scholarly journals A Medium for Facilitating Hepatitis B Virus Detection and Replication of the Virus 

Author(s):  
İ. Afşin KARİPER ◽  
Tuğba Demir ◽  
Armağan Caner ◽  
Selma Gökahmetoğlu ◽  
Yusuf Tutar ◽  
...  

Abstract In this study, we tested a chemical mixture causing hepatitis virus to grow in excessive amounts. We obtained this chemical mixture by disintegrating albumin and invertase enzymes separately in supercritical environments, using acidic methanol as solvent. Then we used this mixture as a medium for hepatitis B virus. The chemical mixture caused the hepatitis B virus to multiply almost 80-100 times after 120 minutes. The real surprising result of the study occurred after carrying out bacterial tests with the same chemical mixture, its effect on cancer cells were also investigated. But no effect was observed neither on cancer cells, nor on bacteria. In other word, we developed a specific medium for the hepatitis virus. In this way, we decided that a method that can reduce the detection limit of PCR device to 1 IU/mL could be developed. This study will be an important milestone in developing vaccines against viruses as well.

2000 ◽  
Vol 44 (7) ◽  
pp. 1964-1969 ◽  
Author(s):  
Karl Y. Hostetler ◽  
James R. Beadle ◽  
William E. Hornbuckle ◽  
Christine A. Bellezza ◽  
Ilia A. Tochkov ◽  
...  

ABSTRACT Acyclovir triphosphate is a potent inhibitor of hepatitis B virus DNA polymerase, but acyclovir treatment provides no benefit in patients with hepatitis B virus infection. This is due in part to the fact that hepatitis B virus, unlike herpes simplex virus, does not code for a viral thymidine kinase which catalyzes the initial phosphorylation of acyclovir. We synthesized 1-O-octadecyl-sn-glycero-3-phospho (3-P)-acyclovir and found that it was highly active in reducing hepatitis B virus replication in 2.2.15 cells, while acyclovir was inactive. The greater antiviral activity of 1-O-octadecyl-sn-glycero-3-P-acyclovir appeared to be due to liver cell metabolism of the compound to acyclovir monophosphate (K. Y. Hostetler et al., Biochem. Pharmacol. 53:1815–1822, 1997). However, a closely related compound without a hydroxyl group at the sn-2 position of glycerol, 1-O-hexadecylpropanediol-3-P-acyclovir, was more active and selective in 2.2.15 cells in vitro. In this study, we treated woodchucks chronically infected with woodchuck hepatitis virus with increasing oral doses of 1-O-hexadecylpropanediol-3-P-acyclovir and assessed the response to therapy versus acyclovir or a placebo. At a dosage of 10 mg/kg of body weight twice a day, the test compound significantly inhibited viral replication in vivo, as indicated by a 95% reduction in serum woodchuck hepatitis virus DNA levels and by a 54% reduction in levels of woodchuck hepatitis virus replicative intermediates in the liver. Higher doses were somewhat less effective. In contrast, 20 mg of acyclovir/kg twice daily, a 5.3-fold-higher molar dosage, had no demonstrable activity against woodchuck hepatitis virus. Oral 1-O-hexadecylpropanediol-3-P-acyclovir appeared to be safe and effective in chronic woodchuck hepatitis virus infection.


1998 ◽  
Vol 72 (6) ◽  
pp. 5085-5092 ◽  
Author(s):  
John E. Donello ◽  
Jonathan E. Loeb ◽  
Thomas J. Hope

ABSTRACT The hepatitis B virus posttranscriptional regulatory element (HBVPRE) is a cis-acting RNA element that partially overlaps with enhancer I and is required for the cytoplasmic accumulation of HBV surface RNAs. We find that the closely related woodchuck hepatitis virus (WHV), which has been shown to lack a functional enhancer I, also contains a posttranscriptional regulatory element (WPRE). Deletion analysis suggests that the WPRE consists of three independent subelements. Comparison of the bipartite HBVPRE and tripartite WPRE activities reveals that the tripartite WPRE is two to three times more active than the bipartite HBVPRE. Mutation of a single WPRE subelement decreases WPRE activity to the level of the HBVPRE. Bipartite and tripartite chimeras of the WPRE and HBVPRE possess activities which suggest that elements containing three subelements are posttranscriptionally stronger than those containing two. These data demonstrate that the posttranscriptional regulatory element is conserved within the mammalian hepadnaviruses and that its strength is determined by the number of subelements within the RNA.


1998 ◽  
Vol 42 (12) ◽  
pp. 3209-3217 ◽  
Author(s):  
E. V. Genovesi ◽  
L. Lamb ◽  
I. Medina ◽  
D. Taylor ◽  
M. Seifer ◽  
...  

ABSTRACT Daily oral treatment with the cyclopentyl 2′-deoxyguanosine nucleoside BMS-200475 at doses ranging from 0.02 to 0.5 mg/kg of body weight for 1 to 3 months effectively reduced the level of woodchuck hepatitis virus (WHV) viremia in chronically infected woodchucks as measured by reductions in serum WHV DNA levels and endogenous hepadnaviral polymerase activity. Within 4 weeks of daily therapy with 0.5 or 0.1 mg of BMS-200475 per kg, endogenous viral polymerase levels in serum were reduced about 1,000-fold compared to pretreatment levels. Serum WHV DNA levels determined by a dot blot hybridization technique were comparably decreased in these treated animals. In the 3-month study, the sera of animals that had undetectable levels of WHV DNA by the dot blot technique were further analyzed by a highly sensitive semiquantitative PCR assay. The results indicate that BMS-200475 therapy reduced mean WHV titers by 107- to 108-fold, down to levels as low as 102 to 103 virions/ml of serum. Southern blot hybridization analysis of liver biopsy samples taken from animals during and after BMS-200475 treatment showed remarkable reductions in the levels of WHV DNA replicative intermediates and in the levels of covalently closed circular viral DNA. WHV viremia in BMS-200475-treated WHV carriers eventually returned to pretreatment levels after therapy was stopped. These results indicate that BMS-200475 should be evaluated in clinical trials for the therapy of chronic human hepatitis B virus infections.


1988 ◽  
Vol 16 (24) ◽  
pp. 11834-11834 ◽  
Author(s):  
J.M. Casacuberta ◽  
R. Jardi ◽  
M. Bulti ◽  
P. Puigdoménech ◽  
B. San Segundo

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