Increased neutralization potency and breadth elicited by a SARS-CoV-2 mRNA vaccine forming virus-like particles.

Vaccines have played a fundamental role in the control of infectious diseases. We previously developed a messenger RNA (mRNA) vaccine against HIV-1 that forms virus-like particles (VLPs) through coexpression of the viral envelope with Gag. Here, we applied the same principle to the design of a VLP-forming mRNA vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To promote cognate interaction with simian immunodeficiency virus (SIV) Gag, we engineered different chimeric proteins encompassing the ectodomain and the transmembrane region of the SARS-CoV-2 Spike protein from the Wuhan-Hu-1 strain fused to the gp41 cytoplasmic tail of either HIV-1 (strain WITO) or SIV (strain mac239) with or without a partial truncation at amino acid 745 to enhance membrane expression. Upon cotransfection with SIV gag mRNA, the Spike-SIVCT.745 (SSt) chimera yielded the highest level of cell-surface expression and extracellular VLP release. Immunization of BALB/c mice with SSt+gag mRNA at 0, 4, and 16 wk induced higher titers of Spike-binding and autologous neutralizing antibodies at all time points compared to SSt mRNA alone. Furthermore, mice immunized with SSt+gag mRNA developed neutralizing antibodies effective against different variants of concern. These data demonstrate that the Gag/VLP mRNA platform can be successfully applied to vaccines against different agents for the prevention of infectious diseases of global relevance.

A naphthyl thiourea-based effective chemosensor for fluorescence detection of Ag+ and Zn2.

A naphthyl thiourea-based effective chemosensor HNC, (E)-2-(2-hydroxy-3-methoxybenzylidene)-N-(naphthalen-1-yl)hydrazine-1-carbothioamide, was synthesized. HNC showed quick responses toward Ag+ and Zn2+ through marked fluorescence turn-on in different solvent conditions, respectively. Binding proportions of HNC to Ag+ and Zn2+ were found to be 2:1 and 1:1, respectively. Detection limits of HNC for Ag+ and Zn2+ were calculated as 3.82 and 0.21 µM. Binding processes of HNC for Ag+ and Zn2+ were represented using Job's plot, DFT, 1 H NMR titration, and ESI-MS.

Environmental impacts and consumer preference for sustainably cultivated Japanese mustard spinach, komatsuna.

Sustainable agriculture is spreading in Japan in response to growing concerns about the environmental burden of the agriculture sector, but less than 1% of the total crop area for each vegetable in Japan is grown sustainably. Environmentally friendly agricultural products are produced by using organic and low-input farming techniques; low-input farming aims to reduce chemical inputs, such as fertilizers, pesticides, and herbicides, by half. Here, we used komatsuna (Japanese mustard spinach, Brassica rapa var. perviridis) as a model vegetable to study the environmental impact of low-input farming and ways to promote the purchase of organically and low-input farmed vegetables. We first assessed greenhouse gas emissions resulting from organic, low-input, and conventional farming of komatsuna. We also evaluated the effectiveness of providing consumers with detailed farm management and seasonality information to market organically and low-input farmed vegetables. We estimated marginal willingness-to-pay (MWTP) using choice-based conjoint analysis, based on attributes of price, fertilizer use, pesticide use, and region of origin. For seasonality, the questionnaire incorporating these attributes was conducted twice: once assuming purchasing in season, the other out of season. The greenhouse gas emissions of organic farming per area (196.7 kg CO2-eq/10 a/year) and per yield (72.3 kg CO2-eq/t/year) were less than those of low-input (322.6 kg CO2-eq/10 a/year, 120.7 kg CO2-eq/t/year) and conventional (594.0 kg CO2-eq/10 a/year, 220.7 kg CO2-eq/t/year) farming. MWTPs were highest for pesticide-free komatsuna (76.9 yen out of season, 66.2 yen in season), followed by full organic fertilizer (66.0 yen out of season, 63.4 yen in season), half organic fertilizer (35.8 yen out of season, 19.8 yen in season), and half pesticide (29.2 yen out of season, 21.0 yen in season). Consumers showed greater preference for organically and low-input farmed komatsuna out of season than in season. Consumers were more interested in pesticide information than in fertilizer and region of origin information. Our findings suggest that providing detailed cultivation and seasonality information would be a beneficial consumer communication tool to increase the market for sustainable agricultural products.

Regional Variation in National Healthcare Expenditure and Health System Performance in Central Cities and Suburbs in Japan.

The increasing national healthcare expenditure (NHE) with the aging rate is a significant social problem in Japan, and efficient distribution and use of NHE is an urgent issue. It is assumed that comparisons in subregions would be important to explore the regional variation in NHE and health system performance in targeted municipalities of the metropolitan area of Tokyo (central cities) and the neighboring municipalities of Chiba Prefecture (suburbs). This study aimed to clarify the differences of the socioeconomic factors affecting NHE and the health system performances between subregions. A multiple regression analysis was performed to extract the factors affecting the total medical expenses of NHE (Total), comprising the medical expenses of inpatients (MEI), medical expenses of outpatients (MEO), and consultation rates of inpatients (CRI) and outpatients (CRO). Using the stepwise method, dependent variables were selected from three categories: health service, socioeconomic, and lifestyle. Then, health system performance analysis was performed, and the differences between regions were clarified using the Mann-Whitney U test. The test was applied to 18 indicators, classified into five dimensions referred to in the OECD indicators: health status, risk factors for health, access to care, quality of care, and health system capacity and resources. In the central cities, the number of persons per household was the primary factor affecting Total, MEI, MEO, and CRO, and the number of persons per household and the percentage of the entirely unemployed persons primarily affected CRI. In the suburbs, the ratio of the population aged 65-74 and the number of hospital beds were significantly positively related to Total, MEI, and CRI, but the number of workers employed in primary industries was negatively related to Total and MEI. The ratio of the population aged 65-74 was significantly positively related to MEO and CRO. Regarding health system performance, while risk factors for health was high in the central cities, the others, including access to care, quality of care, and health system capacity and resources, were superior in the suburbs, suggesting that the health system might be well developed to compensate for the risks. In the suburbs, while risk factors for health were lower than those in the central cities, access to care, quality of care, and health system capacity and resources were also lower, suggesting that the healthcare system might be poorer. These results indicate a need to prioritize mitigating healthcare disparities in the central cities and promoting the health of the elderly in the suburbs by expanding the suburbs' healthcare systems and resources. This study clarified that the determinants of NHE and health system performance are drastically varied among subregional levels and suggested the importance of precise regional moderation of the healthcare system.

A distinct freshwater-adapted subgroup of ANME-1 dominates active archaeal communities in terrestrial subsurfaces in Japan.

Anaerobic methane-oxidizing archaea (ANME) are known to play an important role in methane flux, especially in marine sediments. The 16S rRNA genes of ANME have been detected in terrestrial freshwater subsurfaces. However, it is unclear whether ANME are actively involved in methane oxidation in these environments. To address this issue, Holocene sediments in the subsurface of the Kanto Plain in Japan were collected for biogeochemical and molecular analysis. The potential activity of the anaerobic oxidation of methane (AOM) (0.38-3.54 nmol cm⁻³ day⁻¹) was detected in sediment slurry incubation experiments with a (13) CH(4) tracer. Higher AOM activity was observed in low-salinity treatment compared with high-salinity condition (20‰), which supports the adaptation of ANME in freshwater habitats. The 16S rRNA sequence analysis clearly revealed the presence of a distinct subgroup of ANME-1, designated ANME-1a-FW. Phylogenetic analysis of the mcrA genes also implied the presence of the distinct subgroup in ANME-1. ANME-1a-FW was found to be the most dominant active group in the archaeal communities on the basis of 16S rRNA analysis (75.0-93.8% of total archaeal 16S rRNA clones). Sulfate-reducing bacteria previously known as the syntrophic bacterial partners of ANME-1 was not detected. Our results showed that ANME-1a-FW is adapted to freshwater habitats and is responsible for AOM in terrestrial freshwater subsurface environments.

Current Utilization and Research Status of Traditional East Asian Herbal Medicine Treatment for Multiple Sclerosis: A Scoping Review.

Background: Multiple sclerosis (MS) is a chronic immune-mediated inflammatory disease of the central nervous system that is gradually increasing in prevalence. The etiology of MS remains unknown; however, it is assumed to be caused by a deterioration of autoimmune regulation. Although immunomodulatory agents are a standard treatment option in patients with MS, there is insufficient evidence about their clinical efficacy in symptomatic treatment, and many MS patients resort to complementary and alternative medicine. For this reason, we conducted a scoping review to investigate the current status of the clinical evidence related to traditional East Asian herbal medicine treatment for MS and to inform future research and treatment strategies. Method: A scoping review is an emerging methodology for knowledge synthesis that adopts the Arksey and O'Malley framework. The research question was, "What has been studied about the herbal medicine treatments administered to patients with MS?" Articles published until 2019 were identified in six databases (PubMed, Embase, Cochrane, KoreaMed, NDSL, and OASIS) in March of 2020. Data from the included studies were charted and descriptively analyzed in relation to the study's research questions. Results: Of the 1,445 articles identified, 14 studies were included in this review. Single and serial case reports constituted the majority (42.86%), with 57.14% of studies conducted in China. A total of 20 prescriptions containing 95 herbs were used in the intervention and observational studies. Herbal medicines were effective at improving clinical symptoms of MS and reducing recurrence frequency. The main cause of MS was presumed to be oxidative stress, which enhances inflammation and, consequently, causes neuronal death. Conclusion: Herbal medicines were determined to improve the symptoms of MS and to reduce the frequency of recurrences. This study suggests that herbal medicines are promising and worth pursuing further studies but the state of current evidence is poor. Thus, further, high-quality studies included larger randomized trial are required.

Traditional East Asian Herbal Medicine for Amyotrophic Lateral Sclerosis: A Scoping Review.

This study aimed to analyze and summarize the existing evidence regarding herbal medicine treatments for amyotrophic lateral sclerosis (ALS). Studies on herbal medicine treatment in patients with ALS were searched within English, Chinese, Japanese, and Korean databases up to July 31, 2021. In the selected studies, we collected the following information: the first author, year of publication, country, language, study methodology, sample size, demographic characteristics of the study participants, disease duration, diagnostic criteria, treatment method, treatment periods, evaluation tools, results, and side effects. The organized data were classified and analyzed narratively. This study included 59 studies. The first clinical study on the effect of herbal medicine was published in 1995; moreover, most studies were conducted in China. Among the 59 selected studies, 47.5% were observational studies, including case reports and case series. Moreover, there was one meta-analysis. The El Escorial criteria were the most commonly used diagnostic criterion for ALS; moreover, the ALS functional rating scale was the most common evaluation tool. Buzhongyiqitang, Sijunzitangjiawei, and Jianpiyifeitang were the most commonly used herbal medicines, with anti-inflammatory, protein aggregation, and anti-oxidant effects. There remain evidence of gaps in the effectiveness of herbal medicine for ALS. To allow effective treatment of patients with ALS using herbal medicine, large-scale and rigorously designed high-quality clinical studies should be performed.

Successful treatment of restless leg syndrome with the traditional herbal medicines Dangguijakyak-san and Shihogyeji-tang: A case report (CARE-compliant).

RATIONALE: Dopamine replacement is currently the standard treatment for restless leg syndrome (RLS); however, various adverse effects are associated with long-term therapy, and the benefits disappear upon discontinuation. To overcome these limitations, interest in traditional East Asian medicine has increased. PATIENT CONCERNS: A 72-year-old Asian woman originally admitted for an intracerebral hemorrhage presented with complaints of an unpleasant sensation throughout the body that appeared at night. DIAGNOSES: The patient was diagnosed with chronic persistent RLS based on the 2012 Revised International Restless Leg Syndrome Study Group Diagnostic Criteria. INTERVENTIONS: The patient was treated with extracts of the traditional herbal medicines Dangguijakyak-san (DS) and Shihogyeji-tang (ST). After 47 days of therapy, all herbal medicines were discontinued, and symptoms had not returned by the last follow-up 244 days after the initial treatment. OUTCOMES: One week after initiating herbal treatment with DS and ST, the RLS symptoms began to improve, and the total hours of sleep had increased from 2 to 9 hours by day 21, with a Korean version of the international restless legs scale score of 11 points. On day 36, ST was discontinued, given the continued improvement of symptoms. On day 47, symptoms had disappeared (Korean version of the international restless legs scale score: 0), and sleep disturbances caused by RLS had completely resolved. After day 47, DS was also discontinued. There were no adverse effects associated with the administration of DS and ST, and the symptoms had not recurred by the last follow-up on day 244. LESSONS: In this case, RLS related symptoms, which had been present for approximately 60 years, were improved using only the traditional herbal medicines DS and ST (without dopamine replacement), and no symptoms recurred for 244 days. This case suggests that if replacement therapy is difficult or not desired, herbal medicinal therapies may be an effective alternative. This also suggests that the effect of herbal medicine on RLS might be semi-permanent. Further investigations, including clinical trials, are needed to confirm these effects.

Herbal medicines for the prevention and treatment of cerebral vasospasm after subarachnoid hemorrhage: A protocol for systematic review and meta-analysis.

BACKGROUND: Despite the rapid advances in medical technology, including endovascular interventions and medications, cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) is still one of the major threats to the lives of patients with SAH. In East Asian countries, various types of herbal medicines have been used to treat cerebrovascular diseases, including SAH. In this review, we aim to evaluate the efficacy and safety of herbal medicines for the prevention and treatment of CVS after SAH. METHODS AND ANALYSIS: Seven databases will be searched for relevant studies from inception to the present date "June 2020". Only randomized controlled trials (RCTs) that assess the effect and safety of herbal medicines for the prevention and treatment of CVS after SAH will be included. The methodological quality will be evaluated using the Cochrane risk of bias assessment tool. After selecting the appropriate studies, a meta-analysis of the RCTs will be performed. RESULTS: This study will provide a high-quality synthesis of current evidence of herbal medicines for CVS after SAH. CONCLUSION: Our systematic review will provide evidence to judge whether herbal medicines are effective interventions for patients with CVS after SAH. ETHICS AND DISSEMINATION: Ethical approval is not required, as this study is based on a review of published research. This review will be published in a peer-reviewed journal and disseminated electronically and in print. TRIAL REGISTRATION NUMBER: Research registry reviewregistry923.

Actinomycetal community structures in seawater and freshwater examined by DGGE analysis of 16S rRNA gene fragments.

The actinomycetal community structures in marine and freshwater environments (the Pacific Ocean, East China Sea, Tokyo Bay, and Arakawa River) were investigated by a culture-independent molecular method to clarify spatial and seasonal distributions. Deoxyribonucleic acid (DNA) was extracted from environmental water samples, and a community analysis was carried out on polymerase chain reaction-amplified 16S ribosomal DNA. The amplified DNA fragments were analyzed by denaturing gradient gel electrophoresis (DGGE) and nonmetric multidimensional scaling analysis, followed by sequencing analysis. The actinomycetal community structures were different at each station in the Pacific Ocean, the East China Sea, Tokyo Bay, and Arakawa River, and different populations predominated in each area. There were vertical variations in actinomycetal communities in the Pacific Ocean and East China Sea between the surface and 100-m depth, but communities were similar from 200- to 1,000-m depths. There were also distinct seasonal variations in communities in Tokyo Bay. Phylogenetic analysis of DNA fragments recovered from DGGE bands revealed that most of the predominant actinomycetal strains were uncultured and were quite different from well known culturable strains, such as the Streptomyces, Micromonospora, Microbispora, Salinispora, and Actinoplanes groups. These results suggest that the marine environment is an attractive target for discovering new actinomycetal populations producing bioactive compounds and that sampling depth and season are important considerations for isolating various populations effectively.

Microbial responses using denaturing gradient gel electrophoresis to oil and chemical dispersant in enclosed ecosystems.

Microbial responses to the addition of oil with or without a chemical dispersant were examined in mesocosm and microcosm experiments by using denaturing gradient gel electrophoresis of bacterial ribosomal DNA and direct cell counting. When a water-soluble fraction of oil was added to seawater, increases in cell density were observed in the first 24h, followed by a decrease in abundance and a change in bacterial species composition. After addition of an oil-dispersant mixture, increases in cell density and changes in community structure coincided, and the amount of bacteria remained high. These phenomena also occurred in response to addition of only dispersant. Our results suggest that the chemical dispersant may be used as a nutrient source by some bacterial groups and may directly or indirectly prevent the growth of other bacterial groups.

Effectiveness of lorazepam-assisted interviews in an adolescent with dissociative amnesia: A case report.

To facilitate gathering information during a psychiatric interview, some psychiatrists advocate augmenting the interview using drugs. Rather than barbiturates, benzodiazepines have been used for drug-assisted interviews. Dissociative amnesia is one of the indications for these interviews. Herein, we present the case of a 15-year-old female who was diagnosed as having dissociative amnesia because of conflicts with her friends. She was administered a lorazepam-assisted interview to aid recovery of her memories. In this case, a small dose of lorazepam was sufficient to recover her memories without any adverse effects.