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.
Posible interference of vamin with the activity of several antibiotics against E. coli was evaluated in vitro. In MBS- glucose medium, significant growth delay was induced by 8 ug/ml of terramycin (oxytetracycline- polymyxin B) and bactrim (trimethoprim-sulphamethoxazole), and by 16 ug/ml of refocin, lincomycin, and chloramphenicol. Rapid growth inhibition was induced by 32 ug/ml of all an- tibiotic tested separately. Significant inactivation of up to 64 ug/ml of licomycin and bactrim was in- duced by the addition of vamin at a concentration of 1:20 v/v of the medium. This effect was found to be due to the presence of specific amino acids in vamin. Among them is valine, leucine, isoleucine tyrosine, tryptophan, phenylalanine, cysteine, meth
... Show MorePolycyclicacetal was prepared from the reaction of PEG with aldehyde derived from Erythro-ascorbic acid (pentulosono-ɣ-lactone-2,3-enedianisoate).All these compounds were characterized by Thin Layer Chromatography (TLC) and FTIR spectra and aldehyde was also characterized by (U.V-Vis), 1HNMR, 13CNMR, and mass spectra.The inhibitory effect of prepared polymer on the activity of human serum AcetylCholinesterase has been studied in vitro. The polymer showed a remarkable activity at low concentration (4.7x10-3 – 4.7x10-8M).
200 samples collected Adrar of patients with urinary tract infection were investigating the types of bacteria most local isolates showed high resistance to antibiotics penicillin c ??????? Amoxicillin Beracelin
This study was aimed to produce bacteriocin from Bacillus. licheniformis isolated from local soil of corn and sunflower fields and using as antimicrobial agent . Fourteen of local isolates of Bacillus sp. were obtained and ability of these isolates for growth on Brain heart infusion agar (BHI) at 550C were tested. Isolate C4 was revealed high growth density in comparison with other isolates. Isolate C4 was identified as Bacillus licheniformis according to morphological, cultural and biochemical tests, Moreover genetic analysis for 16S rRNA gene and given accession number MT192715.1 in GenBank of NCBI . Production of bacteriocin from this isolate was carried out in Luria Broth (LB) and partially purified by precipitation with 30-70 % saturat
... Show MoreIn present study 74 specimens of urine were collected from patients suffering from urinary tract infections.Fifty (67.56%) isolates were identified as Escherichia coli. 78% of isolates were identified as extendedspectrum beta lactamases (ESBL) producer. Antibiotic susceptibility t est was done and ceftazidime wasselected to complete this study by implying stress at sub-MIC on isolate harbor high number of resistancegenes (N11) and compared with sensitive isolate (S). Only four β-lactamase coding genes were detected;blaTEM, blaPER, blaVIM and blaCTX-M-2 and N11 had blaTEM, blaPER, and blaVIM. It was found that the resistantisolate did not form biofilm when compared with the sensitive one, which formed moderate biofilm. Inaddition, ceftazidi
... Show MoreBackground: Common and persistent isolate ina the teeth following failed therapy of the root canal is the gram-positive facultative bacterium Enterococcus faecalis and Escherichia coli, which develop biofilm through a complicated process that results in the formation of a biofilm. Enterococcus faecalis and Escherichia coli are significant factors that cause chronic periradicular lesions after root canal therapy. Aim: This study aimed to treat the root canal tooth infected with Escherichia coli and Enterococcus faecalis Methods: In this study biofilm formation was done for Escherichia coli in growth phase cultured in a brain heart broth Enterococcus faecalis and Escherichia coli cultured in Luria-Bertani (LB) infusion medium for 18 hrs. Then
... Show MoreMulti-drug-resistant uropathogenic Escherichia coli (UPEC) is considered a significant challenge due to its ability to resist antibiotics and form biofilms. UPEC biofilm formers are well protected and largely inaccessible to antibiotics, which leads to persistent infections and evasion of the host immune system. Understanding how ciprofloxacin and trimethoprim/sulfamethoxazole affect biofilm formation is essential for improving treatment strategies for urinary tract infections (UTIs). A total of 76 UPEC isolates were obtained from Iraqi patients and identified using morphological and biochemical characteristics, as well as the Vitek®-2 Compact system. Minimum inhibitory concentrations (MICs) were determined using the Vitek®-2 system, whic
... Show MoreOne hundred twelve urine samples were collected from Baghdad hospitals and examined by different identification techniques. Seventy isolates (62.5%) were diagnosed as Escherichia coli after microscopic and cultural identifications. The result of PCR product electrophoresis on the isolates showed that thirteen isolates (18.57%) have Pap E gene which are uropathogenic E. coli. Antibiotic susceptibility test was done, and four high resistant strains were mixed with aqueous extract of Quercus infectoria plant in 96 well ELISA plate and incubated for different times. After 0, 6, and 12 hr. of incubation, the effect of the plant extract on the bacterial growth was determined by ELISA reader, and the effect on the expression of P
... Show MoreHuman Adenosine deaminase is an essential enzyme for modulating the bioactivity of thyroid hormones, and It is important for the maturation and differentiation of lymphocytes, although its clinical importance in thyroid diseases have yet to be identified. Objective: The aim of the current study is to determine the Adenosine deaminase concentration in healthy controls, and in autoimmune thyroid diseases such as Graves' Disease, and Hashimoto's Thyroiditis. Patients and methods: A total of 183 serum specimens of 103 female patients with autoimmune thyroid diseases and 80 healthy control groups were included in this study and collected from the Baghdad Medical City, Iraq. Quantitative Human Adenosine Deaminase ELISA kits were used to estimate
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