Mona M. Fathy

Molecular copro-prevalence of Cryptosporidium in Egyptian children and evaluation of three diagnostic methods

ObjectiveTo determine molecular prevalence of Cryptosporidium in a cohort of Egyptian children and compare three diagnostic tests.MethodsStool samples from children with diarrhea (n=150) and from apparently healthy children (n=100) were examined for Cryptosporidium using microscopy, enzyme linked immuosorbant assay (ELISA) and nested polymerase chain reaction (nPCR).ResultsnPCR detected Cryptosporidium in 22.4% of children. Acid-fast stain and ELISA showed false negativity but 100% specificity with nPCR as gold standard.ConclusionCryptosporidium is a common cause of diarrhea in children in Egypt.

Significance of plasma osteopontin in diagnosis of hepatitis C virus-related hepatocellular carcinoma.

BACKGROUND AND STUDY AIMS Alfa fetoprotein (AFP) is widely used as a surveillance test for hepatocellular carcinoma (HCC) among patients with liver cirrhosis (LC). However, the clinical use of AFP has been shown to present some important limitations in sensitivity and specificity. Osteopontin (OPN) is a secreted matrix glycoprotein that is emerging as a significant protein in the biology of HCC. The aim of this study was to assess the diagnostic value of plasma OPN compared with that of AFP in the diagnosis of HCC among hepatitis C virus (HCV)-related LC. PATIENTS AND METHODS Plasma levels of OPN and AFP were measured in 69 Egyptian patients with HCV-related LC (35 with HCC and 34 without HCC) and 20 healthy controls. RESULTS Both median AFP and OPN levels were significantly higher in the HCC group compared to LC and healthy control groups (p<0.001 in each) and in LC compared to the control group (p<0.001). In the HCC group, both OPN and AFP levels were significantly higher in patients with Child-Pugh class C and B compared to class A (p<0.05 in each). There was no correlation between OPN and AFP levels. The OPN level was significantly higher in patients with multiple focal lesions than in those with single lesions (p<0.05) and in patients with portal vein invasion compared to patients without portal vein invasion (p<0.05). Receiver operator characteristic (ROC) curves showed that the area under the curve (AUC) for OPN and AFP was 0.824 and 0.730, respectively. CONCLUSION OPN is a promising tumour marker which could be used as a screening test for the diagnosis of HCC in patients with LC and, hence, improves the prognosis and survival rate of these patients. The association of OPN with the multiplicity of focal lesions and portal vein invasion suggests an additional prognostic value.

Impact of CYP3A5 and MDR-1 gene polymorphisms on the dose and level of tacrolimus among living-donor liver transplanted patients: single center experience

Abstract Aim of work: To assess the impact of Cytochrome P450 3A5 (CYP3A5) and multidrug resistance-1 gene (MDR-1) single nucleotide polymorphisms on the dose and blood level of tacrolimus among liver transplanted patients. Patients and methods: We enrolled a prospective study of 41 liver transplanted patients. Dose-adjusted trough blood concentration (C/D ratio) was calculated. Polymerase chain reaction-restriction fragment length polymorphism followed by sequencing was done for genotyping of CYP3A5*3 (6986A > G). Results: At 1 week, 1 and 3 months C/D ratio were significantly lower in CYP3A5 expressers *1/*1 patients compared to non-expressers *3/*3. Conclusion: CYP3A5 (6986A > G) genotype, rather than MDR-1 (2677G > A/T) variant, has an impact on tacrolimus pharmacokinetics.

Assessment of interleukin 28B genotype as a predictor of response to combined therapy with pegylated interferon plus ribavirin in HCV infected Egyptian patients

BACKGROUND AND AIM Single nucleotide polymorphisms (SNPs) of interleukin 28B (IL28B) gene is associated with spontaneous clearance and variable response to combined therapy with pegylated interferon (PEG-IFN) and ribavirin (RBV) in chronic hepatitis C virus (HCV) infected patients. This study aimed at assessing the value of IL28B rs8099917 gene polymorphism in predicting sustained virological response (SVR) among HCV infected Egyptian patients treated with PEG-IFN and RBV. METHODS Our study was conducted on 153 chronic HCV infected patients treated with PEG-IFN and RBV. Genotyping of rs8099917 near the IL-28B gene was performed by Real Time PCR using Taq-Man probe assay. RESULTS The overall SVR was achieved in 49.6% of patients. Patients with TT genotype showed significantly higher SVR rate than minor allele (TG/GG) carriers (74% vs. 26%, P=0.004). Logistic regression analysis revealed that TT carriers had 2.8 higher chance for SVR achievement than G allele carriers TG/GG (OR=2.8, 95% CI=1.4-5.6, P=0.004). Younger age, male sex and low activity grading were significant predictors of SVR (P=0.003, P=<0.001 and P<0.001 respectively). High pretreatment AST levels and advanced liver fibrosis were negative predictors of SVR (P=0.04 and P<0.001 respectively). CONCLUSION IL28B genotype is a significant pre-treatment predictor of response to PEG-IFN/RBV in HCV infected Egyptian patients.

COPRO -MOLECULAR CHARACTERIZATION OF CRYPTOSPORIDIUM SPP. AND GENOTYPES AMONG EGYPTIAN CHILDREN

Stool samples from 182 diarrhoeic (symptomatic) children and 100 apparently healthy (asymptomatic) children, matched for age, from Aboul-Reesh Cairo University Pediatrics Hospital were examined by ELISA and by nPCR (targeting COWP gene) for the detection of Cryptosporidium. The demographic and environmental data of the diarrhoeic group was recorded. The PCR amplified product of positive cases was then subjected to RFLP by digesting it with the restriction enzyme RsaI. The obtained fragments were resolved by electrophoresis and the bands were visualized and characterized versus a standard. ELISA results demonstrated a prevalence rate of 13.2% (24/182) among diarrhoeic group, and 8% (8/100) among non-diarrheic group, with overall detection rate of 11.3% (32/282). Higher rates of detection were obtained by nested PCR assay among diarrhoeic group 25.8% (47/182) and 16% (16/100) among non-diarrhoeic group with overall detection rate of 22.3% (63/282). Considering nPCR as the reference method, ELISA had a sensitivity of 47.6% and a specificity of 99.1%. RsaI digestion of nPCR product of COWP gene revealed the presence of 2 genotypes: genotype 1 with 4 bands (34, 106, 125 and 285) and genotype 2 in which 3 bands (34, 106 and 401). Among the 63 cases with cryptosporidiosis, 53 (88.3%) had genotype 1, and 7 (11.7%) had genotype 2. The higher prevalence of genotype 1 suggests a relatively greater risk of human source of infection than zoonosis.

Molecular characterization of exons 6, 8 and 9 of ABCB4 gene in children with Progressive Familial Intrahepatic Cholestasis type 3.

Abstract Aim of work: To estimate the frequency of mutations involving exons 6, 8 and 9 of Adenosine triphosphate-binding cassette, subfamily B, member 4 (ABCB4) gene among children with progressive intrahepatic cholestasis with high γ-GT activity (PFIC3). Subjects and methods: Cross sectional study was conducted on 30 children with PFIC3. Genotyping was performed by sequencing analysis of exons 6, 8 and exon 9 of ABCB4 gene. Results: Heterozygous synonymous polymorphic variant was detected in exon 6 (rs 1202283) and in exon 8 (rs 2109505). No mutations in studied exons were detected. Conclusion: Exons 6, 8 and 9 mutations of ABCB4 gene are not common among Egyptian children with PFIC3.

RETRACTED ARTICLE: Combined blood and pleural levels of mesothelin and osteopontin for the diagnosis of malignant pleural mesothelioma

This article has been retracted by the Editor-in-Chief of Comparative Clinical Pathology due to similarities to a previously published work; Wafaa M. Ashour, Hassan Amin, Irin M. Sabri, Aisha M. Samir, Dalia A.H. Zaki, Mona M. Fathy: Combined blood and pleural levels of mesothelin and osteopontin for the diagnosis of malignant pleural mesothelioma. Egyptian Journal of Chest Diseases and Tuberculosis, Volume 61, Issue 3, July 2012, Pages 121–128. http://dx.doi.org/10.1016/j.ejcdt.2012.10.016; ht tp: / /www.sciencedirect .com/science/ar t ic le /pi i / S0422763812000179.

The slow activity-associated genotype of microsomal epoxide hydroxylase-1 (m-EPHX-1) and low serum paraoxonase-1 (PON1) activity are associated with increased susceptibility to chronic obstructive pulmonary disease (COPD) in smokers

Oxidative stress is believed to play an important role in the pathogenesis of smoking-induced chronic obstructive pulmonary disease (COPD). We hypothesized that low serum activity level of paraoxonase-1 (PON1) that play a protective role in the lungs by metabolizing lipid peroxides and genetic polymorphism of antioxidant enzymes that detoxify cigarette smoke products such as microsomal epoxide hydroxylase-1 (m-EPHX-1) would be associated with increased susceptibility to COPD in smokers. The study was conducted on patients admitted to Chest Diseases Department, Kasr Al-Aini Hospital, Cairo University. Sixty subjects were divided into three groups—25 COPD patients, 25 chronic smokers without COPD, and 10 healthy nonsmokers. Pulmonary function tests were done for confirmation of COPD. Serum PON1 activity assay using spectrophotometric kinetic method and PCR-RFLP for genetic polymorphism of m-EPHX-1 exon 3 113 T>C were done to all participants. The significantly low median value for serum PON1 activity was found in COPD and smoker groups compared to the nonsmoker group (P = 0.015). Homozygote mutant genotype (HH) of m-EPHX-1 was present only in the COPD group compared to the other two groups with a frequency of 20%, 0%, and 0% respectively [P = 0.013; odds ratio (OR), 4.26; P = 0.008]. The OR between COPD and nonsmoker group was 14.222 (P = 0.008). The frequency of TH genotype was highest in the COPD group (44%, P = 0.013). While the wild genotype (TT) was more frequent in the nonsmoker group compared to the smokers and COPD groups (80%, 64%, and 36% respectively; P = 0.013). The slow m-EPHX-1 exon 3 polymorphisms (HH and HT) and the reduced serum PON1 activity are associated with higher susceptibility to COPD in smokers.