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.

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.

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.

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

OBJECTIVE: To determine molecular prevalence of Cryptosporidium in a cohort of Egyptian children and compare three diagnostic tests. METHODS: Stool 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). RESULTS: nPCR detected Cryptosporidium in 22.4% of children. Acid-fast stain and ELISA showed false negativity but 100% specificity with nPCR as gold standard. CONCLUSION: Cryptosporidium is a common cause of diarrhea in children in Egypt.

Assessment of the role of paraoxonase gene polymorphism (Q192R) and paraoxonase activity in the susceptibility to atherosclerosis among lead-exposed workers.

BACKGROUND AND OBJECTIVE: Lead exposure is a well known cause of cardiovascular damage, including atherosclerosis. Paraoxonase 1 (PON1), a high-density lipoprotein-associated antioxidant enzyme, is capable of hydrolyzing oxidized lipids and thus it protects against atherosclerosis. The mechanism by which heavy metals inhibit serum PON1 activity is still not clear. Our aim was to detect the association between lead exposure and serum PON1 activity and lipid profile and also to study the polymorphism of the PON1 gene. DESIGN AND SETTING: A case-control, cross-sectional study conducted from June 2008 until May 2009. SUBJECTS AND METHODS: Male workers (n=100) in a lead battery manufactory were recruited for this study. They were compared with 100 male age-matched workers not exposed to lead (control group). Serum lipid profile, paraoxonase activity and lead were measured in blood samples. The DNA was extracted for detecting the Q192R polymorphism of the PON1 gene by polymerase chain reaction followed by restriction fragment length polymorphism. RESULTS: There was significant difference in triglycerides, total cholesterol and high-density lipoprotein cholesterol (HDL-C) (P=.01, .05 and .04, respectively) between cases and controls. Multiple linear regression analysis showed that blood lead levels were significantly associated with decreased serum paraoxonase activity (P=.03) in lead workers. The paraoxonase genotype QR was the most prevalent in 34/53 subjects (64%) among the lead-exposed groups, while the genotype QQ was more prevalent in the control group, in 15/25 subjects (60%), with a significant difference between the control and other groups (P<.05). CONCLUSION: Lead exposure is associated with increased triglycerides, total cholesterol and low-density lipoprotein cholesterol and decreased HDL-C. Because of the protective role of PON1 in the development of atherosclerosis, a decrease in serum PON1 activity due to lead exposure may render individuals more susceptible to atherosclerosis.