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.
Two locally isolated microalgae (Chlorella vulgaris Bejerinck and Nitzschia palea (Kützing) W. Smith) were used in the current study to test their ability to production biodiesel through stimulated in different nitrogen concentration treatments (0, 2, 4, 8 gl ), and effect of nitrogen concentration on the quantity of primary product (carbohydrate, protein ), also the quantity and quality of lipid. The results revealed that starvation of nitrogen led to high lipid yielding, in C. vulgaris and N. palea the lipid content increased from 6.6% to 40% and 40% to 60% of dry weight (DW) respectively.Also in C. vulgaris, the highest carbohydrate was 23% of DW from zero nitrate medium and the highest protein was 50% of DW in the treatment 8gl. Whil
... Show MoreTwelve albino mice was divided randomly into four groups comprising A through D injected with ceftazidime at sub MIC, Escherichia.. coli 11, Escherichia.. coli 11 with ceftazidime solution, and standard strain, respectively. Histopathological sections did not show any changes in respect to group A. however, group C suffered signs of infection less than those appeared in group B sections. Simultaneously, group D suffered intense histpathological changes more than other groups infected with resistant isolate.
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 MoreUropathogenic E. coli (UPEC) is problematic and still the leading cause of urinary tract infections worldwide. It is developed resistance against most antibiotics. The investigation, surveillance system, and efficient strategy will facilitate selecting an appropriate treatment that could control the bacterial distribution. The present study aims to investigate the epidemiology and associated risk factors of uropathogenic E. coli and to study their antibiotic resistance patterns. 1585 midstream urine specimens were collected from symptomatic urinary tract infections (UTI) patients (225 males and 1360 females) admitted to Zakho emergency hospital, Zakho, Kurdistan Region, Iraq from January 2016 until the end of December 2
... Show MoreBiofilm formation represents one of the biggest problems facing scientists because of this phenomenon linkage with virulence of bacteria and other clinical environmental problems. In the present study, two clinical isolates,
Escherichia coli, and Staphylococcus aureus were exposed to the non thermal plasma for different intervals of time (1, 2, 4, 8, and 16 min). The biofilm was measured post exposing. It was found that 2 min. exposing to non-thermal plasma reduced the biofilm formation by both clinical isolates significantly. It can be concluded that the ability of S. aureus to form biofilm higher than E. coli and exposing for 2 min to non-thermal plasma sufficient to reduce the biofilm formati
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
Twenty-seven S. aureus isolates were obtained from patients referring various hospitals in Baghdad. Only 17 isolates produced DNase. SNase was extracted and purified from Saphylococcus aureus 3 isolate since it produces the largest zone of clearance on DNase agar. Nevertheless, only those phenotypically-producer of DNase harboured nuc gene. Present study revealed that the crude enzyme had a specific activity of 50.66 unit/mg; while it reached 241 unit/mg after ion exchange chromatography using carboxymethyl cellulose column. SDS-PAGE showed a single sharp band with an approximately 16.8 kDa molecular weight. A matter indicates that the enzyme is consistently pure. Results proved that SNase was able to significantly (P< 0.05) reduce th
... Show MoreReactive arthritis (ReA) has been as joint developing after infection, it belongs to spongylo arthritis (SpA). The etiology of this disease was multi factorial, the combination between genetic and environmental factors for triggering this disease. This study included 75 Iraqi Arab patients and 39 healthy control. Urine samples and blood were collected from each subject. The results showed that Escherichia coli bacteria (E. coli) was isolated from 32% of urine samples. HLA-B*27 allele frequencies was higher in ReA patients infected with E. coli. This lead to suggest that E. coli may be trigger factor in ReA patients with UTI which had HLA-B*27 positive.
Objectives. This study aimed to evaluate the impact of nonnutritive sucking habits on the presence of oral Escherichia coli. Methods. One hundred and twenty children aged 3–5 years old were enrolled in the present case-control study, as follows: 60 children with continuous pacifier and thumb sucking habits (study group) and 60 children without any sucking habits (control group). The children in the two groups were matched in terms of age and gender. Information was gathered from the parents concerning their children using a special sheet. Sterile swabs were taken from both groups and cultured on agar plates. Then, they were subjected to further biochemical tests to identify E. coli species. The mean of the E. coli count was determ
... Show MoreA total of (25) stool samples were collected from children and adults (2- 4) years old suffering from diarrhea to isolate E. coli strains that produce heat-stable enterotoxin a (STa), and after performing microscopic examination, cultural characterization and biochemical identification only (11) isolates showed positive E. coli. STa activity was estimated by using suckling mouse assay (SMA) and from these (11) isolates only (5) showed STa activity and the one with the highest STa activity was selected for large scale production of STa, which was followed by partial purification using ion-exchange chromatography (normal phase) using DEAE sephadex A-50 column. After purification and determination of protein concentration by using the standard
... Show More