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 CIGS/CdS p-n junction thin films were fabricated and deposited at room temperature with rate of deposition 5, and 6 nm secG1 , on ITO glass substrates with 1mm thickness by thermal evaporation technique at high vacuum pressure 2×10G5 mbar, with area of 1 cm2 and Aluminum electrode as back contact. The thickness of absorber layer (CIGS) was 1 µm while the thickness of the window layer CdS film was 300 nm. The X-ray Diffraction results have shown that all thin films were polycrystalline with orientation of 112 and 211 for CIGS thin films and 111 for CdS films. The direct energy gaps for CIGS and CdS thin films were 1.85 and 2.4 eV, respectively. Atomic Force Microscopy measurement proves that both films CIGS and CdS films have nanostru
... Show MoreCorona 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)
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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
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The Experiment was carried out to determine the level vibration transfer in three axes Horizontal X, Lateral Y and Vertical Z direction to seat driver tractor, Vector sum of vibration and Daily Vibration Exposure (8 hours) in seat driver tractor, and vibration in steering wheel tractor, Heart Rate, Systolic and Diastolic blood pressure and temperature were measure to all Drivers before and after used Chisel plow in operation tillage. Statistical analysis system was used, Split-Split Plot Design under Randomized Complete Block Design, Three factors were used in this experiment included Two types of Soil Moist and Dry soil which represented main plot, Three Velocity Tractor was second factor included 1.6,3.5 and 5.4 km/hr and Three Drivers Tr
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Blood samples of One hundred and twenty patients from different hospitals in Baghdad infected with hydatidosis in different sites of the body (Liver, Lung, multiorgans and kidney) were collected for this study. On the other hand, 30 healthy individuals were included as a control group. This study was conducted to evaluate the effect of this disease on the serum protein profile of the patients using electrophoresis. The results revealed four different protein banding patterns with difference in number of bands and their molecular weights in comparison to the control group, and these differences depended on the site of infection. However the data showed a presence of the same band in all patients with different site of infection.