ハナノキ ニ カッショクマルハンビョウ ヲ ヒキオコス Phyllosticta minima ノ シュトクイテキ プライマー ヲ モチイタ PCRホウ ニ ヨル ケンシュツ

ハナノキはカエデ属の日本固有種で，愛知県，岐阜県および長野県の限られた地域に自生している。本樹木は，個体数が減少していることから環境省レッドリストの絶滅危惧II類に選定され，その種の存続が危ぶまれている。植物病原菌Phyllosticta minimaによって引き起こされる褐色円斑病は，ハナノキに深刻な被害を与え，天然更新の阻害要因となっていることが考えられている。本研究では，ハナノキ罹病組織から病原菌を特異的に検出し，病害防除の上で重要となる感染経路を特定する目的で，種特異的プライマーを設計した。種特異的プライマーPmiFおよびPmiRは，P. minimaのrDNA ITS領域の塩基配列から設計され，分離菌株由来の全DNAを用いたPCR法で，その特異性が確認された。愛知県名古屋市で採集されたハナノキ組織から病原菌の検出を試みたところ，罹病葉のみならず潜在的に感染している無病徴の葉からも病原菌の検出が確認された。さらに，東京都八王子市に植栽されたハナノキの葉，葉柄，小枝，冬芽および腋芽について調査した結果，それぞれの組織から病原菌が検出された。P. minimaは6月から7月頃に罹病葉から腋芽（冬芽）へ感染，潜伏して，翌年の第一次感染源となることが明らかとなった。Acer pycnanthum is a maple tree indigenous to Japan, which grows in limited areas of the Aichi, Gifu, and Nagano Prefectures. A. pycnanthum is currently an endangered species, with a declining population. This tree is considered to be a vulnerable species according to the Red List of the Ministry of the Environment. Leaf spot disease caused by Phyllosticta minima, a phytopathogenic fungus, inflict severe damage on A. pycnanthum. These fungi secrete inhibitory factors, which have been suggested to repress natural regeneration. In the present study, we designed species-specific primers with the aim of specifically detecting pathogens in tissue samples of A. pycnanthum, thereby identifying infection routes crucial for disease control. The species-specific primers PmiF and PmiR, which were designed based on the sequence of the P. minima rDNA Internal Transcribed Spacer region, were verified for their specificity in PCR tests using the total DNA of isolated strains. In A. pycnanthum tissue samples from Hachioji, Tokyo and Nagoya, Aichi Prefecture, pathogenic fungi were isolated not only from affected but also asymptomatic leaves. Furthermore, in trees planted at Hachioji, Tokyo, pathogens were observed in tissue samples of leaves, petioles, twigs, winter buds, and axillary buds. These results suggest that P. minima infects axillary (winter) buds from June to July via previously infected leaves, thus becoming latent and finally triggering primary infections in the following year

プラスミド性キノロン耐性遺伝子を保有するIMP-6産生腸内細菌科細菌でのイノカラムサイズ効果の比較

Almost all cases of carbapenemase-producing Enterobacteriaceae infections in Japan are caused by blaIMP-positive Enterobacteriaceae (especially blaIMP-6) and infections caused by other types of carbapenemase-producing Enterobacteriaceae are quite rare. We examined drug resistance genes co-harboring with blaIMP-6 and their inoculum size effects. We screened β-lactamase genes, plasmid-mediated quinolone resistance (PMQR) genes, and aminoglycoside-modifying enzyme genes by PCR and performed sequencing for 14 blaIMP-6-positive Enterobacteriaceae. Further, all PMQR-positive isolates were submitted to conjugation and inoculum effect evaluation. Our data showed that 13 of the 14 isolates harbored CTX-M-2 and one co-harbored CTX-M-2 and CTX-M-1 as extended-spectrum β-lactamases. All isolates carried one or more PMQRs; aac(6’)-Ib-cr was the most prevalent (92.8%), and was followed by oqxA (64.3%), qnrS (50%), oqxAB (21.4%), and qnrB (14.3%). However, Klebsiella pneumoniae contains chromosomal OqxAB. Inoculum size effects were significant in all strains for meropenem, 13 strains for imipenem, 7 for levofloxacin, and 3 for amikacin. We observed that 11 of the experimental strains (100%), 8 strains (72.7%), and 1 strain showed inoculum size effects for meropenem, imipenem, and amikacin, respectively. However, four strains harbored qnr genes and two strains harbored qnr genes and QRDR mutations concurrently; no inoculum size effect was seen for levofloxacin. The blaIMP-6-positive Enterobacteriaceae that we studied was found to harbor at least one plasmid-mediated drug resistance gene. The inoculum size effect for carbapenems was thought to be mainly due to IMP-6-type metallo-β-lactamase; however qnrB and qnrS also had a minimal impact on the inoculum size effect for levofloxacin.博士（医学）・乙第1463号・令和2年6月30日Copyright: © 2019 Ogawa et al. This is an open access article distributed under the terms of the Creative Commons Attribution License(https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

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/)

Detection of DR antigen on leukemic cells from a patient suffering from adult T-cell leukemia and progressive systemic sclerosis.

This report concerns an unusual case of adult T cell leukemia (ATL) complicated with progressive systemic sclerosis (PSS). The surface markers of peripheral blood mononuclear cells (PBM) and lymph node cells, both of which mainly consisted of leukemic cells, were examined. The effect of these cells on the pokeweed mitogen (PWM)-induced IgG synthesis by normal PBM also was studied. The leukemic cells formed rosettes with sheep red blood cells (SRBC; E) and expressed T cell antigen, Leu-1, and DR antigen. The detection of cell surface antigens was carried out by employing monoclonal antibodies against these antigens. We diagnosed this case as DR positive ATL. In terms of the immunoregulatory function of these leukemic cells, the co-culture experiments showed that these cells had some suppressive effect on the PWM-induced IgG production by allogeneic normal PBM.</p

Liquid-crystalline phase transition in organophosphazenes

Organophosphazenes with a similar mesogenic moiety were prepared and their mesogenicity was studied by differential scanning calorinetry (DSC) measurements and polarizing microsope observations. In cyclotriphosphazenes with alk-­oxybiphenyl and Schiff base moieties, mesomorphic phase transitions were observed, but no mesomorphic phase was observed for the corresponding cyclotetraphosphazenes. In polyphos-­phazenes with an alkoxybiphenyl moiety, no mesomorphic phase was observed. The molecular structure of cyclotriphosphazenes facilitated the formation of a mesomorphic layer structure; in contrast, the formation of a mesomorphic layer structure did not occur in cyclotetraphosphazenes and polyphosphazenes, even though they bore a similar mesogenic moiety. Moreover, in cyclotriphosphazenes with an optically active alkoxybiphenyl group, a smectic C* phase was observed. The spontaneous polarization of the compound was −190 μ C m −2 at 436 K in 25 μ in cell using the triangular-wave method. © 1998 John Wiley & Sons, Ltd.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/38314/1/785_ftp.pd

Crucial roles of RSK in cell motility by catalysing serine phosphorylation of EphA2

Crosstalk between inflammatory signalling pathways and receptor tyrosine kinases has been revealed as an indicator of cancer malignant progression. In the present study, we focus on EphA2 receptor tyrosine kinase, which is overexpressed in many human cancers. It has been reported that ligand-independent phosphorylation of EphA2 at Ser-897 is induced by Akt. We show that inflammatory cytokines promote RSK-, not Akt-, dependent phosphorylation of EphA2 at Ser-897. In addition, the RSK-EphA2 signalling pathway controls cell migration and invasion of metastatic breast cancer cells. Moreover, Ser-897-phosphorylated EphA2 co-localizes with phosphorylated active form of RSK in various human tumour specimens, and this double positivity is related to poor survival in lung cancer patients, especially those with a smoking history. Taken together, these results indicate that the phosphorylation of EphA2 at Ser-897 is controlled by RSK and the RSK-EphA2 axis might contribute to cell motility and promote tumour malignant progression

3 Tesla MRI detects accelerated hippocampal volume reduction in postmenopausal women

Purpose To clarify age-related structural changes specific to hippocampal volume by hierarchizing according to age, gender, and menopausal status. Many studies report the neuroprotective effects of estrogen and age-related brain volume changes; however, there are no studies regarding age-related change specific to hippocampal volume in terms of age, gender, and menopausal status. Materials and Methods T1-weighted MR images were obtained in 412 healthy adults divided into eight groups according to age and gender, to analyze brain volume change focusing on hippocampal volume. Results Voxel-based morphometry (VBM) revealed significantly smaller gray matter volume in the hippocampus bilaterally in females aged in their fifties (51 of 59 females were at menopause) compared with females in their forties (3 of 46 females were at menopause). No significant difference was found, however, between female groups in their fifties versus sixties, or sixties versus seventies; or between male groups in their forties versus fifties, fifties versus sixties, or sixties versus seventies. In addition, VBM revealed significant hippocampal volume reduction bilaterally in all postmenopausal women compared with all premenopausal women. Conclusion The results of the current study suggest that the menopause may be associated with hippocampal volume reduction. © 2010 Wiley-Liss, Inc.Thesis of Goto,Masami / 後藤 政実 博士学位論文(金沢大学 / 大学院医薬保健学総合研究科