Sahar Sharaf

Hepatitis C Virus (HCV) Vertical Transmission in 12-Month-Old Infants Born to HCV-Infected Women and Assessment of Maternal Risk Factors

We summarize the results of HCV RNA testing of 12 month old infants born to HCV infected mothers in Cairo, Egypt. We used real-time PCR testing and demonstrated a transmission rate of 14.3%.

Inborn errors of metabolism detectable by tandem mass spectrometry in Egypt: The first newborn screening pilot study.

Objectives To estimate the burden of metabolic disorders detectable by tandem mass spectrometry in Egypt, through a pilot expanded newborn screening programme at Cairo University Childrens Hospital in 2008, and examining the results of 3,900 clinically at-risk children, investigated at Cairo University Children’s Hospital for the same disorders over the past 7 years using the same technology. Methods Dried blood spots of 25,276 healthy newborns from three governorates in Upper, Middle, and Lower Egypt were screened, to give a representative sample of the Egyptian newborn population. Based on the pilot study outcomes and the results of clinically suspected children, we estimated the total birth prevalence of tandem mass spectrometry detectable metabolic disorders, and the relative frequency of several individual disorders. Results Among the healthy newborns, 13 metabolic disorder cases (five phenylketonuria [1:5,000], two methylmalonic acidemia, and isovaleric acidemia [1:12,500], one each of maple syrup urine disease, propionic acidemia, β-ketothiolase deficiency, and primary carnitine deficiency [1:25,000]) were confirmed, giving a total birth prevalence of 1:1944 live births. Among the clinically suspected children, 235 cases were diagnosed, representing a much wider disease spectrum. Conclusions Egypt has one of the highest reported birth prevalence rates for metabolic disorders detectable by tandem mass spectrometry. Early diagnosis and management are crucial for the survival and well-being of affected children. A nationwide NBS programme by tandem mass spectrometry is recommended.

Vitamin D Status in Egyptian Children and Adolescents with Type 1 Diabetes Mellitus

Introduction: Vitamin D is an important factor for bone health especially in type 1 DM (T1DM). Thus, optimal supply of vitamin D may be of particular importance for bone health in children and adolescents with T1DM. Objective: To assess the vitamin D status in children and adolescents with T1DM and to study related factors that may influence serum vitamin D level. Methods: Sixty (30 prepubertal, 30 pubertal) Egyptian children and adolescents with T1DM were enrolled in this cross-sectional study, the mean age for the prepubertal group was 6.85 ± 1.64 years and for the pubertal group was 14.43 ± 1.524 years. Detailed history and complete physical examination laying stress on the sun exposure, skin color, exercise and detailed dietetic history. Dietary content of calcium (mg/day) and vitamin D (IU/day) were calculated. Laboratory assessment included serum calcium, phosphorus, 25-hydroxy-vitamin D (25OHD) and parathormone (PTH). Results: Most of the study group (91.67 %) was vitamin D deficient. There was no significant correlation between serum vitamin D and serum calcium, phosphorus, parathormone, anthropometric measures, duration of diabetes, mean HbA1c, insulin dose, and sun exposure. Despite the high prevalence of vitamin D deficiency, there was a low prevalence of secondary hyperparathyroidism (11.67%) in the study group. Conclusion: Prevalence of vitamin D deficiency in diabetic children and adolescents is very high but underestimated. Therefore, screening and supplementation of vitamin D should be considered.

Human telomerase reverse transcriptase messenger RNA (TERT mRNA) as a tumour marker for early detection of hepatocellular carcinoma.

BACKGROUND AND STUDY AIMS Hepatocellular carcinoma (HCC) is the fifth most common malignancy in the world. Although tumour markers such as α-foetoprotein (AFP) are widely used and important for HCC detection in clinical scenes, they still do not provide a satisfactory solution to detect HCC at the early stage. The aim of our study was to illustrate the significance of serum human telomerase reverse transcriptase messenger RNA (hTERT mRNA) as a novel biomarker for early detection of HCC. PATIENTS AND METHODS Thirty-five patients with HCC, 15 patients with liver cirrhosis, and 10 healthy subjects were sex and age matched. History taking, full physical examination, and laboratory investigations including liver function tests, hepatitis markers, AFP, and quantification of serum human telomerase reverse transcriptase mRNA (hTERT mRNA) using real-time reverse transcription polymerase chain reaction (RT-PCR) were conducted. Ultrasonography (US), triphasic computed tomography (CT), and liver biopsy were carried out. RESULTS The hTERT was above the cutoff point (>144 copies/ml) in 27 HCC patients with a sensitivity of 77.14% and a specificity of 100% and with a positive predictive value (PPV) of 100% and a negative predictive value (NPV) of 65.2%. AFP was above the cutoff point (>50 ng/ml) in 23 HCC patients with a sensitivity of 65.71% and a specificity of 96% and with a PPV of 96.3% and an NPV of 53.8%. Our study also showed a statistically significant relationship between the size of the tumour in HCC patients and both AFP and hTERT, and hTERT appears to be more correlated with the size of the tumour than AFP. There is no direct correlation between hTERT or AFP and the number of focal lesions with p value>0.05. CONCLUSION Serum hTERT mRNA is more sensitive and specific than AFP in the early detection of HCC and its level correlates with the size of the tumour.

Spontaneous Viral Load Decline and Subsequent Clearance of Chronic Hepatitis C Virus in Postpartum Women Correlates With Favorable Interleukin-28B Gene Allele

Background Postpartum hepatitis C viral (HCV) load decline followed by spontaneous clearance has been previously described. Herein we identify predictors for viral decline in a cohort of HCV-infected postpartum women. Methods Pregnant women at Cairo University were screened for anti-HCV antibodies and HCV RNA, and viremic women were tested for quantitative HCV RNA at 3, 6, 9, and 12 months postpartum. Spontaneous clearance was defined as undetectable viremia twice at least 6-months apart. Associations between viral load and demographic, obstetrical, HCV risk factors, and interleukin-28B gene (IL28B) polymorphism (rs12979860) were assessed. Results Of 2514 women, 97 (3.9%) had anti-HCV antibodies, 54 (2.1%) were viremic and of those, 52 (2.1%) agreed to IL28B testing. From pregnancy until 12 months postpartum, IL28B-CC allele women had a significant viral decline (P = .009). After adjusting, the IL28B-CC allele had a near significant difference compared to the CT allele (odds ratio [OR], 0.75; 95% confidence interval [CI], 0.75,1.00; P = .05), but not the TT allele (OR, 0.91; 95% CI, 0.61,1.38; P = .64). All 14/52 (26.9%) women who subsequently cleared were among the 15 with undetectable viremia at 12 months, making that time point a strong predictor of subsequent clearance (sensitivity = 100%, specificity = 97.4%, positive predictive value = 93.3%, negative predictive value = 100%). Conclusions IL28B-CC genotype and 12-month postpartum undetectable viremia were the best predictors for viral decline and subsequent clearance. These 2 predictors should influence clinical decision making.

Gene mutations in Wilson disease in Egyptian children: Report on two novel mutations

BACKGROUND AND STUDY AIMS Wilson disease (WD) is an autosomal recessive disorder, caused by defects in copper-transporting P-type adenosine triphosphatase (ATPase) encoded by the ATP7B gene, resulting in the deposition of copper in the liver and brain with significant disability or death if left untreated. An available regimen of treatment gives hope to those predisposed to the disease if diagnosed early. The objective of this study was to determine the frequency of the most common European mutation (p.H1069Q) in Egyptian children with WD, in addition to screening for previously reported mutations in the Egyptian patients in our selected group. PATIENTS AND METHODS Direct DNA sequencing was applied to exons (13, 14, 18, and 19) of the ATP7B gene for 19 patients previously diagnosed with WD. Then DNA sequencing and pedigree analysis were performed in the families of the patients showing variations in their results for the purpose of family screening and carrier detection. Six out of 19 patients were studied with their families (three families). RESULTS We identified five variants of which two were novel among the studied patients. One of the novel variants was synonymous substitution (p.A1074A) in 16% of patients and the other was predicted to be missense disease-causing mutations (p.T1076I) in 16% of patients, and three previously published mutations p.H1069Q were detected in 5% of patients, p.P1273Q in 10% of patients, and a silent variant p.A1003A in 26% of patients. CONCLUSION Screening for the two exons 14 and 18 of the ATP7B gene is important in Egyptian patients especially in suspected patients without hepatic manifestations.

Identification of insulin gene variants in neonatal diabetes.

Abstract Objectives: Permanent neonatal diabetes (PNDM) is caused by mutations in the genes responsible for the synthesis of different proteins that are important for the normal behavior of beta cells in the pancreas. Mutations in the insulin gene (INS) are considered as one of the causes of diabetes in neonates. This study aimed to investigate the genetic variations in the INS gene in a group of Egyptian infants diagnosed with PNDM. Methods: We screened exons 2 and 3 with intronic boundaries of the INS gene by direct gene sequencing in 30 PNDM patients and 20 healthy controls. A detailed clinical phenotyping of the patients was carried out to specify the diabetes features in those found to carry an INS variant. Results: We identified five variants (four SNPs and one synonymous variant), c(0).187 + 11T > C, c.-17-6T > A, c.*22A > C, c.*9C > T, and c.36G > A (p.A12A), with allelic frequencies of 96.7%, 80%, 75%, 5%, and 1.7%, respectively. All showed no statistically significance difference compared with the controls, with the exception of c.*22A > C. Conclusion: Genetic screening for the INS gene did not reveal an evident role in the diagnosis of PNDM.

Effect of interleukin-10 gene promoter polymorphisms -1082 G/A and -592 C/A on response to therapy in children and adolescents with chronic hepatitis C virus infection

BACKGROUND AND AIM Studying predictors of response to therapy for hepatitis C virus (HCV) infection in children may help avoid the inappropriate use of currently available costly therapy associated with numerous adverse effects. We tested the hypothesis that inheritance of single nucleotide polymorphisms (SNPs) of the interleukin-10 (IL-10) promoter gene might influence response to HCV treatment. PATIENTS AND METHODS The impact of SNPs, -1082 G/A and -592 C/A, in the promoter region of IL-10 gene, on response to HCV therapy was assessed in a cohort of 40 children treated with a combination of pegylated interferon (Peg-IFN) α2b and ribavirin. RESULTS Sustained virological response was achieved in 48.7%. High viral load was associated with non-response to therapy. There was no association between histopathological degree of inflammation or fibrosis and response to therapy. There was no direct statistically significant association between polymorphisms in the IL-10 gene (-1082G/A and -592 C/A) as regards inflammation or response to therapy in children. As for the SNP -592 C/A; there was a statistically significant association with the score of fibrosis (P<0.004), concluding that the A allele was protective from moderate and severe fibrosis. Meanwhile the SNP -1082G/A did not show any association with the fibrosis score. CONCLUSION We could not associate response to therapy for HCV with IL-10 polymorphisms -1082 G/A and -592 C/A. For the SNP -592 C/A, the A allele protected from moderate and severe fibrosis.

Potential genetic markers for prediction of treatment response in Egyptian children infected with HCV genotype 4

BACKGROUND Egypt has a high prevalence of hepatitis C virus (HCV) infection. Limitations of the current HCV treatment in children are low rate of sustained virological response, significant side effects and high expenses, making prediction of treatment response crucial. AIM This study aimed to investigate association of single nucleotide polymorphisms (SNPs) in interleukins (IL) 10, 28 and 29 genes in predicting the response to therapy in HCV infected children. METHODS Sixty-six Egyptian children infected with HCV genotype 4, receiving pegylated interferon alpha 2b and ribavirin, were included. Genotyping of six SNPs in interleukin 10, 28B and 29 gene as well as HCV genotype were analyzed by real-time polymerase chain reaction. RESULTS The CC genotype in IL28B; rs12979860 had 8.547 folds higher chance to develop sustained virological response than CT and TT genotypes (P=0.014). Genotype distribution of rs8099917 in IL28B gene (TG and GG genotypes) was found to be 3.348 more likely not to respond to treatment than the TT genotype (P=0.018). In multivariate analysis, interleukin 28 gene single nucleotide polymorphisms rs 12979860, interleukin 10 single nucleotide polymorphisms -592A > C and basal viral load were independent variables that significantly improved prediction of response to HCV therapy. CONCLUSION This association can be translated into clinical decision making for HCV treatment.

CYP21A2 genetic profile in 14 Egyptian children with suspected congenital adrenal hyperplasia: a diagnostic challenge

CYP21A2 genotyping remains an important element in the diagnosis and management of congenital adrenal hyperplasia, and establishing accurate genotype–phenotype correlations has facillitated adequate genetic counseling and prenatal management for at‐risk families. Despite extensive efforts to establish a clear genotype–phenotype correlation, some discordance remains. Establishing a diagnosis of congenital adrenal hyperplasia on the basis of biochemical and clinical data is occasionally challenging, and the identification of CYP21A2 mutations may help confirm the diagnosis. We review the diagnostic challenges despite an extensive genetic evaluation for 14 patients with a suspected clinical and biochemical diagnosis of congenital adrenal hyperplasia. Other diagnostic entities should be considered in the absence of convincing genetic data.