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.
(22)
(18)
(8)
(7)
Corona Virus Disease-2019 (COVID-19) is a novel virus belongs to the corona virus's family. It spreads very quickly and causes many deaths around the world. The early diagnosis of the disease can help in providing the proper therapy and saving the humans' life. However, it founded that the diagnosis of chest radiography can give an indicator of coronavirus. Thus, a Corner-based Weber Local Descriptor (CWLD) for COVID-19 diagnostics based on chest X-Ray image analysis is presented in this article. The histogram of Weber differential excitation and gradient orientation of the local regions surrounding points of interest are proposed to represent the patterns of the chest X-Ray image. Support Vector Machine (SVM) and Deep Belief Network (DBN)
... Show More
(3)
(2)
BackgroundCarcinoma of the larynx represent 10% of head & neck malignancies. The treatment of advanced carcinoma of larynx may include partial or total laryngectomy, with or without laser , radiotherapy,and or chemotherapy . Carcinoma of the larynx usually affect old age , heavy smoker with possible risk of pulmonary diseases & ischemic heart disease , which add risk to the general anaesthetic complication operative & postoperative - Objectives this study was designed to assess the feasibility of total laryngectomy under local anaesthesia in medically unfit patients for general anaesthesia & to re-establish practice doing total laryngectomy under local anaesthesia in those patients.
Methods a prospective study on 12 pa
BackgroundCarcinoma of the larynx represent 10% of head & neck malignancies. The treatment of advanced carcinoma of larynx may include partial or total laryngectomy, with or without laser , radiotherapy,and or chemotherapy . Carcinoma of the larynx usually affect old age , heavy smoker with possible risk of pulmonary diseases & ischemic heart disease , which add risk to the general anaesthetic complication operative & postoperative - Objectives this study was designed to assess the feasibility of total laryngectomy under local anaesthesia in medically unfit patients for general anaesthesia & to re-establish practice doing total laryngectomy under local anaesthesia in those patients.
Methods a prospective study on 12 pa
(21)
(17)
(3)
Celiac disease (CD) is the most common genetically - based disease in correlation with food intolerance. The aim of this study is to measure the activity of ALT enzyme and purify enzyme from sera women with celiac disease. Alanine aminotransferase (ALT) activity has been assayed in (30) women serum samples with celiac disease, age range between (20-40) year and (30) serum of healthy women as control group, age range between (22-38) year. In the present study, the mean value of ALT activity was significantly higher in patients with celiac disease than healthy group (p<0.01). The ALT enzyme was partial purified from sera women with celiac disease by dialysis, gel filtration using Sephadex G- 50 and ion exchange chromatography using DEAE- cell
... Show More
(5)
Celiac disease (CD) is the most common genetically - based disease in correlation with food intolerance. The aim of this study is to measure the activity of ALT enzyme and purify enzyme from sera women with celiac disease. Alanine aminotransferase (ALT) activity has been assayed in (30) women serum samples with celiac disease, age range between (20-40) year and (30) serum of healthy women as control group, age range between (22-38) year. In the present study, the mean value of ALT activity was significantly higher in patients with celiac disease than healthy group (p<0.01). The ALT enzyme was partial purified from sera women with celiac disease by dialysis, gel filtration using Sephadex G- 50 and ion exchange chr
... Show More
(5)