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 research demonstrates new species of the games by applying separation axioms via sets, where the relationships between the various species that were specified and the strategy of winning and losing to any one of the players, and their relationship with the concepts of separation axioms via sets have been studied.
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The tunnel’s stability during construction is a very important matter. Some methods have been proposed for stability evaluation, but the hazard warning levels (HWLs) are more applicable among these methods. Despite monitoring and applying HWLs, several collapses in Shibli twin tunnels in Iran have cast doubts on the accuracy of this criterion in the presence of water. In this study, the critical strains under different water contents were measured through uniaxial compressive strength tests on 11 different shale and marl samples. A comparison of laboratory tests and numerical results shows that the influence of the moisture content on the critical strain is negligible. In addition, the results show that there is no dir
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In this paper, we consider a new approach to solve type of partial differential equation by using coupled Laplace transformation with decomposition method to find the exact solution for non–linear non–homogenous equation with initial conditions. The reliability for suggested approach illustrated by solving model equations such as second order linear and nonlinear Klein–Gordon equation. The application results show the efficiency and ability for suggested approach.
This study aims to reach the right of governorates that are not organized in a region to impose local legislation, including tax legislation, and the extent of the constitutionality of this legislation and its consistency with constitutional texts and legal rules. The imposition of local taxes finds its constitutional and legal basis in the Iraqi constitution for the year 2005 and the law of governorates not organized in a region.The imposition of local taxes corresponds to the principle of tax legality, which is reflected in the necessity of issuing tax laws from a competent authority, whether this authority is federal, regional or local. Rather, it is sufficient that it be competent
The local asphalt concrete fracture properties represented by the fracture energy, J-integral, and stress intensity factor are calculated from the results of the three point bending beam test made for pre notches beams specimens with deformation rate of 1.27 mm/min. The results revealed that the stress intensity factor has increased by more than 40% when decreasing the testing temperature 10˚C and increasing the notch depth from 5 to 30mm. The change of asphalt type and content have a limited effect of less than 6%.
Inhibition of T cell derived cytokine production could help suppress osteoclast differentiation in inflammatory skeletal disorders. Bisphosphonates are typically prescribed to prevent inflammatory bone loss but are not tolerated by all patients and are associated with an increased risk of osteonecrosis of the jaw. In light of this other anti-resorptives such as phytoestrogens are being considered. However the effect of phytoestrogens on T cell-induced osteoclast formation is unclear. The effect of genistein and coumestrol on activated T cell-induced osteoclastogenesis and cytokine production was therefore examined. Concentrations of genistein and coumestrol (10−7 M) previously shown to directly inhibit osteoclast formation also suppressed
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Background: Squamous cell carcinoma of the oral
cavity (OSCC) is a highly invasive neoplasm. Many
MMPs play role in human cancer invasion and
metastases.
Aim: Estimating The MMp-7 expression level in
HPV-16 positive and HPV-16 negative OSCC
paraffin embedded sections.
Method: Biopsies from thirty three patients with oral
squamous cell carcinoma (OSCC) were obtained and
investigated for the presence of HPV-16 RNA with
the application of ISH and the MMP-7 expression
level using IHC .
Results: Expression level of MMP-7 found to be high
in OSCC sections 29 (87.8%) cases with no
significant difference in its expression level between
HPV-16 positive and HPV-16 negative OSCC cases
p= 1.00.
Conc
The increasing level of residents’ requirements of the local community led to the necessity for sufficient local funding to satisfy the residents’ requirements and services of the local units affiliated with the decentralized administrative systems on the one hand, and to the role of local financing in the financial independence of local units on the other hand.With the presence of local financing, the financial independence of local units is achieved and is considered one of the conditions for financial independence, which is the provision of local financing to the units away from central support. The study focused in this research to clarify the concept of local financing for local units with a statement of its conditions and importan
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