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.
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We investigated at the optical properties, structural makeup, and morphology of thin films of cadmium telluride (CdTe) with a thickness of 150 nm produced by thermal evaporation over glass. The X-ray diffraction study showed that the films had a crystalline composition, a cubic structure, and a preference for grain formation along the (111) crystallographic direction. The outcomes of the inquiry were used to determine these traits. With the use of thin films of CdTe that were doped with Ag at a concentration of 0.5%, the crystallization orientations of pure CdTe (23.58, 39.02, and 46.22) and CdTe:Ag were both determined by X-ray diffraction. orientations (23.72, 39.21, 46.40) For samples that were pure and those that were doped with
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New evidence on nanotechnology has shown interest in the creation and assessment of nanoparticles for cancer treatment. Worldwide, a wide range of tumor-targeted approaches are being developed to reduce side effects and boost the efficacy of cancer therapy. One strategy that shows promise is the use of metallic nanoparticles to increase the radio sensitization of the cancer cells while reducing or maintaining the normal tissue complication probability during radiation therapy. In this study, atmospheric plasma was created using argon gas to create Au NPs using the plasma jet scheme, and their ability to induce apoptosis as an anticancer mechanism was tested. Aqueous gold tetrachloride salts (HAuCl4·3H2O) ere used to produce gold nanopartic
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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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The angiotensin converting enzyme (ACE) I\D gene polymorphism influences the blood ACE enzyme activity. Renoprotective effect of ACE inhibitors (ACEIs) varies among patients due to genetic variation, particularly in Renin-Angiotensin-Aldosterone System genes. This study investigates the genetic variations of ACE I\D and AGT1RA1166C gene polymorphisms in the antiproteinuric effect of ACEI therapy in type 2 diabetes mellitus (T2DM) patients. This is a cross-sectional study that included 76 T2DM patients who are ACEI users, divided into two groups: T2DM without diabetic kidney disease (DKD) included 31 patients, and T2DM with DKD included 45 patients. Urine samples were taken for measurement of urine albumin and creatinine, then calcul
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This study involves the investigation of the effect of nitrogen laser with 337.1 nm wavelength on the sensitivity of Staphylococcus aureus bacteria by using local therapeutic due to burns. Thirty six isolate of Staphylococcus aureus bacteria were isolated from 25 patients suffering from sever burns, each isolate of bacteria was irradiated with nitrogen laser at (5, 10, 15 and 30) pulses/second repetition rates for 1, 5, 10, 20 and 30 minutes for each repetition rate. The effects of nitrogen laser on the local therapeutics sensitivity of bacteria were obtained using Kirby Baur method. Changes in the sensitivity of bacteria to local therapeutics (Tetracyclin, Chloramphenicol, Flumizin and Fucidin) occur at high repetition rate(30 pulses/seco
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?Natural Occurrence of aflatoxin Bi and B2 was investigated monthly in paddy rice stored for duration of six months. Aflatoxin B| was detected at the beginning of storage. The concentration of aflatoxin Bi in paddy and polished rice was found to be (0.3 and ()) ppb respectively while aflatoxin B2 W'as not detected in both paddy and polished rice afler six months of storage, concentration of aflatoxin Bi ‘ ’ ' reached 3 ppb in paddy rice and 0.9 ppb in polished rice, while the study proved that the concentration of aflatoxin in paddy rice was higher than polished rice as a result of processing which reduce the toxin
This dissertation explores the role of Iraqi E-press in crystallizing the orientations or directions of the public towards the local political issues like (demonstrations, parliamentarian elections, provincial elections, the public budget and its consequences, Iraq’s relations with neighboring countries, Iraqi HOR and its sessions, and the relation of the federal government with KRG). The dissertation’s main problem revolves around a central inquiry: what is the role of the Iraqi E-press in crystalizing the direction of the Iraqi public towards the local political issues?
The dissertation included a number of assumptions; the first assumes the relation of demographic variants (gender, age, social status, education, working status