farmers used asset liquidation as a climate risk management strategy, whereas poor farmers resorted to engaging in spiritual activities such as prayers, sacrifices, and divine consultations.

Keywords: smallholder, households, climate risk, variables, typologies

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Authors: Asma Rashid, Shazia Iram, Iftikhar Ahmad

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Mango sudden death is an emerging problem in Pakistan. As its prevalence is observed in almost all mango growing areas and severity varied from 2-5% in Punjab and 5-10% in Sindh. Symptoms on affected trees include bark splitting, discoloration of the vascular tissue, wilting, gummosis and at the end rapid death. Total of n= 45 isolates were isolated from different mango growing areas of Punjab and Sindh. Pathogenicity of these fungal isolates was tested through artificial inoculation method on different hosts (potato tubers, detached mango leaves, detached mango twigs and mango plants) under controlled conditions and all were proved pathogenic with varying degree of aggressiveness in reference to control. The findings of the present study proved that out of these four methods, potato tubers inoculation method was the most ideal as this fix the inoculums on the target site. Increased fungal growth and spore numbers may be due to soft tissues of potato tubers from which Ceratocystis isolates can easily pass. Lesion area on potato tubers was in the range of 7.09-0.14 cm2 followed by detached mango twigs which were ranged from 0.48-0.09 cm2). All pathological results were proved highly significant at P<0.05 through ANOVA but isolate to isolate showed non-significant behaviour but they have the positive effect on lesion area. Re-isolation of respective fungi was achieved with 100 percent success which results in the verification of Koch’s postulates. DNA of fungal pathogens was successfully extracted through phenol chloroform method. Amplification was done through ITS, b-tubulin gene, and Transcription Elongation Factor (EF1-a) gene primers and the amplified amplicons were sequenced and compared from NCBI which showed 99-100 % similarity with Ceratocystis manginecans. Fungus Ceratocystis manginecans formed one of strongly supported sub-clades through phylogenetic tree. Results obtained through this work would be supportive in establishment of relation of isolates with their region and will give information about pathogenicity level of isolates that would be useful to develop the management policies to reduce the afflictions in orchards caused by mango sudden death.

Keywords: artificial inoculation, mango, Ceratocystis manginecans, phylogenetic, screening

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Authors: Hosam Abdelhady

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Filming the behaviors of individual DNA molecules in their environment when they interact with individual medicinal nano-polymers in a molecular scale has opened the door to understand the effect of the molecular shape, size, and incubation time with nanocarriers on optimizing the design of robust genetic Nano molecules able to resist the enzymatic degradation, enter the cell, reach to the nucleus and kill individual cancer cells in their environment. To this end, we will show how we applied the 4D AFM as a guide to finetune the design of genetic nanoparticles and to film the effects of these nanoparticles on the nanomechanical and morphological profiles of individual cancer cells.

Keywords: AFM, dendrimers, nanoparticles, DNA, gene therapy, imaging

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Authors: Thais O. de Paula, Marjorie R. A. Sarmiento, Francis M. Borges, Alessandra B. Ferreira-Machado, Juliana A. Resende, Dioneia E. Cesar, Vania L. Silva, Claudio G. Diniz

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Obesity is currently a worldwide public health threat, being considered a pandemic multifactorial disease related to the human gut microbiota (GM). Add to that GM is considered an important reservoir of antimicrobial resistance genes (ARG) and little is known on GM and ARG in obesity, considering the altered physiology and xenobiotics intake. As regional and social behavior may play important roles in GM modulation, and most of the studies are based on small sample size and various methodological approaches resulting in difficulties for data comparisons, this study was focused on the investigation of GM bacterial diversity in obese (OB), overweight (OW) and eutrophic individuals (ET) considering their nutritional, clinical and social characteristics; and comparative screening of AGR related to their physiology and xenobiotics intake. Microbial community was accessed by FISH considering phyla as a taxonomic level, and PCR-DGGE followed by dendrograms evaluation (UPGMA method) from fecal metagenome of 72 volunteers classified according to their body mass index (BMI). Nutritional, clinical, social parameters and xenobiotics intake were recorded for correlation analysis. The fecal metagenome was also used as template for PCR targeting 59 different ARG. Overall, 62% of OB were hypertensive, and 12% or 4% were, regarding the OW and ET individuals. Most of the OB were rated as low income (80%). Lower relative bacterial densities were observed in the OB compared to ET for almost all studied taxa (p < 0.05) with Firmicutes/Bacteroidetes ratio increased in the OB group. OW individuals showed a bacterial density representative of GM more likely to the OB. All the participants were clustered in 3 different groups based on the PCR-DGGE fingerprint patterns (C1, C2, C3), being OB mostly grouped in C1 (83.3%) and ET mostly grouped in C3 (50%). The cluster C2 showed to be transitional. Among 27 ARG detected, a cluster of 17 was observed in all groups suggesting a common core. In general, ARG were observed mostly within OB individuals followed by OW and ET. The ratio between ARG and bacterial groups may suggest that AGR were more related to enterobacteria. Positive correlations were observed between ARG and BMI, calories and xenobiotics intake (especially use of sweeteners). As with nutritional and clinical characteristics, our data may suggest that GM of OW individuals behave in a heterogeneous pattern, occasionally more likely to the OB or to the ET. Regardless the regional and social behaviors of our population, the methodological approaches in this study were complementary and confirmatory. The imbalance of GM over the health-disease interface in obesity is a matter of fact, but its influence in host's physiology is still to be clearly elucidated to help understanding the multifactorial etiology of obesity. Although the results are in agreement with observations that GM is altered in obesity, the altered physiology in OB individuals seems to be also associated to the increased xenobiotics intake and may interfere with GM towards antimicrobial resistance, as observed by the fecal metagenome and ARG screening. Support: FAPEMIG, CNPQ, CAPES, PPGCBIO/UFJF.

Keywords: antimicrobial resistance, bacterial diversity, gut microbiota, obesity

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