研究者詳細 - 黒沼　幸治

論文 - 黒沼　幸治

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Rapid and Integrated Bacterial Evolution Analysis unveils gene mutations and clinical risk of Klebsiella pneumoniae.

Kojiro Uemura, Toyotaka Sato, Soh Yamamoto, Noriko Ogasawara, Jirachaya Toyting, Kotaro Aoki, Akira Takasawa, Masayuki Koyama, Atsushi Saito, Takayuki Wada, Kaho Okada, Yurie Yoshida, Koji Kuronuma, Chie Nakajima, Yasuhiko Suzuki, Motohiro Horiuchi, Kenichi Takano, Satoshi Takahashi, Hirofumi Chiba, Shin-Ichi Yokota

Nature communications 16 ( 1 ) 2917 - 2917 2025年03月 [国際誌]

Bacteria continually evolve. Previous studies have evaluated bacterial evolution in retrospect, but this approach is based on only speculation. Cohort studies are reliable but require a long duration. Additionally, identifying which genetic mutations that have emerged during bacterial evolution possess functions of interest to researchers is an exceptionally challenging task. Here, we establish a Rapid and Integrated Bacterial Evolution Analysis (RIBEA) based on serial passaging experiments using hypermutable strains, whole-genome and transposon-directed sequencing, and in vivo evaluations to monitor bacterial evolution in a cohort for one month. RIBEA reveals bacterial factors contributing to serum and antimicrobial resistance by identifying gene mutations that occurred during evolution in the major respiratory pathogen Klebsiella pneumoniae. RIBEA also enables the evaluation of the risk for the progression and the development of invasive ability from the lung to blood and antimicrobial resistance. Our results demonstrate that RIBEA enables the observation of bacterial evolution and the prediction and identification of clinically relevant high-risk bacterial strains, clarifying the associated pathogenicity and the development of antimicrobial resistance at genetic mutation level.

DOI PubMed

JANISでは把握が困難な中小規模病院における緑膿菌の薬剤耐性の状況

藤谷好弘, 富樫篤生, 齋藤充史, 黒沼幸治, 高橋聡

日本感染症学会総会・学術講演会・日本化学療法学会学術集会合同学会プログラム・抄録集 ( 日本感染症学会・日本化学療法学会 ) 98回・72回 np255 - np255 2024年05月

結核の診断に時間を要した肺外結核の2例

小林拓海, 齋藤充史, 小玉賢太郎, 高橋守, 黒沼幸治, 千葉弘文

日本呼吸器学会誌 ( (一社)日本呼吸器学会 ) 13 ( 増刊 ) 395 - 395 2024年03月

当院改修時におけるアスペルギルス対策の評価

黒沼幸治, 中江舞美, 韮澤慎也, 富樫篤生, 藤谷好弘, 齋藤充史, 高橋聡

感染症学雑誌 ( (一社)日本感染症学会 ) 98 ( 2 ) 281 - 281 2024年03月

間質性肺疾患における気管支肺胞洗浄液中の肺胞マクロファージの代謝活性についての検討

萬谷峻史, 齋藤充史, 佐藤達也, 山口美樹, 亀田優美, 佐久間裕司, 黒沼幸治, 千葉弘文

日本呼吸器学会誌 ( (一社)日本呼吸器学会 ) 13 ( 増刊 ) 341 - 341 2024年03月

当院改修時におけるアスペルギルス対策の評価

黒沼幸治, 中江舞美, 韮澤慎也, 富樫篤生, 藤谷好弘, 齋藤充史, 高橋聡

感染症学雑誌 ( (一社)日本感染症学会 ) 98 ( 2 ) 281 - 281 2024年03月

The impact of respiratory reactance in oscillometry on survival in patients with idiopathic pulmonary fibrosis.

Tatsuru Ishikawa, Hirotaka Nishikiori, Yuki Mori, Keiko Fujino, Atsushi Saito, Mamoru Takahashi, Koji Kuronuma, Shiro Hinotsu, Hirofumi Chiba

BMC pulmonary medicine 24 ( 1 ) 10 - 10 2024年01月 [国際誌]

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a progressive disease with a poor prognosis. Pulmonary function tests (PFTs) aid in evaluating the disease status of IPF. The clinical significance of oscillometry measurements in interstitial lung diseases has recently been reported. Our previous study showed that respiratory reactance (Xrs) measured by oscillometry reflected disease severity and predicted subsequent lung capacity decline in patients with IPF. However, the direct impact of Xrs on survival needs to be determined, and there are currently no reference values in oscillometry to predict prognosis. Therefore, this study aimed to investigate the association between oscillometry measurements, particularly Xrs, and survival in patients with IPF and to determine the cutoff values of Xrs that predict 3-year survival. METHODS: We analyzed the relationship between the measured values of PFT and oscillometry derived from 178 patients with IPF. Univariate and multivariate Cox proportional hazards analyses were performed to investigate the relationships between clinical indices at the time of the first oscillometry and survival. We performed the time-dependent receiver operating characteristic (ROC) curve analysis to set the optimized cutoff values of Xrs for 3-year survival prediction. We examined the discriminating power of cutoff values of Xrs on survival using the Kaplan-Meier method and the log-rank test. RESULTS: Xrs components, especially in the inspiratory phase (In), significantly correlated with the PFT values. In the multivariate analyses, Xrs (all of reactance at 5 Hz [X5], resonant frequency [Fres], and low-frequency reactance area [ALX] in the inspiratory phase) had a significant impact on survival (X5, p = 0.003; Fres, p = 0.016; ALX, p = 0.003) independent of age, sex, and other prognostic factors derived from the univariate analysis. The area under the ROC curve was 0.765, 0.759, and 0.766 for X5 In, Fres In, and ALX In, with cutoff values determined at - 0.98, 10.67, and 5.32, respectively. We found significant differences in survival after dividing patients using each of the cutoff values of Xrs. CONCLUSIONS: In patients with IPF, Xrs measured by oscillometry significantly impacted survival. We also determined the cutoff values of Xrs to discriminate patients with poor prognoses.

DOI PubMed

手洗器の水栓口や給水システムの汚染による緑膿菌の感染伝播

藤谷好弘, 中江舞美, 佐藤勇樹, 韮澤慎也, 富樫篤生, 齋藤充史, 黒沼幸治, 高橋聡

感染症学雑誌 ( (一社)日本感染症学会 ) 98 ( 1 ) 119 - 119 2024年01月

Factors affecting the sensitivity of quantitative severe acute respiratory syndrome coronavirus 2 antigen test.

Yuki Sato, Ryosei Murai, Ryo Kobayashi, Atsuo Togashi, Yoshihiro Fujiya, Koji Kuronuma, Satoshi Takahashi

Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy 29 ( 8 ) 754 - 758 2023年08月 [国際誌]

INTRODUCTION: The accuracy of nucleic acid amplification tests (NAATs) is affected by various factors; however, studies examining the factors affecting the accuracy of quantitative severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen test (QAT) are limited. METHODS: A total of 347 nasopharyngeal samples were collected from patients with coronavirus disease 2019 (COVID-19), and the date of onset was obtained from the electronic medical records. The SARS-CoV-2 antigen level was measured using Lumipulse Presto SARS-CoV-2 Ag (Presto), while NAAT was performed using the Ampdirect 2019-nCoV Detection Kit. RESULTS: Presto had a sensitivity rate of 95.1% (95% confidence interval: 92.8-97.4) in detecting the SARS-CoV-2 antigen in 347 samples. The number of days from symptom onset to sample collection was negatively correlated with the amount of antigen (r = -0.515) and sensitivity of Presto (r = -0.711). The patients' age was lower in the Presto-negative samples (median age, 39 years) compared with that in the Presto-positive samples (median age, 53 years; p < 0.01). A significant positive correlation was observed between age (excluding teenagers) and Presto sensitivity (r = 0.764). Meanwhile, no association was found between the mutant strain, sex, and Presto results. CONCLUSION: Presto is useful for the accurate diagnosis of COVID-19 owing to its high sensitivity when the number of days from symptom onset to sample collection is within 12 days. Furthermore, age may affect the results of Presto, and this tool has a relatively low sensitivity in younger patients.

DOI PubMed

COVID-19患者における血清レクチン値についての検討

竹中遥, 齋藤充史, 黒沼幸治, 千葉弘文

日本肺サーファクタント・界面医学会雑誌 ( 日本肺サーファクタント・界面医学会 ) 54 28 - 29 2023年08月

COVID-19患者における血清レクチン値についての検討

竹中遥, 齋藤充史, 黒沼幸治, 千葉弘文

日本肺サーファクタント・界面医学会雑誌 ( 日本肺サーファクタント・界面医学会 ) 54 28 - 29 2023年08月

ゲノム解析は同時期に複数病棟で発生したCOVID-19クラスター事例の全体像の把握に有用である

藤谷好弘, 中江舞美, 中村広士, 富樫篤生, 齋藤充史, 黒沼幸治, 高橋聡, 大久保和洋, 川代愛梨, 田宮和真

日本環境感染学会総会プログラム・抄録集 ( (一社)日本環境感染学会 ) 38回 271 - 271 2023年06月

COVID-19の濃厚接触者が陽性となる要因の検討

中江舞美, 藤谷好弘, 富樫篤生, 中村広士, 斎藤充史, 黒沼幸治, 高橋聡

日本環境感染学会総会プログラム・抄録集 ( (一社)日本環境感染学会 ) 38回 382 - 382 2023年06月

Pseudo-outbreak of Mycobacterium lentiflavum at a general hospital in Japan.

Yutaro Nagano, Koji Kuronuma, Yasuo Kitamura, Kanami Nagano, Hayato Yabe, Sayaka Kudo, Toyotaka Sato, Shinya Nirasawa, Mami Nakae, Motohiro Horiuchi, Shin-Ichi Yokota, Yoshihiro Fujiya, Atsushi Saito, Satoshi Takahashi, Hirofumi Chiba

Infection control and hospital epidemiology 1 - 7 2023年04月 [国際誌]

BACKGROUND: Mycobacterium lentiflavum is a slow-growing nontuberculous mycobacterium that is widely distributed in soil and water systems, but it is sometimes pathogenic to humans. Although cases of M. lentiflavum infections are rare, 22 isolates of M. lentiflavum were identified at a single hospital in Japan. We suspected a nosocomial outbreak; thus, we conducted transmission pattern and genotype analyses. METHODS: Cases of M. lentiflavum isolated at Kushiro City General Hospital in Japan between May 2020 and April 2021 were analyzed. The patient samples and environmental culture specimens underwent whole-genome sequencing (WGS). Additionally, we retrospectively collected clinical data from patient medical records. RESULTS: Altogether, 22 isolates of M. lentiflavum were identified from sputum and bronchoalveolar lavage samples. Clinically, the instances with M. lentiflavum isolates were considered contaminants. In the WGS analysis, 19 specimens, including 18 patient samples and 1 environmental culture from the hospital's faucet, showed genetic similarity. The frequency of M. lentiflavum isolation decreased after we prohibited the use of taps where M. lentiflavum was isolated. CONCLUSIONS: WGS analysis identified that the cause of M. lentiflavum pseudo-outbreak was the water used for patient examinations, including bronchoscopy.

DOI PubMed

Klebsiella pneumoniaeにおける遺伝子変異頻度と病原性の関連性評価

上村幸二朗, 佐藤豊孝, 黒沼幸治, 高橋聡, 横田伸一

日本化学療法学会雑誌 ( (公社)日本化学療法学会 ) 71 ( 2 ) 179 - 179 2023年03月

SARS-CoV-2ワクチン接種後の長期的抗体価推移と次期接種のタイミングの検討

藤谷好弘, 中江舞美, 富樫篤生, 齋藤充史, 黒沼幸治, 高橋聡

日本感染症学会総会・学術講演会・日本化学療法学会学術集会合同学会プログラム・抄録集 ( 日本感染症学会・日本化学療法学会 ) 97回・71回 O - 127 2023年03月

間質性肺疾患を有する難治性咳嗽に対してのP2X3受容体拮抗薬の有効性の検討

高橋知之, 齋藤充史, 萬谷峻史, 池田貴美之, 黒沼幸治, 千葉弘文

日本呼吸器学会誌 ( (一社)日本呼吸器学会 ) 12 ( 増刊 ) 216 - 216 2023年03月

Analysis of diagnostic performance and factors causing nonspecific reactions in SARS-CoV-2 rapid antigen detection tests.

Natsuki Narumi, Takashi Kondo, Yuki Sato, Yuki Katayama, Shinya Nirasawa, Masachika Saeki, Yuki Yakuwa, Yoshihiro Fujiya, Koji Kuronuma, Satoshi Takahashi

Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy 29 ( 2 ) 157 - 162 2023年02月 [国際誌]

INTRODUCTION: Early diagnosis and appropriate infection control are important to prevent the transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, we aimed to assess the diagnostic performance of SARS-CoV-2 rapid antigen detection (RAD) tests and the factors that cause nonspecific reactions. METHODS: Nasopharyngeal swab specimens (n = 100), sputum specimens (n = 10), and lithium-heparin plasma samples (n = 100) were collected. We evaluated Espline®SARS-CoV-2 (Espline) and SARS-CoV-2 Rapid Antigen Test that also known as STANDARD Q® (STANDARD Q), with reverse transcription-polymerase chain reaction (RT-PCR) and Lumipulse® Presto SARS-CoV-2 Ag as reference tests. In addition, we investigated the effects of inadequate pretreatment methods and five potential causes of nonspecific reactions. RESULTS: The sensitivities of Espline and STANDARD Q were 60% and 57%, respectively, and their specificity was 100%. It was confirmed that the judgment line for the positive insufficiently mixed specimens was faint. A false-positive result was observed with STANDARD Q when sputum was used as a specimen to investigate judgment the effect of viscosity. CONCLUSIONS: Espline and STANDARD Q show good sensitivity for specimens with Ct values less than 25, but specimens collected within 9 days of symptom onset may still give false negatives. The test should be performed carefully, and the results should be judged comprehensively, taking into account clinical symptoms and patient background.

DOI PubMed

Protective effect of Bifidobacterium longum BB536 against nausea caused by pirfenidone in a mouse model of pellagra.

Koji Kuronuma, Natsumi Susai, Tomohiro Kuroita, Takeshi Yoshioka, Atsushi Saito, Hirofumi Chiba

Bioscience of microbiota, food and health 42 ( 3 ) 195 - 202 2023年 [国内誌]

Pellagra is caused by abnormal intake and/or use of nicotinic acid and is known in part to be induced by the use of medications such as isoniazid or pirfenidone. We previously investigated atypical phenotypes of pellagra, such as nausea, using a mouse model of pellagra and found that gut microbiota play an important role in the development of these phenotypes. Here, we investigated the effect of Bifidobacterium longum BB536 on pellagra-related nausea caused by pirfenidone in our mouse model. Our pharmacological data indicated that pirfenidone (PFD) causes modulation of the gut microbiota profile, which appeared to play an important role in the development of pellagra-related nausea. A gut microbiota-mediated protective effect of B. longum BB536 against nausea caused by PFD was also identified. Finally, the urinary ratio of nicotinamide/N-methylnicotinamide was shown to be a biomarker of pellagra-like adverse effects induced by PFD, and it may contribute to the prevention of these effects in patients with idiopathic pulmonary fibrosis.

DOI PubMed

The Soluble Lectin Families as Novel Biomarkers for COVID-19 Pneumonia.

Haruka Takenaka, Atsushi Saito, Koji Kuronuma, Keigo Moniwa, Hirotaka Nishikiori, Satoshi Takahashi, Hirofumi Chiba

In vivo (Athens, Greece) 37 ( 4 ) 1721 - 1728 2023年 [国際誌]

BACKGROUND/AIM: The lung-specific soluble lectins, SP-A and SP-D have been clinically used to diagnose interstitial lung disease, but their clinical significance in COVID-19 remains controversial. This study was undertaken to determine their association with other lectins (MBL and FCN1), disease severity, and radiographs in COVID-19 patients. PATIENTS AND METHODS: A total of 131 patients with COVID-19 admitted in the Sapporo Medical University Hospital between May 22 and September 19, 2021, were enrolled in the study. Data including demographics, medical history, symptoms, signs, laboratory findings, and radiological images were collected from the patients' medical records. Chest computed tomography (CT) scanning was performed at admission. Serum levels of surfactant protein A and D (SP-A and SP-D), mannose-binding lectin (MBL) and ficolin1 (FCN1) were measured using enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: Compared to the control group, the COVID-19 group had significantly higher serum SP-A and FCN1 levels on admission (SP-A: 59.60±38.89 vs. 35.61±11.22 ng/ml; p<0.01, FCN1: 542.45±506.04 vs. 250.6±161.1 ng/ml; p<0.01). The severe group in COVID-19 had significantly higher serum SP-D and lower MBL levels than the non-severe group (SP-D: 141.7±155.7 vs. 61.41±54.54 ng/ml; p<0.01, MBL: 1,670±1,240 vs. 2,170±1,140 ng/ml; p<0.05). SP-D strongly reflected the degree of imaging findings, whereas SP-A showed a significant correlation, albeit slightly weaker than SP-D. Conversely, MBL and FNC1 were not significantly correlated with imaging findings. CONCLUSION: Among soluble serum lectins, SP-A and SP-D may be more sensitive to CT findings than reported disease biomarkers such as IL-6, LDH, and CRP due to their lung-specific characteristics.

DOI PubMed

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