This research was aimed to the purification and characterization of cytosine deaminase as a medically important enzyme from locally isolated Escherichia coli; then studying its cytotoxic anticancer effects against colon cancer cell line. Cytosine deaminase was subjected to three purification steps including precipitation with 90% ammonium sulfate saturation, ion exchange chromatography on DEAE-cellulose column, and gel filtration chromatography throughout Sephadex G-200 column. Specific activity of the purified enzyme was increased up to 9 U/mg with 12.85 folds of purification and 30.85% enzyme recovery. Characterization study of purified enzyme revealed that the molecular weight of cytosine deaminase produced by E. coli was about 48 KDa, the highest enzyme activity at pH 8.5, and is most stable at pH 7.5 - 9, the enzyme also showed a full activity at a range of temperatures between 45-60 0C. Enzyme activity was strongly inhibited in the presence of mercuric chloride and copper sulphate, when added individually at a constant concentration. However, calcium chloride, manganese chloride and ferric chloride caused a little increase in enzyme activity while sodium azide had no effect on enzyme activity. Upon cytotoxic effect study through micro-cultured tetrazolium assay (MTT) against Caco-2 cell line. Purified cytosine deaminase was found to inhibit the growth of Caco-2 cancer cell line with an IC50 of 242.5 ?g/ml in a comparison to an IC50 of 1864 ?g/ml for crude enzyme. Besides, the enzyme didn’t show significant effect on WRL normal cell line.
In this study, out of 50 isolates of some nosocomial infections from some Baghdad hospitals, only 13 (26%) were identified as Escherichia coli. Depending on selective media, morphological and biochemical tests the species was then confirmed by molecular methods. Later on antimicrobial resistance test was performed by the Kirby-Bauer method. The molecular characterization of blaTEM and blaCTX-M genes in different clinical isolates of E. coli was done through polymerase chain reaction (PCR) by utilizing special primers. These genes were positive to only 4 (30.7%) isolates. The sequence of nucleotides of positive genes was carried out for four isolates. The results showed that there was no vari
... Show MoreThiazoles are heterocyclic nitrogenous compounds, which were given a great attention due to their antibacterial effect, antihypertensive, stimulating glucose absorption (like antibiotics), acts as cytoprotective agent. The present work is conducted to evaluate the degree of antibacterial activity on the growth activity of Escherichia Coli (E.Coli) and Pseudomonas aeruginosa (P. aeruginosa) by incubation with 2-methyl-4-(3,4 dihydroxy phenyl)-Thiazole compound (MDHPT). Optical density (O.B) was measured by turbidmetric method, any increase in O.D would represent the increase in the bacterial growth .The results showed that there was a 75.7 % inhibition when using 190.4 ïg/ml (MDHPT) solution and 64.3% was obtained when using concentr
... Show MoreThe research aim was to observe the distribution pattern of
This study includes isolation, purification, and identification of algae from different aquatic environments in Baghdad city. Nine unialgal cultures were obtained. These algal cultures included 6 species of blue-green algae (Microcystis aeruginosa, Microcystis flos-aquae, Oscillatoria limnetica, Nostoc carneum, Westillopesis prolifica, Mastigocluds lamiosus), and 3 species of green algae (Mougeotia sclaris, Scenedesmus dimorphus and Chlorella vulgaris). In addition that aerial parts from Convolvulus arvensis were collected. Terpens, alkaloids and phenols were extracted of mentioned plant, and the antialgal activity of extracts types were evaluated in 3 concentrations (5, 10, and 20 mg/ml) by wells and diffused in the agar media. Results
... Show MoreBackground: One of the most dominant bacterial infections is urinary tract infection (UTI), both in the community and in the hospital settings Since drug resistance become one of the predominant problems of health worldwide, it is necessary to use new methods to overcome drug-resistant bacteria. In this regard, medicinal plants (Medicago sativa ) are considered one of the richest sources to produce antibiotics.
Aim of the study: evaluate the antibacterial effect of the fraction of ethyl acetate of Medicago Sativa extract against Escherichia Coli isolated from urine samples of patients with urinary tract infections.
Material and methods:
... Show MoreThe guava plant, Psidium guajava L., serves as proof of the abundant donations of nature, providing a delicious guava fruit; this plant is rich in groups of medicinal and nutritional benefits. Guava belonging to the Myrtaceae family, many previous studies reported many phytochemical constituents in its leaves that have many pharmacological activities and medicinal properties; this study focuses on the isolation, structural elucidation and calculation concentration of flavonoids, assessment of the cytotoxic activityof hyperin from Psidium guajava leaves newly cultivated in Iraq. The isolation process involved the use of thin-layer chromatography (TLC) and preparative high-performance liquid chromatography (PHPLC) and structural eluci
... Show MoreThe synthesis of zeolite NaX from locally available kaolin has been studied. The operating conditions for zeolite NaX production from kaolin with good crystallinity were as follows; a gel formation step of metakaolin in alkaline medium in presence of additional silica to crystallize the zeolite was achieved at 60 oC for 1 hr,and with stirring. In ageing step of the reactants at room temperature for 5 days and crystallization step at 87±2 oC for 24 hr. The catalytic activity of catalyst prepared from local kaolin was studied by using cumene cracking as a model for catalytic cracking and compared with standard HY zeolite and HX zeolite catalysts. The activity test was carried out in a laboratory continuous flow unit with fixed bed reactor
... Show MoreInterferon’s plays a role in innate immune responses through upregulation of costimulatory molecules and induction of proinflammatory cytokines. Interferon alpha (IFN α) type of Interferons. The present study characterized IFNα cDNA . The interferon’s play a great role in protection from infections, caused by microorganisms, and have powerful antiproliferative and immunomodulation activity. In this study DNA was isolated from bovine blood leukocyte, which was used in the quality of matrix for amplification of α-interferon gene with the use of PCR, and isolation of gene α-interferon and transformation in vector pUC18 and expression vector pET24b (+). All plasmids contained an additional DNA fragment size corresponding to the gene
... Show MoreThe aldol condensation of 2-acetylnaphthalene with 9-anthracene carboxaldehyde afforded α, β-unsaturated keton (1) . New heterocyclic compounds containing: cyclohexenone[2], indazole[3], pyrimidinethion [4], thiazolo fused pyrimidine[5], isoxazoline[6], substituted pyrazoline[7]a-d and pyrimidinone[8] rings system were synthesized from α, β-unsaturated keton[1]. Cyclization of [1] with ethylacetoacetate gave the mentioned heterocycle cyclohexanone [2]. The cyclo condensation of [2] with hydrazine gave the new indazole derivative [3]. furthermore, the reation of [1]with thiourea gives thiopyrmidine derivative [4]. The cyclo condensation of [4] with chloroacetic acid gave the fused rings [5]. Then reacted compound[1] with hydroxy
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