Prevalence of hepatitis C virus infection and human immunodeficiency virus in a cohort of Egyptian hemophiliac children

The risk of blood-borne infections, especially hepatitis C virus [HCV] and human immunodeficiency virus [HIV] infection still remains in developing countries among children receiving blood products as hemophiliacs, but the risk is not known in Egypt. The objective of this study was to detect the prevalence of HCV and HIV infection among hemophiliac children to know the magnitude of the problem and determine potential risk factors. This was a cross-sectional study conducted on 100 hemophiliac children that assessed the liver clinically and by laboratory tests. All children were screened for HCV and HIV antibodies by enzyme-linked immunosorbent assay. Those with positive HCV antibody titre were tested by polymerase chain reaction [HCV-PCR]. Forty were positive for HCV antibodies with 19 children [47.5%] HCV-PCR positive as well. The mean age, average frequency of bleeds/year, dose of replacement therapy/year and alanine aminotransferase [ALT] levels were significantly high in HCV-antibody and PCR positive patients as compared to HCV antibody and PCR negative ones. None of our patients had clinical evidence of hepatic involvement or was co-infected with HIV HIV infection does not appear to be a current health problem in Egyptian hemophiliac children though the prevalence of HCV infection is still high

Genetic polymorphisms of DNA repair gene XRCC1 and susceptibility to acute leukemia

Defective DNA repair has been reported to be a risk factor for various malignancies. Polymorphisms of DNA repair genes could alter protein structure and may impair DNA repair capacity. Genetic polymorphisms of XRCC1 gene could lead to defective base excision repair [BER] pathway resulting in impaired DNA repair capacity and increased risk of acute leukemia. To determine the possible effect of XRCC1 gene polymorphisms 194Arg to Trp and 399Arg to Gin on the risk of development of acute leukemia in a group of Egyptian patients. The study was also extended to evaluate the association between these polymorphisms and disease outcome. Polymorphisms of XRCC1 codon 194 [Arg to Trp] and codon 399 [Arg to Gin] were genotyped in 35 patients with acute lymphoblastic leukemia [ALL], 35 patients with acute myeloid leukemia [AML] and 70 healthy controls using polymerase chain reaction-restriction fragment length polymorphism [PCR-RFLP] method. Individuals with heterozygous XRCC1 194 Arg/Trp variant demonstrated a significant increased risk of AML than controls [Odds Ratio [OR] 3.5, 95% confidence interval [CI], 1.3-9.5]. The frequency of homozygous XRCC1 399 Gin/Gin variant was statistically higher in ALL patients than controls [OR 3.69, 95% CI, 1.19-11.4]. Stratification for sex with regard to codon 194Trp carriers showed that males had 3.2-fold increased risk of ALL than females with borderline significance. In case of codon 399Gln polymorphism, a highly significant risk of ALL among females was observed with 7.5-fold increased risk. The frequency of XRCC1 haplotype A [399Gln carriers and 194Trp carriers] was significantly higher in both ALL and AML patients than controls [OR5.2, 95% CI, 1.6-16.7, p-value <0.01 for ALL] [OR 3.2, 95% CI, 0.9-11.1, p-value=0.055 for AML]. The polymorphic variant of XRCC1 194Trp has a significant unfavorable effect on disease outcome among ALL and AML patients [p-value 0.002 and 0.05 respectively]. Acute lymophoblastic leukemia patients carrying the 399Gln allele experienced a significant unfavorable outcome than ALL patients carrying the wild-type allele [p-value<0.01]. An increased risk of AML among carriers of XRCC1 194Trp and an increased risk of ALL among patients with XRCC1 399Gln variant genotypes were observed. Combined presence of XRCC1 194Trp and 399Gln variants [haplotype A] had significantly higher risk of both ALL and AML. The polymorphic variants of XRCC1 codons 194 and 399 had significant unfavorable effect on disease outcome of both AML and ALL

Molecular screening for G6PD gene mutations in children with glucose-6-phosphate dehydrogenase deficiency

Glucose-6-phosphate dehydrogenase [G6PD] deficiency is a heterogeneous enzyme abnormality with a high frequency among people of African, Mediterranean and Southeast Asian origins. In almost every group studied in the Middle East, only three to four different G6PD mutations were detected. Among these, the G6PD Mediterranean mutation [563C -> T] was by far the most common. Apart from this mutation, little is known about the genetic heterogeneity of G6PD deficiency in Egypt. To screen for G6PD gene mutations in a group of Egyptian children with G6PD-deficiency who were previously screened for the Mediterranean [563C -> T] mutation. This work was conducted on twenty-one unrelated Egyptian children [17 males and 4 females] presenting with G6PD deficiency previously screened for the G6PD Mediterranean mutation [563C -> T]. Carefully-preserved DNA of patients refrigerated at -20°C and DNA of 21 age-matched normal subjects extracted from blood leukocytes by saltingout technique were screened for mutations in the G6PD gene by PCR-single strand conformation polymorphism [SSCP] analysis followed by DNA sequencing. In addition to the G6PD Mediterranean mutation 563C -> T previously identified by PCR-RFLP analysis in 6/21 male patients [28.6%], a further of 2 different mutations; G6PD A- mutation and G6PD Chatham were observed in 2/21 [9.5%] and 1/21 [4.8%] patients respectively. Twelve patients [57.1%] remained uncharacterized at the genetic level, a normal African G6PD A genotype was detected in one patient of them. Patients with G6PD Mediterranean mutation were more susceptible to hemolysis than were patients with G6PD A- and G6PD Chatham mutations. The lower prevalence of G6PD Mediterranean mutation in our patients and the finding of three different mutations in a relatively small number of G6PD-deficient subjects reflect the considerable genetic heterogeneity of G6PD deficiency of the Egyptian population

Monocytes expressing tissue factor as a diagnostic marker for neonatal sepsis

In neonatal sepsis, several clinical and laboratory parameters have been proposed for its diagnosis, however, with variable sensitivity and specificity. The bacterial products in sepsis including endotoxin induce the production of proinflammatory cytokines that evoke the expression of tissue factor [TF] on monocytes and endothelial cells. To estimate the percentage of monocytes expressing tissue factor [TF%] by flowcytometry in patients with neonatal sepsis and to delineate its significance to diagnose neonatal sepsis. Twenty-seven neonates with neonatal sepsis and positive blood culture were recruited and evaluated clinically for their risk factors. Laboratory investigations including complete blood picture, C-reactive protein [CRP] and estimation of the monocytes TF expression by flowcytometry were done. Twenty-four normal newborns were included as a control for the laboratory data. The monocytes expressing TF% of the studied patients was significantly higher than that of the controls, p-value = 0.0001. The level of TF% was significantly influenced positively by premature rupture of membrane [PROM], Multiplicity, WBC count, staff/segment ratio, CRP and negatively by gestalional age, body weight, and platelet count. The sensitivity and overall accuracy of the TF% were higher than those of the staff/segment ratio and the WBC count for diagnosing neonatal sepsis. The areas under the receiver operating characteristic curve [AUC] of TF%, staff/segment ratio and WBC count were 0.84, 0.79 and 0.60 respectively, 95% confidence interval]. The monocytes expressing TF% is a promising diagnostic and prognostic marker of infection in neonatal sepsis with high sensitivity and overall accuracy. Adding the estimation of monocytes expressing TF% to the sepsis screen may improve the diagnosis of neonatal sepsis

PCR and immunocytochemistry detection of p27[kip1] in chronic lymphocytic leukemia

Chronic B-cell lymphocytic leukemia [B-CLL] is a clonal expansion of B-cells with low proliferative activity in which the cells are arrested in G[0]/G[1] phase of cell cycle. p27[KIP1] is one of the KIP/CIP family of cyclin-dependent kinase inhibitors [CKIs] which inhibit all cyclin dependent kinases by direct binding to cdk complexes. It is highly expressed when cells are arrested in G[0]/G[1]and its expression declines as cells progress towards S phase. In B-CLL the non-physiological increase in p27[KIP1] appears to be of clinical relevance since high protein levels correlate with poorer survival of patients. To study the expression of p27[KIP1] in B-CLL patients and correlate these results with the clinical and laboratory data of patients. p27[KIP1] expression was determined at the mRNA level by semi-quantitative reverse transcriptase polymerase chain reaction [RT-PCR] and at the protein level by immunocytochemistry in 35 patients with de novo B-CLL and 30 healthy age- and sex-matched control subjects. p27[KIP1] mRNA levels by RT-PCR was significantly higher among CLL patients compared to the control subjects [p<0.001] and was significantly higher among group II CLL patients [lymphocyte count >30 x 10[3]/L] compared to group I CLL patients [lymphocyte count