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 n-type Au thin films of 500nm thickness was evaporated by thermal evaporation method on p-type silicon wafer of [111] direction to formed Au/Si heterojunction solar cell. The AC conductivity, C-V and I-V characteristics of fabricated c-Au/Si diffusion heterojunction-(HJ) solar cell, has been studied. The first methods demonstrated that the AC conductivity due to with diffusiontunneling mechanism, while the second show that, the heterojunction profile is abrupt, the heterojunction parameters have been played out, such as the depletion width, built-in voltage, and concentration. And finally the third one show that the c-Au/Si HJ has rectification properties, and the solar cell yielded an open circuit voltage of (Vic) 0.4V, short circuit c
... Show MoreCoffee bean contains bioactive compounds including caffeine and chlorogenic acid (CGA) that have a stimulant effect and are used for combating fatigue and drowsiness, and enhancing alertness. However, when the coffee bean was processed in the form of green coffee bean (GCB) extract, it has an unpleasant flavour and limitations instability, activity, and bioavailability. This study aimed to produce microcapsules of the GCB (Coffea canephora) ethanolic extract containing considerable amounts of the bioactive compounds for nutraceutical supplements. The GCB ethanolic extract was microencapsulated by spray drying using a whey protein concentrate (WPC) biopolimer. The particle size (PSA), morphology (SEM), and physicochemical charact
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Molasse medium containing different concentrations of (NH4)2 SO4, (NH4)3 PO4, urea, KCI, and P2O5 were compared with the medium used for commercial production of C. utilis in a factory south of Iraq. An efficient medium, which produced 19. 16% dry wt. and 5. 78% protein, was developed. The effect of adding various concentrations of micronutrients (FeSO4, 7T20, MnSO4. 7H20, ZnSO4. 7E20) was also studied. Results showed that FeSo4. 7H20 caused a noticeable increase in both dry wt. and protein content of the yeast.
Sera samples were collected from 60 children aged 4-60 months, all were clinically and serologically proven cases of visceral leishmaniasis, as well as from 10 healthy children, all were seronegative with no history of parasitic infection who serve as a control during the study. Serum total protein and albumin were measured and compared between the control and visceral leishmaniasis patients. Serum protein profiles have been investigated using the conventional sodium dodecyl sulphate – polyacrylamide gel electrophoresis (SDS-PAGE). Serum of control group showed the specific protein pattern with five protein bands, while serum protein profile in visceral leishmaniasis pat
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Assessment of Salivary Macrophage Inflammatory Protein-1 Alpha Level in Different Stages of Periodontitis, Riyam Muthanna Muhammed*, Hadeel Mazin Akram
Multiple sclerosis (MS) is a neurodegenerative disorder with various clinical types. Glial fibrillary acidic protein (GFAP) is significantly elevated in the cerebrospinal fluid (CSF) of MS patients compared with that of healthy controls. The aim of this study is to evaluate serum levels of GFAP in relation to disease activity in relapsing-remitting MS patients and to compare them with those of healthy controls. Method: This study involved 58 MS patients of relapsing-remitting MS (RRMS) type, 22 in an active stage of the disease and 36 in remission, and 50 healthy individuals as age- and sex-matched controls. Blood samples were taken from the patients at the MS Clinic of the Baghdad Teaching Hospital, and the serum levels of GFAP were determ
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Focal adhesion kinase (FAK), ephrin receptor type A4 (EphA4), and adiponectin (ADPN) are important indicators in inflammation, tumor growth, migration, and angiogenesis in some cancers. The predictive impact of their concentrations in acute myeloid leukemia (AML) patients to be identified remains. The research sought to explore the effect of FAK, EphA4, and ADPN as prognostic biomarkers, and their influence on patient survival, and to look for any potential correlation between their levels with hematological parameters in AML patients.
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Purpose: To assess the antioxidant and antineoplastic effects of Hibiscus sabdariffa Linn. on oral squamous cell carcinoma cells. Materials and Methods: Human squamous cell carcinoma HSCC cells were tested for cytotoxicity by a methanol extract of Hibiscus sabdariffa (MEHSP). After 24, 48, and 72 ...
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This study aimed to determine the optimal conditions for extracting basil seed gum in addition to determine the chemical components of basil seeds. Additionally, the study aimed to investigate the effect of the mixing ratio of gum to ethanol when deposited on the basis of the gum yield which was1:1, 1:2, 1:3 (v/v) respectively. The best mixing ratio was one size of gum to two sizes of ethanol, which recorded the highest yield. Based on the earlier, the optimal conditions for extracting basil seed gum in different levels which included pH, temperature, mixing ratio seeds: water and the soaking duration were studied. The optimal conditions were: pH 8, temperature of 60°C, mixing ratio seeds: water 1:65 (w/v) and soaking duration of 30 min
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