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.
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Newcastle disease virus (NDV) is a wide-spectrum anti-tumor agent. The oncolytic selectivity of NDV, a family of Paramyxoviridae, depends on the differential type of inducing different death pathways. This work was conducted to further understand the oncolytic effect of LaSota strain. A mouse breast cancer model (Murine mammary adenocarcinoma cell line AMN3) was used in this study. Methyl Thiazolyl Tetrazolium (MTT) viability assay tested different NDV multiplicity of infection (MOI) values on mouse mammary adenocarcinoma cells incubated for 72 hours post-infection. The IC50 values and anti-tumor activity of LaSota strain against AMN3 cell line were determined. Following Hematoxylin and Eosin Stain, we examined t
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Gelatin-grafted N- proflavine acryl amide was synthesized through two steps; firstly the Gelatin was grafted with acrylic acid free radically using Ammonium per-sulfate at 60℃, Then it was modified to its corresponding acyl chloride derivation, second step included the substitution with amino group of proflavine, in this research Gelatin was used as a natural nontoxic, water soluble polymer as a drug carrier. The prepared pro drug polymer was characterized by FTIR and 1H-NMR spectroscopies, Controlled drug release was studied in different pH values at 37℃. Many advantages were obtained comparing with other known methods.
Given an exterior algebra over a finite dimension vector space v, and let , where is a graded ideal in . The relation between the algebra and regarding to -quadratic and - quadratic will be investigated. We show that the algebra is - quadratic if and only if is - quadratic. Furthermore, it has been shown that the algebra is - quadratic if and only if is - quadratic.
In this work, the effect of variation of semi-angle of the conical part on the vibration characteristics of cylindrical-conical coupled structure is investigated. The shell is made of polyester resin reinforced by continuous E-glass fibers. The case is analyzed experimentally and numerically for orthotropic shell structures. The experimental program is conducted by exciting the fabricated structure by an impact hammer and monitoring the response using an attached accelerometer for different semi-angles of the conical part.
Software named SIGVIEW is used to perform the signal processing on the acquired signal in order to measure the natural frequencies and the corresponding mode shapes. The numerical investigation is achieved using ANS
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This work is an experimental investigation for single basin-single slope solar still coupled with an evacuated tube solar collector. The work is carried out under the climatic conditions of Baghdad city (33.2456º North and East latitude, 44.3337º longitude) through certain days of the months of the year 2019 to study the impact of using evacuated tube solar collector on the daily productivity and efficiency under the outdoors climatic conditions. It was found that using the evacuated tube solar collector increase daily productivity from 2.175 kg/ to 2.95 kg/ for 9 hours (35.63 %) for clear days, also an enhancement about 10.97 % in daily efficiency.
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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
Three hundred and twelve (312) local fungal isolates were isolated from sixty four (64) different contaminated soil samples with oil wastes at different periods, using potato dextrose agar (PDA).the fungal isolates were tested for its ability to degrade naphthalene .Primary and secondary screening were done using solid (MSM) and liquid (MSM) with 100ppm naphthalene and pH 7 respectively. Results from Primary screening showed that 25 isolates gave good growth, 47 gave moderate growth, 66 gave weak growth and 174 were never growing. According to above results 25 fungal isolates were tested for its ability to degredade naphthalene using liquid mineral media (MSM) pH7,100ppm naphthalene and incubated at 30 0C 120rpm for 7 days. Reduction of
... Show MoreBackgroundCarcinoma 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