FAS and BCL-2 oncoproteins as an apoptotic markers in children with chronic Hepatitis C virus infection

The objective to evaluate the factor of apoptotic signal [FAS] and B-cell leukemia lymphoma [BCL-2] oncoprotein as markers of apoptosis in children with chronic hepatitis C . The study included 100 children with chronic hepatitis C, divided into three groups. Group I [control, n =20] HCVantibodies and RNA-PCR negative, Group II; [n=60] HCVantibodies positive and HCV- RNA positive without liver cirrhosis and Group III; [n=20] HCVantibodies positive and HCV-RNA positive with liver cirrhosis

Blood samples were subjected to: Complete blood count, liver function tests, alpha fetoprotein assay by ELISA method, FAS [CD95] antigen, and intracellular BCL-2 oncoprotein on lymphocytes by flow cytometry

The Results showed that FAS was significantly higher in group II and group III as compared to group I [P=0.0 for both]. FAS was significantly higher in group III than group II [P=0.021]. Positive correlation between FAS and ALT, AST, and AFP was found [r =0.39, 0.47 and 0.39, respectively, P = 0.0 for all]. Combining groups II and III, negative correlations between FAS and platelets and albumin [r= -0.52 and -0.46, respectively, P = 0.0 for both]. BCL-2 was significantly higher in group II and group III as compared to group I [P=0.001 and 0.0, respectively]. BCL-2 was higher in group III than group II [P= 0.0]. BCL-2 was positively correlated to ALT, AST, and AFP [r= 0.29, 0.39 and 0.41, respectively, P= 0.0 for all]. BCL-2 was negatively correlated to platelets and albumin in the combined diseased group [r= -0.53 and -0.41 respectively and P= 0.0 for both]. A positive correlation was found between FAS and BCL-2 in the diseased group [r=0.321 and p=0.007]

In Conclusion: FAS and BCL-2 as an apoptotic markers play an important role in the pathogenesis and further outcome of chronic liver diseases

Cytomegalovirus frequency in neonates with biliary atresia

Biliary atresia [BA] is the most severe hepatic disorder in newborns and its etiopathogenesis remains unknown. Viral involvement has been proposed, including the human cytomegalovirus [HCMV]. The aims of the study were to use the polymerase chain reaction [PCR] to screen the liver tissue of neonates with biliary atresia or cholestasis due to non atretic causes for HCMV-DNA and to correlate the results with serological antibodies to CMV and plasma PCR for CMV-DNA. This study was carried on sixty patients suffered from cholestasis, thirty patients suffered from BA [BA group] and thirty patients suffered from cholstasis without BA [Non Atretic cholestasis group]. Patients with BA were divided according to the result of the tissue PCR for CMV-DNA in the liver biopsies into two groups, group I, BA patients with positive tissue PCR and group II, BA patients with negative tissue PCR. All groups were subjected to complete history take, clinical biopsy, CMV [IgG and IgM] antibodies [ELIZA], Plasma-PCR for CMVDNA by Cobas Amplicator Analyzer and tissue PCR for CMV-DNA. Results showed that liver tissues PCR for CMV in BA patients was positive in 43.3%, however its incidence was very low in patients with non Atretic cholestaesis [3.3%], we found also that CMV IgG was positive in [73.3%], CMV IgM was positive in [16.7%], plasma PCR-CMV was positive in [13.3%] in BA group. On other hand IgG was positive in 33.3%, IgM was positive in 10%, plasma PCR was positive in 3.3% in non atretic group with significant differnces p< 0.05. So these results indicate an elevated frequency of CMV in neonates with BA which was more prevalent in females. No significant correlation between groups I and II of BA group as regard clinical picture, laboratory results and pathological changes p>0.05. The accuracy of plasma PCR was 45.9%, CMV IgM was 47.9%and CMV IgG was 46.1% when consider tissue PCR as golden test. In Conclusions, there was high frequency of CMV in neonates with BA which was more prevalent in females. The accuracy of serological testing for CMV using ELISA techniques and plasma CMV-DNA PCR are not reliable and indicate that virological assessment by tissue PCR should be carried out to prove the role of the CMV infection in etiology BA cases

Early infections after living donor liver transplantation in Egyptian children [single center experience]

Liver transplantation [LT] has become an established therapeutic option for a large variety of fulminant and chronic liver diseases. Postoperative infections are the major cause of morbidity and the leading cause of mortality. The aim of this work was to investigate the incidence, types and possible predisposing factors for infection after living donor liver transplantation [LDLT] in children occurring early in the post operative period. We enrolled 22 children [10 boys [45.5%] and 12 girls[54.5%]] who underwent LDLT from 2003 to 2010 at National Liver Institute, Menofiya University, Egypt, the mean age +/- SD was 5.59 +/- 4.45 years [range, 7 months to 17yr] . We retrospectively investigated the proven episodes of bacterial, viral, and fungal infections. There were 62 infections episodes in 18 [81.2%] of 22 patients [2.8 infections/patient], two or more infections occurred in 14 out of 22 transplanted children [63.6%], bacterial infections accounted for 41 episodes of infection [66.1%], Viral infections accounted for 10 episodes of infection [16.1%] while fungal infections accounted for 11 episodes of infection [17.8%].The most common sites of bacterial infection were chest [24.4%], bile ducts [cholangitis] [17.1%] the blood stream [bacteremia] [12.2%] ,and abdomen either peritonitis or intra abdominal abscess [9.8%]. Most of the bacterial infections were polymicrobial mainly with gram negative bacilli occurred within the first month after LDLT. The most common causes of viral infection were cytomegalovirus in 4 patients, Epstein-Barr virus in 2 patients, Herpes Simplex virus types I and II presented in 4 patients. The mortality rate due to infection was 18.2% mainly caused by bacterial infection. This study found that infection is one of the important causes of morbidity and mortality after LDLT, so careful monitoring and management of infections is crucial for improving the outcome of LDLT in children

Screening for phenylketonuria and galactosemia among Egyptian newborns in Menoufiya governorate

Was to study the prevalence of phenylketonuria and galactosemia in Menoufiya Governorate newborns. Among 3000 newborns, their mean ages were 9.3 +/- 2.43 days; mean weight was 3.1 +/- 0.82 Kg. Among them 1800[60%] males and 1200 [40%] females who attended the central hospital and medical units for BCG vaccination in the duration from March 2005 to May 2008. The results showed that the mean of phenylalanine level was 3.19 +/- 1.82 mg/dl and the mean total galactose level was 3.34 +/- 2.23mg/ dl, among the 3000 neonates, 2183 [72.8.%] had phenylalanine levels ranging from 2-5 mg /dl, 705 [23.5%] had levels ranging from 5-7 mg/dl, 111[3.7%] had levels ranging from 7-10 mg/dl and one newborn [0.033%] had phenylalanine level of 22 mg/dl. The results for galactosemia screening assay showed that 2528 neonates [84.3%] had galactose levels ranging from 2-6 mg/dl, 450 [15%] had levels ranging from 6-8 mg/dl, 21[0.7%] had levels ranging from 8-12 mg/dl and one newborn [0.033%] had galactose level of 19 mg/dl. The child was reassayed and was found to be true hypergalactosemia l20mg/dl. We concluded that the prevalence of each of phenylketonuria and galactosemia in Menoufiya Governorate in the 3000 newborn tested was 1/3000 [0.03%]. So, we estimate that about 333 neonates are affected every year with PKU and 333 with galactosemia as one million babies are born yearly, which could be prevented. The prevention of such treatable disorders depends on planning an efficient screening programme especially within three weeks after birth. So we recommend multicenter studies to encourage national neonatal screening programmes specialy for these treatable diseases

Plasma endothesin-1 and renin as predictor factors in schistosomal liver disease of children

Endotheh-1 [ET-1] is a mitogenic and a potent vasoconstrictor peptide. It affects the hemodynamics of the liver and plays a role in fibro genesis through activation of stellate cells. Also renin-angiotensin system, which is activated in liver cirrhosis, may play a role in the pathogenesis of portal hypertension associated with schistosomal liver disease. The role of plasma ET-1 and renin in schistosomal liver disease and their clinical significance remain largely under investigated particularly in children. This study included 20 patients attending Pediatric Department of National Liver Institute, Menoufyia University, or Pediatric Department of Tanta University Hospital. Their mean age was 10.2 +/- 2.1 years. A control group of 20 healthy children of matched age and sex was included in the study. For all study children, history, physical examination, urine and stool analysis, CBC, kidney function tests and liver function tests were undertaken. Rectal snip biopsy, abdominal ultrasound examination and liver biopsy were utilized to diagnose our patients. Plasma ET-4 and renin were measured by radioimmunoassay. The mean level of ET-1 in the control group was 0.83 +/- 0.20 pg/ml with a range of 0.49 to 0.99 pg/ml, In contrast, the mean ET-1 level in the patients group was 1,37 +/- 0.6 pg/ml with a range of 0.49 to 2.24 pg/ml. The difference was statistically significant [p=0.001]. ET-1 was found to be elevated [2 SD above the mean control value] in 9 of 20 patients [45 %] with mean of 7.8 +/- 0.37 pg/ml and a range of 1.49 to 2.24 pg/ml. ET-1 was elevated in 2/11 [19%] of patients with hepatomegaly, in 2/3 [67%] of patients with hepatosplenomegaly, in the only case with shrunken liver 1/1 [100%] and in 4/5[80%] of patients with ascites. The relation of ET-1 elevation to the stage of disease was statistically significant [p=0.026]. Elevated ET-1 was found in 6/1 [86%] of patients with elevated ALT, compared with 3/13 [23%] of patients with normal ALT. This relation was statistically significant [p=0.017]. Plasma ET-1 levels appeared to correlate with duration and severity of the disease. Unlike in cirrhotic patients previously reported, renin was normal in all studied cases with no significant difference between patients of all disease stages versus controls. in the control group the mean plasma renin level was 2.52 +/- 1.23 ng/L and ranged from 1.23 to 3.81 ng/L while in the patients group it was 2.09 +/- 1.06 ng/L and ranged from 0.72 to 3.45 ng/L. This might be attributed to the stable hemodynamics and normal renal function of our study cases

Conclusions: the present study suggests that ET-7 level might be elevated in cases of schistosomal liver disease of Egyptian children particularly advanced cases. Elevated ET-1 might be involved in the aggravation of the disease or development of complications like portal hypertension 3rd ascites. So, monitoring ET-1 level along the course of the disease may be useful as a predictor for the development of these complications and may be helpful in the prevention planning. Plasma renin measurement probably has no important role in following up schistosomal liver disease during childhood

Hepatitis C virus [HCV] core antigen in children with hepatitis C virus infection

Recently, the availability of an anti-core antigen [Ag] monoclonal antibody allowed the development of an enzyme linked immunosorbent assay [ELISA] that showed the presence of hepatitis C virus [HCV] core Ag in peripheral blood of HCV- infected patients. The aim of our study was to determine the specificity and sensitivity of HCV-core antigen assay in serum compared to serum HCV-RNA by qualitative and quantitative PCR in diagnosis of HCV infection in children. Our study was performed on fifty children complaining of acute and chronic liver diseases, attending the pediatric department, National Liver Institute, Menoufiya University from May 2002 to October 2003. They were divided into two groups. Group one consists of 25 children with HCV infection, 20 of them with chronic infection and the other 5 with acute infection. Group two consists of 25 children with liver diseases other than HCV as a control group [to with autoimmune hepatitis, 4 with HBV infection, 4 with hepatosplenic shistosomiasis, 4 with fulminant hepatic failure, 2 with venous outflow block and one with congenital hepatic fibrosis]. In our study HCV core antigen assay showed a specificity of 100 % [25/25] compared to PCR in the diagnosis of HCV infection. However, HCV-antibody testing showed 72 % specificity compared to PCR. While sensitivity of HCV core antigen was 88% among all cases, 80 % [44 in diagnosis of acute HCV cases and 90 % [18/20] in diagnosis of chronic HCV cases in children and showed total validity of 94 % in comparison to that of PCR. The 3 negative cases defected by PCR and not by HCV-core Ag were with low viremia with the highest one being 11000 IU/ml. In comparison, HCV- Ab detected one out of the 5 acute cases and 19 out of the 20 chronic ones. In our study, by applying the ROC curve it was found that the best cutoff value of viremia is 15 X 10[3] IU/ml. At that level of HCV- RNA, the sensitivity and specificity of HCV- core Ag assay are 100 %

Conclusion: HCV core antigen assay may obviate many of the problems encountered with the serologic assay of HCV infection and be used in diagnostic settings as if shows comparable specificity and sensitivity to PCR. Moreover, it is simple, of less cost and not prone to contamination as PCR technique. It is also of special value in the window phase of HCV infection before seroconversion. Further study on a larger number of children with HCV infection is recommended to prove its value and its relation to the level of HCV viremia

Leptin, insulin-like growth factor I and its binding proteins I and 3 as indicators of pre-natal and post-natal growth potentials

The study was conducted on 50 newborn infants [20 females and 30 males] who were born in Menoufiya University Hospital or Tanta University Hospital. Among these infants 25 cases were full term appropriate for gestational age [group I or controls] while the other 25 cases were low birth weight small for gestational age [LB W-SGA] patients [group II]. Cases with maternal pre-eclampsia and infants of diabetic mothers were excluded. The study also included the mothers of these two groups [50 mothers]. The aims were to evaluate cord serum IGF-I, IGFBP-3, IGFBP-1 and leptin as indicators of prenatal and postnatal growth potentials and to correlate between maternal and fetal concentrations of these parameters. History and clinical examination including anthropometric measurements [birth weight, length, head circumference and chest circumference] were done and cord serum leptin, IGF-l, IGFBP-I and IGFBP-3 were measured. For mothers of studied groups, full history and through clinical examination were done including general nutritional status, blood pressure, weight, height, parity and any complication. Also their serum leptin, IGF-l, IGFBP-I and IGFBP-3 were measured. We followed up our cases with intrauterine growth retardation, [IUGR] at birth [group II] who were observed after 6 months whether catch up of weight and length has occurred [catch up subgroup] or not [non-catch up subgroup]. Cord serum leptin, IGF-I and IGFBP-3 were significantly lower while IGFBP-I higher in LBW SGA infants [group II] than that in normal controls [group I]. These parameters correlated well with birth anthropometric measurements in the studied cases. On the other hand, maternal serum levels of leptin, IGF-I, IGFBP-I and IGFBP-3 did not correlate significantly with their levels in cord serum of the studied infants nor with birth anthropometric measurements. On follow up after 6 months, infants with successful catch up of growth showed higher birth weights and cord serum levels of IGF-I than those without growth catch up. Cord serum leptin, IGF-I, IGFBP-1 and IGFBP-3 could be useful in evaluating preterm small gestational age [SGA] infants and correlate with their anthropometric parameters at birth. Cord serum IGF-l and birth weight can be predictive of the future catch up potential of growth in IUGR infants. Maternal anthropometric parameters and maternal serum leptin, IGF-I, IGFBP-1 and IGFBP-3 did not correlate to the infant's parameters at birth [anthropometric or cord serum levels] nor to the potential for successful catch up of growth during the following six months