A Polymorphism in the Microsomal Triglyceride Transfer Protein Can Predict the Response to Antiviral Therapy in Egyptian Patients with Chronic Hepatitis C Virus Genotype 4 Infection

BACKGROUND/AIMS: A polymorphism in the microsomal triglyceride transfer protein (MTP) is associated with hepatic fibrosis, and carriers showed higher levels of steatosis, higher levels of hepatitis C virus (HCV) RNA and advanced fibrosis. The aim of this study was to study MTP expression pattern in HCV patients and impact of the MTP polymorphism on the response to antiviral therapy. METHODS: One hundred consecutive naive HCV genotype 4 patients were recruited to receive antiviral therapy, and 40 control subjects were also recruited. Demographic, laboratory, and histopathology data were collected. DNA was isolated, and the samples were subjected to polymerase chain reaction analysis and genotyping for MTP by restriction fragment length polymorphism analysis. RESULTS: Patients and controls were age- and sex-matched (male/female, 56/44, age, 39.2+/-7.8 years for patients with HCV; male/female, 18/22, age, 38.1+/-8.1 years for controls). MTP single nucleotide polymorphisms (SNPs) (GG, GT, TT) and alleles (G, T) in the patients versus the controls were 70%, 21%, 9% & 80.5%, 19.5% versus 10%, 87.5%, 2.5% & 53.8%, 46.3%, respectively (p=0.0001). The sustained viral response (SVR) of the patients was 60%. SNPs in MTP genotypes (GG, GT, and TT) and alleles (G and T) in the responders and nonresponders were 71.7%, 25%, 3.3% & 84.2%, 15.8% versus 67.5%, 15%, 17.5% & 75%, 25% (p=0.038 and p=0.109, respectively). A multivariate analysis showed that the GT genotype was an independent predictor of SVR (area under the curve 90% and p=0.0001). CONCLUSIONS: MTP could be a new predictor for SVR to antiviral therapy in patients with HCV genotype 4 infection.

Hepatic fibrosis and serum alpha-fetoprotein [AFP] as predictors of response to HCV treatment and factors associated with serum AFP normalisation after treatment

Elevated levels of alpha-fetoprotein [AFP] can be seen in patients with chronic hepatitis C [CHC] and liver cirrhosis without hepatocellular carcinoma and were negatively associated with treatment response. However, factors associated with its changes are not identified. We aimed in this study to verify a cut-off value for AFP as a predictor of response to standard of care [SOC] antiviral therapy in Egyptian chronic hepatitis C virus [HCV]-infected patients and identify factors associated with its changes post treatment. A total of 175 chronic non-cirrhotic HCV-infected patients were evaluated for baseline serum AFP and liver biopsy were classified according to Ishak scoring system of hepatic fibrosis. All patients were scheduled to receive SOC antiviral therapy for 48 weeks and had been followed up to week 72. Reassessment of AFP and repeated liver biopsy at week 72 were feasible only in 79 patients. High baseline AFP levels were observed in non-respondents [non-sustained virological respondents [non-SVRs]] [P< 0.01]; the AFP level decreased in all patients post treatment [P= 0.01], especially in the SVRs [P < 0.01]. In multivariate analysis, hepatic fibrosis was a predictor of response to treatment [P=0.02], while body mass index [BMI] [25-30 kg mr[2]], hepatic activity [A2], hepatic fibrosis stage [F2-F4] and fibrosis improvement were predictors of AFP difference [P = 0.007, 0.01, 0.012, <0.001, 0.030, and 0.018], respectively. The diagnostic performance to predict the HCV treatment response was best by adding both AFP and hepatic fibrosis stage factors; the best cut-off value for AFP was 3.57 ng dr1 with 50% sensitivity and 68% specificity with area under the curve [AUC] of 0.55 and for hepatic fibrosis stage was 3, with a sensitivity of 88%, a specificity of 30% with an AUC of 0.58. In chronic HCV-infected patients, serum AFP below 3.57 ng dl[-1] and hepatic fibrosis stage 3 are expected to have good response to treatment; BMI [25-30 kg m[-1]], A2, fibrosis >2 and fibrosis improvement predict AFP change post treatment

Evaluation of serum LINE-1 hypomethylation as a prognostic marker for hepatocellular carcinoma

Global hypomethylation is one of the most consistent epigenetic changes in cancer. Development of hepatocellular carcinoma [HCC] must be understood as a multistep process with accumulation of genetic and epigenetic alterations. In the last decades, in addition to genetic alterations, epigenetic changes have been recognized as an important and alternative mechanism in tumourigenesis. We investigated the clinical implications of global hypomethylation in the sera of patients with hepatocellular carcinoma [HCC]. PCR was used to assess the methylation status of long interspersed nuclear element type 1 [LINE-1] repetitive sequences in genomic DNA derived from sera of 50 patients with HCC, 20 patients with cirrhosis, 20 patients with chronic hepatitis C and 10 healthy subjects. Serum genome hypomethylation was significantly increased in patients with HCC [p < 0.001]. The levels of serum LINE-1 hypomethylation at initial presentation correlated significantly with tumour size, tumour number and alpha-foetoprotein level. Moreover high serum LINE-1 hypomethylation correlates significantly with poor survival. Serum LINE-1 hypomethylation may serve as a prognostic marker for patients with HCC

Effect transfusional of iron overload on hepatic fibrosis in Egyptian beta-thalassemia major patients

Multitransfused beta-thalassemia patients constitute a population with high prevalence of Hepatitis C virus [HCV] infection, because of transmission of HCV from infected blood donors. Increased hepatic iron is assumed to potentiate progression towards liver fibrosis in chronic HCV infection. The aim of the present work is to evaluate the potentiating effect of marked hepatic iron overload and chronic HCV infection on hepatic fibrosis in Thalassemia patients. Sixty eight patients, previously diagnosed to have homozygous beta-thalassemia and followed up at the Hematology Clinic of the New Children's Hospital of Cairo University [44 hepatitis C positive and 24 hepatitis C negative patients], were selected to participate in this study after signing a written informed consent. Their age ranged between 6 and 27 years with a mean age of 9.7 +/- 2.1 years and compared to a group of 42 non thalassemic chronic HCV patients whose age ranged between 7 and 27 years with a mean age of 10.9 +/- 1.5 years [control group]. Liver Biopsies were done for all patients for estimation of stage of hepatic fibrosis and liver iron content [LIC]. The results were then correlated to liver function tests and serum ferritin. The stage of fibrosis and LIC were significantly higher in beta-thalassemia patients than the non thalassemia HCV patients [p = 0.005, p<0.0001 respectively]. There was no significant difference between the two groups of thalassemia as regards staging of fibrosis. The degree of hepatic fibrosis was significantly correlated to the LIC in hepatitis negative thalassemic group while it was significantly correlated to serum ferritin in thalasemic patients with positive HCV. Hepatic iron overload has a potentiating effect on hepatic fibrogenesis in beta-thalassemia major. The proper use of chelating agents is of great importance in delaying progression of liver disease in these patients

Molecular cloning and sequencing of disintegrin like domain from Cerastes cerastes venom gland

Disintegrin-like domain was cloned and sequenced from Cerastes cerastes venom gland tissue. Nested RT-PCR was performed using initial primers designed based on the homology of disintegrins from Trimeresurus flavoviridis, Giodius halys, Agkistrodon halys and Trimeresurus macrosquamatus. The homology was reached using BLAST searching tool. The primers were selected using doprimers algorithm. Nested primers were those reported by Yamada et al [1999] for Trimeresurus flavoviridis. The nested PCR product was approximately 150 bP as determined by agarose gel electrophoresis Cloning of the PCR product was performed using Qiagen PCR cloning system. The construct in pDrive cloning vector was introduced into competent JM109 bacterial cells [Promega]. Plasmid DNA minipreps were prepared having the 150bp insert. Sequence analysis of the insert gave 124 bp which is in extensive sequence homology with many snake venoms disintegrin domain. Translation of the nucleotides sequence and searching the protein database revealed amino acid sequence of disintegrin with 70% consensus identity. Prosite motif search gave two phosphorylation sites [kinase c and casein kinase 1]. One myristoylation site was also identified. Four cysteines seems to form disulfide bonds

Impact of 10 years of hepatitis B vaccination on the frequency of acute symptomatic hepatitis B at a major urban referral center in Egypt

This study aimed to determine the impact of 10 years of universal hepatitis B immunization on the prevalence of acute symptomatic viral hepatitis B in Egypt compared with previous results reported by the same authors in 1983. Two hundred consecutive patients with acute symptomatic viral hepatitis, diagnosed clinically and biochemically, were enrolled from Embaba Fever Hospital [EFH], Giza Governorate, Egypt in the period from December 2001 to September 2002. Serological tests were done using ABBOTT AXSYM [Abbott laboratory, Abbott Park, III] for hepatitis A virus [anti-HAV IgM], hepatitis B virus [HBsAg, anti-HBc total and IgM, anti HBs], hepatitis C virus [anti-HCV IgM and total, in addition to PCR], hepatitis D virus [anti-HDV IgM] and hepatitis E [anti-HEV IgM]. In addition, the patients were screened for IgM Ab of cytomegalovirus [CMV], Ebstein barr virus [EBV] and PCR for HGV and transfusion transmitted virus [TTV]. In the present study, the overall acute HBV infection was accounted for 31.5% of the acute viral hepatitis cases, with a male predominance of 62.4% and a significant increased prevalence in adolescent and adults. Although the clinical presentation of HBV did not differ from those of the various causes of hepatitis, the mean ALT levels were significantly higher in acute HBV when compared with acute HAV infection. The frequency of acute hepatitis B was decreased from 43.4% in 1983 to 31.5% in 2002, which was particularly evident in children and there were no significant changes in the rate of acute hepatitis among HBsAg carriers [12.3% and 9%, respectively]. Despite the lower coverage rate of hepatitis vaccination in those <9 years [37.9%], there was virtual absence of acute hepatitis B. The use of multiple seromarkers decreased the prevalence of the undiagnosed cases in those suffering from acute hepatitis to 6.5%

community-based study of viral hepatitis infection in Giza Governorate, Egypt: Seroprevalence, risk factors and associated morbidity

Hepatitis viruses are major causes of acute and chronic liver diseases in Egypt. The aim of this study was to investigate the seroprevalence, risk factors and associated morbidity of viral hepatitis in Giza Governorate, Egypt. The study was conducted in 4 rural and 4 semiurban communities and included 2305 subjects selected by a cluster r and om method. They underwent complete clinical and abdominal ultrasonographic [US] assessment and laboratory tests including stool and urine examination, hemoglobin, ALT estimation and viral hepatitis markers by enzyme immunoassay. The latter included anti-HAV IgG, HBsAg, anti-HBc, anti-HBs, anti-HCV and anti-HEV IgG. Subsets of sera were tested for HBV DNA and HGV RNA by specific PCR. The overall prevalence of anti-HCV was 20.9% [age-adjusted prevalence = 24.5%; CI: 22.7-26.3%] and was significantly rising with age from 10% below age of 20 years to 40% among those above 50 years. The infection rate of hepatitis B virus [HBV] was 57.1% with HBsAg carrier rate of 3.6%. Seropositivity of anti-HBs and anti-HBc was very high [45.8% and 44.7%, respectively] and correlated positively with age with no sex-related difference. Combined HCV and HBV infection was evident in 13% of subjects. HBV DNA was detected in 86% of HBsAg-positive cases and in 20% of HBsAg-negative anti-HBc- and anti-HCV-positive cases. Seromarkers for hepatitis B and C were significantly commoner in semiurban than in rural communities. Dental manipulation and previous parenteral antischistosomal therapy were significant risk factors for hepatitis B and C infection. Anti-HAV was positive in 99.7% whereas anti-HEV was positive in HBV= Hepatitis B virus. CLD = Chronic liver disease. HCC = Hepatocellular carcinoma. US = Ultrasonography. ALT = Alanine transaminase. OR = Odds ratio. CI = Confidence interval. 9.2%. HGV RNA was detected in 16.5% of the studied samples. It was always associated with HBV and /or HCV infection. History of hematemesis was recorded in 1.2% of individuals. US examination revealed hepatomegaly in 19.2% of subjects, splenomegaly in 8.2%, bright liver in 31%, coarse liver texture in 10%, periportal fibrosis in 20% and ascites in 2%. These findings were significantly more common in anti-HCV- and HBsAg-positive subjects. ALT elevation was commonest and highest in individuals with positive HBsAg or with HBV-HCV coinfection. Hepatitis B and C infection and associated morbidity still constitute a great health problem in Egypt. Hepatitis A is holoendemic and hepatitis E is endemic. Hepatitis G is always associated with HBV or HCV infection. Occult hepatitis B should be considered in future studies

Pravastatin and its effects on renal reperfusion injury in an experimental model in dogs

The aim of this study was to determine if pretreatment with pravastatin would attenuate acute renal dysfunction that occurs following ischemic reperfusion injury in an experimental model in dogs. Twenty mongrel dogs were randomized into two groups [n = 10 per group]. In addition, a control group was also included in which bilateral renal ischemic arterial clamping was applied for one hour before which blood samples were taken for baseline biochemical evaluation and tumor necrosis factor alpha [TNF alpha] estimation. Declamping was done for 15 min and another blood sample for TNF alpha estimation was taken. Then, the animal's abdomen was closed and the animal awakened from anesthesia. At 48 hours later, the animal was re-operated upon and the left kidney was harvested for histopathologic examination and blood samples were taken for the previously mentioned tests. Another group of ten dogs underwent the same procedure after five days preoperative treatment with pravastatin [0.5 mg/kg/day for 5 days]. The results showed that pravastatin may play a role in modulating renal impairment following aortic, renovascular or transplantation surgery allowing early reduction form an ischemic reperfusion injury

Herpes group viruses diagnosis by polymerase chain reaction in Guillain bare syndrome and transverse myelitis

This study included 35 patients, their ages ranged from 11 months to 7 years. 20 cases presented with clinical manifestation of Guillain Barre syndrome [GBS] and 15 cases with clinical manifestations of transverse myelitis [TM]. All cases were subjected to general and neurological examinations and CSF analysis, computed tomography of dorsolumbar region for 15 cases with transverse myelitis. Polymerase chain reaction [PCR] was done for both CSF and blood of all cases for detection of cytomegalovirus-DNA, herpes simplex DNA and Epstein-Barr- DNA. Preceding illness in the form of upper respiratory infection, gastroenteritis and exanthem were detected in 65% and 39.99% among cases of GBS and TM, respectively. Total outcome among all 35 patients [8 weeks after onset of neurological manifestations] showed complete recovery in 45.71%, recovery with residual squelae in 40% and death in 14.29%. Cytomegalo-virus DNA PCR positive signal was the most commonly detected marker of viral infection in this study