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 study involved isolation and characterization of E.coli from patient’s infected with diarrhea , in order to study the ability of the bacteria to produce cytosine deaminase (CD). Result showed eight isolates of E.coli which showed adifference in the production of (CD) and the isolate of E. coli E33 was the beast of its production of CD than the other’s and the value of the specific activity was 4.920 u/mg protein , when grown in the medium which contains 1% glycerol ,3% peptone as a source of Carbon and Nitrogen respectively with pH 8. The optimum cultural condition‘s for the production of CD from E. coli E33 was studied the result‘s showed that the isolate gave the
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Seventy-six urine specimens were collected from of patients suffering from recurrent
urinary tract infections (UTIs). Specimens were bacteriologically analyzed, fifty
(65.8%) of isolated bacterial strains were belonged to E.coli. 100% of isolated
uropathogenic E.coli (UPEC)strains displayed a biofilm positive phenotype under
optimized condition using microtiter plate assay. 21 of E.coli strains classified as highly
positive biofilm producers (42%), and 29 (58%) as weakly positive biofilm producers.
In this study, only four isolates produce CNF1 from 76 isolatesof uropathogenic Escherichia coli.cnf1 gene was detected by using PCR technique, while cytotoxic necrotizing factor 1(CNF1) was determined by Immunoblotting assay.
A significant increase in the incidence of non-O157 verotoxigenic Escherichia coli (VTEC) infections have become a serious health issues, and this situation is worsening due to the dissemination of plasmid mediated multidrug-resistant microorganisms worldwide. This study aims to investigate the presence of plasmid-mediated verotoxin gene in non-O157 E. coli. Standard microbiological techniques identified a total of 137 E. coli isolates. The plasmid was detected by Perfectprep Plasmid Mini preparation kit. These isolates were subjected to disk diffusion assay, and plasmid curing with ethidium bromide treatment. The plasmid containing isolates were subjected to a polymerase chain reaction (PCR) for investigating
... Show MoreThree hospitals were chosen for the present (Maternity hospital, Raperin hospital and Rhizgari hospital) survey within Erbil city, 36 water samples were collected at regular monthly interval periods beginning at January to December 2012. Microbial analysis was done by selective medium and biochemical tests and the isolated bacteria from those hospitals were Eshcerichia coli, Acinetobacter lowffii, Klebsiella pneumoniae, Moraxilla spp., Salmonella Typhi, Citrbtobacter freundii, Vibrio fluvials, Acinetobacter haemolyticus, Weeksella zoohelcum, Pasteurella multicida, and Pseudomonas aeroginosa. E. coli isolates were subjected to antimicrobial susceptibility testing. In vitro activities of 10 different antibiotics against E. coli isolates we
... Show MoreThe severity of UTI produced by E. coli is due to the expression of a wide
spectrum of virulence factors. In this study the role of E. coli virulence determinants
in the pathogenesis of UTI in urinary catheterized and non-catheterized patients has
been evaluated. The isolates were recovered from 129 patients admitted to the
hospital. Virulence genes of E. coli were detected by polymerase chain reaction
analysis for the prevalence of these virulence factors. The targeted genetic
determinants were those coding for Type 1 fimbriae, Pyelonephritis-Associated Pili
(PAP), Antigen 43 (Ag43), α-Hemolysin and Aerobactin siderophores among the
studied isolates. The prevalence of genes fimH, papC, ang43, hlyA and iutA were<
This work aimed to investigate the effect of Diode laser 805 nm on plasmid DNA and RNA
contents of some Gram negative bacteria represented by Escherichia coli and Proteus mirabilis isolates
.Plasmid extraction was done using two methods (Salting out and CTAB method).Different powers and
pulse repetition rates for 805 nm Diode Laser were used to study this effect. Results revealed that the
plasmid profile of the two species were highly affected using (2, 3) W at different frequencies including
5and 10 kHz as compared with 1 kHz while plasmids were gradually disappeared at 1W, 10 kHz. In the
same time the shining of RNA was also decreased gradually then disappeared with increasing powers
especially at 2W and 10 kHz cau
This study was established to discover and determine multidrug-resistant Escherichia coli and Klebsiella pneumoniae from women suffering urinary tract infections, specifically in Mosul city. A total of 62 E. coli and 32 K. pneumoniae bacterial isolates were considered for this study. All isolates were characterized using standard bacterial culture methods, including culture on MacConkey agar, Eosin Methylene Blue agar and biochemical tests. Also antibiotic sensitivity test using standard disc method for different antibiotics and also special discs to detect ESBL activity were carried out, in addition to PCR as molecular identification tool. The results showed that most isolated E. coli
... Show MoreBackground: Urinary tract infections (UTIs) and their complications such as Bladder cancer (Bl. C.) are a health growing problem worldwide. Objective: To shed light on this subject, present study was done to investigate relationship between recurrent urinary tract infection (RUTI) due to Escherichia coli (E. coli) and Bl. C.Type of study: Cross-sectional study. Methods: This study included 130 patients with RUTI, 50 patients with Bl. C. and 50 control of both sexes (aged 7-85 years) attending Al-Zahra Teaching Hospital in Al-Kut/Wassit governorate and Al-Harery Teaching Hospital of specialized surgeries/Baghdad. The patients were divided into two groups: the first group (n=130) included those who were suffering from recurrent UTI without
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