Ribavirin (RBV), a guanosine analogue, has been suggested to exert an antiviral action against hepatitis C virus (HCV) by causing lethal mutations and suppressing RNA polymerase in vitro, but the mechanism of its clinical therapeutic effects is currently unknown. To test the hypothesis that RBV could act both as an RNA mutagen and inhibit viral RNA synthesis in vivo, we studied the evolution of the nucleotide sequences of HCV RNA at the nonstructural (NS) 5B region in patients receiving RBV, placebo, or interferon alfa (IFN-alpha) monotherapy. The RBV group showed a slightly more accelerated evolution rate of HCV RNA quasispecies than either the IFN-alpha or placebo group. RBV caused preferentially A-to-G and U-to-A mutations. Interestingly, an NS5B amino acid 415 Phe-to-Tyr (F415Y) mutation emerged in all (5 of 5) patients infected with HCV genotype 1a during the RBV treatment. Subsequently, the parental 415F strain reemerged in some patients after the treatment was discontinued. The effect of the amino acid substitution at NS5B415 on HCV RNA replication was then investigated using an HCV subgenomic replicon in Huh7 cells. We showed that treatment of replicon cells with RBV reduced the HCV RNA level of NS5B415F replicon, but not NS5B415Y, in a dose-dependent manner. Thus, NS5B F415Y mutation represents an RBV-resistant variant. The 3-dimensional modeling and structure analysis of NS5B protein revealed that the 415th amino acid is located at the P helix region of the thumb subdomain, which may interact with the minor groove of the template-primer duplex in the putative RNA-binding cleft. In conclusion, RBV could work as a weak mutagen for HCV RNA in HCV-infected patients. Furthermore, the selection of an RBV-resistant variant with a single amino acid substitution in NS5B suggested that RBV may directly interact with HCV RNA polymerase, thus interfering with its enzymatic activity.

The effect of initial electrolyte loading (IEL) on cell performance in a coin-type molten carbonate fuel cell (MCFC) was investigated in this work. Since the material of MCFC depends on the manufacturer, optimisation requires experimental investigation. In total, four IEL values, 1.5, 2.0, 3.0, and 4.0 g, were used, corresponding to a pore filling ratio (PFR) of 38, 51, 77, and 102%, respectively. The cell performance with respect to the PFR was analysed via steady-state polarisation, step-chronopotentiomtery, and impedance methods. The electrochemical analyses revealed that internal resistance and overpotential of the cell decreased with increasing PFR, and a large overpotential was observed when the PFR was 102%, probably due to the flooding phenomenon. After operation, cross-section of the cell was analysed via surface analysis of SEM and EDS methods, and the remaining electrolyte was estimated by dissolution of the cell in 10 wt% acetic acid. A linear relationship between IEL and the weight reduction ratio by dissolution was obtained. Thus, the remaining amount of electrolyte could be measured after operation. The results of SEM and EDS showed that a PFR of 38 and 102% showed a lack and flooding of electrolytes at the cell, respectively, which led to a large overpotential. This work reports that MCFC performance is allowed only in the narrow range of PFR.

The effect of initial electrolyte loading (IEL) on cell performance in a coin-type molten carbonate fuel cell (MCFC) was investigated in this work. Since the material of MCFC depends on the manufacturer, optimisation requires experimental investigation. In total, four IEL values, 1.5, 2.0, 3.0, and 4.0 g, were used, corresponding to a pore filling ratio (PFR) of 38, 51, 77, and 102%, respectively. The cell performance with respect to the PFR was analysed via steady-state polarisation, step-chronopotentiomtery, and impedance methods. The electrochemical analyses revealed that internal resistance and overpotential of the cell decreased with increasing PFR, and a large overpotential was observed when the PFR was 102%, probably due to the flooding phenomenon. After operation, cross-section of the cell was analysed via surface analysis of SEM and EDS methods, and the remaining electrolyte was estimated by dissolution of the cell in 10 wt% acetic acid. A linear relationship between IEL and the weight reduction ratio by dissolution was obtained. Thus, the remaining amount of electrolyte could be measured after operation. The results of SEM and EDS showed that a PFR of 38 and 102% showed a lack and flooding of electrolytes at the cell, respectively, which led to a large overpotential. This work reports that MCFC performance is allowed only in the narrow range of PFR.

This study evaluated the performance characteristics of varied electrolyte species and amounts in a molten carbonate fuel cell (MCFC). Coin-type MCFCs were used at the condition of 650 o C and 1 atm. In order to measure the effects of varied electrolyte species and amounts, electrolytes of (Li+K) 2 CO 3 and (Li+Na) 2 CO 3 were selected and the amounts of 1.5 g, 2.0 g, 3.0 g, and 4.0 g were used. Insignificant performance differences were observed in the cell using different electrolytes, but the cell performance was sensitive to the amount of the electrolyte used. The pore-filling ratio (PFR), a ratio of pore filling in the components by the liquid carbonate electrolytes, was used to determine the optimum performance range. Consequently, 77% PFR demonstrated the optimum performance for both electrolytes. Thus, the MCFC had a permissible but narrow optimum performance range. The remaining amounts of electrolyte in the cells were determined using the weight reduction ratio (WRR) method after several hours of cell operation. The WRR used the relationship between the initial loaded amount of electrolyte and weight reduction of components in 10 wt% acetic acid. The relationships were linear and identical between the two electrolyte species.

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