本邦で分離されたカルバペネマーゼ産生肺炎桿菌の分子遺伝学的解析

Carbapenemase-producing Enterobacteriaceae represent a serious public health threat worldwide. Carbapenemase genes, harbored on a transferable plasmid, have been isolated globally with distinct geographical features. Klebsiella pneumoniae, included in Enterobacteriaceae, also produces carbapenemase and often shows hypervirulence. Overlapping carbapenem resistance and hypervirulence in K. pneumoniae have been reported, but such strains have not yet been found in Japan. Here, we screened 104 carbapenemase-producing K. pneumoniae isolates collected from 37 hospitals and outpatient clinics in Japan between September 2014 and July 2015. PCR and DNA sequencing demonstrated IMP-1 in 21 isolates and IMP-6 in 83 isolates, 77 of which coharbored CTX-M-2. Most of the isolates showed low MICs toward imipenem and meropenem but high MICs toward penicillin and cephalosporins. Conjugation experiments with an Escherichia coli J53 recipient showed that most of the plasmids in IMP-6 producers were transferable, whereas only one-half of the plasmids in IMP-1 producers were transferable. PCR-based replicon typing and multiplex PCR identified five isolates belonging to the CG258 non-tonB79 cluster and no isolate belonging to the CG258-tonB79 cluster or sequence type 307 (ST307). Four K1-ST23 isolates, 10 K2-ST65 isolates, and 7 K2-ST86 isolates were detected that harbored virulence genes. The resistance genes in 85 isolates were transferable, but the virulence genes were not transferred. These results demonstrate the acquisition of IMP-type carbapenemase genes and CTX-M-type genes among hypervirulence isolates in Japan, warranting further attention and countermeasures. In this study, we have determined the molecular characteristics and epidemiology of IMP-6 producers that coharbored various CTX-M genes in Japan.IMPORTANCE Carbapenems serve as a last resort for the clinical treatment of multidrug-resistant infections. Therefore, the rapid spread of carbapenemase-producing strains represents a serious public health threat, further limiting antibiotic choices. The current findings of hypervirulent carbapenemase-producing Klebsiella pneumoniae clinical isolates in Japan demonstrate the potential broad spread and transfer of these genes, necessitating close surveillance.博士（医学）・乙第1509号・令和3年3月15日Copyright © 2020 Yonekawa et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license(https://creativecommons.org/licenses/by/4.0/)

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements

STCW-MANILA 2010 OCH DESS INVERKAN PÅ ÅLANDS SJÖSÄKERHETSCENTRUM

Abstrakt Meningen med detta arbete har varit att se på vilket sätt de så kallade Manilaändringarna påverkar kurscentrum och deras utbildning. Fokus har varit att se i vilken omfattning Manilaändringarna leder till omarbetande av kurser och då speciellt de kurser som anordnas på Ålands sjösäkerhetscentrum. Ålands sjösäkerhetscentrum är skribentens arbetsplats. Innehållet i dessa kurser behandlas och beskrivs också. Jag gör en kort tillbakablick gällande STCW konventionens historia och dess uppkomst, samt en överblick gällande själva Manilaändringarna och deras implementerings period och faser. I bilagorna hittas intervjuer med två anställda på Ålands sjösäkerhetscentrum, den ansvariga chefen och en instruktör. Där berörs frågorna rörande hur Manilaändringarna har påverkat själva centrets verksamhet och de anställda.The purpose of this thesis has been to look at the way the so called Manila Amendments has affected course centers and their training. I have studied how the Manila Amendments affect the courses, and then in particular the courses Ålands sjösäkerhetscenter provides. Ålands sjösäkerhetscenter is also the author´s current place of work. The content of the affected courses is also addressed. I make a brief description of the history of the STCW Convention and how it was founded, as well as an overview on the Manila Amendments and their implementation period and phases. In the annexes I include two interviews made with the responsible Manager and one instructor from Ålands sjösäkerhetscentrum. In the interviews issues relating to how the Manila Amendments haves had an impact on the operation of the center and its employee

Tyrosine Kinase Inhibitor Profiling Using Multiple Forskolin-Responsive Reporter Cells

We have developed a highly sensitive promoter trap vector system using transposons to generate reporter cells with high efficiency. Using an EGFP/luciferase reporter cell clone responsive to forskolin, which is thought to activate adenylate cyclase, isolated from human chronic myelogenous leukemia cell line K562, we found several compounds unexpectedly caused reporter responses. These included tyrosine kinase inhibitors such as dasatinib and cerdulatinib, which were seemingly unrelated to the forskolin-reactive pathway. To investigate whether any other clones of forskolin-responsive cells would show the same response, nine additional forskolin-responsive clones, each with a unique integration site, were generated and quantitatively evaluated by luciferase assay. The results showed that each clone represented different response patterns to the reactive compounds. Also, it became clear that each of the reactive compounds could be profiled as a unique pattern by the 10 reporter clones. When other TKIs, mainly bcr-abl inhibitors, were evaluated using a more focused set of five reporter clones, they also showed unique profiling. Among them, dasatinib and bosutinib, and imatinib and bafetinib showed homologous profiling. The tyrosine kinase inhibitors mentioned above are approved as anticancer agents, and the system could be used for similarity evaluation, efficacy prediction, etc., in the development of new anticancer agents

一次線毛動態制御による新たな膵癌治療法の開発

application/pdf術前放射線化学療法 (NCRT) を施行した局所進行膵癌の細胞において、一旦消失した一次線毛がCRT後に再度認められる現象を確認した。一次線毛が形成されると細胞増殖抑制効果があると考えられ、今後は、三重大学でCRTを施行し手術に至った患者の膵癌摘出標本を用いて、膵癌細胞における一次線毛の有無、Hhシグナル伝達分子の発現を検討する。In cells of locally advanced pancreatic cancer that underwent preoperative chemoradiotherapy (CRT), it was confirmed that the primary cilia which had once disappeared were re-recognized after CRT. The formation of primary cilia is thought to have an inhibitory effect on cell proliferation. The presence or absence of primary cilia in pancreatic cancer cells will be determined and will be examined expression of Hh signaling, using pancreatic cancer excised specimens of patients who underwent CRT and underwent surgery.2018年度～2020年度科学研究費補助金(基盤研究(C))研究成果報告書18K0793

UDP-glucosyltransferase UGT84B1 regulates the levels of indole-3-acetic acid and phenylacetic acid in Arabidopsis

Auxin is a key plant growth regulator for diverse developmental processes in plants. Indole-3-acetic acid (IAA) is a primary plant auxin that regulates the formation of various organs. Plants also produce phenylacetic acid (PAA), another natural auxin, which occurs more abundantly than IAA in various plant species. Although it has been demonstrated that the two auxins have distinct transport characteristics, the metabolic pathways and physiological roles of PAA in plants remain unsolved. In this study, we investigated the role of Arabidopsis UDP-glucosyltransferase UGT84B1 in IAA and PAA metabolism. We demonstrated that UGT84B1, which converts IAA to IAA-glucoside (IAA-Glc), can also catalyze the conversion of PAA to PAA-glucoside (PAA-Glc), with a higher catalytic activity in vitro. Furthermore, we showed a significant increase in both the IAA and PAA levels in the ugt84b1 null mutants. However, no obvious developmental phenotypes were observed in the ugt84b1 mutants under laboratory growth conditions. Moreover, the overexpression of UGT84B1 resulted in auxin-deficient root phenotypes and changes in the IAA and PAA levels. Our results indicate that UGT84B1 plays an important role in IAA and PAA homeostasis in Arabidopsis.</p

Role of Arabidopsis&nbsp;INDOLE-3-ACETIC ACID CARBOXYL METHYLTRANSFERASE 1&nbsp;in auxin metabolism

The phytohormone auxin regulates a wide range of developmental processes in plants. Indole-3-acetic acid (IAA) is the main auxin that moves in a polar manner and forms concentration gradients, whereas phenylacetic acid (PAA), another natural auxin, does not exhibit polar movement. Although these auxins occur widely in plants, the differences between IAA and PAA metabolism remain largely unknown. In this study, we investigated the role of Arabidopsis IAA CARBOXYL METHYLTRANSFERASE 1 (IAMT1) in IAA and PAA metabolism. IAMT1 proteins expressed in Escherichia coli could convert both IAA and PAA to their respective methyl esters. Overexpression of IAMT1 caused severe auxin-deficient phenotypes and reduced the levels of IAA, but not PAA, in the root tips of Arabidopsis, suggesting that IAMT1 exclusively metabolizes IAA in&nbsp;vivo. We generated iamt1 null mutants via CRISPR/Cas9-mediated genome editing and found that the single knockout mutants had normal auxin levels and did not exhibit visibly altered phenotypes. These results suggest that other proteins, namely the IAMT1 homologs in the SABATH family of carboxyl methyltransferases, may also regulate IAA levels in Arabidopsis