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.
Background: Squamous cell carcinoma is a disease of elderly peopleand it is uncommon in people with less than 40 years old; however many literatures revealed that tumor developing in patients younger than forty years appears more aggressive at the time of diagnosis. The purpose of the present study was to focus on the clincopathological features of the oral SCC in different age groups. Material and methods: In this study thirty five cases of paraffin embedded tissue blocks of oral squamous cell carcinoma were studied. The age range was from 16 to 80 years. The clinicopathological data were recorded for evaluating the tumor characters according to age of patients. Results : The age was not significantly correlated to the clinicopathological
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Copper doped Zinc oxide and (n-ZnO / p-Si and n-ZnO: Cu / p-Si) thin films thru thickness (400±20) nm were deposited by thermal evaporation technique onto two substrates. The influence of different Cu percentages (1%,3% and 5%) on ZnO thin film besides hetero junction (ZnO / Si) characteristics were investigated, with X-ray diffractions examination supports ZnO films were poly crystal then hexagonal structural per crystallite size increase from (22.34 to 28.09) nm with increasing Cu ratio. The optical properties display exceptional optically absorptive for 5% Cu dopant with reduced for optically gaps since 3.1 toward 2.7 eV. Hall Effect measurements presented with all films prepared pure and doped have n-types conductive, with a ma
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Bacteriocin is an important antimicrobial peptide that can be used in industrial and medical fields due to its characteristics of antibacterial, food preservation and anticancer activities. Fifty isolates of Bacillus sp were collected from different soil samples which were already recognized via morphological and biochemical identification process. The isolates were screened for bacteriocin production effective against Staphylococcus spp in order to select the highest producing isolate. The isolate NK16 showed the maximum bacteriocin production (80 AU/ml) which was further characterized as Bacillus subtilis NK 16 through using API identification system (API 20E and API 50CHB). Then, next step was to detect the optimal conditions for maximum
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The current study deals with estimating the protein concentration and the effect of fish weight on protein concentration values in red and white muscles in two different regions ( R1 : Anterior region lies 2 cm behind the head and R2: posterior region lies 2cm from caudal fin (in two types of bony fish, namely common carp (Cyprinus carpio) and Nile tilapia (Oreochromis niloticus). Samples were collected from Karmat Ali river- north of Basrah between October 2019 and February 2020. The protein was extracted using protein extraction buffer, the current study show that the average of protein concentration in red muscles of Nile tilapia ranged between 7.74-7.4 mg / ml and ( 6.8-8.85 mg / ml) in R1 and R2 region re
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Abstract: Recombinant human bone morphogenetic protein-2 (rhBMP-2) and platelet-rich fibrin (PRF) bioactive materials have been used to enhance healing and improve dental implant stability. This study aimed to compare the effect of rhBMP-2 and PRF bioactive materials on dental implant stability at different intervals and to evaluate the correlation of implant length and diameter with implant stability. Two bioactive materials were compared to evaluate their effect on dental implant stability. A total of 32 patients (102 dental implants) were divided into 3 groups: 24 dental implants with bone morphogenetic protein (BMP), 27 dental implants with PRF, and 51 dental implants without BMP or PRF (control group). Data were statistically analyzed
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Gingival crevicular fluid (GCF) may reflect the events associated with orthodontic tooth movement. Attempts have been conducted to identify biomarkers reflecting optimum orthodontic force, unwanted sequallea (i.e. root resorption) and accelerated tooth movement. The aim of the present study is to find out a standardized GCF collection, storage and total protein extraction method from apparently healthy gingival sites with orthodontics that is compatible with further high-throughput proteomics. Eighteen patients who required extractions of both maxillary first premolars were recruited in this study. These teeth were randomly assigned to either heavy (225g) or light force (25g), and their site specific GCF was collected at baseline and aft
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This study aimed to evaluate the IHC expression of CDX2 protein in HGC patients and control groups and also to study the correlation between IHC expression of the CDX2 and different clinicopathological variables such as: age, gender, histopathological subtype, grade, and stage of the tumor in HGC cases. the retrospectively sectional study for the period from 2014 to 2018 included a total of 60 formalin fixed paraffin embedded blocks of the HGC tissue (partial or total gastrectomy specimens) that collected from the archived materials of the Department of Pathology of Baghdad Teaching Hospital and the Center of Gastrointestinal and Hepatic Diseases, and also some samples were collected from other private laboratories. The IHC expression of th
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The effect of using grinded rocks of (quartzite and porcelanite) as powder of (10 and 20) % replacement by weight of cement for self-compacting concrete slabs was investigated in this study. Five slabs with 15 concrete cubes were tested experimentally at 28 days to study the compressive strength, ultimate load, ultimate deflection, ductility, crack load and steel strain. The test results show that, the compressive strength improvement when replacement of local rock powder reached to (7.3, 4.22) % for (10 and 20) % quartzite powder and (11.3, 16.1) % for (10 and 20) % porcelanite powder, respectively compared to the reference specimen. The ultimate load percentage increase for slabs with (10 and 20) % rep
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