長期ホルマリン固定により失活したProliferating Cell Nuclear Antigen (PCNA) の免疫反応性回復条件の基礎的検討 ―マイクロウェーブ、オートクレーブの影響について―

Using paraffin-embedded tissue sections of liver cancer obtained from autopsy which had been preserved in 10% buffered formalin solution for 6 months while PCNA immunoreactivity was lost, we examined the effects of heat processing by either microwave(MW) and autoclave(AC) in the presence of various processing solution. It appeared that AC processing took shorter time period than MW irradiation to restore equal immunoreactivity. With regard to immunoreactivity retrieval by MW irradiation,however, variation of the degree of retrieval depending on processing time was smaller than in AC, and so the stable consequences were obtained. Although AC processed tissues tended to be stained deep, prolonged processing time presented strong background staining and blurred nuclear margins which made it difficult to estimate the positive cell count. As for the effects of processing solution, there was little difference in retrieval of PCNA among 0.01 M citrate buffer (pH 6.0), saturated solution of lead thiocyanate and distilled water, but the least background staining was observed with distilled water. These observations suggest that MW irradiation of which effect of retrieval is less dependent of processing time and with the least background stainability, is superior to AC processing for PCNA immunoreactivity retrieval on formalin-fixed tissues.10％緩衝ホルマリンに6ヶ月間浸漬していた剖検材料（肝臓癌）のパラフィン包埋後の組織切片を用いて、ホルマリンの固定作用により失活したPCNAの免疫反応性の回復にマイクロウェーブ（MW）及びオートクレーブ（AC）による熱処理とその時用いる処理溶液が、どのような影響を与えるかについて検討した。その結果、同等の免疫反応性を回復するのには、AC処理の方がMW照射より短時間でよいことが解った。しかし、MW照射による免疫反応性回復では、処理時間による影響がAC処理に比べて少なく、安定した結果が得られた。また、PCNAの染色所見については、AC処理の方が濃く染まる傾向が見られた。しかし、処理時間が長くなるとバックグラウンドの染色性が高くなる、核の周囲ににじみ現象が見られる等の所見があり判定に困難をきたした。一方、処理溶液についてはクエン酸（0.01M pH6.0）、チオシアン酸鉛飽和溶液、蒸留水について検討を行った結果、PCNAの回復には差は認められなかったが、バックグラウンドの染色については蒸留水が最も少なかった。以上の結果により、ホルマリン固定により失活したPCNAの免疫反応性を回復するには、MW照射の方がAC処理より処理時間に関係なく安定した染色性が得られ、なおかつバックグラウンドの染色性が少ない等の点で優れていることが示唆された

High Time-Resolution Monitoring of Free-Tropospheric Sulfur Dioxide and Nitric Acid at the Summit of Mt. Fuji, Japan

This is the first paper that describes the atmospheric sulfur dioxide (SO2) and nitric acid (HNO3) monitored with a good time-resolution at the summit (3,776 m a.s.l.), which is located in the free troposphere, and southeastern foot (1,284 m a.s.l.) of Mt. Fuji. Japan. During the summer of 2012, two analytical systems consisting mainly of a parallel-plate wet denuder and ion chromatograph operated simultaneously at both the sampling sites. All the samples collected at both the sampling sites contained detectable levels of sulfate from gas-phase SO2 while the nitrate from gas-phase HNO3 was detectable in 97.8% of air samples at the southeastern foot and 88.4% at the summit. The average concentrations of SO2 and HNO3 were, respectively, 0.061 ± 0.071 and 0.031 ± 0.020 ppbv at the summit (n = 672), and 0.347 ± 0.425 and 0.146 ± 0.070 ppbv at the southeastern foot (n = 1344) of Mt. Fuji. Both the acidic gases at the southeastern foot and the HNO3 at the summit showed a diurnal pattern with daytime maxima and nighttime minima. Meanwhile, the SO2 at the summit did not show a distinct shift, which indicates the SO2 concentrations at the summit would be principally controlled by the advection of air parcel in the free troposphere

Partially functional outer arm dynein in a novel Chlamydomonas mutant expressing a truncated γ heavy chain

The outer dynein arm of Chlamydomonas flagella contains three heavy chains (α, β, and γ), each of which exhibits motor activity. How they assemble and cooperate is of considerable interest. Here we report the isolation of a novel mutant, oda2-t, whose γ heavy chain is truncated at about 30% of the sequence. While the previously isolated γ chain mutant oda2 lacks the entire outer arm, oda2-t retains outer arms that contain α and β heavy chains, suggesting that the N-terminal sequence (corresponding to the tail region) is necessary and sufficient for stable outer-arm assembly. Thin-section electron microscopy and image analysis localize the γ heavy chain to a basal region of the outer-arm image in the axonemal cross section. The motility of oda2-t is lower than that of the wild type and oda11 (lacking the α heavy chain) but higher than that of oda2 and oda4-s7 (lacking the motor domain of the β heavy chain). Thus, the outer-arm dynein lacking the γ heavy-chain motor domain is partially functional. The availability of mutants lacking individual heavy chains should greatly facilitate studies on the structure and function of the outer-arm dynein

Isolation and characterization of cancer stem cells derived from human glioblastoma

Cancer stem cell (CSC) is considered as a cause of cancer recurrence and metastasis. Simultaneously CSCs are responsible for the heterogeneous population in tumor tissues due to their differentiation potential. However, the characterizations of CSCs are still not enough and cancer stem cell lines widely available is desired to be established for the advancement of cancer research. In this study, we tried to isolate and characterize stem like cells from human glioblastoma cell line U-251MG cells. U-251MG P1 cells, which was previously condensed in the presence of hyaluronic acid as CD44 positive population were subjected to single cell isolation procedure. Although 5 clones were isolated, only one clone exhibited high expression of CD44, Nanog, OCT3/4 and SOX2, and named U-251MGSC1. The sphere forming ability of U-251MGSC1 cell was significantly higher than the parental U-251MG cells. Tumorigenicity of U-251MG-SC1 cells were higher than that of U-251MG cells. U-251MGSC1 cells exhibited higher expression of CD44, SOX2, Nestin and A2B5 than U-251MG cells in vitro and in vivo. The expression of GFAP and NF-M was enhanced when the cells were treated with the conditioned medium of U-251MG cells indicating the potential of differentiation. Sphere forming ability was more efficient than that of U-251MG cells and was enhanced in the presence of hyaluronic acid, which enhanced the cell growth as well. U-251MGSC1 cells exhibited rapid growth tumor in nude mice and efficient metastatic ability in transmembrane assay when compared with U-251MG cells. As the result, we concluded U-251MGSC1 cell was a glioblastoma CSC line derived from the parental U-251MG cells. U-251MGSC1 cells will be a good tool to develop effective therapeutic agents against CSCs and to elucidate the properties of glioma derived CSCs and the mechanism of tumor development in brain

Human mast cell activation through Fc receptors and Toll-like receptors

ABSTRACTMast cells express high-affinity IgE receptors (FcεRI) on their surface and can be activated to secrete a variety of biologically active mediators by cross-linking of receptor-bound IgE. Recent studies in animal models indicate that mouse mast cells may play a protective role in host defense against bacteria through the production of tumor necrosis factor-α, mainly as a result of Toll-like receptor (TLR) 4- or CD48-mediated activation. Moreover, several recent observations in animal models have indicated that mast cells may also play a pivotal role in coordinating the early phases of autoimmune diseases, particularly those involving auto-antibodies. We recently identified functional TLR4 and FcγRI on human mast cells, in which their expression had been upregulated by interferon-γ. We compared each of the receptor-mediated gene expression profiles with the FcεRI-mediated gene expression profile using high-density oligonucleotide probe arrays and discovered that human mast cells may modulate the immune system in a receptor-specific manner

Nutrient Profiling of Japanese Dishes: The Development of a Novel Ajinomoto Group Nutrient Profiling System

Government agencies and private companies have supported the development of nutrient profiling (NP) systems to facilitate the selection of nutrient-dense foods by consumers, promote nutritious food development, and limit excessive advertising of products with low nutritional value. While most NP models were developed to assess individual foods, the Ajinomoto Group Nutrient Profiling System (ANPS) was developed to assess the overall nutritional value of cooked dishes that are culturally specific to Japan. Based on the national dietary recommendations and nutritional surveys, target values were created for 13 dish categories, while considering the combinations of meal units. For the ANPS, the four evaluating elements were protein and vegetables, which should be encouraged, and sodium and saturated fatty acids, which should be limited. The ANPS algorithm for dishes was the sum of the scores of individual elements, with a maximum of 10 points per serving. The sum of scores was then multiplied by 2.5 to convert to the 100-point scale. Convergent validity was tested using the nutrient-rich food index (NRF) score of 6.3. In total, 1,089 popular Japanese dishes were evaluated using the ANPS, and the median score of ANPS was 70.0 points (interquartile range, 55–78.8), and the average score was 67.7 (standard deviation, 16.5) points. Since salt intake is a major health risk in Japan, this tool was designed to evaluate sodium content with high sensitivity, and low-salt dishes significantly improved sodium and ANPS scores compared with regular dishes. The Pearson’s correlation coefficient between the total score of NRF 6.3 and ANPS in 1,089 dishes was r = 0.452 (p < 0.0001). This newly developed ANPS could be used to evaluate culture-specific cooked dishes per serving size. It can determine the nutritional values of dishes, with a high sensitivity to sodium content, a major Japanese nutritional issue. Further research is needed to determine the accuracy and usefulness of the ANPS as a system that would lead to changes in eating behavior nationwide

Endothelin suppresses cell migration via the JNK signaling pathway in a manner dependent upon Src kinase, Rac1, and Cdc42

AbstractCell migration is a complex phenomenon that is stimulated by chemoattractive factors such as chemokines, a family of ligands for G protein-coupled receptors (GPCRs). In contrast, factors that suppress cell migration, and the mechanism of their action, remain largely unknown. In this study, we show that endothelin, a GPCR ligand, inhibits cell motility in a manner dependent upon signaling through the c-Jun N-terminal kinase (JNK) pathway. We further demonstrate that this effect is dependent upon Src kinase and small GTPases Rac1 and Cdc42. These findings provide new insight into GPCR-mediated regulation of cell migration

Multiple Physical Symptoms Are Useful to Identify High Risk Individuals for Burnout: A Study on Faculties and Hospital Workers in Japan

Healthcare workers have a high risk of burnout. This study aimed to investigate if the numbers of physical symptoms are associated with burnout among healthcare workers. We conducted a cross-sectional survey at a large university in Tokyo, Japan, in 2016. Participants were 1080: 525 faculties and 555 hospital workers. We investigated 16 physical symptoms perceived more than once per week and examined the association between the number of physical symptoms and Copenhagen Burnout Inventory (CBI); work-related (WBO), personal (PBO), and client-related (CBO) burnout. All CBI scores were higher among hospital workers than among faculties: WBO (43 vs. 29), PBO (50 vs. 33), CBO (33 vs. 29). Moreover, the higher the number of physical symptoms perceived, the higher the degree of burnout scores became (trend p-values < 0.001), except for CBO among faculties. Job strain (all except for CBO among hospital workers) and work-family conflict were associated with an increased risk of burnout. Being married (WBO and CBO among faculties), having a child (except for PBO and CBO among faculties), and job support (faculty and hospital workers with WBO and faculties with PBO) were associated with a decreased risk of burnout. Multiple physical symptoms might be useful for identifying high risk individuals for burnout

看護学生の看護の認識

看護学生は入学初期から学習する看護学や，日常生活の中で多様な体験や経験を積むことにより，看護に対する見方・考え方を変えながら，自らの看護観を形成していく．今回，看護学生が入学時と，１年間看護学やその関連科目，教養科目などを学習した１年終了時における看護の見方・考え方について学生の記述内容を分析した．その結果，看護学生は入学後１年間の学習や経験などが加味され，看護に関する認識を新たにしたり深めたりしていた．この認識の変化には，専門科目である看護学概論や看護理論の影響が大きいと考えられた．なぜならば看護学生は，１年間に学習した看護学概論や看護理論の科目の中で捉えた看護の考え方をベースに看護の認識を深めていた．Nursing students develop their own view of nursing through learning of nursing and accumulating diverse experiences during daily living after entering nursing school. The present study was undertaken to analyze the result of the descriptive survey designed to investigate changes in the students awareness of nursing during the first year of the nursing school (i.e., after one year of learning nursing, nursing-related subjects and subjects for general education). This survey revealed that the nursing students accumulated knowledge and experience and deepened thir awareness of nursing during the one year period. These changes in their awareness seem to be particularly related to education of nursing-specific subjects (introduction and theories of nursing). The students deepened their awareness of nursing on the basis of the views they learned from these nursing-specific subjects