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Abstract Background Hepatitis C virus (HCV) infection is a major public health problem with more than 170 million cases of chronic infections worldwide. There is no protective vaccine currently available for HCV, therefore the development of novel strategy to prevent chronic infection is important. We reported earlier that a recombinant human antibody clone blocks viral NS3 helicase activity and inhibits replication of HCV 1b virus. This study was performed further to explore the mechanism of action of this recombinant antibody and to determine whether or not this antibody inhibits replication and infectivity of a highly efficient JFH1 HCV 2a virus clone. Results The antiviral effect of intracellular expressed antibody against the HCV 2a virus strain was examined using a full-length green fluorescence protein (GFP) labeled infectious cell culture system. For this purpose, a Huh-7.5 cell line stably expressing the NS3 helicase gene specific IgG1 antibody was prepared. Replication of full-length HCV-GFP chimera RNA and negative-strand RNA was strongly inhibited in Huh-7.5 cells stably expressing NS3 antibody but not in the cells expressing an unrelated control antibody. Huh-7.5 cells stably expressing NS3 helicase antibody effectively suppressed infectious virus production after natural infection and the level of HCV in the cell free supernatant remained undetectable after first passage. In contrast, Huh-7.5 cells stably expressing an control antibody against influenza virus had no effect on virus production and high-levels of infectious HCV were detected in culture supernatants over four rounds of infectivity assay. A recombinant adenovirus based expression system was used to demonstrate that Huh-7.5 replicon cell line expressing the intracellular antibody strongly inhibited the replication of HCV-GFP RNA. Conclusion Recombinant human anti-HCV NS3 antibody clone inhibits replication of HCV 2a virus and infectious virus production. Intracellular expression of this recombinant antibody offers a potential antiviral strategy to inhibit intracellular HCV replication and production.

Abstract Background The antiviral action of interferon alpha targets the 5' untranslated region (UTR) used by hepatitis C virus (HCV) to translate protein by an internal ribosome entry site (IRES) mechanism. Although this sequence is highly conserved among different clinical strains, approximately half of chronically infected hepatitis C patients do not respond to interferon therapy. Therefore, development of small interfering RNA (siRNA) targeted to the 5'UTR to inhibit IRES mediated translation may represent an alternative approach that could circumvent the problem of interferon resistance. Results Four different plasmid constructs were prepared for intracellular delivery of siRNAs targeting the stem loop II-III of HCV 5' UTR. The effect of siRNA production on IRES mediated translation was investigated using chimeric clones between the gene for green fluorescence protein (GFP) and IRES sequences of six different HCV genotypes. The siRNA targeted to stem loop II effectively mediated degradation of HCV IRES mRNA and inhibited GFP expression in the case of six different HCV genotypes, where as siRNAs targeted to stem loop III did not. Furthermore, intracytoplasmic expression of siRNA into transfected Huh-7 cells efficiently degraded HCV genomic RNA and inhibited core protein expression from infectious full-length infectious clones HCV 1a and HCV 1b strains. Conclusion These in vitro studies suggest that siRNA targeted to stem-loop II is highly effective inhibiting IRES mediated translation of the major genotypes of HCV. Stem-loop II siRNA may be a good target for developing an intracellular immunization strategy based antiviral therapy to inhibit hepatitis C virus strains that are not inhibited by interferon.

Background: To reduce airway and circulatory responses during extubation, various pharmacological and non-pharmacological methods have been used such as opioids, inhalational agents, local anaesthetics, vasodilators, alpha blockers, beta blockers, and calcium channel blockers. Present study was aimed to compare the hemodynamic effects of esmolol and lidocaine in attenuation of haemodynamic responses to tracheal extubation in neurosurgical patients. Material and Methods: Present study was single centric, observational, prospective, comparative study, conducted in patients with ASA 1 & 2, undergoing neurosurgical procedure (elective and emergency) with invasive blood pressure monitoring. Patients were divided in two group as Group 1 (Esmolol, n=30) & Group 2 (Lignocaine, n=30). Results: No significant difference was seen in the age wise distribution, gender wise distribution, mean weight & ASA grade distribution of patients enrolled in both groups. No significant difference in heart rate was seen among patients in both the group at baseline and at pre reversal stage. Average heart rate was significantly higher among patients in lignocaine after pre reversal stage from min 1 to 15 min. No significant difference in SBP was seen among patients in both the group at baseline and at pre reversal stage. Average SBP was significantly higher among patients in lignocaine after pre reversal stage from min 1 to 15 min. Nausea - vomiting were the two most common side effect seen among patients in both the group. No significant difference was seen among the reported side effect. Majority of patients in both the group has no coughing at extubation. No significant difference in the quality of extubation was seen among patients in both group Conclusion: No difference in the side effect was seen among patients in both group & majority of patients has smooth extubation with no coughing episode. [ABSTRACT FROM AUTHOR]

Background: Hepatitis C virus (HCV) infection is a major public health problem with more than 170 million cases of chronic infections worldwide. There is no protective vaccine currently available for HCV, therefore the development of novel strategy to prevent chronic infection is important. We reported earlier that a recombinant human antibody clone blocks viral NS3 helicase activity and inhibits replication of HCV 1b virus. This study was performed further to explore the mechanism of action of this recombinant antibody and to determine whether or not this antibody inhibits replication and infectivity of a highly efficient JFH1 HCV 2a virus clone. Results: The antiviral effect of intracellular expressed antibody against the HCV 2a virus strain was examined using a full-length green fluorescence protein (GFP) labeled infectious cell culture system. For this purpose, a Huh-7.5 cell line stably expressing the NS3 helicase gene specific IgG1 antibody was prepared. Replication of full-length HCV-GFP chimera RNA and negative-strand RNA was strongly inhibited in Huh-7.5 cells stably expressing NS3 antibody but not in the cells expressing an unrelated control antibody. Huh-7.5 cells stably expressing NS3 helicase antibody effectively suppressed infectious virus production after natural infection and the level of HCV in the cell free supernatant remained undetectable after first passage. In contrast, Huh-7.5 cells stably expressing an control antibody against influenza virus had no effect on virus production and high-levels of infectious HCV were detected in culture supernatants over four rounds of infectivity assay. A recombinant adenovirus based expression system was used to demonstrate that Huh-7.5 replicon cell line expressing the intracellular antibody strongly inhibited the replication of HCV-GFP RNA. Conclusion: Recombinant human anti-HCV NS3 antibody clone inhibits replication of HCV 2a virus and infectious virus production. Intracellular expression of this recombinant antibody offers a potential antiviral strategy to inhibit intracellular HCV replication and production. [ABSTRACT FROM AUTHOR]

Dash S, Prabhu R, Hazari S, Bastian F, Garry R, Zou W, Haque S, Joshi V, Regenstein FG, Thung SN. Interferonsα,β,γ each inhibit hepatitis C virus replication at the level of internal ribosome entry site-mediated translation.Liver International 2005: DOI: 10.1111/j.1478-3231.2005.01082.x.© Blackwell Munksgaard 2005Interferon (IFN)-α is the standard therapy for the treatment of chronic hepatitis C, but the mechanisms underlying its antiviral action are not well understood. In this report, we demonstrated that IFN-α, -β and -γ inhibit replication of the hepatitis C virus (HCV) in a cell culture model at concentrations between 10 and 100 IU/ml. We demonstrated that the antiviral actions each of each these IFNs are targeted to the highly conserved 5′ untranslated region of the HCV genome, and that they directly inhibit translation from a chimeric clone between full-length HCV genome and green fluorescent protein (GFP). This effect is not limited to HCV internal ribosome entry site (IRES), since these IFNs also inhibit translation of the encephalomyocardititis virus (EMCV) chimeric mRNA in which GFP is expressed by IRES-dependent mechanisms (pCITE-GFP). These IFNs had minimal effects on the expression of mRNAs from clones in which translation is not IRES dependent. We conclude that IFN-α, -β and -γ inhibit replication of sub-genomic HCV RNA in a cell culture model by directly inhibiting two internal translation initiation sites of HCV- and EMCV-IRES sequences present in the dicistronic HCV sub-genomic RNA. Results of thisin vitrostudy suggest that selective inhibition of IRES-mediated translation of viral polyprotein is a general mechanism by which IFNs inhibits HCV replication. [ABSTRACT FROM AUTHOR]

Recent studies suggest that Epstein–Barr virus (EBV) may act as a helper virus for the development of hepatocellular carcinoma by promoting replication of the hepatitis C virus (HCV) in the infected liver. Detection of EBV DNA in a high percentage of HCV-positive human hepatocellular carcinomas (HCC) from Japanese patients has supported this concept. In order to determine whether EBV infection is associated with HCC, we examined paraffin-embedded tissues from 31 cases of non-cirrhotic livers with hepatocellular carcinoma for the presence of EBV, HCV and hepatitis B virus (HBV) infection. RNA prepared from tumor samples were used as a template for reverse transcription followed by double-nested PCR with primers for the 5′ untranslated region (NT) of HCV. DNA extracts of tumor samples were tested by single polymerase chain reaction for the detection of EBV and HBV (X- and/or S-gene) DNA sequences. To control for nucleic acid integrity, all tumor samples were amplified for human β-globin DNA by polymerase chain reaction and subjected to Southern blot hybridization. None of the cases was found to be positive for EBV. Ten HCC cases (32%) tested positive for HCV and 12 HCC cases (38%) tested positive for HBV. Six of the surveyed patients had nucleic acids of both HCV and HBV in their tumor tissue. All HCC tumor samples were positive for β-globin. Our study shows that HCV and HBV infections, but not EBV infection, are associated with hepatocarcinogenesis in non-cirrhotic livers. Other unknown risk factors seem to be in effect in the development of hepatocellular carcinoma in non-cirrhotic livers. [Copyright &y& Elsevier]

Background: It is known fact that pre-oxygenation with positive end-expiratory pressure (PEEP) improves the Partial pressure of oxygen (PO (2)). In this regard not many studies have been done in pregnant women to know its effect on foetus. In this randomised double blind controlled study, we analysed effect of pre-oxygenation with PEEP during caesarean section on foetal umbilical venous PO (2). PATIENTS #ENTITYSTARTX00026;, Methods: 40 term pregnant women, ASA I or II, undergoing elective Caesarean section under spinal anaes-thesia were randomly divided into PEEP and Non-PEEP groups of 20 each. PEEP group received oxygen flow of 6 L/minute with PEEP of 5 cmH (2)O using a modified Mapleson A circuit with fixed unidirectional PEEP valve at the expiratory port during pre-oxygenation and Non PEEP group received same fresh gas flow of oxygen using same breathing circuit without PEEP. Maternal arterial blood samples were collected before applying PEEP and at the end of 5 minutes of facemask application for oxygen analysis. Immediately after baby was delivered umbilical venous samples were taken for blood gas analysis., Results: Both groups were comparable in terms of maternal baseline oxygen saturation (Spo (2)) and base line Po (2). After 5 minutes PO (2)was higher in PEEP Group than non PEEP group (491.65 + 49.96 vs. 452.08 + 77.61). Umbilical venous Po (2)in PEEP group was higher than non PEEP group (34.22 + 6.50 vs. 28.29 + 6.10 mm of Hg)., Conclusion: Application of PEEP during pre-oxygenation for spinal anaesthesia can increase foetal umbilical venous PO(2).