Effects of the Cultivation Method on the Characteristics or Gene Expression Profiles of Aspergillus oryzae Using mCD or DPY Media

We used modified Czapek-Dox (mCD) or dextrin-peptone-yeast extract (DPY) media to cultivate a filamentous fungus, Aspergillus oryzae IAM 2706 by three different cultivation methods, i.e., shaking-flask culture (SFC), agar-plate culture (APC), and membrane-surface liquid culture (MSLC), to identify the differences in cultivation behaviors and gene transcriptional profiles. The fungi cultivated by APC or MSLC secreted a greater number of different proteins/enzymes in larger quantities compared with fungi cultivated by SFC, particularly when DPY medium was used. In particular, the amounts of protease secreted by fungi cultivated via MSLC or APC were much greater compared with SFC. When mCD medium was used, α-amylase activity was barely detectable in all cultures while the activity was detected in MSLC and APC in a quantity that was several times higher than that in SFC using DPY medium. SDS-PAGE analysis and N-terminal amino acid sequences confirmed 6 proteins in the culture supernatants when DPY medium was used. Among these proteins oryzin (an alkaline protease) and α-amylase were detected at much higher levels in APC and MSLC compared with SFC, which was consistent with the measured activity of the secreted enzymes. However, when mCD medium was used, only oryzin was detected in significant amounts in MSLC and APC. Microarray analyses of the fungi cultivated by SFC, APC or MSLC using either mCD or DPY media indicated that the gene transcriptional profile of the MSLC sample was similar to that of the APC sample but different from that of the SFC sample. When mCD medium was used, most of the genes that were up-regulated 10-folds or greater in the MSLC sample relative to the SFC sample were unknown or predicted proteins. Transcription of the oryzin gene was only slightly up-regulated in the MSLC sample while transcription of the α-amylase gene was slightly down-regulated. On the other hand, when DPY medium was used, many known genes including the oryzin gene were up-regulated in the MSLC sample versus the SFC sample

Impact of the cefazolin shortage on the selection and cost of parenteral antibiotics during the supply disruption period in Japan: A controlled interrupted time series analysis

[Background] A serious shortage of cefazolin (CEZ) occurred in Japan in 2019. We compared the impact of the CEZ shortage on the selection of parenteral antibiotics at affected and non-affected hospitals. [Methods] The data were extracted from a nationwide Japanese administrative database and included all hospitalized cases between April 2016 and December 2020. We defined ‘hospitals with shortage’ as those hospitals with a statistically significant decrease in the use of CEZ during the supply disruption period compared to the same months of the previous year; other hospitals as ‘hospitals without shortage’. We determined the proportion of each selected parenteral antibiotic use to the sum of all selected antibiotic use in the two groups of hospitals during the supply disruption period and during the same months of the previous year. A controlled interrupted time series (CITS) analysis was conducted to estimate the impact of the CEZ shortage on each antibiotic use and the cost of all parenteral antibiotics per patient day in hospitals with shortage as compared to those without shortage. [Results] In the hospitals with shortage, the proportion of CEZ use to the sum of all selected antibiotics decreased (23.5–11.1%). The decrease in CEZ use was mainly offset by the use of ceftriaxone, ceftriaxone, and ampicillin/sulbactam. The CITS analysis showed a statistically significant increase in the use of broader-spectrum beta-lactams and clindamycin during the supply disruption period (flomoxef up 58.1%, cefotiam up 63.1%, cefmetazole up 14.5%, ceftriaxone up 13.9%, and clindamycin up 20.1%). The analysis showed no statistically significant change in the cost of all parenteral antibiotics per patient day. [Conclusions] During the CEZ supply disruption, there was a statistically significant increase in the use of broader-spectrum beta-lactams and clindamycin in hospitals with shortage compared with those without shortage

Hospitalization for ischemic stroke was affected more in independent cases than in dependent cases during the COVID-19 pandemic: An interrupted time series analysis

[Background] The pandemic of the coronavirus disease 2019 (COVID-19) has affected health care systems globally. The aim of our study is to assess the impact of the COVID-19 pandemic on the number of hospital admissions for ischemic stroke by severity in Japan. [Methods] We analysed administrative (Diagnosis Procedure Combination—DPC) data for cases of inpatients aged 18 years and older who were diagnosed with ischemic stroke and admitted during the period April 1 2018 to June 27 2020. Levels of change of the weekly number of inpatient cases with ischemic stroke diagnosis after the declaration of state of emergency were assessed using interrupted time-series (ITS) analysis. The numbers of patients with various characteristics and treatment approaches were compared. We also performed an ITS analysis for each group (“independent” or “dependent”) divided based on components of activities of daily living (ADL) and level of consciousness at hospital admission. [Results] A total of 170, 294 cases in 567 hospitals were included. The ITS analysis showed a significant decrease in the weekly number of ischemic stroke cases hospitalized (estimated decrease: −156 cases; 95% confidence interval (CI): −209 to −104), which corresponds to −10.4% (95% CI: −13.6 to −7.1). The proportion of decline in the independent group (−21.3%; 95% CI: −26.0 to −16.2) was larger than that in the dependent group (−8.6%; 95% CI: −11.7 to −5.4). [Conclusions] Our results show a marked reduction in hospital admissions due to ischemic stroke after the declaration of the state of emergency for the COVID-19 pandemic. The independent cases were affected more in proportion than dependent cases

Crystallization characteristics of amorphous trehalose dried from alcohol

Trehalose forms a glass that can be used to preserve labile substances under desiccation. The crystallization characteristics, namely crystallization temperature (Tcry) and isothermal crystallization behavior of amorphous trehalose, dried from alcohol (methanol, ethanol), was analyzed and the results were compared with those for the amorphous trehalose freeze-dried from water. The use of alcohol as a solvent lowered the Tcry from 184 ± 6 °C for the case of an aqueous solvent to 103 ± 5 °C/methanol and 120 ± 8 °C/ethanol. The formation of multiple forms of crystals and partial melting were suggested by the thermal analysis. Isothermal crystallization experiments showed that the alcohol-originated amorphous trehalose was eventually exclusively converted into β-form crystals. The induction period (tind) before the start of isothermal crystallization was markedly shortened when alcohol was used as the solvent compared to water. The tind values for various amorphous sugar samples including the alcohol-originated ones could be correlated with difference between Tcry and the sample temperature

Case Report: Acute Eosinophilic Myocarditis With a Low-Flow Heart Failure With Preserved Ejection Fraction Phenotype

Eosinophilic myocarditis is a rare subtype of myocarditis characterized by myocardial eosinophilic infiltration, and it is potentially fatal if left untreated. Although endomyocardial biopsy (EMB) is a cornerstone for the histological diagnosis of acute eosinophilic myocarditis (AEM), as it is an invasive procedure and has a low diagnostic accuracy, the diagnosis of AEM with hemodynamic instability remains challenging. We describe a case of AEM presenting as low-flow heart failure with preserved ejection fraction (HFpEF), with rapid progression to cardiogenic shock. The constellation of peripheral eosinophilia, increased left ventricular wall thickness, and HFpEF raised the suspicion of AEM. Contrast-enhanced computed tomography (CT) scan revealed heterogeneous hypoenhancement localized in the basal-to-mid septal and mid anterolateral walls of the left ventricle, strongly suggestive of acute inflammation. Based upon these findings, we performed CT-guided EMB, which lead to a definitive diagnosis. Subsequent high-dose corticosteroids allowed a rapid and dramatic recovery and normalization of cardiac structure and function. This case highlights the clinical importance of assessing AEM as a rare cause of HFpEF and the usefulness of CT-guided EMB in patients with hemodynamic instability

A promoter haplotype of the interleukin-18 gene is associated with juvenile idiopathic arthritis in the Japanese population

Recently, we reported that genetic polymorphisms within the human IL18 gene were associated with disease susceptibility to adult-onset Still's disease (AOSD), which is characterized by extraordinarily high serum levels of IL-18. Because high serum IL-18 induction has also been observed in the systemic type of juvenile idiopathic arthritis (JIA), we investigated whether similar genetic skewing is present in this disease. Three haplotypes, S01, S02, and S03, composed of 13 genetic polymorphisms covering two distinct promoter regions, were determined for 33 JIA patients, including 17 with systemic JIA, 10 with polyarthritis, and 6 with oligoarthritis. Haplotypes were also analyzed for 28 AOSD patients, 164 rheumatoid arthritis (RA) patients, 102 patients with collagen diseases, and 173 healthy control subjects. The frequency of individuals carrying a diplotype configuration (a combination of two haplotypes) of S01/S01 was significantly higher in the JIA patients, including all subgroups, than in the healthy controls (P = 0.0045, Fischer exact probability test; odds ratio (OR) = 3.55, 95% confidence interval (CI) = 1.55–8.14). In patients with systemic JIA, its frequency did not differ statistically from that of normal controls. Nevertheless, it is possible that haplotype S01 is associated with the phenotype of high IL-18 production in systemic JIA because the patients carrying S01/S01 showed significantly higher serum IL-18 levels compared with patients with other diplotype configurations (P = 0.017, Mann-Whitney U test). We confirmed that the frequency of the diplotype configuration of S01/S01 was significantly higher in AOSD patients than in healthy control subjects (P = 0.011, OR = 3.45, 95% CI = 1.42–8.36). Furthermore, the RA patients were also more predisposed to have S01/S01 (P = 0.018, OR = 2.00, 95% CI = 1.14–3.50) than the healthy control subjects, whereas the patients with collagen diseases did not. In summary, the diplotype configuration of S01/S01 was associated with susceptibility to JIA as well as AOSD and RA, and linked to significantly higher IL-18 production in systemic JIA. Possession of the diplotype configuration of S01/S01 would be one of the genetic risk factors for susceptibility to arthritis in the Japanese population

Hospitalization of mild cases of community-acquired pneumonia decreased more than severe ones during the COVID-19 epidemic

COVID-19流行が市中肺炎の緊急入院に与えた影響を検証. 京都大学プレスリリース. 2021-05-10.[Objective] The coronavirus disease 2019 (COVID-19) pandemic has affected all healthcare systems. This study aimed to assess the impact of the COVID-19 pandemic on the number and severity of cases of community-acquired pneumonia (CAP) in Japan. [Methods] Using claims data from the Quality Indicator/Improvement Project (QIP) database, urgent cases of inpatients for CAP from 01 August 2018 to 30 July 2020 were included. The monthly ratios of inpatient cases were compared from August 2018 to July 2019 and August 2019 to July 2020 as a year-over-year comparison. These ratios were also compared according to the “A-DROP” severity score, and an interrupted time series (ITS) analysis was performed to evaluate the impact of the COVID-19 pandemic on the monthly number of inpatient cases. [Results] This study included a total of 67, 900 inpatient cases for CAP in 262 hospitals. During the COVID-19 pandemic (defined as the period between March and July 2020) the number of inpatient cases for CAP drastically decreased compared with the same period in the previous year (–48.1%), despite a temporary reduction in the number of other urgent admissions. The number of inpatient cases decreased according to the severity of pneumonia. Milder cases showed a greater decrease in the year-over-year ratio than severe ones: mild –55.2%, moderate –45.8%, severe –39.4%, and extremely severe –33.2%. The ITS analysis showed that the COVID-19 pandemic significantly reduced the monthly number of inpatient cases for CAP (estimated decrease: –1233 cases; 95% CI –521 to –1955). [Conclusions] This study showed a significant reduction in the number of inpatient cases for CAP during the COVID-19 pandemic in Japan. The milder cases showed a greater decrease in the year-over-year ratio of the number of inpatient cases

チョウコウネツ シゲンキン Thermococcus kodakaraensis KOD1 ノ トウ タイシャ カンレン コウソ ニ カンスル ケンキュウ

京都大学0048新制・課程博士博士(工学)甲第10228号工博第2291号新制||工||1278(附属図書館)UT51-2003-H649京都大学大学院工学研究科合成・生物化学専攻(主査)教授 青山 安宏, 教授 木村 俊作, 教授 村上 正浩学位規則第4条第1項該当Doctor of EngineeringKyoto UniversityDA