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.
Rheumatoid arthritis (RA), is an autoimmune, and inflammatory disease that is closely related to the destruction of cartilage and bone. DC-SIGN are important types of C-type lectin receptors (CLRs), expressed on dendritic cells and macrophages, and have a central role in regulating innate and adaptive immunity, function as pattern recognition receptors, and as cell adhesion molecules. Recent evidence has demonstrated that DC-SIGN is involved in the pathophysiological of chronic inflammation, so DC-SIGN has been linked to several autoimmune and may play an essential indicator in the pathogenesis and progression of RA. Therefore, the purpose of this study is to determine the serum level of DC-SIGN in RA patients, as well as the level of DC
... Show MoreColorectal cancer (CRC) is the most common disease and cause of death globally. The aim of the study is investigation and detection of some bacterial interfering with CRC occurrence and progression. The study conducted between September 2022 till February 2023, a total of 50 specimens were collected from confirmed CRC patients. In addition, 50 stool specimens were collected from Healthy volunteers, considers as control group. Isolation and identification of bacteria in all collected specimens were done by using cultural and differential media (blood agar, macconkey agar and Pfizer agar), as well as the VITEK- 2 compact system. The bacterial species, in the specimens of control were ( Escherichia coli 50 (86.20%), Klebsiella Pneumoni
... Show MoreBackground: Tumor associated tissue eosinophilia (TATE) has been described in a variety of neoplasms. In regard to squamous cell carcinoma, some studies worldwide done to assess stromal eosinophilia in oral and cervical squamous cell carcinoma. The objectives of this study is to evaluate the association of stromal eosinophilic infiltration of cutaneous squamous cell carcinoma and to detect the significance of this association. Aim of the study The aim of our study is to establish the relationship between the degree of stromal eosinophilia and the level of invasion and the histological grade in cutaneous squamous cell carcinoma. Methods: In this retrospective study done at the histopathology department of al Wasity teaching hospital for orth
... Show MoreBackground: Tumor associated tissue eosinophilia (TATE) has been described in a variety of neoplasms. In regard to squamous cell carcinoma, some studies worldwide done to assess stromal eosinophilia in oral and cervical squamous cell carcinoma. The objectives of this study is to evaluate the association of stromal eosinophilic infiltration of cutaneous squamous cell carcinoma and to detect the significance of this association.
Aim of the study
The aim of our study is to establish the relationship between the degree of stromal eosinophilia and the level of invasion and the histological grade in cutaneous squamous cell carcinoma.
Methods: In this retrospective study
... Show MoreMany people take protein supplements in an effort to gain muscle. However, there is some controversy as to whether this is really effective. There is evidence suggesting that consuming high level s of protein may in fact have negative side effects for health. The current study included 29 young Iraqi building muscles in two different groups (taken and not protein supplements) (age range=17-31 years), the cases were selected from family, friends, college students, and Gyms), from November 2014 to March 2015. A careful history was obtained from each volunteer including age, duration of sports, type of supplements, and family history of diseases. Some biochemical parameters like (glucose, urea, uric acid, creatinine, bilirubin, serum protei
... Show MoreThis 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
... Show MoreThe investment budget represents a stage of the investment decision in service units, and the preparation and implementation needs to be a complement of the same planning part, because the planning does not end with the development of the plan, but includes a follow-up implementation, so it has to be effective and efficient oversight of the estimates and procedures for disbursement of funds approved for investment projects, The problem with research in that local governments suffer from the presence of Allkaat and problems facing the implementation of the investment budget projects due to the adoption budget items which can not be measured the efficiency of the performance of these units of government by, and shortcomings in the control
... Show MoreBackground and Aim: The aryl hydrocarbon receptor (AhR) plays a pivotal role in spermatogenesis through its regulatory functions in redox balance and gene expression. This study aimed to investigate the effects of resveratrol (RES), a polyphenolic AhR modulator, and CH223191, a selective AhR antagonist, on male reproductive function in rats by assessing sperm quality, oxidative stress, testicular histopathology, and AhR gene expression. Materials and Methods: Forty adult male rats were randomly divided into four groups: (i) Control, (ii) dimethyl sulfoxide (vehicle), (iii) RES (100 mg/kg i.p., twice weekly), and (iv) AhR⁻ (CH223191, 10 mg/kg i.p., twice weekly), treated for 60 days. Post-treatment, sperm motility, survival, viabi
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