The development of fluorescent protein tracing vectors for multicolor imaging of clinically isolated Staphylococcus aureus

Recent advances in fluorescent protein technology provide a wide variety of biological imaging applications; however current tools for bio-imaging in the Gram-positive bacterium Staphylococcus aureus has necessitated further developments for fluorescence intensity and for a multicolor palette of fluorescent proteins. To enhance the expression of multicolor fluorescent proteins in clinical S. aureus strains, we developed new fluorescent protein expression vectors, containing the blaZ/sodp promoter consisting of the β-lactamase gene (blaZ) promoter and the ribosome binding site (RBS) of superoxide dismutase gene (sod). We found S. aureus-adapted GFP (GFPsa) driven by the blaZ/sodp promoter was highly expressed in the S. aureus laboratory strain RN4220, but not in the clinical strains, MW2 and N315, harboring the endogenous blaI gene, a repressor of the blaZ gene promoter. We therefore constructed a constitutively induced blaZ/sodp promoter (blaZ/sodp(Con)) by introducing substitution mutations into the BlaI binding motif, and this modification allowed enhanced expression of the multicolor GFP variants (GFPsa, EGFP, mEmerald, Citrine, Cerulean, and BFP) as well as codon-optimized reef coral fluorescent proteins (mCherry and AmCyan) in the S. aureus clinical strains. These new fluorescent probes provide new tools to enhance expression of multicolor fluorescent proteins and facilitate clear visualization of clinical S. aureus strains.This study was supported by Grant-in-Aid for Young Scientists (B) Grant Number JP25861744 and Grant-in-Aid for Scientific Research (C) Grant Number JP25460533 from the Japan Society for the promotion of Science (JSPS). A confocal laser scanning microscopy was performed at the Analysis Center of Life Science, Natural Science Center for Basic Research and Development, Hiroshima University

Unconscious Biases Affect Women and Mothers

The purpose of this study is to sketch unconscious biases that affect women and mothers. As a result of this study, the following points become clear: (1) Unconscious biases also inform social norms and roles in women’s dress and makeup. Women accept them unconsciously. (2) The unconscious bias that “childcare is the responsibility of women” is still strong today. Therefore, mothers tend to feel guilt about their children. Mothers also hesitate to leave their children at daycare as a rest for themselves. (3) The unconscious bias that “childcareis the responsibility of women” causes various forms of maternal guilt. However, previous studies have not shown that mothers who work have a negative effect on their children’s development. These unconscious biases have been created by the social environment and culture and have an impact on the formation of our values and stereotypes. We need to think about the issues that women and mothers face by recognizing our unconscious biases.本稿は，日本発達心理学会第30回大会ラウンドテーブルで行った話題提供がもとになっている

Why is the Temporary Childcare System Difficult for Japanese Parents to Use for Parental Relief?

This study clarifies the reasons Japanese parents experience difficulties in using the temporary childcare system for parental relief. Three factors were identified following a literature review of previous research related to temporary childcare systems and information from the official websites of each municipality. First, it was difficult to identify the little information provided about institutional procedures. When parents attempt to use the temporary childcare system for parental relief, they need to collect information and procedures from over one month prior and it is necessary to organize with the childcare provider in advance. Local governments may not provide enough information and may not maintain or manage temporary childcare systems. Second, temporary childcare was difficult to use for parental relief because of unconscious bias. For example, the unconscious bias, i.e., “childcare is the mother’s responsibility,” means that principals may not respond positively to the parents’ need to use the temporary childcare system for relief. Third, the mothers’ guilt makes them hesitate using the temporary childcare systems. Mothers also hold the same unconscious bias that childcare is the mother’s responsibility; therefore, they may hesitate using the temporary childcare system. Furthermore, these behaviors and actions also interfere with other mothers’ use of the temporary childcare systems for parental relief

A Comparison of SCAT and Ueno’s Qualitative Analysis as the Qualitative Data Analysis: A Study of Interview Data from a Kindergarten Director

The purpose of this study is to compare the characteristics of SCAT (Steps for Coding and Theorization) and Ueno’s qualitative analysis as a form of qualitative data analysis applied to interview data from a kindergarten director. The subject and methods of the study are as follows. (1) We interviewed the director of Kindergarten A. (2) The research question asked why he started to run the child-based kindergarten. (3) We analyzed the interview data using SCAT. (4) We also analyzed the interview data using Ueno’s qualitative analysis. (5) We compared their characteristics. The analyses made the following points clear. First, the SCAT is characterized by the decontextualisation of the segmented text, step by step. By contrast, Ueno’s qualitative analysis is marked by its recontextualization through the mapping and charting of metadata. Second, when we use SCAT, we need to understand the theory of qualitative inquiry and must read the text over and over, analyzing it over time. However, the intellectual excitement when we discover the inherent meaning in the text is great. However, Ueno’s qualitative analysis is superior in terms of cost performance, time saved, and energy savings, though it does not convert qualitative data into words. With this approach, the intellectual excitement we felt when we discovered the meaning inherent in a text was not as great as when using SCAT

津波減災のための粘り強い海岸堤防の提案と避難施設周辺の津波挙動の評価

学位の種別: 論文博士審査委員会委員 : (主査)東京大学教授 佐藤 愼司, 東京大学教授 古関 潤一, 東京大学教授 田島 芳満, 東京大学教授 佐竹 健治, 東京大学講師 下園 武範University of Tokyo(東京大学

The Possibilities and Challenges of Multi-vocal Visual Ethnography as a Research Methodology

The purpose of this study is to provide an overview of how multi-vocal visual ethnography is applied in the world and to examine its possibilities and problems as a research methodology. Twenty-three academic articles using multi-vocal visual ethnography were collected and analyzed. The analysis results revealed the following: (1) multi-vocal visual ethnography is used as a tool for understanding not only the “visible,” but also the “invisible.” (2) The technique is used not only in comparative cultural research, but also as a tool for capturing different perspectives within a single culture. (3) Photographs and websites are used as cues, in addition to videos. (4) Cues consist of a variety of content. Unlike conventional ethnography, multi-vocal visual ethnography is a research methodology that has the potential to promote dialogues between researchers and people in and outside the field

The carboxy-terminal fragment of α1A calcium channel preferentially aggregates in the cytoplasm of human spinocerebellar ataxia type 6 Purkinje cells

Spinocerebellar ataxia type 6 (SCA6) is an autosomal dominant neurodegenerative disease caused by a small polyglutamine (polyQ) expansion (control: 4–20Q; SCA6: 20–33Q) in the carboxyl(C)-terminal cytoplasmic domain of the α1A voltage-dependent calcium channel (Cav2.1). Although a 75–85-kDa Cav2.1 C-terminal fragment (CTF) is toxic in cultured cells, its existence in human brains and its role in SCA6 pathogenesis remains unknown. Here, we investigated whether the small polyQ expansion alters the expression pattern and intracellular distribution of Cav2.1 in human SCA6 brains. New antibodies against the Cav2.1 C-terminus were used in immunoblotting and immunohistochemistry. In the cerebella of six control individuals, the CTF was detected in sucrose- and SDS-soluble cytosolic fractions; in the cerebella of two SCA6 patients, it was additionally detected in SDS-insoluble cytosolic and sucrose-soluble nuclear fractions. In contrast, however, the CTF was not detected either in the nuclear fraction or in the SDS-insoluble cytosolic fraction of SCA6 extracerebellar tissues, indicating that the CTF being insoluble in the cytoplasm or mislocalized to the nucleus only in the SCA6 cerebellum. Immunohistochemistry revealed abundant aggregates in cell bodies and dendrites of SCA6 Purkinje cells (seven patients) but not in controls (n = 6). Recombinant CTF with a small polyQ expansion (rCTF-Q28) aggregated in cultured PC12 cells, but neither rCTF-Q13 (normal-length polyQ) nor full-length Cav2.1 with Q28 did. We conclude that SCA6 pathogenesis may be associated with the CTF, normally found in the cytoplasm, being aggregated in the cytoplasm and additionally distributed in the nucleus

急速に増大する腫瘤影を呈した肺Mycobacterium avium症の1例

症例は66歳,男性.慢性閉塞性肺疾患とい草塵肺で経過観察をしていた.6カ月前の胸部CTでは明らかな異常を認めなかったが,新たに左上葉の気腫性病変周囲に腫瘤性病変を認めた.気管支鏡検査にて,局所検体からM.avium が検出されたものの生検で肉芽腫病変を認めなかったため,CTガイド下肺生検を実施した結果,肺MAC症と最終診断した.近年,孤立性腫瘤形成型肺MAC症の症例を散見するようになってきているが,本症例のごとく短期間で急速に増大することもあることから,抗酸菌を含めた肺感染症に対する積極的な検査が必要と思われる.A 66-year-old man was admitted to our hospital for follow-up on chronic obstructive pulmonary disease with a recent-showing abnormal chest shadow. He had received a periodic chest computed tomography (CT) six months prior due to a past history of COPD and Igusa pneumoconiosis. Although there was no mass shadow on the chest CT six months ago, a solitary tumorous shadow appeared surrounding the emphysematous lesions in the left upper lobe. M. avium was detected from local specimens viabronchoscopic examination, but because a granulomatous lesion was not observed, we performed a CT-guided lung biopsy and made a final diagnosis of pulmonary MAC disease. We recently observed that pulmonary MAC disease presents as a solitary tumorous shadow. However, as there are cases of pulmonary MAC disease presenting as a rapidly growing tumorous shadow within a short time, it is necessary to perform aggressive examinations for infectious diseases including an acid-fast bacilli examination