日本における基質特異性拡張型β-ラクタマーゼ産生肺炎桿菌の分子遺伝学的特徴：CTX-M-15の優位性と高病原性クローンの出現

Objective: To provide data on the molecular characteristics of extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae clinical isolates in Japan. Methods: A total of 100 clinical isolates of ESBL-producing K. pneumoniae collected throughout Japan between June and July 2018 were studied. ESBL genes were analyzed using PCR and DNA sequencing. Transferability of ESBL genes was investigated by conjugation experiments. Plasmid replicon types, virulence genes (rmpA, rmpA2, iucA, iroB, and peg-344) associated with hypervirulent K. pneumoniae (hvKp), and capsule types were detected using PCR. Genotyping was performed using multilocus sequence typing. Results: All ESBL-producing isolates carried blaCTX-M genes. The most predominant CTX-M-type identified was CTX-M-15 (n=55). We identified 24 sequence types (STs) among the CTX-M-15 producers, with ST25 (n=8) being the most common. Most of the transconjugants carrying blaCTX-M-15 contained the FIIk replicon. Of the 100 ESBL-producing isolates, 31 were hvKp defined by the presence of the virulence genes. These ESBL-producing hvKp isolates belonged to eight STs (STs 23, 25, 36, 65, 86, 268, 412, and 4492), with five capsule types (K1, K2, K20, K57, and undefined). Conclusions: CTX-M-15 was the predominant ESBL among K. pneumoniae isolates from Japan. This study shows that ESBL-producing hvKp strains comprising various clones are emerging in Japan.博士（医学）・甲第769号・令和3年3月15日© 2020 The Authors. Published by Elsevier Ltd on behalf of International Society for Infectious Diseases. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Structured review of the literature of type 2 diabetes self-management: Toward the development of a patients' characteristics-based assessment tool

2型糖尿病患者の特性に応じた教育・支援を行うためのアセスメントツールの開発を目指して,自己管理に影響する要因を文献から抽出した.検索方法は,2型糖尿病の自己管理に関連したキーワードを設定し,MEDLINE,CINAHL,医学中央雑誌のデータベース過去10年間を検索した.ヒット件数は,各々1162,860,691件であった.次いで,ADA(American Diabetes Association)Evidence guideline(2002年)の水準を参考に研究デザインを吟味した.その結果,該当する文献は71件で,国内文献が20件あった.有意差を認めた因子を整理したところ86因子あり,「知識・教育」「心理面」「満足度」「個人的要因」「治療」「収入・保険」「関係」の7カテゴリーに分類できた.しかし,介入方法の分類に結びつく因子(特性)や因子間の関係性は不明であり,効果的な介入を導くためのアセスメントツールを開発するためには,これらを検討する必要性が示唆された.To develop an assessment tool for education and follow-up for type 2 diabetes patients related to their characteristics, a structured literature review was conducted. The method of the review was by key words related to self-management/self-care for type 2 diabetes taken from the database for the past 10 years of MEDLINE, CINAHL, and Japan Medical Abstract Society. One-thousand one- hundred and sixty-two articles were obtained from MEDLINE, 860 from CINAHL, and 691 from the Japan Medical Abstract Society. Research designs were then examined, based on the ADA (American Diabetes Association) Evidence guideline (2002) evidence level. As a result, 71 articles, of which 20 were domestic, were retained. Eighty-six factors were found statistically significant, and were categorized in groups under the headings “knowledge/education", “psychological aspects", “level of satisfaction", “personal factors",“therapy/treatment", “income/insurance", and “family/relationship". Factors/characteristics lead toselecting interventional strategies. However, their overall relations have not been examined yet, whichsuggests that these need to be tested to develop an assessment tool which leads to an effectiveintervention program

奈良における侵襲性GBS感染症における臨床的特徴と分子疫学的特徴(2007～2016年)

Invasive Streptococcus agalactiae (GBS) infections are increasingly common among neonates and the elderly. Therefore, GBS surveillance for better antibiotic treatment and prophylaxis strategies are needed. We retrospectively evaluated the clinical aspects of invasive infections and the phenotypic and genetic diversity of infectious isolates from Nara, Japan, collected between 2007 and 2016, by using information from hospital records. GBS strains collected from the blood and cerebrospinal fluid cultures were evaluated for capsular types, multi-locus sequence typing (MLST), antibiotic susceptibility, antibiotics resistance gene, and pulsed-field gel electrophoresis. Forty GBS isolates (10 from children and 30 from adults) were analyzed, and the distribution of molecular serotype and allelic profiles varied between children and adults. We found the rates of early-onset disease in neonates with birth complications to be higher than that of previous reports, indicating that there could be relevance between complications at birth and early-onset disease. Standard antibiotic prophylaxis strategies may need to be reconsidered in patients with birth complications. In adults, the mean age of the patients was 68 years (male: 63%). Primary bacteremia was the most common source of infection. In the neonates, six had early-onset diseases and four had late-onset diseases. The most frequently identified strains were molecular serotype Ia ST23 (40%) and molecular serotype Ib ST10 (20%) in children and molecular serotype Ib ST10 (17%), molecular serotype VI ST1 (13%), and molecular serotype V ST1 (13%) in adults. Levofloxacin-resistant molecular serotype Ib strains and molecular serotypes V and VI ST1 were common causes of GBS infection in adults but were rarely found in children. Furthermore, pulsed-field gel electrophoresis in our study showed that specific clone isolates, that tend to have antibiotics resistance were widespread horizontally for a decade. Continuous surveillance and molecular investigation are warranted to identify the transmission route and improve antibiotic treatment strategies.博士（医学）・甲第773号・令和3年3月15日Copyright: © 2020 Hirai 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

Trends of correlations between serum levels of growth hormone and insulin-like growth factor-I in general practice

Serum levels of growth hormone (GH) and insulin-like growth factor (IGF)-I are crucial in the diagnosis and management of GH-related diseases. However, these levels are affected by nutritional and metabolic status. To elucidate the correlations between GH and IGF-I in various conditions, a retrospective analysis was performed for adult patients in which GH levels were examined by general practitioners during the period from January 2019 to December 2021. Of 642 patients, 33 patients were diagnosed with acromegaly, 21 were diagnosed with GH deficiency (GHD), and 588 were diagnosed with non-GH-related diseases (NGRD). In contrast to the positive correlations found between the levels of GH and IGF-I in patients with acromegaly (R=0.50; P<0.001) and patients with GHD (R=0.39; P=0.08), a negative correlation was found in the NGRD group (R=-0.23; P<0.001). In that group, the results of multivariable analysis showed that GH levels were predominantly influenced by gender and body mass index (BMI), whereas IGF-I levels were modulated by albumin in addition to age and GH. Of note, in the NGRD group, there was an enhanced negative correlation between GH and IGF-I under conditions of BMI < 22 and albumin < 4.0 g/dL (R=-0.45; P<0.001), and the negative correlation between GH and IGF-I was reinforced by excluding patients with other pituitary diseases and patients taking oral steroids (R=-0.51; P<0.001 and R=-0.59; P<0.001, respectively). Collectively, the results indicate that attention should be given to the presence of a negative correlation between serum levels of GH and IGF-I, especially in lean and low-nutritious conditions

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/)

Age-Specific Prevalence of Glaucoma is Determined by the Presence of Refractive Errors Among Japanese Workers

To develop appropriate glaucoma mass screening programs for occupational health among Japanese workers, we estimated the prevalence of glaucoma and the increase rate by age. A total of 10,579 Japanese general workers (men/women = 9292/1287) underwent frequency doubling technology (FDT) perimetry testing. Visual field abnormalities (VFA) were identified by the FDT-based glaucoma screening protocol (FDT-VFA). Subjects with FDT-VFA were ophthalmologically diagnosed and classified as "normal," "glaucomatous VFA" (preperimetric, suspicious, and definitive glaucoma) or "other ocular diseases." Prevalence of FDT-VFA and positive predictive values for "glaucomatous VFA" and "definitive glaucoma" were calculated by five-year age intervals, and then the prevalence of "glaucomatous VFA" and "definitive glaucoma" in each age interval was estimated. Prevalence of "glaucomatous VFA" and "definitive glaucoma" in workers younger than 30 years old was approximately 1.5% and 0.5%, respectively. Interestingly, the increase in prevalence of glaucoma by age was significantly different between workers with and without refractive errors (RE). From ages 30 to 55 years, the estimated prevalence of "definitive glaucoma" linearly increased with a regression coefficient (%/age in years) that was 2.5-fold higher in subjects with RE than in those without RE {regression coefficient = 0.131 [95% confidence interval (CI) = 0.109, 0.152; R2 = 0.980]vs. 0.047 [95% CI = 0.026, 0.068; R2 = 0.869]for subjects with RE vs. those without RE, respectively}. Further, among workers older than 55 years, the prevalence of glaucoma continued increasing in workers with RE, whereas it plateaued in those without RE. From these estimates, we propose that FDT testing should be conducted as follows: 1) once in workers under the age of 30 years, 2) according to both age and the presence of RE in 30-55 years old, and 3) by age only in those over 55 years old

Generation of Embryonic Stem Cell Lines from Immature Rabbit Ovarian Follicles

In mammalian ovaries, many immature follicles remain after the dominant follicles undergo ovulation. Here we report the successful production of rabbit embryonic stem cells (ESCs) from oocytes produced by in vitro culture of immature follicles and subsequent in vitro maturation treatment. In total, we obtained 53 blastocysts from oocytes that received intracytoplasmic sperm injection followed by in vitro culture. Although only weak expression of POU5f1 was observed in the inner cell masses of in-vitro-cultured follicle-derived embryos, repeated careful cloning enabled establishment of 3 stable ESC lines. These ESC lines displayed the morphological characteristics of primed pluripotent stem cells. The ESC lines also expressed the pluripotent markers Nanog, POU5f1, and Sox2. Further, these ESCs could be differentiated into each of the 3 different germ layers both in vitro and in vivo. These results demonstrate that immature follicles from rabbits can be used to generate ESCs. Moreover, the use of rabbit oocytes as a cell source provides an experimental system that closely matches human reproductive and stem cell physiology.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/140197/1/scd.2012.0300.pd