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.
ABSTRACT
Ticagrelor is an orally administered antiplatelet medicine, direct-acting P2Y12-receptor antagonist. Ticagrelor binds reversibly and noncompetitively to the P2Y12 receptor at a site distinct from that of the endogenous agonist adenosine diphosphate (ADP). Inhibition of platelet aggregation stimulated by ADP is a commonly used pharmacodynamic parameter for P2Y12-receptor antagonists.
Ticagrelor is a crystalline powder with an aqueous solubility of approximately 10?g/mL at room temperature.
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Nanoparticles of copper sulfide have been prepared by simple reaction between using copper nitrate with different concentrations ratio 0.1, 0.3, and 0.5 mM, thiourea by a simple chemical route. The prepared Nano powders have been deposited onto glass substrates by casting method at 60°C. The structure of the product Nano- films has been studied by x-ray diffraction, where the patterns showed that all the samples have a hexagonal structure of covellite copper sulfide with the average crystalline sizes 14.07- 16.51 nm. The morphology has been examined by atomic force microscopy, and field emission scan electron microscopy. The AFM images showed particles with almost spherical and rod shapes with average diameter sizes of 49.11- 90.64 nm.
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Attitude is an important subject that has attracted researchers and thinkers in organizational theory and organizational behavior because of its great importance in various field studies. Field evidence suggests that individuals in their daily lives or careers respond to specific events through a set of internal tendencies Internal tendencies are negative or positive and systematic. These trends tend to be invisible, not limited to certain boundaries, and involve a significant number of individuals within organizations or society.
The research aims to identify the impact of trends in the ethics of public service workers for individuals, through a sample of (72) doctors and (60) administrative staff
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Abstract A descriptive correlation study which was utilizing an assessment approach, was carried out from November 19th, 2002 through April 30, 2004 in order to assess the psychosocial domain of the quality of life for the infertile men. A purposive sample of (200) men with infertility was selected from the High Institute for Embryo Research and Infertility Treatment and Alsamaraee Hospital in Baghdad city. A questionnaire was adoapted and developed of the World Health Organization quality of life scale for the purpose of the study. The questionnaire (WHOQOL) (1998) Reliability and validity of the questionnair
There are a lot of measures for Post Traumatic Stress Disorder (PTSD), but what distinguishes it from other measures is the number of short paragraphs and concise depending on the classification of American Psychiatric Association (APA), which can be applied by the researchers on a large number, Because the Iraqi society suffered from psychological trauma due to wars, conflicts and occupation and the resulting psychological disorders of the members of Iraqi society, especially post-traumatic stress disorder, as mentioned by many studies and researches.
Therefore, the aim of this study was to standardize the PTSD scale for students at Baghdad University. For the Psychologist Fua . E. which was applied in many foreign studies.
This paper has investigated experimentally the dynamic buckling behavior of AISI 303 stainless steel Aluminized and as received long columns. These columns, hot-dip aluminized and as received, are tested under dynamic buckling, 22 specimens, without aluminizing (type 1), and 50 specimens, with hot-dip aluminizing at different aluminizing conditions of dipping temperature and dipping time (type 2), are tested under dynamic compression loading and under dynamic combined loading (compression and bending) by using a rotating buckling test machine. The experimental results are compared with Perry Robertson interaction formula that used for long columns. Greenhill formula is used to get a mathematical model that descripts the buckling behavior
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The research problem is dedicated to investigate reservoirs irrational economic behavior adopted by the ruling elites in developing countries about the investment methodology of human capital and operating policies is based on the terms of reference of economic theory and standards governing the market, which led to a chronic structural imbalance in the workforce structure and lack of consistency with different production structure, in turn, which had a reported effects in the emergence of the phenomenon of unemployment and that they involved a certain privacy, as has become the issues of unemployment and employment in the various countries of the world are issues more important due to the presence of large numbers of the workforce in th
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Eighty five samples were taken from patients suffering from pneumonia. Seventy-eight isolates were diagnosed as following: Staphylococcus aureus (23), klebsiella pneumoniae (29), Streptococcus pneumoniae (15), Serratia sp. (4), Haemophilus influenzae (4) and Pseudomonas aeruginosa (3). The clinical isolates were tested for antibiotics sensitivity. They appeared highly resistance to penicillin G and Ampicillin at percentage 89.7 and 84.6% respectly while the results showed highly sensitivity to streptomycin at percentege of (12.8%). To study the antibacterial activity of Alium sativum, Eucalyptus microtheca leaves and Cydonia oblonga seeds extracts, five multi resistant strains were used by using agar well diffusion and disk methods at c
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Out of a total of fifty samples, thirty-five isolates were identified as Serratia marcescens. Thesediverse clinical samples were collected over a three-month period, from October 2023 to December 2023, fromseveral hospitals in Baghdad, including Fatima Al-Zahraa Hospital, Al-Sader Hospital, Ibn Al-Balady Hospital,and Al-Imam Ali Hospital. The clinical samples primarily included urine from patients with urinary tractinfections (UTIs). All isolates were cultured on nutrient agar, MacConkey agar, and blood agar, and theiridentities were confirmed through biochemical testing and the Vitek 2 compact system. Based on phenotypicvirulence factors, the S. marcescens isolates showed varying positive patterns: 32 out of 35 (91.42%) forprotease
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