Phospholipase C Produced by Clostridium botulinum Types C and D:Comparison of Gene, Enzymatic, and Biological Activities with Those of Clostridium perfringens Alpha-toxin

Clostridium botulinum type C and D strains recently have been found to produce PLC on egg yolk agar plates. To characterize the gene, enzymatic and biological activities of C. botulinum PLCs (Cb-PLCs), the cb-plc genes from 8 strains were sequenced, and 1 representative gene was cloned and expressed as a recombinant protein. The enzymatic and hemolytic activities of the recombinant Cb-PLC were measured and compared with those of the Clostridium perfringens alpha-toxin. Each of the eight cb-plc genes encoded a 399 amino acid residue protein preceded by a 27 residue signal peptide. The protein consists of 2 domains, the N- and C-domains, and the overall amino acid sequence identity between Cb-PLC and alpha-toxin was greater than 50%, suggesting that Cb-PLC is homologous to the alpha-toxin. The key residues in the N-domain were conserved, whereas those in the C-domain which are important in membrane interaction were different than in the alpha-toxin. As expected, Cb-PLC could hydrolyze egg yolk phospholipid, p-nitrophenylphosphorylcholine, and sphingomyelin, and also exhibited hemolytic activity;however, its activities were about 4- to over 200-fold lower than those of alpha-toxin. Although Cb-PLC showed weak enzymatic and biological activities, it is speculated that Cb-PLC might play a role in the pathogenicity of botulism or for bacterial survival

Promotion of clinical trials before / after CTA

Background : Enforcement of the 2018 Clinical Trials Act (CTA) in Japan resulted in strict and complicated regulations surrounding intervention studies. Few Japan-specific measures have been developed to promote intervention studies in Japan despite concerns about CTA’s negative influence on such studies. Therefore, this study examined the changes in academic investigators’ interest in conducting clinical studies before and after enforcement of the CTA to determine measures to promote specified clinical trials. Methods : We conducted a questionnaire survey with investigators belonging to the Institute of Biomedical Sciences, Tokushima University Graduate School, before and after enforcement of the CTA. Results : Investigators had lesser interest in intervention studies in the post-questionnaire survey (post) group than in the pre-questionnaire survey (pre) group. Their desire for “project management” was significantly higher in the post-group than in the pre-group. Their desire for “support for preparing documents when conducting specified clinical trials” was significantly higher in the group interested in conducting specified clinical trials than that in the not-interested group. Conclusion : We revealed that investigators were highly interested in “project management” and “support for preparing documents when conducting specified clinical trials” after enforcement of the CTA. Measures for these desires may promote specified clinical trials

Adaptive sampling dual terahertz comb spectroscopy using dual free-running femtosecond lasers

Terahertz (THz) dual comb spectroscopy (DCS) is a promising method for high-accuracy, high-resolution, broadband THz spectroscopy because the mode-resolved THz comb spectrum includes both broadband THz radiation and narrow-line CW-THz radiation characteristics. In addition, all frequency modes of a THz comb can be phase-locked to a microwave frequency standard, providing excellent traceability. However, the need for stabilization of dual femtosecond lasers has often hindered its wide use. To overcome this limitation, here we have demonstrated adaptive-sampling THz-DCS, allowing the use of free-running femtosecond lasers. To correct the fluctuation of the time and frequency scales caused by the laser timing jitter, an adaptive sampling clock is generated by dual THz-comb-referenced spectrum analysers and is used for a timing clock signal in a data acquisition board. The results not only indicated the successful implementation of THz-DCS with free-running lasers but also showed that this configuration outperforms standard THz-DCS with stabilized lasers due to the slight jitter remained in the stabilized lasers

Environmental radioactive contamination in settlements around the former Soviet Union’s Semipalatinsk nuclear test site: Sarzhal, Karaul and Kainar settlements, and some settlements located south of the Pavlodar Region

More than 450 nuclear atomic explosions including atmospheric, aboveground and underground nuclear tests had been conducted by the former Soviet Union (USSR) at the Semipalatinsk Nuclear Test Site (SNTS) over the past 40 years. Since 1994, we have concentrated our energies on assessing the human health and environmental impacts of nuclear testing. Especially, residual long-lived radionuclides such as 137Cs and Pu isotopes have been measured for soil samples from various areas. The studies focused on the evaluation of radioactive contamination in the settlements where the Semipalatinsk historical cohort resided. In October 2005, detailed soil sampling was carried out around Dolon settlement to estimate the width and position of center-axis of the fallout plume associated with the first USSR atomic bomb on 29 August 1949, and provided useful information on the efforts to estimate radiation dose in Dolon settlement as reliably as possible. Such studies were, next, devoted more time and attention to the Sarzhal and Karaul settlements located in the southeastern direction from the SNTS. The local fallout from the fist thermonuclear event on 12 August 1953 (30 m in height, 400 kt in scale) heavily affected those settlements. Here, laying stress on Sarzhal and Karaul settlements, the accumulated levels and spatial distribution of soil contamination by 137Cs and 239,240Pu have been excessively investigated since 2007. Besides, the radionuclides 236U and 237Np derived from bomb materials themselves were also measured together with the induced radionuclides, 152Eu and 60Co, in some soil samples. Based on those data obtained, radiation doses in air in two settlements were first reconstructed. As a result, adsorbed doses in air in case of the evacuation (non-evacuation) for inhabitants in Sarzhal and Karaul settlements were evaluated to be 20〜100 mGy (210〜530 mGy) and 10〜75 mGy (120〜380 mGy), respectively, as our tentative estimate for 1-year cumulative external doses mainly due to the local fallout from the first USSR hydrogen bomb test in August 1953. Apart from the above researches, in the Kainar village, one of the villages where the historical cohort resides, located south of the SNTS, and in the Pavlodar District located in the adjust north direction of the SNTS in which data were very limited, accumulated levels of 137Cs and Pu isotopes in soils have been measured for evaluating 137Cs and Pu inventories and their spatial distributions. The inventories of 137Cs found out both inside and outside of the Kainar village were in a wide range of 130〜7,500 Bq/m2. A clear shape similar to a Gaussian function along the center-axis of the radioactive plume, which has been found in Dolon, Sarzhal and Karaul settlements, was not observed in their spatial distributions, indicating a wider and uniformly close contamination of this region. On the other hand, in Pavlodar District, the 137Cs inventories ranged from 1,000 to 3,000 Bq/m2 for soil samples. As a whole, Pavlodar areas that the surveys have been conducted were found not to be contaminated so highly, compared with the northeastern areas including Dolon settlement contaminated heavily by the first nuclear atomic explosion in August 1949. The obtained data were also compared with all the data from other areas where we have surveyed until now. The obtained data will be useful for future residents' exposure dose assessmentAbstract… 1 Ⅰ．はじめに… 2 Ⅱ．サルジャール及びカラウル集落… 3 Ⅲ．カイナル集落及びパブロダール州南部の集落…16 Ⅳ．これまでに調査したセミパラチンスク全地域の137Cs及び239,240Pu蓄積量と 　　239,240Pu/137Cs及び238Pu/239,240Pu放射能比の比較… 24 Ⅴ．全体のまとめ…24 参考文献…26 Appendix…29本研究は、長期に渡る文部省の科学研究費助成金（基盤研究（A,B）: 海外、代表者 : 星 正治、山本政儀、川野徳幸） No.22404004、19404005、23406002、20406002、26257501、24310044、23310183、15H03137の助成を受けて行われた

An Exploratory Trial of EPI-589 in Amyotrophic Lateral Sclerosis (EPIC-ALS) : Protocol for a Multicenter, Open-Labeled, 24-Week, Single-Group Study

Background: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder, with its currently approved drugs, including riluzole and edaravone, showing limited therapeutic effects. Therefore, safe and effective drugs are urgently necessary. EPI-589 is an orally available, small-molecule, novel redox-active agent characterized by highly potent protective effects against oxidative stress with high blood-brain barrier permeability. Given the apparent oxidative stress and mitochondrial dysfunction involvement in the pathogenesis of ALS, EPI-589 may hold promise as a therapeutic agent. Objective: This protocol aims to describe the design and rationale for the EPI-589 Early Phase 2 Investigator-Initiated Clinical Trial for ALS (EPIC-ALS). Methods: EPIC-ALS is an explorative, open-labeled, single-arm trial that evaluates the safety and tolerability of EPI-589 in patients with ALS. This trial consists of 12-week run-in, 24-week treatment, and 4-week follow-up periods. Patients will receive 500 mg of EPI-589 3 times daily over the 24-week treatment period. Clinical assessments include the mean monthly change of Amyotrophic Lateral Sclerosis Functional Rating Scale–Revised total score. The biomarkers are selected to analyze the effect on oxidative stress and neuronal damage. The plasma biomarkers are 8-hydroxy-2′-deoxyguanosine (8-OHdG), 3-nitrotyrosine (3-NT), neurofilament light chain (NfL), phosphorylated neurofilament heavy chain (pNfH), homocysteine, and creatinine. The cerebrospinal fluid biomarkers are 8-OHdG, 3-NT, NfL, pNfH, and ornithine. The magnetic resonance biomarkers are fractional anisotropy in the corticospinal tract and N-acetylaspartate in the primary motor area. Results: This trial began data collection in September 2021 and is expected to be completed in October 2023. Conclusions: This study can provide useful data to understand the characteristics of EPI-589

Development of a Qualitative Research Method Using Family Image Technique (FIT)

This article suggests a method to use FIT (Family Image Technique) in qualitative studies. This method could find ""the rich data"" easier, and is a mew approach that has characteristics of quantitative and qualitative methods

Beam and SKS spectrometers at the K1.8 beam line

High-resolution spectrometers for both incident beams and scattered particles have been constructed at the K1.8 beam line of the Hadron Experimental Facility at J-PARC. A point-to-point optics is realized between the entrance and exit of QQDQQ magnets for the beam spectrometer. Fine-pitch wire chamber trackers and hodoscope counters are installed in the beam spectrometer to accept a high rate beam up to 107 Hz. The superconducting kaon spectrometer for scattered particles was transferred from KEK with modifications to the cryogenic system and detectors. A missing-mass resolution of 1.9 ± 0.1 MeV/c2 (FWHM) was achieved for the ∑ peaks of (π±, K+) reactions on a proton target in the first physics run of E19 in 2010