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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Background: In spite of all efforts, Non-small cell lung cancer (NSCLC) is a fatal solid tumor with a poor prognosis as of its high metastasis and resistance to present treatments. Tyrosine kinase inhibitors (TKI) such as erlotinib are efficient in treating NSCLC but the emergence of chemoresistance and adverse effects substantially limits their single use. Objective: in this study, the combination treatments of either 2-deoxy-D-glucose (2DG) or cinnamic acid (CINN) with erlotinib (ERL) were tested for their possible synergistic effect on the proliferation and migration capacity of NSCLC cells. Methods: In this study, NSCLC model cell line A549 was used to investigate the effects of single compounds and their combination on cell gro
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Neuroendocrine differentiation has been mentioned in many cancers of non-neuroendocrinal organs, involving the gastrointestinal tract. In contrast, the correlation of focally diffused neuroendocrine differentiation in colorectal adenocarcinoma with neuroendocrine cell hyperplasia has not been somewhat reported. The objective of this research is to study the relationship between neuroendocrine cell hyperplasia and neuroendocrine differentiation in colorectal adenocarcinoma and to find the correlation of neuroendocrine differentiation and VEGF expression with clinicopathological parameters of colorectal adenocarcinoma. Methods employed in the current study were including eighty-one patients with colorectal cancer. Formalin fixed paraffin e
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This effort is related to describe and assess the performance of the Iraqi cement sample planned for oil well-cementing jobs in Iraq. In this paper, major cementing properties which are thickening time, compressive strength, and free water in addition to the rheological properties and filtration of cement slurry underneath definite circumstances are experimentally tested. The consequences point to that the Iraqi cement after special additives encounter the requests of the API standards and can consequently is used in cementing jobs for oil wells. At this research, there is a comparative investigation established on experimental work on the effectiveness of some additives that considered as waste materials which are silica fume, baux
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Gaining the relationship between the central government and local governments is great importance especially after the expanded functions of the central government and became unable to carry out their works without all that delegate or transfer parts of its powers to lower levels.
The aims of the research is to determine the relationship of the central government with the local government in accordance with the principle of administrative decentralization with the presentation of the competent authorities, the control of local governments, according to the Provincial Council Act No. 21 of 2008 amended.
The research has a problem that has been the relationship of the central
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This effort is related to describe and assess the performance of the Iraqi cement sample planned for oil well-cementing jobs in Iraq. In this paper, major cementing properties which are thickening time, compressive strength, and free water in addition to the rheological properties and filtration of cement slurry underneath definite circumstances are experimentally tested. The consequences point to that the Iraqi cement after special additives encounter the requests of the API standards and can consequently is used in cementing jobs for oil wells. At this research, there is a comparative investigation established on experimental work on the effectiveness of some additives that considered as waste materials which are silica fume, bauxite,
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