The aim of study To purify GPCR from a local strain of S. cerevisiae using Ion exchange and gel filtration chromatography techniques , by packing materials for columns which will be chosen of low cost comparing to the already used in published researches, which depend on the costly affinity chromatography and other expensive methods of purification. 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 on Yeast Extract Pepton Dextrose Broth (YEPDB) and incubated at 30 C˚ for 24 h .Loop fully of the yeast culture was transferred to (10ml) of yeast extract peptone glucose agar (YEPGA) slant , then incubated at 30C˚for 24h , after that it was stored at 4C˚ ,the yeast cultures were reactivated and persevered after each two weeks period. S.cerevisiae was identified by morphological, microscopic characterization and biochemical test . The GPCR that extract from whole cell of S.cerevisiae was purified by ion exchange chromatography using DEAE-Sepharose ,the bound proteins (negatively charged) were then eluted using gradient concentration of NaCl ranged between( 0.1 -0.5M). Gel filtration chromatography using Sepharose 6B was applied as a second step of purification. The optical density for each fraction was measured at 280 nm by UV-VS spectrophotometer then the GPCR concentration was determined by using ELISA Kit . The fractions which gave the highest absorbance and concentration of GPCR were collected .The molecular weight of GPCR was determined by gel filtration chromatography using blue dextrin solution. Standard curve was plotted between log of molecular weight for standard protein and the ratio of Ve/Vo of GPCR . The purity of the GPCR that extracted and purified from whole cell of S, cerevisiae were carried out by using SDS-PAGE electrophoresis . In ion exchange 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 of sodium chloride, while no protein peaks appeared in the washing fractions. The GPCR concentration was measured in the fractions of these two protein peaks, data indicated that GPCR located in the first protein peak (eluted at 0.1M of NaCl) at fraction numbers between 3 and 9, the maximum concentration of GPCR was 9.281 with specific activity 71.58(ng/mg)protein , 3.125 purification folds and72.9(%) yield while the second peaks (eluted at 0.4 M of NaCl) don't give any concentration for GPCR, thus its neglected. Gel filtration chromatography was used as second step of purification which applied by using sepharose 6B. Results show single active protein peaks appeared that identical with the peak of GPCR at fractions numbers(29-35). The maximum concentration of GPCR was 9.082 (ng/ml)was observed in these fractions. The specific activity for these fractions was 151.37 (ng/mg) protein with 6.608 purification folds and 39.64 (%) 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.
The definition of the role of any institution in society is achieved through its objectives, The same is true for the military and how to deal with security threats in the humanitarian field ,Terrorism, which has almost replaced the traditional pattern of war, has waged a street war and intimidated individuals, families and society ,On the other hand, he found someone to meet him from a popular crowd of volunteers to defend their homeland from different sects, sects and religions, Thus, our study will be exposed to the role of popular mobilization in human security from a sociological point of view in Samarra, a field study of 100 male and female respondents.
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has caused enormous issues worldwide and is the most infectious pandemic. This study included 50 subjects (evenly distributed between sexes) and their range of ages starting from 2 to 67 years. According to the study's result, the ages and genders of subjects include susceptibility to COVID-19. Males were found to be more infected than females, and the ages of 36 to 67 were more common than other age ranges. Also, BMI calculations revealed that male patients with COVID-19 have the highest percentage of obesity. The clinical parameter results have been found serum C‐reactive protein (CRP) as an essential indicator that changes significantly in infection with COVID‐19 an
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Present study was conducted to evaluate the different levels of energy to protein ratios (EPR) using food waste and black soldier fly larvae meal (FWBSFL) on growth performance and nutrient digestibility of broilers. A total of 160 one-day old broiler chicks were divided randomly to four groups and each group had 8 replicates with 5 chicks per replicate. The control diet was formulated using conventional feed ingredients with EPR of 154 for the starter period and 167 for the finisher period. The other treatments were diets with normal, low, and high EPR (154,143, and 166 for the starter period; 167, 155, and 177 for the finisher period) using FWBSFL. Feed consumption and body weight gain as well as digestibility of crude protein, cr
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Here, a high sensitive method for biomarker identification according to nanostructure, using enzyme-linked immunosorbent assays (ELISAs), called Nano-ELISA, was presented. Different shapes of gold nanostructures (star and sphere; GNSs and GNPs) with a particle size of 40 nm for sphere particles were altered with a monoclonal antibody (Ab) as a detector Ab. To amplify the optical signal, gold nanostructures were employed as carriers of the signaling specific antibody against insulin growth factor binding protein- 3 (IGFBP-3). The substrate was catalytically oxidized by the Horseradish Peroxidase (HRP) conjugated gold nanostructure, and HRP also enhanced the optical signals, reflecting the amount of the targeting IGFBP-3. In comparison to t
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Diabetic nephropathy (DN) is the most common microvascular complication that may lead to chronic renal failure in diabetic patients. Till now microalbuminuria, with its restrictions, is the early marker of DN, appeared after the disease exacerbation. Thus, new biomarkers are required to predict the early onset of DN before the appearance of microalbuminuria. The aim of this study is to investigate the possible use of uVDBP in the early prediction of DN. Fifty diabetic patients with DN and 40 diabetic patients without DN for both types of diabetes were enrolled in this study. All patients were tested for uACR, uVDBP (measured by ELISA), and blood HbA1c. The results demonstrated a highly significant elevation of uAC
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Elevated C-Reactive Protein (CRP) level in serum is a risk factor for type 2 diabetes ,this relationship is likely to be the cause it means elevated CRP leads to T2D in future . Our objective was to examine CRP in male Type 2 Diabetes(T2D) patients in different age ,we studied 120 male subjects divided to two groups according to their age. First group A age (31 - 40) year old ,60 person )30 control & 30 T2D patients(,3 person for each same age: second group B age (41 – 50) years old ,60 person )30 control & 30 T2D patients(,3 person for each same age. We examined blood sugar ,cholesterol and CRP in each group. and we toke the mean of samples in the same age in each data in all the 4 groups. Our data shows that CRP
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This study looked at how the synthetic chitosan-AgNPs-Doxorubicin-folic acid combination affected the A549 cell line in terms of cytotoxicity and anticancer activity. By reducing silver nitrate (AgNO3) and biodegradable chitosan, silver nanoparticles were biosynthesized. The produced conjugate was examined by using FT-IR spectroscopy, atomic force microscopy (AFM), and field emission scanning electron microscopy (FE-SEM). The cytotoxicity assay for the viability of A549 cells revealed that the combination of chitosan, AgNPs, doxorubicin, and folic acid decrease cell viability in a dose-determined by method over 48 hours, which direct to a dependent reduce in the activity of A549 cells. The mechanism analysis of the impacted living cells lea
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Background: Red clover oil (Trifolium pratense) has isoflavones bunches which have estrogen-like exercises and may establish an option in contrast to hormone substitution treatment. The present study investigated the impact of Red clover oil on bone healing in rats by histomorphometric study. Materials and Methods: Intra bony defect was performed in right femur of thirty six healthy male albino rats. Then these rats were randomly divided into three groups (12 rats each): one control and 2 experimental groups. For control, the bony defect was left for normal healing; for the experimental (S) group, the defect was treated with hemostatic absorbable gelatin sponge; and for experimental (RS) group, the bony defect was treated with 0.2 ml red
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In this paper, Cu2S/CdS solar cells were prepared with different thickness of CdS layer, these layers were prepared by using chemical spray pyrolysis technique. The chemical spray solution was prepared by mixing cadmium chloride CdCl2 and thiourea CS(NH2)2 of molar concentration 0.1 M/L, the CdS layer was formed after the solution was sprayed on hot Aluminum substrate at temperature 400°C. Experimentally the type of CdS film was found as n–type depending on the results of Hall Effect, the value of the Hall factor (RH) is about – 1.348 x 10–6 m3/C and the density of the majority charge carriers (N) is about 4.64x1018 cm–3. The prepared film was tested by using X-Ray
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