Worldwide, ~71 million people are infected with hepatitis C virus (HCV). Claudin-1 (CLDN1) and occludin (OCLN), both tetraspanins of epithelial tight junctions, are entry receptors for HCV. Previously, we generated anti-CLDN1 and anti-OCLN monoclonal antibodies (mAbs), both of which strongly inhibit HCV entry into hepatocytes. However, the relevance of CLDN1 and OCLN in persistent HCV infection remains unclear. In the present study, we evaluated the involvement of CLDN1 and OCLN in persistent HCV infection using the mAbs against CLDN1 (clone 3A2) and OCLN (clone 1-3). Interestingly, both mAbs significantly reduced intracellular HCV RNA levels in a cell culture system. Additionally, the anti-OCLN mAb reduced serum HCV levels in chronic HCV-infected human liver chimeric (PXB) mice (often used as an in vivo HCV infection model), whereas the anti-CLDN1 mAb did not have this effect. These results suggest that the OCLN molecule contributes to maintaining persistent HCV infection in vivo. In further investigation, we determined whether combinations of NS5B inhibitor, nesbuvir, and the anti-OCLN mAb had anti-HCV effects on persistent HCV-infected PXB mice. Administration of nesbuvir and control IgG caused a breakthrough of serum HCV levels in all mice, whereas nesbuvir and anti-OCLN mAb combinations caused the breakthrough at a later phase in only one of three mice. Thus, anti-OCLN mAb seems to suppress the occurrence of resistant viruses against nesbuvir. Based on these results, we suggest that the anti-OCLN mAb, which could be combined with direct-acting antiviral agents, might be a potential candidate antiviral agent in HCV therapeutics.
Miso is a traditional Japanese fermented food made by fermenting steamed soybean with koji (fermented cereals with Aspergillus oryzae). Many types of miso are produced in Japan, including miso with rice and/or barley depending on the region where it is produced. In this study, we used 1H NMR metabolomic analysis to compare the characteristics of the components (metabolites) of miso with different ingredients. Three types of miso were compared: soybean miso, rice miso, and barley miso. After measuring the 1H NMR of the aqueous solution of each miso, multivariate analysis of the spectral integration data was performed to compare the characteristic metabolites. Principal component analysis (PCA) showed a separation between soybean miso and rice, barley miso. Orthogonal Projection to Latent Structure Discriminant Analysis (OPLS-DA) extracted ethanol, saccharides such as glucose, and amino acids as metabolites contributing to the separation. Ethanol and glucose were higher in rice and barley miso, especially glucose in barley miso and ethanol in rice miso. Soybean miso was characterized by its high content of amino acids, including branched-chain amino acids. It was suggested that the characteristics of these ingredients were influenced not only by differences in ingredients, but also by the fermentation period and other factors. Although the number of metabolites that can be analyzed by 1H NMR metabolomic analysis is smaller than that by GC/MS or LC/MS, it does not require any pretreatment and is easy to measure, so it can be applied to the comparison of food components and quality control, as in the analysis of miso components.
N-methyl-D-aspartate (NMDA) receptor has important role in synapse function and neurotransmission by the high permeability of Ca2+. In Alzheimer’s disease (AD), NMDA mediated neurotransmission is impaired by decreasing NMDA receptor subunits and increasing glutamate, which is an NMDA receptor ligand. Therefore, the NMDA antagonist (memantine) is used as a therapeutic drug of AD. Huperzia serrata is a traditional Chinese herbal medicine, and the constitute huperzine A has been reported to inhibit NMDA receptor. In this study, we clarified the effect of Huperzia serrata and the constitute huperzine A on MK-801-induced cognitive dysfunction. Memantine, Huperzia serrata and huperzine A were administered orally once a day and Y-maze test was performed at day 7 to investigate cognitive function. After Y-maze test, mice brains were collected and evaluated the expression levels of glutamate receptors and Ca2+ signalling associated protein. Memantine (5 mg/kg, p.o.), Huperzia serrata (1000 mg/kg, p.o.) and huperzine A (0.7 mg/kg, p.o.) were improved collect alternation behaviour in Y-maze test. Treatment of memantine, Huperzia serrata and huperzine A also improved the total arm entries increased by MK-801. The expression level of NMDA receptor subunit NR2A was increased by Huperzia serrata and huperzine A treatment. In addition, the decreased expression level of PKCα and phosphorylation of Erk1/2 by MK-801 were improved. These results suggest that Huperzia serrata and the constitute huperzine A improve cognitive dysfunction through NMDA receptor function and glutaminergic signaling pathway.
ICR and C57BL/6J mice have been widely used in several research fields. The reproductive toxicology parameters, such as fertilization rate, which may differ between the two strains, are well known. However, the details of the sperm quality parameters are not well known. To reveal these, we compared the sperm morphology of the two strains. Eosin-stained sperm smears from adult ICR and C57BL/6J mice were analyzed. We observed that 79.6 ± 1.2 and 49.5 ± 1.7% of ICR and C57BL/6J mice sperm, respectively, showed a normal form. Furthermore, abnormal sperm samples were classified into ten types based on their defective sites. The percentage of abnormal sperm with an amorphous head, bent head, no head, hairpin loop, short tail, and two tails in ICR mice was significantly lower than that in C57BL/6J mice. In contrast, the percentage of coil-tailed sperm in ICR mice was significantly higher than that in C57BL/6J mice. These results suggest that C57BL/6J mice have a limited ability to remove the cytoplasm during spermiation and ICR mice have fewer sperm abnormalities than C57BL/6J mice. The characteristics of male reproductive traits among mouse strains should be taken into consideration in sperm analysis, as the negligence of this could generate an increased potential for a misleading in toxicology evaluation.
The gastrointestinal absorption of heavenly blue anthocyanin derived from morning glory (Pharbitis nil L.) was examined in rats. Ingested heavenly blue anthocyanin was directly absorbed from the gastrointestinal tract and detected in its original polyacylated form in rat blood plasma. The maximum plasma concentration and area under the plasma concentration curve during 8-h post-administration after a single oral dose of 0.0569 mmol/kg heavenly blue anthocyanin were 0.143 ± 0.023 μM and 23.30 ± 3.76 μM·min, respectively. Heavenly blue anthocyanin, which contains asymmetrical branched chains with glucosyl terminals, was absorbed at a similar level to that of total ternatin. By contrast, the plasma amount of heavenly blue anthocyanin was approximately 6-fold higher than that in a previous report of peonidin 3-O-β-(6′′-O-caffeoyl)-sophoroside-5-O-β-D-glucopyranoside with an attached caffeoyl terminal on 6′′-position.
The impact of renal impairment on the drug-drug interaction between azathioprine and allopurinol that causes myelosuppression and hepatotoxicity remains unclear. This case series study investigated adverse effects caused by azathioprine owing to drug-drug interaction considering renal impairment. Patients who started the combination therapy of azathioprine and allopurinol at Mie University Hospital between January 2013 and February 2021 were enrolled. The outcome of adverse events associated with azathioprine was assessed according to Common Terminology Criteria for Adverse Events version 5.0. The Drug Interaction Probability Scale was used to determine the probability of drug-drug interaction. Of the three patients, two were identified as exhibiting drug-drug interaction with the Drug Interaction Probability Scale > 5 points. They experienced grade 3 myelosuppression or hepatotoxicity with fatigue, after initiation of azathioprine (1.28 and 0.44 mg/kg once daily) and allopurinol (50 mg once daily). They received appropriate dose-adjusted allopurinol according to renal function. Additionally, both patients had the estimated glomerular filtration rate < 60 mL/min/1.73 m2. Thus, renal impairment might reduce the excretion of oxypurinol, an active metabolite of allopurinol, which certainly enhances the side effects of azathioprine.