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.
This study focused on extracting the outer membrane nanovesicles (OMVs) from Escherichia coli BE2 (EC- OMVs) by ultracentrifugation, and the yield was 2.3mg/ml. This was followed by purification with gel filtration chromatography using Sephadex G-150, which was 2mg/ml. The morphology and size of purified EC-OMVs were confirmed by transmission electron microscopy (TEM) at 40-200 nm. The nature of functional groups in the vesicle vesicle was determined by Fourier transforms infrared spectroscopy (FT-IR) analysis. The antitumor activity of EC-OMVs was conducted in vitro by MTT assay in human ovarian (OV33) cancer cell line at 24,48 and 96hrs. The cytotoxicity test showed high susceptibility to the vesicles in ovarian compared to normal
... Show MoreIn this research, new series of Furo-2-quinolone [FQ] compounds have been synthesized. These novel [FQ] compounds were prepared from coumarin derivatives (Furocoumarins: psoralen and isopsoralen).Identifications of these FQ compounds were performed by using infrared spectrum (I.R), Ultraviolet spectrum (U.V) and Nuclear Magnetic Resonance spectrum (H1-NMR) besides some physical data. The cytotoxic screening involves ;using HEP-2 cell line which gave differential responses against tested compounds : 4,6 Dimethyl furo[2, 3-g] coumarin (C1), 1-(2`, 4`, Dimethoxy benzylideneimino)-2,6-dimethyl Furo [2, 3-g] quinoline-2-one (C3) and the angular psoralen of the same derivative
... Show MoreEighty one bacterial isolates were obtained from 53 soil samples of different plants rhizosphere. All the isolated bacterial were screened for antifungal effect against Fusarium oxysporum . Three isolates gave antifungal activity with inhibition zone ranged between (0.5-2.5 cm). Two isolates (Bd1 and Bd2) were Brevundimonas diminuta, while the third (Pf1) was Pseudomonas fluorescence . B. diminuta (Bd1) which used in this study isolated from Raphanus sativus gave the highest inhibition zone against F. oxysporum. Cell free supernatant of B.diminuta(Bd1) was better in antifungal activity than bacterial cells against F. oxysporum. The highest antifungal substance production was obtained from mineral salt broth containing 1% peptone after in
... Show MoreThe molecular structures of acetophenonylidine-4-aminopyridine (I), 2, 6-dihydroxyacetophenonlidine-4-aminopyridine (II), 2, 4, 6- trihydroxyaceto phenonylidine-4-aminopyridine (III) and 2, 6-dihydroxyacetophenonylidine-2-aminopyridine (IV) have been investigated by IR and UV-visible spectrophotometry. The IR data indicate that the hydroxyl groups of these Schiff bases exist as tautomeric mixtures of free and bonded with the azomethine groups. The electronic spectra, effect of polar and nonpolar solvents, and the effect of acidity and basicity on the electronic spectra were studied and discussed. Their charge-transfer (CT) complexes with chloranil in chloroform solvent were also investigated; these complexes absorb light at 398-533 nm. T
... Show MoreThis study was designed to investigate the hepatoprotective activity and antioxidant enzymes of purified Bauhinia variegate leaves extract and purified flowers extract were administered (200 mg/kg, orally once daily) to reduce the effect of carbon tetrachloride-damage in rat’sliver for three weeks. Thereafter, the levels of some serum biochemical factorssuch as alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase,and the activity of three different antioxidant enzymes (glutathione, superoxide dismutase, and catalase) were investigated.Liver homogenate can used to estimated antioxidant parameters: glutathione, superoxide dismutase and catalase. The purified Bauhinia variegate leaves and purified flowers significantly
... Show MoreOne hundred fifty bacterial strains were isolated from patients with urinary tract infections (UTIs). They were belong to ten different species of gram-negative bacteria and to two genera of gram–positive bacteria. E. coli was the major causative agent and comprise 40% of all cases. Klebsiella pneumoniae and Proteus mirabilis were second and third with 18.67% & 18.0% respectively. Other gram-negative bacteria were belong to the genera Enterobacter, Acinitobacter, Pseudomonas, Citrobacter and Serratia. Ten cases (6.67%) were caused by genus Staphylococcus and seven (4.66%) were caused by Streptococcus. Out of the 150 positive cases, 96(64%) were from female patients, while 54(36%) were from males. High percentage of all
... Show MoreOrthodontic treatment has two major problems: being lengthy and costly procedure. The present study was designed to evaluate the clinical efficacy of locally injected vitamin D3 Keywords: Orthodontic, Calcitriol, Local injection, OTM (calcitriol) in accelerating orthodontic teeth movement (OTM) and reducing treatment time and cost in humans. The study was performed on 15 Iraqi adult orthodontic patients within the age range 17-28 years, they are randomly allocated into three groups, each of five patients and treated with either 15 pg, 25 pg, or 40 pg/0.2ml calcitriol diluted with 10% dimethylsulfoxide (DMSO). The maxillary arch of every patient was divided into control (right) and experimental (left) sides. In addition to force application,
... Show MoreDevelopment of improved methods for the synthesis of metal oxide nanoparticles are of high priority for the advancement of material science and technology. Herein, the biosynthesis of ZnO using hydrahelix of beta vulgaris and the seed of abrus precatorius as an aqueaus extracts adduced respectivily as stablizer and reductant reagent. The support are characterized by spectroscopic methods ( Ft-IR, Uv-vis ).The FTIR confirmed the presence of ZnO band. The Uv-visible showed absorption peak at corresponds to the ZnO nanostructures. X-ray diffraction, scaning electron microscopy (SEM), dispersive X-ray spectroscopy (EDX) techniques are taken to investigation the size, structure and composition of synthesised ZnO nanocrystals. The XRD pattern mat
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