Genetic Study for G-Protein Coupled Receptor from Saccharomyces Cerervisiae and From Sera of Patients with Heart Thrombosis

Aim of the present study is Identification of specific gene for GPCR using specific primers .and identification of difference in PCR analysis in patients with heart thrombosis  and compared with healthy, Sequencing of PCR product regarding GPCR compared for all three subject, Identification  the similarity of human GPCR with local strain of yeast fifty healthy control and fifty patients with thrombosis which diagnosed medically with cardiac specific troponin t, troponin 1 levels and electro myocardiogram ECG. The aged for all subjects ranged (39-75) years   patients were lying in cardiac care unit at Ibn- al- Nafees teaching hospital and  Sheikh  Zayed  teaching  hospital. Genomic DNA of whole blood was extracted from buffy coat and cell cultured handbook protocol using Bioneer- kit and  Genomic DNA fungus/yeast  kit was used in isolation  and purification of DNA. patients divided  into three  groups according  to their age: group A (60-75) years , group B (50-59) years ,  group C (39-49) years the  results of genomic  DNA  isolation  from blood cells extracted in pure  form which ensured by the absorbance ratio (260/280 ) was (1.6 – 1.9 ) with a concentration of 50µg/ml and one DNA band with high resolution in gel electrophoresis. The result of  genomic DNA extracted from  the local strain of  S. cerevisiae showed that DNA   extracted with  high   purity   because   the  absorbance  ratio  (260 /280 )was (1.7 to 2.0) with a concentration of 60 µg/ml  and presence one DNA band with high resolution  in gel electrophoresis. primers were designed  depending on the sequence of the gene responsible for the production of GPCR on the chromosome 11 , GPCR  contain  three exons which covered  with six primers to detect a defect in gene sequence among. Results of gel electrophoresis are showed that primer  GPRX1 gave one band for (Control , A,B ) groups but absent amplified band in the patient eight and nine from group C. with molecular weight of this band is 1000 bp. The  GPRX2  primer  used to amplify second exon in the GPCR gene ,the molecular weight of amplified bands are  400 bp were present in all control samples and three groups of thrombosis patientsand yeast. GPRX2A primer that designed to amplify  part two from second exon of GPCR gene by PCR gave one band for all samples which include control and patient, the molecular weight of this band is 500 bp. PCR analysis  showed one amplify band for all control and patients group with molecular weight 500 bp  for GPRX3 primer and 400 bp for GPRX3A ,300 bp for GPRX3B. The specific primers which  designed to covering  GPCR gene used to amplification  genomic DNA  of the local strain  S.cerevisiae by PCR technique. Results showed all six primers which gave  one band with difference molecular weight  for each primer. All samples demonstrate identity planned sizes to control and local strain of S. cerevisiae samples except patient number 8 and 9 in the group(C) that  showed non specialist bands in specific primer  with  first  exon (GPRX1) .So the genetic sequence analysis of these two case based on the sequence  of the  remainder exons to detect the genetic defect in these case. The sequence of the first part for the second exon (X2) was identity standard sequence found on the NCBI web site for case( 8). The  case (9) showed  identity with  the sequence present in the human gene bank but some difference in the first of sequence which neglected because it is in the place link of primer. The  results for  case  8  showed some mutation for Exon X2(part2). but case (9) demonstrate one  deletion and one substitution. The results, also, illustrated that the ether48 thrombosis patients didn't appeared any mutation despite the positive results for ( Troponin) that gives strong indication of thrombosis. The conclusion Primer  GPRX1 gave one band for (Control , A,B ) groups but absent amplified band in the patient eight and nine from group C. The molecular weight of this band is 1000 bp. The amplified band with molecular weight 400 bp were present in all control samples and three groups of thrombosis patients with primer GPRX2  and 500 bp with primer GPRX2A.PCR analysis  showed one amplify band for all control and patients group with molecular weight 500 bp for GPRX3 primer.,400 bp with GPRX3A and 300 bp for primer GPRX3B.Similarity between results given by healthy group and local strain of yeast. Genetic study  showed that there are  only two case of  patients  eight and nine demonstrated mutation in nucleic location on exon two and three from GPCR gene Keyword: GPCR, Genetic Study, Saccharomysis  cerevisiae, Patient with Thrombosis

Purification of G-Protein Coupled Receptor from Whole Cell of Local Strain of Saccharomyces cerevisiae

The aim of study To purify GPCR from a local strain of S. cerevisiae using Ion exchange and gel filtrationchromatography techniques , by packing materials for columns which will be chosen of low cost comparing to the alreadyused in published researches, which depend on the costly affinity chromatography and other expensive methods ofpurification. Local strain of S. cerevisiae chosen for extraction and purification of G-protein coupled receptor (GPCR).The strains were obtained from biology department in Al- Mosul University, Iraq. The isolated colony was activated onYeast Extract Pepton Dextrose Broth (YEPDB) and incubated at 30 C° for 24 h .Loop fully of the yeast culture wastransferred to (10ml) of yeast extract peptone glucose agar (YEPGA) slant , then incubated at 30C°for 24h , after that itwas stored at 4C° ,the yeast cultures were reactivated and persevered after each two weeks period. S.cerevisiae wasidentified by morphological, microscopic characterization and biochemical test . The GPCR that extract from whole cellof S.cerevisiae was purified by ion exchange chromatography using DEAE-Sepharose ,the bound proteins (negativelycharged) were then eluted using gradient concentration of NaCl ranged between( 0.1 -0.5M). Gel filtrationchromatography using Sepharose 6B was applied as a second step of purification. The optical density for each fractionwas measured at 280 nm by UV-VS spectrophotometer then the GPCR concentration was determined by using ELISAKit . The fractions which gave the highest absorbance and concentration of GPCR were collected .The molecular weightof GPCR was determined by gel filtration chromatography using blue dextrin solution. Standard curve was plottedbetween log of molecular weight for standard protein and the ratio of Ve/Vo of GPCR . The purity of the GPCR thatextracted and purified from whole cell of S, cerevisiae were carried out by using SDS-PAGE electrophoresis . In ionexchange chromatography the fraction were collected with 5 ml tube at a flow rate 0.5 ml/ min and eluted with gradient(0.1-0.5M) of sodium chloride solution. Two proteins peaks appeared after eluted by the gradient concentration ofsodium chloride, while no protein peaks appeared in the washing fractions. The GPCR concentration was measured inthe fractions of these two protein peaks, data indicated that GPCR located in the first protein peak (eluted at 0.1M ofNaCl) at fraction numbers between 3 and 9, the maximum concentration of GPCR was 9.281 with specific activity71.58(ng/mg)protein , 3.125 purification folds and72.9(%) yield while the second peaks (eluted at 0.4 M of NaCl) don'tgive any concentration for GPCR, thus its neglected. Gel filtration chromatography was used as second step ofpurification which applied by using sepharose 6B. Results show single active protein peaks appeared that identical withthe peak of GPCR at fractions numbers(29-35). The maximum concentration of GPCR was 9.082 (ng/ml)was observed inthese fractions. The specific activity for these fractions was 151.37 (ng/mg) protein with 6.608 purification folds and39.64 (%) yield. The present study a chive a relatively high purification of GPCR from whole cell of a local strain S.cerevisiae with fold purification 6.608 and a yield of 39.64 % and molecular weight about~33KD.Key words: GPCR purification, S.Cerevisea, whole cel

Multi-ancestry genome-wide association analyses improve resolution of genes and pathways influencing lung function and chronic obstructive pulmonary disease risk

Lung-function impairment underlies chronic obstructive pulmonary disease (COPD) and predicts mortality. In the largest multi-ancestry genome-wide association meta-analysis of lung function to date, comprising 580,869 participants, we identified 1,020 independent association signals implicating 559 genes supported by ≥2 criteria from a systematic variant-to-gene mapping framework. These genes were enriched in 29 pathways. Individual variants showed heterogeneity across ancestries, age and smoking groups, and collectively as a genetic risk score showed strong association with COPD across ancestry groups. We undertook phenome-wide association studies for selected associated variants as well as trait and pathway-specific genetic risk scores to infer possible consequences of intervening in pathways underlying lung function. We highlight new putative causal variants, genes, proteins and pathways, including those targeted by existing drugs. These findings bring us closer to understanding the mechanisms underlying lung function and COPD, and should inform functional genomics experiments and potentially future COPD therapies

Monitoring Studies of Arthropod Complex Fauna Associated with Rapeseed- Mustard Crop Grown in Kashmir Himalayas

Kashmir has quietly undergone a yellow revolution as thousands of hectares of land have been brought under mustard cultivation to double the farmers' income. Earlier, most of the land in Kashmir was used only for growing a single crop -- paddy in most areas but now farmers are rotating crops in Kharif and Rabi seasons just like in other parts of the country. There is still many challlanges which are hurdle in high production goal. Insect biodiversity is associated with the quantity and type of host plants available, environmental factors, and their physiological state. Arthropod complex of mustard was studied and about eight species of insects-pests at Mountain Research centre for field crops (MRCFC) during Rabi 2020 -21 was recorded. It has been observed that the mustard aphid, Lipaphis erysimi Kalt., persists constantly in a sizable population and causes substantial crop damage from the blossoming to the maturity stage. Phyllotreta cruciferae Goeze (flea beetle) occurred at the seedling stage, and Carpocoris sp. (shield bug) caused little harm to the crop from the seedling to the maturity stage. Leaves have been only infested by cabbage aphid, Brevicoryne brassicae, mustard aphid, Lipaphis erysimi (Kalt.), and mustard leaf miner, Chromatomyia horticola, and they been detected in sporadic, low-population occurrences. Among them, Brevicoryne brassicae is found at flowering stage. Pieris brassicae, the cabbage butterfly, occurs sporadically and with high abundance. The status of all eight insect-pests on Brassica species was observed as minor with irregular occurrence while L. erysimi as a regular key pest.six Insect pollinators were recorded during the bloom period of the crop

Screening of French Bean (Phaseolus vulgaris L.) Genotypes against Alternaria Leaf Spot Caused by (Alternaria alternata) under Dryland Conditions of Kashmir

Sixty-three genotypes of  french bean was screened against leaf spot (Alternaria  alternata) in sick plots at Research Farm of Dryland Agriculture Research Srinagar, Rangreth during  Kharif  2018 and 2019. The highest mean disease incidence ranged from 0.00 to 85.00 per cent  with  the mean disease intensity ranged from 0.00 to 53.26 per cent .One genotype namely  ‘Local Pulwama’ was highly susceptible in their disease reaction. Among the screened germplasm, ‘Highly Resistant’ genotypes was SKU-R-601, SKUA-R-105, SKU-R-927, DARS-25, DARS-66, DARS-R-615,  while as ‘Susceptible’ genotypes was  DARS-8, DARS-12,  DARS-11, SKUAST-R-155, SKU-R-928, DARS-7, DARS-R-4, Bhaderwah (L),  Local  Kupwara black and Raj Jawala. Local Pulwama was found to be a highly susceptible (HS) genotype.  Twenty nine genotypes namely., DARS-16, DARS-9, DARS-54, DARS-39, VL-125, DARS-63, ENTO-504, SKUAST-204,SKU-R-925, DARS-60, DARS-109, DARS-43, DARS-44, SKU-R-23, DARS-4, DARS-74, SKU-R-105, DARS-40, DARS-23, DARS-18, SKU-R-71, WB-341, SKU-R-605, Uri local, Shopian (L), SKU-R-23, DARS-71, SSGB-729, DARS-R-19 showed resistant reaction to disease. The selection for resistance was based on the reaction of varieties on leaves

Impact of Weather Parameters on Population Dynamics of Cabbage Aphid (Brevicoryne brassicae L.) on Kale (Brassica oleracea var. acephala)

Studies on “population dynamics of Cabbage Aphid (Brevicoryne brassicae L.) on Kale (Brassica oleracea var. acephala)’’ in relation to weather parameters revealed cabbage aphid (Brevicoryne brassicae L.) as a major pest on Kale. The incidence of cabbage aphid commenced from 16th standard meteorological week (SMW) with a population of 2.02 (aphids/plant). The population of cabbage aphid started increasing and reached its peak in 25th SMW with a population of 137.16 (aphids/plant) after that the population started declining gradually in 26th SMW with a population of 83.14 (aphids/plant). However, the mean population of nymphs during 16th SMW was (2.02 ± 0.46) which gradually started increasing and was highest in 25th SMW (32.68 ± 0.74). The mean population of adults during 17th SMW was (8.24 ± 0.38) and was highest during 25th SMW (62.12 ± 0.50) and the mean population of winged aphids during 18th SMW was (7.10 ± 0.42) which gradually started increasing and attained its peak during 25th SMW (42.36 ± 0.41) respectively. Correlation between maximum and minimum temperature showed positive correlation with cabbage aphid population. Relative humidity of morning showed non-significant negative correlation while relative humidity of evening exhibited positive non-significant correlation. Rainfall were found non-significant positive correlation

Thousands of Qatari genomes inform human migration history and improve imputation of Arab haplotypes

Arab populations are largely understudied, notably their genetic structure and history. Here we present an in-depth analysis of 6,218 whole genomes from Qatar, revealing extensive diversity as well as genetic ancestries representing the main founding Arab genealogical lineages of Qahtanite (Peninsular Arabs) and Adnanite (General Arabs and West Eurasian Arabs). We find that Peninsular Arabs are the closest relatives of ancient hunter-gatherers and Neolithic farmers from the Levant, and that founder Arab populations experienced multiple splitting events 12–20 kya, consistent with the aridification of Arabia and farming in the Levant, giving rise to settler and nomadic communities. In terms of recent genetic flow, we show that these ancestries contributed significantly to European, South Asian as well as South American populations, likely as a result of Islamic expansion over the past 1400 years. Notably, we characterize a large cohort of men with the ChrY J1a2b haplogroup (n = 1,491), identifying 29 unique sub-haplogroups. Finally, we leverage genotype novelty to build a reference panel of 12,432 haplotypes, demonstrating improved genotype imputation for both rare and common alleles in Arabs and the wider Middle East

A population study of clinically actionable genetic variation affecting drug response from the Middle East

Clinical implementation of pharmacogenomics will help in personalizing drug prescriptions and alleviate the personal and financial burden due to inefficacy and adverse reactions to drugs. However, such implementation is lagging in many parts of the world, including the Middle East, mainly due to the lack of data on the distribution of actionable pharmacogenomic variation in these ethnicities. We analyzed 6,045 whole genomes from the Qatari population for the distribution of allele frequencies of 2,629 variants in 1,026 genes known to affect 559 drugs or classes of drugs. We also performed a focused analysis of genotypes or diplotypes of 15 genes affecting 46 drugs, which have guidelines for clinical implementation and predicted their phenotypic impact. The allele frequencies of 1,320 variants in 703 genes affecting 299 drugs or class of drugs were significantly different between the Qatari population and other world populations. On average, Qataris carry 3.6 actionable genotypes/diplotypes, affecting 13 drugs with guidelines for clinical implementation, and 99.5% of the individuals had at least one clinically actionable genotype/diplotype. Increased risk of simvastatin-induced myopathy could be predicted in ~32% of Qataris from the diplotypes of SLCO1B1, which is higher compared to many other populations, while fewer Qataris may need tacrolimus dosage adjustments for achieving immunosuppression based on the CYP3A5 diplotypes compared to other world populations. Distinct distribution of actionable pharmacogenomic variation was also observed among the Qatari subpopulations. Our comprehensive study of the distribution of actionable genetic variation affecting drugs in a Middle Eastern population has potential implications for preemptive pharmacogenomic implementation in the region and beyond. 2022, The Author(s).PVJ is supported by faculty funding from the College of Health & Life Sciences, HBKU. Qatar Biobank and Qatar Genome Program are Research, Development & Innovation's entities within Qatar Foundation for Education, Science and Community Development. Funders had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.Scopu

Multi-ancestry genome-wide association analyses improve resolution of genes and pathways influencing lung function and chronic obstructive pulmonary disease risk

Funder: British Lung Foundation (BLF); doi: https://doi.org/10.13039/501100000351Funder: DH | National Institute for Health Research (NIHR); doi: https://doi.org/10.13039/501100000272AbstractLung-function impairment underlies chronic obstructive pulmonary disease (COPD) and predicts mortality. In the largest multi-ancestry genome-wide association meta-analysis of lung function to date, comprising 580,869 participants, we identified 1,020 independent association signals implicating 559 genes supported by ≥2 criteria from a systematic variant-to-gene mapping framework. These genes were enriched in 29 pathways. Individual variants showed heterogeneity across ancestries, age and smoking groups, and collectively as a genetic risk score showed strong association with COPD across ancestry groups. We undertook phenome-wide association studies for selected associated variants as well as trait and pathway-specific genetic risk scores to infer possible consequences of intervening in pathways underlying lung function. We highlight new putative causal variants, genes, proteins and pathways, including those targeted by existing drugs. These findings bring us closer to understanding the mechanisms underlying lung function and COPD, and should inform functional genomics experiments and potentially future COPD therapies.</jats:p