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 focuses on diagnosis of Candida species causing Vulvovaginal Candidiasis using phenotype and genotype analyzing methods, and frequencies of candida species also using Vulvovaginal Candidiasis patients. 130 samples (100 from patients and 30 from non infected women) were collected and cultured on biological media. Identifying the yeasts, initially some phenotypic experiments were carried out such as germ tube, from motion of pseudohyphae and clamydospores in CMA+TW80 medium, API20 candida and CHROMagar Candida. Genomic DNA of all species were extracted and analyzed with PCR and subsequent Polymerase Chain Reaction - Restriction Fragments Length Polymorphism (PCR-RFLP) methods. Frequency of C. albicans, C. krusei, C. tropicalis , C.
... Show MoreYeasts are distributed in all environments and have been reported as potential biocontrol agents against various phytopathogenic fungi. To investigate their enzymatic and biological activities, 32 yeasts were isolated from 15 date vinegar samples. Evaluation of the antagonistic activities of isolated yeasts against the plant pathogens Fusarium oxysporium, Sclerotinia sclerotiorum, and Macrophomina phaseolina indicated that there are two yeasts had the highest inhibitory effect against plant pathogens, these yeasts identified as Kluyveromyces marxianus and Torulaspora delbrueckii using traditional and molecular methods. These yeast isolates were tested for fungal cell wall degrading enzymes (in vitro), and results indicated that the
... Show MoreP. aeruginosa is one of the complex targets for antimicrobial chemotherapy. Also, it is intrinsically resistant to several antibiotics. It produces β-lactamases enzymes that are responsible for the widespread β-lactam antimicrobial resistance. There are three major groups of β-lactamase enzymes, MBLs and ESBLs forming Pseudomonas is a major issue for the treatment of burns victims. Methods: A total of 28 clinical isolates related to P. aeruginosa have been obtained from the burns specimens from patients attending to AL-Imam hospital/Baghdad-Iraq, through the period from October 2015 to March 2016. Also, all isolates have been recognized as P. aeruginosa via utilizing bacteriological assay and confirmed by Vitek 2. In addition, the suscep
... Show MoreA qualitative chemical test was performed on functional groups extracted from fenugreek plant and its extracts (aqueous, alcoholic and volatile oil). Results revealed that fenugreek seeds contain the main functional groups, while extracts are varied accorodihg to their content of functional groups qualitatively and quantitively. Moreover, inhibition activity was tested for extracts of fenugreek seeds (aqueous, alcoholic and volatile oil). against gram negative (Salmonella typhimurium, Escherichia coli and Pseudomonas aeruginosa) and gram positive (Staphylococcus aureus) by the ager well diffusion method. Data have revealed that inhibition activity was different in accoradance with extract solvent and the tested microorgan. Oil extract (15)%
... Show MoreFrom different hospitals in Baghdad city, 25 clinical isolates of Proteus spp. were collected from different clinical samples, all isolates were identified as Proteus mirabilis by using bacteriological and biochemical assays in addition to Vitek-2 identification system. 15 (60%) isolates were identifying as Proteus mirabilis. The susceptibility of P. mirabilis isolates towards cefotaxime and ceftazidime was (66.6 %), (20%) consecutively; while extended spectrum β-lactamases producing P. mirabilis percentage was (30.7 %). Because blaVEB-1 was documented as an important indicator for increasing risk of extended spectrum beta ßlactamases producing P. mirabilis isolates that began to spread from many geographic area to Far east which inc
... Show MoreA new Azo‐Schiff base ligand L was prepared by reaction of m‐hydroxy benzoic acid with (Schiff base B) of 3‐[2‐(1H–indol‐3‐yl)‐ethylimino]‐1.5‐dimethyl‐2‐phenyl‐2,3‐dihydro‐1H‐pyrazol‐4‐ylamine. This synthesized ligand was used for complexation with different metal ions like Ni(II), Co(II), Pd(II) and Pt(IV) by using a molar ratio of ligand: metal as 1:1. Resulted compounds were characterized by NMR (1H and 13C), UV–vis spectroscopy, TGA, FT‐IR, MS, elemental analysis, magnetic moment and molar conductivity studies. The activation thermodynamic parameters, such as ΔE*, ΔH*, ΔS*, ΔG*and
... Show MoreEighteen new cyclic imides (maleimides) conncted to benzothiazole moiety through sulfonamide group were synthesized via multistep synthesis.The first step involved preparation of two maleamic acids N-phenylmaleamic acid and N-benzylmaleamic acid via reaction of maleic anhydride with aniline or benzyl amine.Dehydration of the prepared amic acids by treatment with acetic anhydride and anhydrous sodium acetate in the second step afforded N-phenylmaleimide and N- benzyl maleimide which in turn were treated with chlorosulfonic acid in the third step to afford 4-(N-maleimidyl) phenyl sulfonyl chloride and 4-(N-maleimidyl) benzyl sulfonyl chloride respectively.In the Fourth step of this work each one of the two prepared maleimidyl sulfonyl chlorid
... Show MoreThis paper deals with the preparation of new monomers and polymers which including heterocyclic unit. The diacid chlorides compounds [1-3] were prepared from the reaction of glutaric acid, adipic acid, terephthalic acid with thionyl chloride. Succinic acid reacted with ethanol to produce compound [4]. Compound [4] reacted with hydrazine hydrate to obtain succinic hydrazide [5].Compound [5] reaction with CS2 and KOH in absolute ethanol to produce compound [6].The polymers [7-12] have been created by reacting diacid chlorides compounds [1-3] with compound[5] or [6] in dry pyridine with some drops of DMF. The topology of produced compounds has characterized through their spectral and analytical data as in FT-IR spectra, Thermal analysis [DSC,
... Show MoreIn this paper we define and study new concepts of fibrewise topological spaces over B namely, fibrewise Lindelöf and locally Lindelöf topological spaces, which are generalizations of will-known concepts: Lindelöf topological space (1) "A topological space X is called a Lindelöf space if for every open cover of X has a countable subcover" and locally Lindelöf topological space (1) "A topological space X is called a locally Lindelöf space if for every point x in X, there exist a nbd U of x such that the closure of U in X is Lindelöf space". Either the new concepts are: "A fibrewise topological space X over B is called a fibrewise Lindelöf if the projection function p : X→B is Lindelöf" and "The fibrewise topological space X over B
... Show MoreLeucine amino peptidases (LAP; EC 3.4.11.1) constitute a diverse set of exopeptidases that catalyze the hydrolysis of leucine residues from the amino-terminal of protein or peptide substrates, (LAP) are present in animals, plants, and microbes. In this study, leucine amino peptidase was purified partial from Arachis hypogaea seeds by using gel filtration chromatography Sephadex G-100. The enzyme was purified 3.965 fold with a recovery of 29.4%. Its pH and temperature optimum were(8.7) and (37oC), respectively. The results show novel properties of LAP from Arachis hypogaea L. or peanut. The Km value for LAP (77 mM), with V max (1538 m mole min-1). We recommend a separate isoenzymeof the enzyme (LAP) from Arachis hypogaea on L. peanut seeds a
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