The chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus, which has infected millions of people in Africa, Asia, Americas, and Europe since it remerged in India and Indian Ocean regions in 2005-2006. The purpose of this study was to evaluate the genetic diversity and evolutionary changes in CHIKV from 2016 to 2018 in Pakistan. Blood specimens were collected and processed following the Centers for Disease Control and Prevention Trioplex Protocol. Sequencing and phylogenetic analysis of complete coding sequence of representative isolates from the CHIKV outbreak was carried out during December 2016 to July 2018, a total of 1549 samples were received, out of which 50% (n = 774) were found positive for CHIKV RNA. Mean age of chikungunya positive patients was 31.8 ± 15.7 years and most affected were between 21 and 40 years of age. The Pakistan CHIKV strains clustered with the Indian Ocean sublineage of East/Central/South African with cocirculation of some variants In the structural proteins region, two noteworthy changes (A226V and D284E) were observed in the membrane fusion glycoprotein E1. Key substitutions in the neutralizing epitopes site and a few changes indicative of adaptive to other insect cells were also detected in Pakistani strains. This study provides the emerging trend of CHIKV in the country for early identification of potential variants of high virulence and preventive measures for vector borne disease especially in the endemic areas.
Background: Non-synchronized pods shattering in the Brassicaceae family bring upon huge yield losses around the world. The shattering process was validated to be controlled by eight different genes in the model plant Arabidopsis thaliana, including SHATTERPROOF1, SHATTERPROOF2, FRUITFULL, INDEHISCENT, ALCATRAZ, NAC, REPLUMLESS and POLYGlACTOURANAZE. To obtain gene family & examine their expression patterns into fresh & mature silique, then completed genome wide identification, characterization, and expression analysis of shattering genes in B. napus and B. juncea.Results: Complete genome analysis of B. napus and B. juncea revealed 32 shattering genes, which were identified and categorize based on protein motif structure, exon-intron organization and phylogeny. The phylogenetic study revealed that these shattering genes contain little duplications that were determined with a distinct chromosome number. Motifs of 32 shattering proteins were also observed where motifs 6 were found to be more conserved. A single motif was observed for other genes like BrnS7, BrnS8, BrjS23 and BrjS26. Comparative genomics for synteny analysis was performed that validated a conserved pattern of blocks among these cultivars. RT-PCR based expressions profile showed higher expression of shattering genes in B. juncea as compared to B. napus. FUL gene was expressed more in the mature silique. ALC gene was not expressed in the fresh silique of B. napus but highly expressed in the mature silique. Conclusion: This study authenticates that shattering genes exist in the local cultivars of Brassica. ALC exhibited strong expression in both the mature and fresh silique of B. juncea. Our results showed that shattering genes expression occurred more in B. juncea as compared to B. napus. It also contributes to the screening of more candidate gene for further investigation and characterization.
Chikungunya virus (CHIKV) is considered a public health problem due to its rapid spread and high morbidity. In 2016–2017 an outbreak of CHIKV was occurred in Pakistan but the data regarding the genomic diversity of CHIKV was not reported. Hence, the current study aimed to determine the genetic diversity of CHIKVs in Pakistan. A cross sectional study was carried out using sera of infected CHIKV patients (n = 1549) during the outbreak in Pakistan (2016–2018). Nucleotide sequencing of non-structural genes of CHIKV from eight isolates were performed followed by phylogenetic analysis using Bayesian method. Phylogenetic analysis suggested that the Pakistani CHIKV strains belonged to Indian Ocean Lineage (IOL) of genotype ECSA and C1.3a clade. Furthermore, the Pakistani isolates showed several key mutations (nsP2-H130Y, nsP2-E145D, nsP4-S55N and nsP4- R85G) corresponding to mutations reported in 2016 Indian strains of CHIKV. The molecular analysis revealed high evolutionary potential of CHIKV strains as well as better understanding of enhanced virulence and pathogenesis of this outbreak. The study highlights the need to continue surveillance in order to understand viral diversity over time and to devise preventive measures to limit diseases transmission in the region.
α-amylase is the key digestive enzyme that has been used widely in food, paper, detergent and textile industries for starch degradation. This study was conducted for the optimization and characterization of α-amylase production from Aspergillus niger SAIB-4. The study further assessed the effect of metal nanowires (NWs) on starch hydrolysis by α-amylase enzyme from A. niger. Copper oxide (CuO) and iron oxide (FeO) NWs were fabricated in anodized aluminum oxide (AAO) templates. Scanning Electron Microscopy (SEM) and Energy Dispersive spectroscopy (EDS) confirmed the diameter and composition of NWs. Different culture conditions were optimized for the production of α-amylase , where optimum production was obtained at incubation time of 60 h, 5% inoculum size, 1% of banana peel, 0.5% ammonium chloride, pH 8 and 30 °C temperature. It was observed that CuO NWs significantly enhanced α-amylase activity at 40 ppm whereas inhibitory affect was observed for FeO NWs at all concentrations. Maximum starch hydrolysis (0.54 µg/ml) was noticed for CuO NWs at 10 ppm concentration while Minimum activity (0.19 µg/ml) was observed at 70 ppm for FeO NWs. Further, molecular docking analysis was performed to endorse the interaction of NWs with α-amylase with improved enzymatic activity for starch hydrolysis. The combination of α-amylase and CuO NWs could be used as better and efficient source for starch hydrolysis with extraordinary industrial applications, particularly in the sugar and detergent industries.
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