Implementation of Guideline-Based HBV Reactivation Management in Patients with Chronic HBV Infections of HBsAg or Resolved HBV Infection Undergoing Immunosuppressive Therapy.

INTRODUCTION: Although patients with HBV have a risk of reactivation after immunosuppressive therapy (IST), the status of their risk management is unclear in Japan. This study aims to describe the proportion of patients who received preventive management of HBV reactivation during ISTs in patients with chronic HBV infection of HBsAg or resolved HBV infection. METHOD: A retrospective cohort study was conducted using the JMDC Japanese claims database from April 2011 to June 2021. Patients with HBV infections of HbsAg who received ISTs or patients who had resolved HBV infections who received ISTs were identified from the database and evaluated for appropriate management to prevent HBV reactivation. RESULTS: In total, 6242 eligible patients were identified. The proportions of patients with appropriate HBV reactivation management, stratified by the HBV reactivation risk level of IST, was 43.1% (276/641) for high-risk, 40.2% (223/555) for intermediate-risk and 14.9% (741/4965) for low-risk patients. When the evaluation period for the outcome calculation was shortened from 360 to 180 days, the proportion for high risk increased to 52.7%. The odds ratios of large hospitals for receiving appropriate management were 2.16 (95% CI 1.12-4.44) in the high-risk, 4.63 (95% CI 2.34-10.25) in the intermediate-risk and 3.60 (95% CI 3.07-4.24) in the low-risk patients. CONCLUSION: HBV reactivation management was tailored according to the reactivation risk associated with IST. However, adherence to HBV reactivation management guidelines was sub-optimal, even among high-risk patients. This is especially the case for ensuring smaller-sized medical institutions, highlighting the need for further educational activities.

The study assesses the implementation of guideline-based management of hepatitis B virus reactivation during immunosuppressive therapy in Japan. The appropriate management of hepatitis B virus treatment involves prophylactic nucleos(t)ide analog (NUC) therapy and regular monitoring of hepatitis B virus DNA. This study aims to assess the extent to which these management practices are implemented in a clinical setting in Japan using a retrospective cohort study using the Japanese Medical Claims Database. The analysis identified 6242 eligible patients and identified whether they received appropriate management to prevent hepatitis B virus reactivation based on the level of risk associated with their immunosuppressive therapy. Based on the guidelines, the proportions of patients receiving appropriate reactivation management were 43.1% for high-risk, 40.2% for intermediate-risk and 14.9% for low-risk immunosuppressive therapy patients. Shortening the evaluation period from 360 to 180 days showed an increase in the proportion of high-risk patients to 52.7%, which indicated the potential challenge for continued monitoring after immunosuppressive therapy administration. The study shows that large hospitals present higher odds of patients receiving appropriate management. Overall, adherence to hepatitis B virus reactivation management guidelines was suboptimal, especially in smaller medical institutions, emphasizing the need for additional educational activities.

Real-world safety and effectiveness of tenofovir alafenamide for 144 weeks in Japanese patients with chronic hepatitis B.

Tenofovir alafenamide (TAF), a prodrug of tenofovir, delivers high levels of active drug to hepatocytes and is given in a lower dose than tenofovir disoproxil fumarate (TDF). TAF reduces viral replication in patients with chronic hepatitis B (CHB) similar to TDF and has shown a lower risk of the renal and bone toxicities associated with TDF use. This post-marketing surveillance study examined the safety and effectiveness of TAF in treatment-naïve and -experienced CHB patients who received TAF for 144 weeks at real-world clinical sites in Japan. Safety assessments included the incidence of adverse drug reactions (ADRs), renal and bone events, and changes in selected laboratory parameters. Effectiveness was based on the proportion of patients with HBV DNA levels below the lower limit of quantitation or <29 IU/mL. This analysis included 580 patients; 18.4% of whom were treatment-naïve. The cumulative incidence of ADRs was 0.21 per 100 person-months, and the incidence of serious ADRs was 0.01 (95% CI, 0.00-0.04) per 100 person-months. There were no ADRs of declines in estimated glomerular filtration rates, renal failure or proximal tubulopathy. The most common ADR was hypophosphataemia in seven (1.2%) patients. Two (0.4%) patients each had decreased blood phosphorus, bone mineral density decreased, dizziness and alopecia. Overall, the proportion of virologically suppressed patients increased from 68.8% at baseline to 97.5% at Week 144. These results confirm the real-world safety and effectiveness of TAF in Japanese patients with CHB and are consistent with the findings of other evaluations of the safety and efficacy of TAF in CHB.

Case fatality rate considering the lag time from the onset of COVID-19 infection to related death from 2020 to 2022 in Japan.

Importance: On an ecological scale, the lag time between coronavirus disease 2019 (COVID-19) infection and related fatality has varied between epidemic waves and prefectures in Japan. The variability in lag time across areas of Japan during the seven distinct waves can help derive a more appropriate estimation of the weekly confirmed case fatality rate (CFR) of COVID-19. Objective: To estimate the 7-day moving average CFR across area block levels in Japan from February 2020 to July 2022 using the lag time between COVID-19 infection and related fatality. Main outcomes and measures: The 7-day moving average CFR of COVID-19 for area blocks in Japan considering the lag time between infection and death (total and subgroup analysis of elderly). Results: Lag time was found to vary substantially among prefectures in Japan from the first wave to the seventh wave of the COVID-19 epidemic. The estimated 7-day moving average CFR based on the lag time reflects the Japanese COVID-19 pandemic and related policy interventions (e.g. vaccination of elderly people) rather than other standard CFR estimations. Conclusions and relevance: The variation in estimated lag time across prefectures in Japan for different epidemic waves indicates that it is inadequate to use the clinical results of the period from the start of infection to death for evaluation of the ecological scale of the CFR. Moreover, the lag time between infection and related fatality was found to be either shorter or longer than the clinically reported period. This revealed that preliminary reports of CFR may be overestimated or underestimated, even if they consider the lag based on clinical reports.

Novel indicator for the spread of new coronavirus disease 2019 and its association with human mobility in Japan.

The Japanese government adopted policies to control human mobility in 2020 to prevent the spread of severe acute respiratory syndrome coronavirus 2, which causes coronavirus disease 2019 (COVID-19). The present study examined the impact of human mobility on COVID-19 cases at the prefectural level in Japan by devising an indicator to have a relationship between the number of infected people and on human mobility. We calculated origin-destination travel mobility within prefectures in Japan from March 1st to December 31st, 2020, using mobile phone data. A cross-correlation function (CCF) was used to examine the relationship between human mobility and a COVID-19 infection acceleration indicator (IAI), which represents the rate of change in the speed of COVID-19 infection. The CCF of intraprefectural human mobility and the IAI in Tokyo showed a maximum value of 0.440 at lag day 12, and the IAI could be used as an indicator to predict COVID-19 cases. Therefore, the IAI and human mobility during the COVID-19 pandemic were useful for predicting infection status. The number of COVID-19 cases was associated with human mobility at the prefectural level in Japan in 2020. Controlling human mobility could help control infectious diseases in a pandemic, especially prior to starting vaccination.

Real-world Safety and Effectiveness of 24-week Sofosbuvir and Ribavirin Treatment in Patients Infected with Rare Chronic Hepatitis C Virus Genotypes 3, 4, 5, or 6 in Japan.

Objectives Real-world evidence on the safety and effectiveness of direct-acting antivirals in patients infected with chronic hepatitis C virus (HCV) genotypes (GTs) 3, 4, 5, or 6 in Japan is limited. This prospective observational study assesses the real-world safety profile and treatment effectiveness among patients prescribed sofosbuvir with ribavirin (SOF+RBV) for HCV GT3-6 infection in Japan. Methods Adults receiving 24-week SOF+RBV treatment for HCV GT3-6 infection were prospectively enrolled and observed through 24 weeks post-treatment for treatment-emergent adverse events (AEs) considered related to SOF and/or RBV by treating physicians and for a sustained virologic response at 12 and 24 weeks post-treatment (SVR12, SVR24). Incidence rates of related AEs and serious AEs (SAEs) were calculated. Proportions of patients experiencing related AEs/SAEs and those achieving SVR12 and SVR24 were assessed overall and by baseline characteristics, including treatment experience and cirrhosis status. Results Among the 50 patients included in the safety analysis, 92% had GT3 infection. The incidence rates of related AEs and SAEs were low overall (1.52 and 0.25 per 100 person-weeks, respectively), with 6.0% and 14.0% patients experiencing AEs related to SOF or RBV, respectively. There were no marked differences in the occurrence of related AEs/SAEs by patient baseline characteristics. SVR12 and SVR24 were achieved in 83.7% (41/49) and 82.2% (37/45) of patients, respectively. Lower effectiveness was observed among treatment-experienced patients and patients with cirrhosis at baseline. Conclusion This study demonstrated that SOF+RBV treatment for HCV GT3-6 infection was safe, effective, and an important treatment option for this difficult-to-treat patient population in Japan.

The impact of declaring the state of emergency on human mobility during COVID-19 pandemic in Japan.

Background/objectives: Japan has responded to the spread of COVID-19 through declaration of a state of emergency to regulate human mobility. Although the declaration was enforced by the government for prefectures, there is limited evidence as to whether the public complied with requests for voluntary stay at home. In this study, we evaluated the impact of declaring a state of emergency on human mobility during the COVID-19 pandemic in Japan. Methods: We utilized daily human mobility data for 47 prefectures in Japan. Data were collected via mobile phone from February 1, 2020 to April 30, 2021. Difference-in-difference analysis was utilized to estimate the effects of the declaration of a state of emergency on prefectures in the Tokyo Metropolitan Area (Tokyo, Kanagawa, Saitama, and Chiba) in comparison to other prefectures where the state of emergency was first lifted (Osaka, Hyogo, Fukuoka, and Aichi). Results: Human mobility was suppressed during the second state of emergency, from January 8 to March 21, 2021. However, the impact was weaker for the second state of emergency compared to the first. Conclusion: In Japan, government requests for stay at home, such as the declaration of a state of emergency, were temporarily able to control human mobility. However, the second state of emergency was not as effective as the first. If additional need to regulate human mobility arises, self-restraint with stronger enforcement should be considered.

Preliminary investigation of aluminium fluoride complexes in aqueous solutions with capillary electrophoresis coupled with electrospray ionization mass spectrometry and with inductively coupled plasma mass spectrometry.

RATIONALE: It is crucial to identify and confirm the original species of aluminium ions (Al3+ ) dissolved in water, since they behave differently. Depending on their species, the toxicity differs. Capillary electrophoresis (CE) coupled with electrospray ionization mass spectrometry (ESI-MS) and CE coupled with inductively coupled plasma mass spectrometry (CE/ICP-MS) were explored to identify and determine simple systems of Al species solutions at pH 3.0. METHODS: The new combinations of techniques, namely, ESI-MS coupled with CE for identification of species and ICP-MS coupled with CE for confirmation, were applied to for the analyses of Al and fluoride (F) solutions. RESULTS: Al monomers, some Al dimers and trimers were detected by CE/ESI-MS. CE/ICP-MS experiments were conducted with the assembled interface. As a result, the calibration line showed R2 = 0.9856, and the detection limits were 35 nL and 0.037 µM. The results were compared with data obtained using MINEQL+. CONCLUSIONS: Most of the Al species detected were monomers; some dimers and trimers were detected by CE/ESI-MS, but they were not detected by CE-ICP-MS, probably owing to extremely low concentrations. The Al speciation technique was improved by CE/ESI-MS, and the Al species present at extremely low concentrations were ascertained by CE/ICP-MS. The use of coupled instruments will be one of the most powerful tools for identifying dissolved metal ions.

Detection of microsatellite alterations in plasma DNA of malignant mucosal melanoma using whole genome amplification.

Malignant mucosal melanoma (MMM) still has the poorest prognosis. There is a paucity of molecular biological studies on MMM of the head and neck. We investigated free-circulating DNA microsatellites with loss of heterozygosity (LOH) in the blood of MMM patients. Cancer-related DNA is found in plasma, with cancer patients showing a higher level of free-circulating DNA than normal subjects. However, it is difficult to obtain sufficient amounts of such DNA for PCR analysis. We have searched for ways to improve all stages of such research, and detected new microsatellite alterations by triplicated whole genome amplification (WGA) and triplicated PCR amplification. In order to achieve a better understanding of the extent of the alterations affecting chromosomes we determined the occurrence of LOH at the following gene loci: D1S243, D6S311, D9S161, and D19S246; only 4 out of the 20 microsatellite markers usually used in MMM were used in this study. We determined LOH in 17 MMM patients.It was possible to confirm LOH on at least one marker in 12 (70.6%) out of the 17 patients. Metastasis or recurrence was confirmed in 3 (17.6%) out of the 17 patients, and all of them were found to have LOH. LOH at microsatellite markers D1S243, D6S311, D9S161 and D19S246 in the plasma of these patients statistically correlated with MMM.The results of this study suggest that these loci are suitable for identifying cancerrelated DNA of MMM, and that analysis of LOH in plasma DNA released into the circulation may be useful as a screening tool.

LOH analysis of free DNA in the plasma of patients with mucosal malignant melanoma in the head and neck.

BACKGROUND: We have conducted charged-particle radiotherapy for mucosal malignant melanoma (MMM) in the head and neck, using carbon ion beams. However, even with the use of carbon ion radiotherapy that is characterized by high local tumor control, a significant number of patients develop metastases after therapy. We conducted research on the assumption that, in MMM, the detection of loss of heterozygosity (LOH) from free DNA in the circulating plasma may be of practical use in the diagnosis of recurrence and metastasis. METHODS: We took blood samples prior to therapy from 17 patients with MMM in the head and neck, and extracted free DNA in plasma. Four types of microsatellite markers were used for LOH detection. RESULTS: LOH was detected in 1 of 5 patients (20%) for D1S243, 2 of 5 patients (40%) for D6S311, 11 of 17 patients (65%) for D9S161, and 1 of 6 patients (17%) for D19S246. CONCLUSION: Evidence based on the irradiated tumor volume suggested a tendency for the group of patients found to have LOH in two loci to have a larger mean tumor volume than the patient groups with no detectable LOH or with LOH detectable in only one locus. Of the 17 patients in this study 4 patients had recurrence and/or metastasis, and all 4 of these patients were found to have LOH in at least one or more loci for any region. LOH analysis of free DNA in plasma may be useful for the early diagnosis of MMM.

Specificity and similarity of functions of the Aux/IAA genes in auxin signaling of Arabidopsis revealed by promoter-exchange experiments among MSG2/IAA19, AXR2/IAA7, and SLR/IAA14.

As indicated by various and some overlapped phenotypes of the dominant mutants, the Aux/IAA genes of Arabidopsis (Arabidopsis thaliana) concomitantly exhibit a functional similarity and differentiation. To evaluate the contributions of their expression patterns determined by promoter activity and molecular properties of their gene products to Aux/IAA function, we examined phenotypes of transgenic plants expressing the green fluorescent protein (GFP)-tagged msg2-1/iaa19, axr2-1/iaa7, or slr-1/iaa14 cDNA by the MSG2 or AXR2 promoter. When driven by the MSG2 promoter (pMSG2), each GFP-tagged cDNA caused the msg2-1 phenotype, that is, the wild-type stature in the mature-plant stage, long and straight hypocotyls in the dark, reduced lateral root formation, relatively mild agravitropic traits in hypocotyls, and a normal gravitropic response in roots. However, development of one or two cotyledonary primordia was often arrested in embryogenesis of the pMSG2::axr2-1::GFP and pMSG2::slr-1::GFP plants, resulting in monocotyledonary or no cotyledonary seedlings. Such defects in embryogenesis were never seen in pMSG2::msg2-1::GFP or the msg2-1, axr2-1, or slr-1 mutant. The MSG2 promoter-GUS staining showed that expression of MSG2 started specifically in cotyledonary primordia of the triangular-stage embryos. When driven by the AXR2 promoter (pAXR2), each GFP-tagged mutant cDNA caused, in principle, aberrant aboveground phenotypes of the corresponding dominant mutant. However, either the axr2-1::GFP or slr-1::GFP cDNA brought about dwarf, agravitropic stems almost identical to those of axr2-1, and the pAXR2::msg2-1::GFP and pAXR2::slr-1::GFP hypocotyls exhibited complete loss of gravitropism as did axr2-1. These results showed functional differences among the msg2-1, axr2-1, and slr-1 proteins, though some phenotypes were determined by the promoter activity.

Inhibition of brassinosteroid biosynthesis by either a dwarf4 mutation or a brassinosteroid biosynthesis inhibitor rescues defects in tropic responses of hypocotyls in the arabidopsis mutant nonphototropic hypocotyl 4.

The nonphototropic hypocotyl 4 (nph4)/auxin response factor 7 (arf7) mutant of Arabidopsis (Arabidopsis thaliana) is insensitive to auxin and has defects in hypocotyl tropism, hook formation, differential leaf growth, and lateral root formation. To understand an auxin-signaling pathway through NPH4, we carried out screening of suppressor mutants of nph4-103 and obtained a dwarf suppressor mutant, suppressor of nph4 (snp2). snp2 had short hypocotyls in the dark condition and dark green and round leaves, short petioles, and more lateral shoots than the wild type in the light condition. The snp2 phenotypes were rescued by adding brassinolide to the growth medium in both light and dark conditions. Genetic mapping, sequence analysis, and a complementation test indicated that snp2 was a weak allele of DWARF4 (DWF4), which functions in brassinosteroid (BR) biosynthesis. snp2, which was renamed dwf4-101, exhibited photo- and gravitropisms of hypocotyls similar to those of the wild type with a slightly faster response in gravitropism. dwf4-101 almost completely suppressed defects in both tropisms of nph4-103 hypocotyls and completely suppressed hyponastic growth of nph4-103 leaves. Treatment with brassinazole, an inhibitor of BR biosynthesis, also partially rescued the tropic defects in nph4-103. Hypocotyls of nph4-103 were auxin insensitive, whereas hypocotyls of dwf4-101 were more sensitive than those of the wild type. dwf4-101 nph4-103 hypocotyls were as sensitive as those of dwf4-101. Auxin inducibility of massugu 2 (MSG2)/IAA19 gene expression was reduced in nph4-103. mRNA level of MSG2 was reduced in dwf4-101 and dwf4-101 nph4-103, but both mutants exhibited greater auxin inducibility of MSG2 than the wild type. Taken together, dwf4-101 was epistatic to nph4-103. These results strongly suggest that BR deficiency suppresses nph4-103 defects in tropic responses of hypocotyls and differential growth of leaves and that BR negatively regulates tropic responses.

Detection of tumor DNA in plasma using whole genome amplification.

Altered microsatellite DNA in the blood of cancer patients may provide a novel means for tumor detection. Such alterations are a major characteristic of many types of tumor especially those associated with head or neck cancer. Moreover, recent evidence suggests that senescent tumor cells release DNA into the circulation, which is subsequently carried by the blood and thus enriched in the serum and plasma. We tested 10 head and neck cancer patients (5 with malignant melanomas (MM) and 5 with adenoid cystic carcinomas (ACC)) by polymerase chain reaction (PCR)-based microsatellite analysis of DNA from white blood cells and paired plasma samples. Our goal was to amplify two microsatellite markers, D1S243 and D19S246, which sometimes show microsatellite alterations in head and neck cancer patients. However amplification of fragments from three loci in the plasma samples proved impossible, probably due to the small amounts of DNA isolated. We used multiple displacement amplification (MDA) to amplify genomic DNA from the plasma samples. Two microsatellite fragments were amplified from whole genome amplified DNA. Among 5 heterozygote samples, 3 showed the same pattern in DNA samples from both blood cells and plasma but 2 showed loss of heterozygosity (LOH). Although further study is necessary to confirm whether the LOH found in this study reflects alteration in circulating tumor cell DNA, application of whole genome amplification may allow DNA analysis from limited amounts of such DNA and provide a minimally invasive diagnostic procedure and useful aid in therapy.