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 urban Gentrification is an inclusive global phenomenon to restructure the cities on the overall levels, the research to propose a specific study about the concept of urban Gentrification in the cities and showcasing its, specifications, and results, and how to deal with the variables that occur on cities through improvements as part of urban renewal projects, then the general axis of the research is shrinked, choosing the urban centers as the most important areas that deal with the urban Gentrification process due to its direct connection with indivisuals and social changes, and to process the specific axis of the research theses and studies will be showcased that discuss the topic in different research directions, and emerged
... Show MoreMalaria is a curative disease, with therapeutics available for patients, such as drugs that can prevent future malaria infections in countries vulnerable to malaria. Though, there is no effective malaria vaccine until now, although it is an interesting research area in medicine. Local descriptors of blood smear image are exploited in this paper to solve parasitized malaria infection detection problem. Swarm intelligence is used to separate the red blood cells from the background of the blood slide image in adaptive manner. After that, the effective corner points are detected and localized using Harris corner detection method. Two types of local descriptors are generated from the local regions of the effective corners which are Gabor based f
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In multivariate survival analysis, estimating the multivariate distribution functions and then measuring the association between survival times are of great interest. Copula functions, such as Archimedean Copulas, are commonly used to estimate the unknown bivariate distributions based on known marginal functions. In this paper the feasibility of using the idea of local dependence to identify the most efficient copula model, which is used to construct a bivariate Weibull distribution for bivariate Survival times, among some Archimedean copulas is explored. Furthermore, to evaluate the efficiency of the proposed procedure, a simulation study is implemented. It is shown that this approach is useful for practical situations and applicable fo
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<span>One of the main difficulties facing the certified documents documentary archiving system is checking the stamps system, but, that stamps may be contains complex background and surrounded by unwanted data. Therefore, the main objective of this paper is to isolate background and to remove noise that may be surrounded stamp. Our proposed method comprises of four phases, firstly, we apply k-means algorithm for clustering stamp image into a number of clusters and merged them using ISODATA algorithm. Secondly, we compute mean and standard deviation for each remaining cluster to isolate background cluster from stamp cluster. Thirdly, a region growing algorithm is applied to segment the image and then choosing the connected regi
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The aim of this study is to investigate the kinetics of copper removal from aqueous solutions using an electromembrane extraction (EME) system. To achieve this, a unique electrochemical cell design was adopted comprising two glass chambers, a supported liquid membrane (SLM), a graphite anode, and a stainless-steel cathode. The SLM consisted of a polypropylene flat membrane infused with 1-octanol as a solvent and bis(2-ethylhexyl) phosphate (DEHP) as a carrier. The impact of various factors on the kinetics constant rate was outlined, including the applied voltage, initial pH of the donor phase solution, and initial copper concentration. The results demonstrated a significant influence of the applied voltage on enhancing the rate of c
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Leishmania species are the causative agent of a tropical disease known as leishmaniasis. Previous studies on the old world species Leishmania major, showed that the amastigotes form which resides inside the macrophage of the vertebrate host, utilize host’s sphingolipids for survival and proliferation. In this study, gene expression of serine palmitoyltransferase (SPT) subunit two (MmLCB2) of the mouse macrophage cell line (RAW264.7), which is the first enzyme in the de novo sphingolipid biosynthesis, was detected in both infected and non-infected macrophages. This was detected under condition where available sphingolipid was reduced, with the new world species Leishmania mexicana. Results of qPCR analysis showed that there was no differen
... Show MoreBackground: Dysfunction of sinoatrial node is a set of abnormal rhythms which are resulted from the sinoatrial node malfunctionof the sinus node, the chief natural cardiac pacemaker. The common, and occasionally, the single method for treatment of heart arrhythmias wasimplantation of pacemaker, which reduce symptoms exactly occurs after implantation. Aim: To detect the association between red cell width diameter (RDW) and some cardiac electrophysiology parameters in sinus node dysfunction in Iraqi patients such as SNRT and AH. Methods: A cross sectional study, was conducted on 59 patient ranging between 20-50 years old and involving 35 female and 24 male patients, suffering from an unexplained symptoms of sinoatrial node dysfunction (SND).
... Show Moreحظيت السمية الخلوية للمستخلصات النباتية باهتمام كبير في السنوات الأخيرة لتقييم نشاطها ضد الأورام. تركز الوقاية الكيميائية من السرطان على السمية الخلوية للمستخلصات النباتية لتكون سامة تجاه خطوط الخلايا السرطانية دون أن تؤثر على الخلايا الطبيعية. الهدف من هذه الدراسة هو تقييم السمية الخلوية لمستخلصي الزنجبيل و أوراق الزيتون ومزيجهما بنسبة 1: 1 تجاه خطوط خلايا RD وتقدير فعالية هذه المسخلصات ضد الاجهاد التا
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Abstract: A novel design of Mach Zehnder Interferometer (MZI) in terms of using special type of optical fiber that has double clad with graded distribution of the refractive index that can be easily implemented practically was suggested and simulated in this work. The suggested design is compact, rapid, and is simple to be modified and tested. The simulated design contains a MZI of 1546.74 nm of central wavelength that is constructed using special type of double clad optical fiber that has two different numerical apertures. The first aperture will supply single mode propagation via its core, while the second numerical aperture supports a zigzag wave propagation (multimode) in the first clad region. The interferometer’s
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