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.
Background: The diagnosis of prostatic pathology may be of challenging , as some difficult and suspected, atypical cases may lack basal cell layer by routine H&E sections . Antibodies against 34BE12(HMW-CK) and p63 aid the diagnosis of such cases , to distinguish benign from malignant prostatic lesions.
Objective: to identify basal cells in atypical prostatic lesions ,and distinguish benign from malignant prostatic lesions.
Type of the study: A retro-spective study.
Methods: 115cases of paraffin embedded prostatic tissue blocks ,diagnosed as : 76 cases were benign prostatic hy
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due to the presence of chemoresistance and the risk of tumor recurrence and metastasis. There is a pressing necessity to develop efficient treatments to improve response for treatment and increase prolong survival of breast cancer patients. Photodynamic therapy (PDT) has attracted interest for its features as a noninvasive and relatively selective cancer treatment. This method relies on light-activated photosensitizers that, upon absorbing light, generate reactive oxygen species (ROS) with powerful cell-killing outcomes. Nuclear factor kappa B (NF-κB), a transcription factor, plays a key role in cancer development by regulating cell proliferation, differentiation, and survival. Inhibiting NF-κB can sensitize tumor cells to chemotherapeuti
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One of these plants utilized in traditional medicine is Lactuca seriolla Linn., which belongs to the Asteraceae family. It goes by a variety of common names in the world, including prickly lettuce, wild lettuce, jagged lettuce, and Kahu and khas. The work aimed to isolate and characterize some bioactive constituent(s) from the aerial part of Lactuca serriola utilizing Combiflash NEXTGEN and high-performance liquid chromatography (HPLC). Lactuca serriola (aerial part) was extracted with 80% ethanol, then fractionated with hexane. Then 250 mg of hexane extract was mixed with 4 g of silica gel and loaded in cartilage, then bounded to the gold column of combi flash using a solvent system comprised of ethyl acetat
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The aim of research is to show the effect of Ferric Oxide (Fe2O3) on the electricity production and wastewater treatment, since 2.5% of Ferric Oxide (Fe2O3) (heated and non heated) nanoparticles has been used. Characterization of nanoparticles was done using X-ray Diffraction (XRD) and Scan Electron Microscopy (SEM). The influence of acidity was also studied on both wastewater treatmenton the Chemical Oxygen demand (COD) and Biological Oxygen Demand (BOD) and voltage output was studied. From the results, it was infused that the dosage of 0.025 g/l and an initial pH 7 were founded to be optimum for the effective degradation of effluents. The results concluded that the treatment of anaerobic sludge wastewater using Ferric Oxide (Fe2O3) in
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CdO:NiO/Si solar cell film was fabricated via deposition of CdO:NiO in different concentrations 1%, 3%, and 5% for NiO thin films in R.T and 723K, on n-type silicon substrate with approximately 200 nm thickness using pulse laser deposition. CdO:NiO/n-Si solar cell photovoltaic properties were examined under 60 mW/cm2 intensity illumination. The highest efficiency of the solar cell is 2.4% when the NiO concentration is 0.05 at 723K.
Present study was conducted to evaluate the different levels of energy to protein ratios (EPR) using food waste and black soldier fly larvae meal (FWBSFL) on growth performance and nutrient digestibility of broilers. A total of 160 one-day old broiler chicks were divided randomly to four groups and each group had 8 replicates with 5 chicks per replicate. The control diet was formulated using conventional feed ingredients with EPR of 154 for the starter period and 167 for the finisher period. The other treatments were diets with normal, low, and high EPR (154,143, and 166 for the starter period; 167, 155, and 177 for the finisher period) using FWBSFL. Feed consumption and body weight gain as well as digestibility of crude protein, cr
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Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has caused enormous issues worldwide and is the most infectious pandemic. This study included 50 subjects (evenly distributed between sexes) and their range of ages starting from 2 to 67 years. According to the study's result, the ages and genders of subjects include susceptibility to COVID-19. Males were found to be more infected than females, and the ages of 36 to 67 were more common than other age ranges. Also, BMI calculations revealed that male patients with COVID-19 have the highest percentage of obesity. The clinical parameter results have been found serum C‐reactive protein (CRP) as an essential indicator that changes significantly in infection with COVID‐19 an
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Here, a high sensitive method for biomarker identification according to nanostructure, using enzyme-linked immunosorbent assays (ELISAs), called Nano-ELISA, was presented. Different shapes of gold nanostructures (star and sphere; GNSs and GNPs) with a particle size of 40 nm for sphere particles were altered with a monoclonal antibody (Ab) as a detector Ab. To amplify the optical signal, gold nanostructures were employed as carriers of the signaling specific antibody against insulin growth factor binding protein- 3 (IGFBP-3). The substrate was catalytically oxidized by the Horseradish Peroxidase (HRP) conjugated gold nanostructure, and HRP also enhanced the optical signals, reflecting the amount of the targeting IGFBP-3. In comparison to t
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Ab – initio restricted Hartree - Fock method within the framework of large unit cell (LUC) formalism is used to investigate the electronic structure of Si and Ge nanocrystals. The surface and core properties are investigated. A large unit cell of 8 atoms is used in the present analysis. Cohesive energy, energy gap, conduction and valence band widths are obtained from the electronic structure calculations. The results are compared with available experimental data and theoretical results of other investigators. The calculated lattice constant is found to be slightly larger than the corresponding experimental value because we use only 8 atoms and we compared the results with that of the bulk crystals, nanoclusters are expected to have str
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