Search results for: genetic diversity and viral proteins

Authors: Jin Hwan Cheong, Mina Hwang, Myung Hoon Han, Je Il Ryu, Young ha Oh, Seong Ho Koh, Wu Duck Won, Byung Jin Ha

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Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) has been reported to play critical roles in the proliferation of various cancer cells. However, the roles of LGR5 in brain tumors and the specific intracellular signaling proteins directly associated with it remain unknown. Expression of LGR5 was first measured in normal brain tissue, meningioma, and pituitary adenoma of humans. To identify the downstream signaling pathways of LGR5, siRNA-mediated knockdown of LGR5 was performed in SH-SY5Y neuroblastoma cells followed by proteomics analysis with 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE). In addition, the expression of LGR5-associated proteins was evaluated in LGR5-inꠓhibited neuroblastoma cells and in human normal brain, meningioma, and pituitary adenoma tissue. Proteomics analysis showed 12 protein spots were significantly different in expression level (more than two-fold change) and subsequently identified by peptide mass fingerprinting. A protein association network was constructed from the 12 identified proteins altered by LGR5 knockdown. Direct and indirect interactions were identified among the 12 proteins. HSP 90-beta was one of the proteins whose expression was altered by LGR5 knockdown. Likewise, we observed decreased expression of proteins in the hnRNP subfamily following LGR5 knockdown. In addition, we have for the first time identified significantly higher hnRNP family expression in meningioma and pituitary adenoma compared to normal brain tissue. Taken together, LGR5 and its downstream sigꠓnaling play critical roles in neuroblastoma and brain tumors such as meningioma and pituitary adenoma.

Keywords: LGR5, neuroblastoma, meningioma, pituitary adenoma, hnRNP

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Authors: Raquel Amanda Villamizar Gallardo, Oscar Orlando Ortíz Rodríguez

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Enteric viruses are cosmopolitan agents present in several environments including water. These viruses can cause different diseases including gastroenteritis, hepatitis, conjunctivitis, respiratory problems among others. Although in Colombia there are not regulations concerning to routine viral analysis of drinking water, an enhanced understanding of viral pollution and resistance to treatments is desired in order to assure pure water to the population. Viral detection is often complex due to the need of specialized and time-consuming procedures. In addition, viruses are highly diluted in water which is a drawback from the analytical point of view. To this end, a fast and selective detection method for detection enteric viruses (i.e. Hepatitis A and Rotavirus) were applied. Micro- magnetic particles were functionalized with monoclonal antibodies anti-Hepatitis and anti-Rotavirus and they were used to capture, concentrate and separate whole viral particles in raw water and drinking water samples from four treatment plants identified as CAR-01, MON-02, POR-03, TON-04 and located in the Northeastern Colombia. Viruses were molecularly by using RT-PCR One Step Superscript III. Each plant was analyzed at the entry and exit points, in order to determine the initial presence and eventual reduction of Hepatitis A and Rotavirus after disinfection. The results revealed the presence of both enteric viruses in a 100 % of raw water analyzed in all plants. This represents a potential health hazard, especially for those people whose use this water for agricultural purposes. However, in drinking water analysis, enteric viruses was only positive in CAR-01, where was found the presence of Rotavirus. As a conclusion, the results confirm Rotavirus as the best indicator to evaluate the efficacy of potable treatment plant in eliminating viruses. CAR potable water plant should improve their disinfection process in order to remove efficiently enteric viruses.

Keywords: drinking water, hepatitis A, rotavirus, virus removal

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Authors: Radhwan Yousif Sedik Al-Jawadi

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Optimization is necessary for finding appropriate solutions to a range of real-life problems. In particular, genetic (or more generally, evolutionary) algorithms have proved very useful in solving many problems for which analytical solutions are not available. In this paper, we present an optimization algorithm called Dynamic Schema with Dissimilarity and Similarity of Chromosomes (DSDSC) which is a variant of the classical genetic algorithm. This approach constructs new chromosomes from a schema and pairs of existing ones by exploring their dissimilarities and similarities. To show the effectiveness of the algorithm, it is tested and compared with the classical GA, on 15 two-dimensional optimization problems taken from literature. We have found that, in most cases, our method is better than the classical genetic algorithm.

Keywords: chromosome injection, dynamic schema, genetic algorithm, similarity and dissimilarity

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Authors: Peter James C. Icalia, Agapita J. Salces, Loida Valenzuela, Kangseok Seo, Geronima Ludan

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This study evaluated polymorphism of 11 microsatellite markers in four local genetic groups of cattle. Batanes cattle which has never been studied using microsatellites is evaluated for its genetic distance from the Ilocos cattle while Brahman and Holstein-Sahiwal are also included as there were insemination programs by the government using these two breeds. PCR products that were genotyped for each marker were analyzed using POPGENEv32. Results showed that 55% (Fst=0.5501) of the genetic variation is due to the differences between populations while the remaining 45% is due to individual variation. The Fst value also indicates that there were very great differences from population to population using the range proposed by Sewall and Wright. The constructed phylogenetic tree based on Nei’s genetic distance using the modified neighboor joining procedure of PHYLIPv3.5 showed the admixture of Brahman and Holstein-Sahiwal having them grouped in the same clade. Batanes and Ilocos cattle were grouped in a different cluster showing that they have descended from a single parental population. This would presumably address the claim that Batanes and Ilocos cattle are genetically distant from other groups and still exist despite the artificial insemination program of the government using Brahman and other imported breeds. The knowledge about the genetic structure of this population supports the development of conservation programs for the smallholder farmers.

Keywords: microsatellites, cattle, Philippines, populations, genetic structure

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Authors: Fatimazahrae Elbakri, Azeddine Ibrahimi, Meryem Lemrani, Dris Belghyti

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Leishmaniasis represents a major public health problem because of the number of cases recorded each year and the wide distribution of the disease. It is a parasitic disease of flagellated protozoa transmitted by the bite of certain species of sandfly, causing a spectrum of clinical pathology in humans ranging from disfiguring skin lesions to fatal visceral leishmaniasis. Cutaneous leishmaniasis due to Leishmania major is a polymorphic disease; in fact, the infection can be asymptomatic, localized, or disseminated. The objective of this work is to determine the genomic diversity that contributes to clinical variability by trying to identify the variation in chromosome number and to extract SNPs and SNPs and InDels; it is based on four sequences (WGS) of Leishmania major available on NCBI in Fastq form, from three countries: Tunisia, Algeria, and Israel, the analysis is set up from a pipeline to facilitate the discovery of genetic diversity, in particular SNP and chromosomal somy.

Keywords: Leshmania major, cutaneous Leishmania, NGS, genomic, somy, variant calling

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Authors: Titanilla Komenda, Viktorio Malisa

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In this paper, the influence of diversity-related factors on the design of collaborative scenarios is analysed. Based on the evaluation, a framework for simulating human-robot-collaboration is presented that considers both human factors as well as the overall system performance. The implementation of the model is shown on a real-life scenario from industry and validated in terms of traceability, safety and physical limitations. By comparing scenarios that consider diversity with those only meeting system performance, an overall understanding of individually adapted human-robot-collaborative workspaces is reached. A diversity-related guideline for human-robot-collaborations provides a summary of the research and aids in optimizing future applications. Finally, limitations and future amendments of the model are discussed.

Keywords: diversity, human-machine system, human-robot collaboration, simulation

Procedia PDF Downloads 271

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: D. l. David, J. A. Wahedi, U. Buba, R. Zakariya

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Between January, 2004 to December, 2005, studies were carried out on the bird species diversity and relative abundance of two lakes, Kiri and Gyawana near Numan using the “Timed Species Count (TSC)” method. 163 species in 53 bird families and 160 species in 55 bird families were recorded at Kiri and Gyawana lakes respectively. There was no significant difference in species diversity within bird families between the two lakes (p > 0.05), whereas in Gyawana Lake, one of the sites qualified as Ramsar site, none strongly qualified as an Important Bird Area (IBA). The significance of these findingsare also discussed.

Keywords: conservation, diversity, lacustrine, wetlands

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Authors: Umairah Natasya Binti Mohd Omeershffudin, Suresh Kumar

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Klebsiella pneumoniae, a Gram-negative enteric bacterium that causes nosocomial and urinary tract infections. Particular concern is the global emergence of multidrug-resistant (MDR) strains of Klebsiella pneumoniae. Characterization of antibiotic resistance determinants at the genomic level plays a critical role in understanding, and potentially controlling, the spread of multidrug-resistant (MDR) pathogens. In this study, drug-resistant Klebsiella pneumoniae strain 825795-1 was investigated with extensive computational approaches aimed at identifying novel drug targets among hypothetical proteins. We have analyzed 1099 hypothetical proteins available in genome. We have used in-silico genome subtraction methodology to design potential and pathogen-specific drug targets against Klebsiella pneumoniae. We employed bioinformatics tools to subtract the strain-specific paralogous and host-specific homologous sequences from the bacterial proteome. The sorted 645 proteins were further refined to identify the essential genes in the pathogenic bacterium using the database of essential genes (DEG). We found 135 unique essential proteins in the target proteome that could be utilized as novel targets to design newer drugs. Further, we identified 49 cytoplasmic protein as potential drug targets through sub-cellular localization prediction. Further, we investigated these proteins in the DrugBank databases, and 11 of the unique essential proteins showed druggability according to the FDA approved drug bank databases with diverse broad-spectrum property. The results of this study will facilitate discovery of new drugs against Klebsiella pneumoniae.

Keywords: pneumonia, drug target, hypothetical protein, subtractive genomics

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Authors: L. K. Davis

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The effects of the evolution force are observable in nature at all structural levels ranging from small molecular systems to conversely enormous biospheric systems. However, the evolution force and work associated with formation of biological structures has yet to be described mathematically or theoretically. In addressing the conundrum, we consider evolution from a unique perspective and in doing so we introduce the “Fundamental Theory of the Evolution Force: FTEF”. We utilized synthetic evolution artificial intelligence (SYN-AI) to identify genomic building blocks and to engineer 14-3-3 ζ docking proteins by transforming gene sequences into time-based DNA codes derived from protein hierarchical structural levels. The aforementioned served as templates for random DNA hybridizations and genetic assembly. The application of hierarchical DNA codes allowed us to fast forward evolution, while dampening the effect of point mutations. Natural selection was performed at each hierarchical structural level and mutations screened using Blosum 80 mutation frequency-based algorithms. Notably, SYN-AI engineered a set of three architecturally conserved docking proteins that retained motion and vibrational dynamics of native Bos taurus 14-3-3 ζ.

Keywords: 14-3-3 docking genes, synthetic protein design, time-based DNA codes, writing DNA code from scratch

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Authors: Xin Chen

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Pre-industrial old-world human cultural diversity and societal complexity exhibits remarkable geographic regularities. Along the latitudinal axis from the equator to the arctic, a descending trend of human ethno-cultural diversity is found to be in coincidence with a descending trend of biological diversity. Along the same latitudinal axis, the pre-industrial human societal complexity shows to peak at the intermediate latitude. It is postulated that human cultural diversity and societal complexity are strongly influenced by collective learning, and that collective learning is positively related to human population size, social interactions, and environmental challenges. Under such postulations the relationship between collective learning and important geographical-environmental factors, including climate and biodiversity/bio-productivity is examined. A hypothesis of intermediate bio-productivity is formulated to account for those latitudinal patterns of pre-industrial human societal complexity.

Keywords: cultural diversity, soetal complexity, latitudinal patterns, biodiversity, bio-productivity, collective learning

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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: Masoud Sheidaei, Mohammad-Reza Kordasti, Fahimeh Koohdar

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Calotropis procera (Aiton) W.T.Aiton, (Apocynaceae), is an economically and medicinally important plant species which is an evergreen, perennial shrub growing in arid and semi-arid climates, and can tolerate very low annual rainfall (150 mm) and a dry season. The plant can also tolerate temperature ran off 20 to30°C and is not frost tolerant. This plant species prefers free-draining sandy soils but can grow also in alkaline and saline soils.It is found at a range of altitudes from exposed coastal sites to medium elevations up to 1300 m. Due to morpho-physiological adaptations of C. procera and its ability to tolerate various abiotic stresses. This taxa can compete with desirable pasture species and forms dense thickets that interfere with stock management, particularly mustering activities. Caloteropis procera grows only in southern part of Iran where in comprises a limited number of geographical populations. We used different population genetics and r landscape analysis to produce data on geographical populations of C. procera based on molecular genetic study using SCoT molecular markers. First, we used spatial principal components (sPCA), as it can analyze data in a reduced space and can be used for co-dominant markers as well as presence / absence data as is the case in SCoT molecular markers. This method also carries out Moran I and Mantel tests to reveal spatial autocorrelation and test for the occurrence of Isolation by distance (IBD). We also performed Random Forest analysis to identify the importance of spatial and geographical variables on genetic diversity. Moreover, we used both RDA (Redundency analysis), and LFMM (Latent factor mixed model), to identify the genetic loci significantly associated with geographical variables. A niche modellng analysis was carried our to predict present potential area for distribution of these plants and also the area present by the year 2050. The results obtained will be discussed in this paper.

Keywords: population genetics, landscape genetic, Calotreropis procera, niche modeling, SCoT markers

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Authors: Sushmita Chaudhuri, Parthiba Basu

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Urbanization leads to habitat degradation and is responsible for the fast disappearance of native butterfly species. Random sampling of rural, suburban and urban sites in an around Kolkata metropolis revealed the presence of 28 species of butterfly belonging to 5 different families in winter (February-March). Butterfly diversity, species richness and abundance decreased with increase in urbanization. Psyche (Leptosia nina of family Pieridae) was the most predominant butterfly species found everywhere in Kolkata during the winter period. The most dominant family was Nymphalidae (11species), followed by Pieridae (6 species), Lycaenidae (5 species), Papilionidae (4 species) and Hesperiidae (2 species). The rural and suburban sites had butterfly species that were unique to those sites. Vegetation cover and flowering shrub density were significantly related to butterfly diversity.

Keywords: butterfly, Kolkata metropolis, Shannon-Weiner diversity index, species diversity

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Authors: Amanda J. Burridge, Keith J. Edwards, Paul A. Wilkinson, Tom Batstone, Erik H. Murchie, Lorna McAusland, Ana Elizabete Carmo-Silva, Ivan Jauregui, Tracy Lawson, Silvere R. M. Vialet-Chabrand

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The rate of genetic progress in wheat production must be improved to meet global food security targets. However, past selection for domestication traits has reduced the genetic variation in modern wheat cultivars, a fact that could severely limit the future rate of genetic gain. The genetic variation in agronomically important traits for the wild relatives and progenitors of wheat is far greater than that of the current domesticated cultivars, but transferring these traits into modern cultivars is not straightforward. Between the elite cultivars of wheat, photosynthetic capacity is a key trait for which there is limited variation. Early screening of wheat wild relative and progenitors has shown differences in photosynthetic capacity and efficiency not only between wild relative species but marked differences between the accessions of each species. By identifying wild relative accessions with improved photosynthetic traits and characterising the genetic variation responsible, it is possible to incorporate these traits into advanced breeding programmes by wide crossing and introgression programmes. To identify the potential variety of photosynthetic capacity and efficiency available in the secondary and tertiary genepool, a wide scale survey was carried out for over 600 accessions from 80 species including those from the genus Aegilops, Triticum, Thinopyrum, Elymus, and Secale. Genotype data were generated for each accession using a ‘Wheat Wild Relative’ Single Nucleotide Polymorphism (SNP) genotyping array composed of 35,000 SNP markers polymorphic between wild relatives and elite hexaploid wheat. This genotype data was combined with phenotypic measurements such as gas exchange (CO₂, H₂O), chlorophyll fluorescence, growth, morphology, and RuBisCO activity to identify potential breeding material with enhanced photosynthetic capacity and efficiency. The data and associated analysis tools presented here will prove useful to anyone interested in increasing the genetic diversity in hexaploid wheat or the application of complex genotyping data to plant breeding.

Keywords: wheat, wild relatives, pre-breeding, genomics, photosynthesis

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Authors: Monika Szymańska-Czerwińsk, Kinga Zaręba-Marchewka, Krzysztof Niemczuk

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Chlamydiaceae are Gram-negative bacteria distributed worldwide in animals and humans. One of them is Chlamydia gallinacea recently discovered. Available data show that C. gallinacea is dominant chlamydial agent found in poultry in European and Asian countries. The aim of the studies was screening of poultry flocks in order to evaluate frequency of C. gallinacea shedding and genetic diversity. Sampling was conducted in different regions of Poland in 2019-2020. Overall, 1466 cloacal/oral swabs were collected in duplicate from 146 apparently healthy poultry flocks including chickens, turkeys, ducks, geese and quails. Dry swabs were used for DNA extraction. DNA extracts were screened using a Chlamydiaceae 23S rRNA real-time PCR assay. To identify Chlamydia species, specific real-time PCR assays were performed. Furthermore, selected samples were used for sequencing based on ompA gene fragments and variable domains (VD1-2, VD3-4). In total, 10.3% of the tested flocks were Chlamydiaceae-positive (15/146 farms). The presence of Chlamydiaceae was confirmed mainly in chickens (13/92 farms) but also in turkey (1/19 farms) and goose (1/26 farms) flocks. Eleven flocks were identified as C. gallinacea-positive while four flocks remained unclassified. Phylogenetic analysis revealed at least 16 genetic variants of C. gallinacea. Research showed that Chlamydiaceae occur in a poultry flock in Poland. The strains of C. gallinacea as dominant species show genetic variability.

Keywords: C. gallinacea, emerging agent, poultry, real-time PCR

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Authors: Lien Gysens, Bert Vanmechelen, Maarten Haspeslagh, Piet Maes, Ann Martens

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Bovine papillomavirus (BPV) types 1 and 2 play a central role in the etiology of the most common neoplasm in horses, the equine sarcoid. The unknown mechanism behind the unique variety in a clinical presentation on the one hand and the host-dependent clinical outcome of BPV-1 infection, on the other hand, indicate the involvement of additional factors. Earlier studies have reported the potential functional significance of intratypic sequence variants, along with the existence of sarcoid-sourced BPV variants. Therefore, intratypic sequence variation seems to be an important emerging viral factor. This study aimed to give a broad insight in sarcoid-sourced BPV variation and explore its potential association with disease presentation. In order to do this, a nanopore sequencing approach was successfully optimized for screening a wide spectrum of clinical samples. Specimens of each tumour were initially screened for BPV-1/-2 by quantitative real-time PCR. A custom-designed primer set was used on BPV-positive samples to amplify the complete viral genome in two multiplex PCR reactions, resulting in a set of overlapping amplicons. For phylogenetic analysis, separate alignments were made of all available complete genome sequences for BPV-1/-2. The resulting alignments were used to infer Bayesian phylogenetic trees. We found substantial genetic variation among sarcoid-derived BPV-1, although this variation could not be linked to disease severity. Several of the BPV-1 genomes had multiple major deletions. Remarkably, the majority of the cluster within the region coding for late viral genes. Together with the extensiveness (up to 603 nucleotides) of the described deletions, this suggests an altered function of L1/L2 in disease pathogenesis. By generating a significant amount of complete-length BPV genomes, we succeeded in introducing next-generation sequencing into veterinary research focusing on the equine sarcoid, thus facilitating the first report of both nanopore-based sequencing of complete sarcoid-sourced BPV-1/-2 and the simultaneous nanopore sequencing of multiple complete genomes originating from a single clinical sample.

Keywords: Bovine papillomavirus, equine sarcoid, horse, nanopore sequencing, phylogenetic analysis

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Authors: Yun-Chin Chung, Nileema Divate, Gen-Hung Chen, Pei-Ru Huang, Rupesh Divate

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Many environmental factors, such as glucose concentration, ethanol, temperature, osmotic pressure and pH, decrease the production rate of ethanol using yeast as a starter. Fermentation starters with high tolerance to various stresses are always demanded for brewing industry. Trehalose, a storage carbohydrate in cell wall of yeast, plays an important role in tolerance of environmental stress by preserving integrity of plasma membrane and stabilizing proteins. Furan aldehydes are toxic to yeast and the growth rate of yeast is significantly reduced if furan aldehydes were present in the fermentation medium. In yeast, aldehyde reductase is involved in the detoxification of reactive aldehydes and consequently the growth of yeast is improved. The aims of this study were to construct a genetic recombinant Saccharomyces cerevisiae or Pichia pastoris with furfural and HMF degrading and high ethanol tolerance capacities. Yeast strains were engineered by genetic recombination for overexpression of trehalose-6-phosphate synthase gene (tps1) and aldehyde reductase gene (ari1). TPS1 gene was cloned from S. cerevisiae by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated with pGAPZαC vector. The constructed vector, pGAPZC-tps1, was transformed to recombinant yeasts strain with overexpression of ari1. The transformants with pGAPZC-tps1-ari1 were generated called STA (S. cerevisiae) and PTA (P. pastoris) with overexpression of tps1, ari1. PCR with tps1-specific primers and western blot with his-tag confirmed the gene insertion and protein expression of tps1 in the transformants, respectively. The neutral trehalase gene (nth1) of STA was successfully deleted and the novel strain STAΔN will be used for further study, including the measurement of trehalose concentration and ethanol, furfural tolerance assay.

Keywords: genetic recombinant, yeast, ethanol tolerance, trehalase, aldehyde reductase

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Authors: Hajar Hosseini Khorami

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Porphyrins are organic, aromatic compounds found in heme, cytochrome, cobalamin, chlorophyll , and many other natural products with essential roles in biological processes that their cationic forms have been used as groups of favorable non-viral vectors recently. Cationic porphyrins are self-chromogenic reagents with a high capacity for modifications, great interaction with DNA and protection of DNA from nuclease during delivery of it into a cell with low toxicity. In order to have high efficient gene transfection into the cell while causing low toxicity, genetically manipulations of the non-viral vector, cationic porphyrin, would be useful. In this study newly modified cationic porphyrin derivative, 5, 10-dihexyl-15, 20bis (N-methyl-4-pyridyl) porphyrin was applied. Cytotoxicity of synthesized cationic porphyrin on Chinese Hamster Ovarian (CHO) cells was evaluated by using MTT assay. This cationic derivative is dose-dependent, with low cytotoxicity at the ranges from 100 μM to 0.01μM. It was uptake by cells at high concentration. Using direct non-viral gene transfection method and different concentration of cationic porphyrin were tested on transfection of CHO cells by applying derived transfection reagent with X-tremeGENE HP DNA as a positive control. However, no transfection observed by porphyrin derivative and the parameters tested except for positive control. Results of this study suggested that applying different protocol, and also trying other concentration of cationic porphyrins and DNA for forming a strong complex would increase the possibility of efficient gene transfection by using cationic porphyrins.

Keywords: cationic porphyrins, gene delivery, non-viral vectors, transfection reagents

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Authors: Ashok Kumar, Parveen Parveen

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India is bestowed with an extremely high population of plant species with medicinal value and even has two biodiversity hotspots. Indian systems of medicine including Ayurveda, Siddha and Unani have historically been serving humankind across the world since time immemorial. About 1500 plant species have well been documented in Ayurvedic Nighantus as official medicinal plants. Additionally, several hundred species of plants are being routinely used as medicines by local people especially tribes living in and around forests. The natural resources for medicinal plants have unscientifically been over-exploited forcing rapid depletion in their genetic diversity. Moreover, renewed global interest in herbal medicines may even lead to additional depletion of medicinal plant wealth of the country, as about 95% collection of medicinal plants for pharmaceutical preparation is being carried out from natural forests. On the other hand, huge export market of medicinal and aromatic plants needs to be seriously tapped for enhancing inflow of foreign currency. Asparagus racemosus Willd., a member of family Liliaceae, is one of thirty-two plant species that have been identified as priority species for cultivation and conservation by the National Medicinal Plant Board (NMPB), Government of India. Though attention is being focused on standardization of agro-techniques and extraction methods, little has been designed on genetic improvement and selection of desired types with higher root production and saponin content, a basic ingredient of medicinal value. The saponin not only improves defense mechanisms and controls diabetes but the roots of this species promote secretion of breast milk, improved lost body weight and considered as an aphrodisiac. There is ample scope for genetic improvement of this species for enhancing productivity substantially, qualitatively and quantitatively. It is emphasized to select desired genotypes with sufficient genetic diversity for important economic traits. Hybridization between two genetically divergent genotypes could result in the synthesis of new F1 hybrids consisting of useful traits of both the parents. The evaluation of twenty seed sources of Asparagus racemosus assembled different geographical locations of India revelled high degree of variability for traits of economic importance. The maximum genotypic and phenotypic variance was observed for shoot height among shoot related traits and for root length among root related traits. The shoot height, genotypic variance, phenotypic variance, genotypic coefficient of variance, the phenotypic coefficient of variance was recorded to be 231.80, 3924.80, 61.26 and 1037.32, respectively, where those of the root length were 9.55, 16.80, 23.46 and 41.27, respectively. The maximum genetic advance and genetic gain were obtained for shoot height among shoot-related traits and root length among root-related traits. Index values were developed for all seed sources based on the four most important traits, and Panthnagar (Uttrakhand), Jodhpur (Rajasthan), Dehradun (Uttarakhand), Chandigarh (Punjab), Jammu (Jammu & Kashmir) and Solan (Himachal Pradesh) were found to be promising seed sources.

Keywords: asparagus, genetic, genotypes, variance

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Authors: Othman E. Othman, Amira M. Nowier, Medhat El-Denary

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Camels play an important socio-economic role within the pastoral and agricultural system in the dry and semidry zones of Asia and Africa. Camels are economically important animals in Egypt where they are dual purpose animals (meat and milk). The analysis of chemical composition of camel milk showed that the total protein contents ranged from 2.4% to 5.3% and it is divided into casein and whey proteins. The casein fraction constitutes 52% to 89% of total camel milk protein and it divided into 4 fractions namely αs1, αs2, β and κ-caseins which are encoded by four tightly genes. In spite of the important role of casein genes and the effects of their genetic polymorphisms on quantitative traits and technological properties of milk, the studies for the detection of genetic polymorphism of camel milk genes are still limited. Due to this fact, this work focused - using PCR-RFP and sequencing analysis - on the identification of genetic polymorphisms and SNPs of two casein genes in Maghrabi camel breed which is a dual purpose camel breed in Egypt. The amplified fragments at 488-bp of the camel κ-CN gene were digested with AluI endonuclease. The results showed the appearance of three different genotypes in the tested animals; CC with three digested fragments at 203-, 127- and 120-bp, TT with three digested fragments at 203-, 158- and 127-bp and CT with four digested fragments at 203-, 158-, 127- and 120-bp. The frequencies of three detected genotypes were 11.0% for CC, 48.0% for TT and 41.0% for CT genotypes. The sequencing analysis of the two different alleles declared the presence of a single nucleotide polymorphism (C→T) at position 121 in the amplified fragments which is responsible for the destruction of a restriction site (AG/CT) in allele T and resulted in the presence of two different alleles C and T in tested animals. The nucleotide sequences of κ-CN alleles C and T were submitted to GenBank with the accession numbers; KU055605 and KU055606, respectively. The primers used in this study amplified 942-bp fragments spanning from exon 4 to exon 6 of camel αS1-Casein gene. The amplified fragments were digested with two different restriction enzymes; SmlI and AluI. The results of SmlI digestion did not show any restriction site whereas the digestion with AluI endonuclease revealed the presence of two restriction sites AG^CT at positions 68^69 and 631^632 yielding the presence of three digested fragments with sizes 68-, 563- and 293-bp.The nucleotide sequences of this fragment from camel αS1-Casein gene were submitted to GenBank with the accession number KU145820. In conclusion, the genetic characterization of quantitative traits genes which are associated with the production traits like milk yield and composition is considered an important step towards the genetic improvement of livestock species through the selection of superior animals depending on the favorable alleles and genotypes; marker assisted selection (MAS).

Keywords: genetic polymorphism, SNP polymorphism, Maghrabi camels, κ-Casein gene, αS1-Casein gene

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Authors: N. M. Sani, E. D. Jatau, O. S. Olonitola, M. Y. Gwarzo, P. Moodley, N. S. Mujahid

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Haemoglobin (HB) indicates anaemia level and by extension may reflect the nutritional level and perhaps the immunity of an individual. Some antiretroviral drugs like zidovudine are known to cause anaemia in People living with HIV/AIDS (PLWHA). A cross-sectional study using demographic data and blood specimen from 218 female commercial sex workers attending antiretroviral therapy (ART) clinics was conducted between December 2009 and July 2011 to assess the effect of zidovudine on haematologic and RNA viral load of female sex workers receiving antiretroviral treatment in north-western Nigeria. Anaemia is a common and serious complication of both HIV infection and its treatment. In the setting of HIV infection, anaemia has been associated with decreased quality of life, functional status, and survival. Antiretroviral therapy, particularly the highly active antiretroviral therapy (HAART), has been associated with a decrease in the incidence and severity of anaemia in HIV-infected patients who have received a HAART regimen for at least 1 year. In this study, result has shown that out of 218 patients, 26 with haemoglobin count between 5.1–10 g/dl were observed to have the highest viral load count of 300,000–350,000 copies/ml. It was also observed that most patients (190) with HB of 10.1–15.0 g/dl had viral load count of 200,000–250,000 copies/ml. An inverse relationship therefore exists, i.e. the lower the haemoglobin level, the higher the viral load count, even though the test statistics did not show any significance between the two (P=0.206). This shows that multivariate logistic regression analysis demonstrated that anaemia was associated with a CD4+ cell count below 50/µL in female sex workers with a viral load above 100,000 copies/mL who use zidovudine. Severe anaemia was less prevalent in this study population than in historical comparators; however, mild to moderate anaemia rates remain high. The study, therefore, recommends that hematological and virologic parameters be monitored closely in patients receiving first line ART regimen.

Keywords: anaemia, female sex worker, haemoglobin, Zidovudine

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Authors: Diana M. Torres, Anngie K. Hernandez, Andrea Villareal, Magda R. Gomez, Sadao Kobayashi

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Colombian Garlic crops exhibited mild mosaic, yellow stripes, and deformation. This group of symptoms suggested a viral infection. Several viruses belonging to the genera Potyvirus, Carlavirus and Allexivirus are known to infect garlic and lower their yield worldwide, but in Colombia, there are no studies of viral infections in this crop, only leek yellow stripe virus (LYSV) has been reported to our best knowledge. In Colombia, there are no management strategies for viral diseases in garlic because of the lack of information about viral infections on this crop, which is reflected in (i) high prevalence of viral related symptoms in garlic fields and (ii) high dispersal rate. For these reasons, the purpose of the present study was to evaluate the viral status of garlic in Colombia, which can represent a major threat on garlic yield and quality for this country 55 symptomatic leaf samples were collected for virus detection by RT-PCR and mechanical inoculation. Total RNA isolated from infected samples were subjected to RT-PCR with primers 1-OYDV-G/2-OYDV-G for Onion yellow dwarf virus (OYDV) (expected size 774pb), 1LYSV/2LYSV for LYSV (expected size 1000pb), SLV 7044/SLV 8004 for Shallot latent virus (SLV) (expected size 960pb), GCL-N30/GCL-C40 for Garlic common latent virus (GCLV) (expected size 481pb) and EF1F/EF1R for internal control (expected size 358pb). GCLV, SLV, and LYSV were detected in infected samples; in 95.6% of the analyzed samples was detected at least one of the viruses. GCLV and SLV were detected in single infection with low prevalence (9.3% and 7.4%, respectively). Garlic generally becomes coinfected with several types of viruses. Four viral complexes were identified: three double infection (64% of analyzed samples) and one triple infection (15%). The most frequent viral complex was SLV + GCLV infecting 48.1% of the samples. The other double complexes identified had a prevalence of 7% (GCLV + LYSV and SLV + LYSV) and 5.6% of the samples were free from these viruses. Mechanical transmission experiments were set up using leaf tissues of collected samples from infected fields, different test plants were assessed to know the host range, but it was restricted to C. quinoa, confirming the presence of detected viruses which have limited host range and were detected in C. quinoa by RT-PCR. The results of molecular and biological tests confirm the presence of SLV, LYSV, and GCLV; this is the first report of SLV and LYSV in garlic plants in Colombia, which can represent a serious threat for this crop in this country.

Keywords: SLV, GCLV, LYSV, leek yellow stripe virus, Allium sativum

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Authors: Sandra Thomas

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This study tries to analyse the level of food security and dietary diversity among households of different socio-economic classes in the urban centers of Thrissur. The study revealed that there is no much difference in purchasing patterns of food articles among the socio-economic classes indicating a very high level of both physical and economic accessibility of food. On analysing the dietary diversity of the households none of the households scored below five and fifty-three per cent of the households scored eleven or twelve indicating higher diversity in diet. It was also found that income and education are the two important factors that influence the level of household food security.

Keywords: food security, dietary diversity, household level, socio-economic classes

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Authors: Imad H. Hameed, Mohammad A. Jebor, Ammera J. Omer

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The aim of this research is to study the mitochonderial coding region by using the Sanger sequencing technique and establish the degree of variation characteristic of a fragment. FTA® Technology (FTA™ paper DNA extraction) utilized to extract DNA. Portion of coding region encompassing positions 11719 –12384 amplified in accordance with the Anderson reference sequence. PCR products purified by EZ-10 spin column then sequenced and Detected by using the ABI 3730xL DNA Analyzer. Five new polymorphic positions 11741, 11756, 11878, 11887 and 12133 are described may be suitable sources for identification purpose in future. The calculated value D= 0.95 and RMP=0.048 of the genetic diversity should be understood as high in the context of coding function of the analysed DNA fragment. The relatively high gene diversity and a relatively low random match probability were observed in Iraq population. The obtained data can be used to identify the variable nucleotide positions characterized by frequent occurrence which is most promising for various identifications.

Keywords: coding region, Iraq, mitochondrial DNA, polymorphic positions, sanger technique

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Authors: Sabitha Rani A., Nagamani V.

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Embelia ribes Burm.f (Family-Myrsinaceae) commonly known as Vidanga or Baibirang, is one of the important medicinal plants of India. The seed extract is reported to be antidiabetic, antitumour, analgesic, anti-inflammatory, antispermatogenic, free radical scavenging activities and widely used in more than 75 Ayurvedic commercial formulations. Among the 100 different species of Embelia, E. ribes is considered as a major source of Embelin, a bioactive compound. Because of high demand and low availability, the seeds of E. ribes are substituted with many cheaper alternatives. Therefore, the present study of RAPD-PCR analysis was undertaken to develop molecular markers for identification of E. ribes. A total of 13 different seed samples of Embelia were collected from different agro-climatic regions of India. The seeds of E.ribes were collected from Kalpetta, Kerala and three different seed samples were collected from traders of Odisha, Madhya Pradesh, Maharastra. The other nine seed samples were collected from local traders which they have collected from different regions of India. Genomic DNA was isolated from different seed samples E. ribes and RAPD-PCR was performed on 13 different seed samples using 47 random primers. Out of all the primers, only 22 primers produced clear and highly-reproducible banding patterns. The 22 selected RAPD primers generated a total of 280 alleles with an average of 12 alleles per primer pair. In the present study, we have identified three RAPD-PCR markers i.e. OPF5_480 bp, OPH11_520 bp and OPH4_530 bp which can be used for genetic fingerprinting of E. ribes. This methodology can be employed for identification of original E. ribes and also distinguishing it from other substitutes and adulterants.

Keywords: Embelia ribes, RAPD-PCR, primers, genetic analysis

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Authors: Dmitry Nosik, Nickolay Nosik, Elli Kaplina, Olga Lobach, Marina Chataeva, Lev Rasnetsov

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Background and aims: Human Immunodeficiency Virus (HIV) infection is very often associated with Herpes Simplex Virus (HSV) infection but HIV patients are treated with a cocktail of antiretroviral drugs which are toxic. The use of an antiviral drug which will be active against both viruses like ferrovir found in our previous studies is rather actual. Earlier we had shown that Fullerene poly-amino capronic acid (FPACA) was active in case of monoinfection of HIV-1 or HSV-1. The aim of the study was to analyze the efficiency of FPACA against mixed infection of HIV and HSV. Methods: The peripheral blood lymphocytes, CEM, MT-4 cells were simultaneously infected with HIV-1 and HSV-1. FPACA was added 1 hour before infection. Cells viability was detected by MTT assay, virus antigens detected by ELISA, syncytium formation detected by microscopy. The different multiplicity of HIV-1/HSV-1 ratio was used. Results: The double viral HIV-1/HSV-1 infection was more cytopathic comparing with monoinfections. In mixed infection by the HIV-1/HSV-1 concentration of HIV-1 antigens and syncytium formations increased by 1,7 to 2,3 times in different cells in comparison with the culture infected with HIV-1 alone. The concentration of HSV-1 increased by 1,5-1,7 times, respectively. Administration of FPACA (1 microg/ml) protected cells: HIV-1/HSV-1 (1:1) – 80,1%; HIV-1/HSV-1 (1:4) – 57,2%; HIV-1/HSV-1 (1:8) – 46,3 %; HIV-1/HSV-1 (1:16) – 17,0%. Virus’s antigen levels were also reduced. Syncytium formation was totally inhibited in all cases of mixed infection. Conclusion: FPACA showed antiviral activity in case of mixed viral infection induced by Human Immunodeficiency Virus and Herpes Simplex Virus. The effect of viral inhibition increased with the multiplicity of HIV-1 in the inoculum. The mechanism of FPACA action is connected with the blocking of the virus particles adsorption to the cells and it could be suggested that it can have an antiviral activity against some other viruses too. Now FPACA could be considered as a potential drug for treatment of HIV disease complicated with opportunistic herpes viral infection.

Keywords: antiviral drug, human immunodeficiency virus (hiv), herpes simplex virus (hsv), mixed viral infection

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Authors: Azizollah Khodakaram- Tafti, Ali Mohammadi, Ghasem Farjanikish

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Bovine viral diarrhea virus (BVDV) is one of the most important viral pathogens of cattle worldwide caused by Pestivirus genus, Flaviviridae family.The aim of the present study was to comparison several diagnostic methods and determine the prevalence of BVDV infection for the first time in dairy herds of Fars province, Iran. For initial screening, a total of 400 blood samples were randomly collected from 12 industrial dairy herds and analyzed using reverse transcription (RT)-PCR on the buffy coat. In the second step, blood samples and also ear notch biopsies were collected from 100 cattle of infected farms and tested by antigen capture ELISA (ACE), RT-PCR and immunohistochemistry (IHC). The results of nested RT-PCR (outer primers 0I100/1400R and inner primers BD1/BD2) was successful in 16 out of 400 buffy coat samples (4%) as acute infection in initial screening. Also, 8 out of 100 samples (2%) were positive as persistent infection (PI) by all of the diagnostic tests similarly including RT-PCR, ACE and IHC on buffy coat, serum and skin samples, respectively. Immunoreactivity for bovine BVDV antigen as brown, coarsely to finely granular was observed within the cytoplasm of epithelial cells of epidermis and hair follicles and also subcutaneous stromal cells. These findings confirm the importance of monitoring BVDV infection in cattle of this region and suggest detection and elimination of PI calves for controlling and eradication of this disease.

Keywords: antigen capture ELISA, bovine viral diarrhea virus, immunohistochemistry, RT-PCR, cattle

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Authors: M. Michael Gromiha, A. Mary Thangakani, Sandeep Kumar, D. Velmurugan

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The main cause of several neurodegenerative diseases such as Alzhemier, Parkinson, and spongiform encephalopathies is formation of amyloid fibrils and plaques in proteins. We have analyzed different sets of proteins and peptides to understand the influence of sequence-based features on protein aggregation process. The comparison of 373 pairs of homologous mesophilic and thermophilic proteins showed that aggregation-prone regions (APRs) are present in both. But, the thermophilic protein monomers show greater ability to ‘stow away’ the APRs in their hydrophobic cores and protect them from solvent exposure. The comparison of amyloid forming and amorphous b-aggregating hexapeptides suggested distinct preferences for specific residues at the six positions as well as all possible combinations of nine residue pairs. The compositions of residues at different positions and residue pairs have been converted into energy potentials and utilized for distinguishing between amyloid forming and amorphous b-aggregating peptides. Our method could correctly identify the amyloid forming peptides at an accuracy of 95-100% in different datasets of peptides.

Keywords: aggregation, amyloids, thermophilic proteins, amino acid residues, machine learning techniques

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Authors: Aizati N. A. Daud, Jorieke E. H. Bergman, Wilhelmina S. Kerstjens-Frederikse, Pieter Van Der Vlies, Eelko Hak, Rolf M. F. Berger, Henk Groen, Bob Wilffert

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Prenatal use of SRIs was previously associated with Congenital Heart Anomalies (CHA). The aim of the study is to explore whether pharmacogenetics plays a role in this teratogenicity using a gene-environment interaction study. A total of 33 case-mother dyads and 2 mother-only (children deceased) registered in EUROCAT Northern Netherlands were included in a case-only study. Five case-mother dyads and two mothers-only were exposed to SRIs (paroxetine=3, fluoxetine=2, venlafaxine=1, paroxetine and venlafaxine=1) in the first trimester of pregnancy. The remaining 28 case-mother dyads were not exposed to SRIs. Ten genes that encode the enzymes or proteins important in determining fetal exposure to SRIs or its mechanism of action were selected: CYPs (CYP1A2, CYP2C9, CYP2C19, CYP2D6), ABCB1 (placental P-glycoprotein), SLC6A4 (serotonin transporter) and serotonin receptor genes (HTR1A, HTR1B, HTR2A, and HTR3B). All included subjects were genotyped for 58 genetic variations in these ten genes. Logistic regression analyses were performed to determine the interaction odds ratio (OR) between genetic variations and SRIs exposure on the risk of CHA. Due to low phenotype frequencies of CYP450 poor metabolizers among exposed cases, the OR cannot be calculated. For ABCB1, there was no indication of changes in the risk of CHA with any of the ABCB1 SNPs in the children and their mothers. Several genetic variations of the serotonin transporter and receptors (SLC6A4 5-HTTLPR and 5-HTTVNTR, HTR1A rs1364043, HTR1B rs6296 & rs6298, HTR3B rs1176744) were associated with an increased risk of CHA, but with too limited sample size to reach statistical significance. For SLC6A4 genetic variations, the mean genetic scores of the exposed case-mothers tended to be higher than the unexposed mothers (2.5 ± 0.8 and 1.88 ± 0.7, respectively; p=0.061). For SNPs of the serotonin receptors, the mean genetic score for exposed cases (children) tended to be higher than the unexposed cases (3.4 ± 2.2, and 1.9 ± 1.6, respectively; p=0.065). This study might be among the first to explore the potential gene-environment interaction between pharmacogenetic determinants and SRIs use on the risk of CHA. With small sample sizes, it was not possible to find a significant interaction. However, there were indications for a role of serotonin receptor polymorphisms in fetuses exposed to SRIs on fetal risk of CHA which warrants further investigation.

Keywords: gene-environment interaction, heart defects, pharmacogenetics, serotonin reuptake inhibitors, teratogenicity

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