Background and aim:
DNA repair represents a protective mechanism against cell injury and cancer. 8-hydroxy-deoxyguanosine (8-OHdG) is the main ROS-induced DNA mutation. The current study aimed to evaluate urinary 8-OHdG levels in patients with chronic hepatitis C virus (HCV) and its related hepatocellular (HCC) and correlate its level to XRCC1 rs25487 G/A and OGG1 rs1052133 C/G gene polymorphisms.
Materials and methods:
Urinary 8-OHdG assays were performed using HPLC technique, and XRCC1 rs25487 G/A and OGG1 rs1052133 C/G gene polymorphisms were analyzed by PCR using confronting two-pair primer method (PCR-CTPP) in 200 subjects allocated into 50 chronic HCV patients, 50 HCV-related HCC patients, and 100 controls.
Results:
There were significantly increased urinary 8-OHdG levels in HCV-related HCC and chronic HCV patients when compared with the controls (
P
<0.05 for all). Urinary 8-OHdG was associated with the tumor spread. Regarding, XRCC1 (Arg399Gln), AA (Gln/Gln) genotype and A-allele were more frequent in HCC and chronic HCV patients than in the controls (
P
<0.05). ORs (95%CI) using the dominant and the recessive genetic models were; 2.1 (1.1–4.1),
P
=0.032 and 1.9 (1–3.6),
P
=0.043 respectively. For OGG1 (Ser326Cys), GG (Cys/Cys) genotype and G-allele were increased significantly in chronic HCV and HCC patients compared to the controls (
P
<0.05). ORs (95%CI) under the dominant and the recessive genetic models were; 2.1 (1.1–4.1),
P
=0.032 and 1.9 (1–3.8),
P
=0.049 respectively. Additionally, XRCC1 (AA) and OGG1 (GG) genotypes had significantly increased urinary 8-OHdG levels among patients (
P
<0.05).
Conclusions:
XRCC1 (AA) and OGG1 (GG) could be considered as possible genotypic risk factors for HCV- related HCC development which were associated with significantly high urinary 8-hydroxy-deoxyguanosine levels, thus urinary 8-OHdG could be considered as non-invasive marker in follow-up chronic HCV progression into HCC.
Background: We aimed to investigate ITGA4 gene expression pattern and to explore its methylation heterogeneity in chronic lymphocytic leukemia (CLL). Patients & methods: Eighty one CLL patients and 75 healthy subjects were enrolled and prognostic evaluation of patients was assessed. ITGA4 q-realtime PCR was performed using Applied Biosystems, TaqMan gene expression assay. ITGA4 gene-specific CpG methylation was investigated in real time using pyrosequencing technology. Results: ITGA4 was differentially expressed in CLL patients. The CpG sites-1, 2 and 3 showed significantly higher mean levels than healthy controls (p = <0.001, 0.007 and 0.009). Significant association between CpG site-1 and CLL has been detected using age-adjusted logistic regression (p < 0.001). Conclusion: Hypermethylation at ITGA4 gene CpG sites (1,2,3) is a characteristic feature in CLL.
AIM:To evaluate the diagnostic performance of MDI and temocillin disk (30 μg) for detection of carbapenem-resistant Enterobacteriaceae in comparison to real-time PCR.MATERIAL AND METHODS:Fifty specimens submitted to the Microbiology Laboratory of Ain Shams University Hospitals and showed resistance to carbapenem drugs through routine culture and susceptibility testing, were assessed by both temocillin disk (30 μg) and MDI set to detect carbapenem-resistant Enterobacteriaceae. Results were compared to real-time PCR for detection of carbapenemase genes blaKPC, blaNDM, blaOXA–48-like, blaVIM, and blaIMP.RESULTS:Our work revealed that most of the CPE isolates were Klebsiella species (62%) followed by E. coli (24%), Serratia (10%) and Citrobacter (4%). Phenotypic detection of carbapenem-resistant classes revealed OXA - 48 in 96% of isolates, followed by MBLs (82%), and KPC (34%). All isolates were negative for AmpC. Detection of the genes by real-time PCR showed that the predominance was for the blaOXA-48 gene (96%) then blaVIM (94%) followed by blaNDM (54%), blaKPC (46%) and finally blaIMP (40%). Evaluation of the MDI set against PCR showed sensitivity (82.1%) and specificity (70%). The temocillin disk had 97.9% sensitivity and 50% specificity. The evaluation of Temocillin disk and MDI in combination for detection of carbapenem-resistant Enterobacteriaceae showed 99.7% sensitivity and 35% specificity.CONCLUSION:Adding Temocillin disk to Mastdisks ID inhibitor combination set provides a simple, easy, rapid and highly sensitive test that can be used for screening and classification of carbapenem-resistant Enterobacteriaceae. However, it still needs confirmation by molecular techniques.
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