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.
The synthesis of ligands with N2S2 donor sets that include imine, an amide, thioether, thiolate moieties and their metal complexes were achieved. The new Schiff-base ligands; N-(2-((2,4-diphenyl-3-azabicyclo[3.3.1]nonan-9-ylidene)amino)ethyl)-2-((2-mercaptoethyl)thio)-acetamide (H2L1) and N-(2-((2,4-di-p-tolyl-3-azabicyclo[3.3.1]nonan-9-ylidene)amino)ethyl)-2-((2-mercaptoethyl)thio) acetamide (H2L2) were obtained from the reaction of amine precursors with 1,4-dithian-2-one in the presence of triethylamine as a base in the CHCl3 medium. Complexes of the general formula K2[M(Ln)Cl2], (where: M = Mn (II), Co(II) and Ni(II)) and [M(Ln)], (where: M = Cu(II), Zn(II) and Cd(II); n =1-2, expect [Cu(HL2)Cl]) were isolated. The entity of ligands and
... Show MoreIn this work, a series of new Nucleoside analogues (D-galactopyranose linked to oxepanebenzimidazole moiety) was synthesized via multisteps synthesis. The first step involved preparation of two benzimidazoles 2-styrylbenzimidazole and 2-(phenyl ethynyl) benzimidazole via reaction of phenylenediamine with cinnamic acid or ?-phenyl propiolic acid. Electrophilic addition of the prepared benzimidazoles by three anhydrides in the second step afforded (4-6) and (14-16) which in turn were treated with 1,2,3,4-di-O-isopropylidene galactopyranose in the third step to afford a series of the desirable protected nucleoside analogues (7-9) ,(17-19)which after hydrolysis in methanolic sodium methoxidein the fourth step afforded the free nucleoside analog
... Show MoreLeucine aminopepotidase (LAP)[EC:3.4.11.1] activity has been assayed in (50) serum samples of patients with diabeties naphrophathy D.N (non-insulin dependent diabetic (NIDD) , and (50)serum sample of healthy individuals without any clinically detectable diseases have been as control group. The aim of this study is to measure leucine aminopeptidase activity and partially purifying the enzyme from sera of patients with diabetes nephropathy The results of this study revealed that Leucine aminopeptidase (LAP) activity of nephropathy patient’s serum shows a high signifiacant increase (p < 0.001) compared to that of the healthy subjects.LAP was purified from the serum of patients with diabetes nephropathy by dialysis and gel filtration (Se
... Show MoreFive derivatives of thiadiazole were prepared with aldehydes and alkyl halides, compoundA: 2-amino-5-thiol-1,3,4- thiadiazole, compound B :2-(o-hydroxybenzylidine)amino-5-thiol-1,3,4-thiadiazole, compoundC: 2(2-butan-lidine)amino-5-thiol-1,3,4-thiadiazole, compound E: 2- amino-5-(2-Propanylthio)-1,3,4-thiadiazol) and compound F:2(o-chlorobenzylamino)-5-(2-propanyl thio)-1,3,4 thiadiazol. All prepared compounds were diagnosed by (IR) and (UV) Spectroscopy. All of those compounds were screened for their anti-microbial activity in vitro. The results show that most of the compounds A, B, C exhibited moderate to good activity against Gram-positive bacteria and the same compound exhibit low to moderate activity on most gram-negative bacte
... Show MoreThe synthesis and characterisation of new macrocyclic binuclear metal(II) complexes derived from dithiocarbamate (DTC) ligand are reported. The reaction of a bis-secondary amine, CS2 and KOH resulted in the formation of the free ligand. Two approaches were implemented to synthesis the macrocyclic bis(dithiocarbamate) complexes; (i) from the reaction of the free ligand with a metal ion, and (ii) via a one-pot reaction. In the free ligand approach, complexes were obtained by the reaction of dithiocarbamate salt with the metal ions; CoII, ZnII and CdII. However, the one-pot reaction is based on the mixing of the bissecondary amine, CS2, KOH and metal(II) chloride. Physico-chemical analyses were implemented to characterise the ligand and its
... Show MoreNon-thermal (low-temperature) plasma may act as an alternative approach to control superficial wound and skin infections when the effectiveness of chemical agents is weak due to natural pathogen or biofilm resistance. In this paper an atmospheric pressure plasma needle jet device which generates a cold plasma jet is used to measure the effectiveness of plasma treatment against different pathogenic bacteria and to test the individual susceptibility of pathogenic bacteria to non-thermal argon plasma. It is found that, Gram-negative bacteria were more susceptible to plasma treatment than Gram-positive bacteria. For the Gram-negative bacteria Pseudomonas aeruginosa, there were no survivors among the initial 1x108C.F.U (Co
... Show MoreThe study aims to investigate the antimicrobial activity of propolis obtained from different regions of Iraq compared with that of propolis obtained from Iran. Samples were investigated for their antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Eschericha coli, Klebsiella pneumoniae, Bacillus cereus , Staphylococcus epidermidis and Candida albicans using standard antimicrobial assays. Marked variations in the antimicrobial activity of the different propolis samples were observed, the method of extraction selected gives the highest antimicrobial activity and the best alcohol concentration using in the extraction of propolis , then the crude extract of propolis showed synergistic effect with some antibiotics in
... Show MoreIntroduction and Aim: Bacteriocins are antimicrobial peptides that have bactericidal and/or bacteriostatic activity against other bacteria. The aim of this study was to assess the antibacterial efficiency of Klebocin a K. pneumoniae bacteriocin, against biofilm formation by clinical isolates of methicillin resistant Staphylococcus aureus MRSA. Materials and Methods: S. aureus isolated from clinical samples was identified according to vitek 2 system Antibiotic susceptibility test was performed according to disc diffusion method. Vitek 2 compact system was also used to detect MRSA strains. Agar well diffusion method was used to evaluate the antibacterial activity of klebocin from K. pneumoniae towards 11 strains of S. aureus by
... Show MoreIn-vitro biological activities of the free new H4L ( indole-7-thiocarbohydrazone) ligand and its Ni(II), Pd(II) , Pt(II), Cu(II), Ag(I), Zn(II) and Cd(II) complexes are screened against two cancerous cell lines, that revealed significant activity only for [Cu2Cl2(H4L)2(PPh3)2] after 72 h treatment by the highest tested concentrations. The Copper(I) complex was characterized by X-ray Crystallography and the NMR spectra, whereas it has been confirmed to have momentous cytotoxicity against ovarian, breast cancerous cell lines (Caov-3, MCF-7). The apoptosis-inducing properties of the Cu(I) complex have been investigated through fluorescence microscopy visualization, DNA fragmentation analysis and propidium iodide flow cytometry.
Background: Cutaneous leishmaniasis (CL) is a neglected disease in tropical countries, including Iraq. Several studies have sought to examine chemotherapies for leishmaniasis treatment but most of them are of toxic and/or undesirable side effect, therefore, the need for investigating new fewer toxic therapies is essential. Aim of study: In this study, the cytotoxic effect of Artemisinin (ART), a novel herbal compound, was screened against the two forms, promastigotes and amastigotes, of the Iraqi isolate of Leishmania tropica, the causative agent of Baghdad boil. Material and methods: Different concentrations (1000, 500, 250, 125, 62.5, 31.25, 15.6 and 7.8) µM of Artemisinin were screened to investigate the leishmanicidal activity of th
... Show More