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.
Water samples were collected from output of water for Al-Wahda plant where located in al-karrada area in Baghdad city to study water contamination with bacteria, fungi and Algae. The study lasted one year started on August, 2016 to July,2017.Results were acquired according to two tests performed, the first is biological test included total coliform,E.coli, pseudomonas aeruginosa, total fungi, Diatom and non Diatom Algae and the second is physiochemical test included temperature, turbidity and residual chlorine. The results of bacteria were within the permitted specification in the Iraqi standards no. 14/2270 for the year 2015 except August was exceeded the permitted standard for total coliform, it was 1.1< cell/100 ml.Total Fungi, Dia
... Show MoreIn this research, the influence of the fermentation treatments and baking in Iraqi’s flour type (Ibaa 99, Al-rashed, Tamus, Abu-grabe) and Turkish flour type (Muaamel) on phytic acid was investigated. In whole wheat flour, the phytic acid was (1500, 1290, 1450, 1230, 1440 ( mg/ 100 g flour respectively, and the inorganic phosphorous was (29.18, 25.15, 23.89, 20.85, 22.83) mg/100 g flour respectively. The dough prepared from flour with a higher phytic acid content also contained higher amount of phytic acid. During fermentation, degradation of phytic acid occurred. The cumulative loss of phytic acid after fermentation in all type of dough was ~ 23, 22, 34, 26 and 27% respectively،While increased of inorganic phosphorous occurred. The c
... Show MoreBackground/Objectives: Nonsurgical periodontal treatment (NSPT) is the gold-standard technique for treating periodontitis. However, an individual’s susceptibility or the inadequate removal of subgingival biofilms could lead to unfavorable responses to NSPT. This study aimed to assess the potential of salivary and microbiological biomarkers in predicting the site-specific and whole-mouth outcomes of NSPT. Methods: A total of 68 periodontitis patients exhibiting 1111 periodontal pockets 4 to 6 mm in depth completed the active phase of periodontal treatment. Clinical periodontal parameters, saliva, and subgingival biofilm samples were collected from each patient at baseline and three months after NSPT. A quantitative PCR assay was us
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Type 2 diabetes mellitus (T2DM) became the most prevalent health problem. Almost half of the world's people are ignorant that have diabetes. Menopause occurs as an important alteration in women through which take place the change in sex hormones, distribution in fat،s body, and metabolism, altogether which participate in the metabolism disease such as type 2 diabetes mellitus. Several studies have appeared the association between the TCF7L2 gene and different diseases like type 2 diabetes mellitus (T2DM). This study aimed to detect the relation of the genetic variation polymorphism for the TCF7L2 gene (rs12255372 G/T) in Iraqi women menopausal with T2DM. The outcomes indicated the increased levels of biochemical characteristics including H
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(8)
Background: Type 2 diabetes mellitus (T2DM) characterized by insulin resistance (IR) and progressive decline in functional beta (β) cell mass partially due to increased β cell apoptosis rate. Pancreatic stone protein /regenerating protein (PSP/reg) is produced mainly by the pancreas and elevated drastically during pancreatic disorder. Beta cells are experiencing apoptosis that stimulate the expression of PSP/reg gene in surviving neighboring cells, and that PSP/reg protein is subsequently secreted from these cells which could play a role in their regeneration.
Objectives: To analyze serum levels of PSP/reg protein in T2DM patients and evaluate its correlation with the microvasc
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(1)
From 144 specimens of tonsillitis which were collected from patient, (children of 3 -12 year olds) there were 70 isolates beta hemolytic and 28 isolates were identified as S. pyogenes. Sensitivity of S. pyogenes isolates to antibiotics was tested, all isolates were sensitive to amoxicillin and cephaloxia while higher resistant were to erythromycin. One isolate whiche was 100 A had a stable characteristics and produce pyrogenic toxin was chosen for study and it was purified and characterized from the cell free supernatant of S. pyrogenes strain.
Ubikh is one of the most important tribes of the Northwest Caucasus and has a long history. Naturally, rebukes are an integral part of the Caucasus region. Their people were able to confront the Russian invasion of their lands like the rest of the Caucasus tribes to defend their existence, freedom and the right to build their independent state, but they were not able to resist the great Russian military progress they achieved on all sides. For the territory that belonged to Persia and the Ottoman Empire and ended the resistance of the Meridian movement in the Northeast Caucasus, the Russians left only the weak resistance of the tribes of the Northwest Caucasus led by the rebuke after Russia managed to eliminate all resistance movements i
... Show MoreThis paper aims at the analytical level to know the security topics that were used with data journalism, and the expression methods used in the statements of the Security Media Cell, as well as to identify the means of clarification used in data journalism. About the Security Media Cell, and the methods preferred by the public in presenting press releases, especially determining the strength of the respondents' attitude towards the data issued by the Security Media Cell. On the Security Media Cell, while the field study included the distribution of a questionnaire to the public of Baghdad Governorate. The study reached several results, the most important of which is the interest of the security media cell in presenting its data in differ
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