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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Steps were taken to obtain the Kojic acid crystals from local fungal isolation A. flavus WJF81 by separating the fermentation products from the fungus mycelium from the production plant at the centrifuge at a speed of 5000 cycles for 10 minutes. The extraction was followed by ethyl acetate then supernatant concentrate by using rotary evaporator, and dried with heat oven 37ºC. Long, yellowish, pristine acid crystals were obtained that examined the optical microscope with a magnification force of 10x and 40x. The melting point of kojic acid was determined between 152.9-153.5 °C Results of the diagnosis of Kojic acid by applying High pressure liquid chromatography HPLC technique showed that the acid was at one peak, which was close to the
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The amount of protein in the serum depends on the balance between the rate of its synthesis, and that of its catabolism or loss. Abnormal metabolism may result from nutritional deficiency, enzyme deficiency, abnormal secretion of hormones, or the actions of drugs and toxins. Renal cancer is the third most common malignancy of the genitourinary system, and accounts for 3% of adult malignancies globally. Total serum proteins were measured in malignant kidney tumor, benign kidney tumors, and non tumoral kidney diseases patient groups, as well as in healthy individuals. A significant decrease (p< 0.001) of total serum protein levels in patients with malignant kidney tumors when compared with those of benign tumors, non tumoral diseases, and hea
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Objective: Per-implantitis is one of the implant treatment complications. Dentists have failed to restore damaged periodontium by using conventional therapies. Tissue engineering (stem cells, scaffold and growth factors) aims to reconstruct natural tissues. The paper aimed to isolate both periodontal ligament stem cells (PDLSCs) and bone marrow mesenchymal stem cells (BMMSCs) and use them in a co-culture method to create three-layered cell sheets for reconstructing natural periodontal ligament (PDL) tissue. Materials and methods: BMMSCs were isolated from rabbit tibia and femur, and PDLSC culture was established from the lower right incisor. The cells were co-cultured to induce BMMSC differentiation into PDL cells. Cell morphology, stem cel
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The aquatic crude extract of Silybum marianum dry grains prepared by melting them in distil water by the method of soak and shake. The effect of Silybum marianum crude extract studied in vitro on three tumor cell line the Hep-2, AMN-3 and RD for 24, 48 and 72 hours of exposure, and one cell line of normal cells REF for 72 hr exposure. The results showed that the prescence of toxic effect of the aquatic crude extract on the cell lines of Hep-2, AMN-3 and RD at 10 and 100 µg/ ml upto the higher concentrations when they exposed to the extract for 48 hr. as compared with the control treatment, and when the exposure period increased to 72 hr. the toxic effect started at low concentrations (5 and 10 µg/ ml) as compared with the control g
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Science occupies great importance in Islamic thought. Science and learning are considered an essential part of Islamic teachings, and this importance appears in several aspects,Among them is thatScience as a means of understanding religion :Science is a means of understanding the teachings of the Islamic religion. Islam encourages thinking and rational research to understand the Holy Quran and the Sunnah of the Prophet, enabling Muslims to direct their lives and actions in accordance with the directives of their religion,And also to encourageResearch :Islam encourages scientific research and the use of reason in understanding the nature of the universe and God’s signs in it. Muslims are encouraged to study the natural and social s
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The levels of circulating angiogenic and anti-angiogenic factors, namely vascular endothelial growth factor–A (VEGF-A) and soluble vascular endothelial growth factor receptor-1 (sVEGFR-1), have been linked to the development of renal dysfunction due to the proliferation of microvasculature within the kidneys of type 2 diabetic (T2DM) patients. The study aims to scrutinize serum levels of VEGF and sVEGFR-1 in a sample of Iraqi diabetic nephropathy patients to support their reliability as markers for the prediction of nephropathy in type 2 diabetes mellitus patients as well as to assess the ACE inhibitor’s effect on the levels of these two markers. Method: The ninety participants of this case-control study were split into three gr
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Objective(s): To measure serum C-reactive protein (CRP) titer as a predictive diagnosis of acute hepatitis C virus (HCV)
infection.
Methodology: Two hundred and ten patients with acute HCV infection and 234 apparently healthy individuals as
control group were enrolled in this study in Baghdad medical city (Teaching Laboratories). The patents include
74(35.2%) females and 136 (64.8%) males with mean age (27±16.5) years. The control group includes 114 (48.7%)
females and 120 (51.3%) males with mean age (26±5.8) years. Blood samples were collected from out patients from
Alfadul in Baghdad city. Sera were separated and stored at 20 0
C. The diagnosis of acute HCV infection was based on
detection of HC Ag and anti- H
The design of this paper is to find the possible correlation of Epstein Barr virus infection ina group of Iraqi women with cervical carcinoma though detection of Latent Membrane Protein 1 (LMP1) in these cervical tissues. Paraffinized blocks of two groups were included. The first sample of 30 cervical carcinomatous tissues and 15 biopsies from an apparently normal cervical tissues. All the samples were sectioned on a positive charged slides with 4 mm – thickness then submitted for immunohistochemical (IHC) staining to detect viral LMP1 expression. Sixty three percentage (19 out of 30) of the studies group showed positive overexpression as shown in with a significant association of the expression with cervical cancer with a significant ass
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