Novel Methods to Study Angiogenesis Using Tissue Explants

Tissue explants of skeletal muscles, brain, kidney, liver and spleen from mice were cultured using collagen gel. Electron microscopic observation revealed that formation of capillary tubes with pericyte-like cells occurred only from the tissue explant of skeletal muscles. The capillary tubes formed in the collagen gel were positive for tomato lectin and platelet/endothelial cell adhesion molecule (PCAM)-1 antibody. Formation of capillary tubes in the rat was more predominant than in the mouse. Plasmalemmal vesicles were clearly observed in the capillary tubes from rat tissue explant. Muscle fiber-type differences were also observed. In the soleus muscle, the formation of capillary tubes was predominant than the tibialis anterior muscle. Using this culture model from the rat soleus muscle, effects of α-isoproterenol (β-adrenergic receptor agonist) and low-frequency electrical stimulation were examined on the formation of capillary tubes and fine structures of skeletal muscle explant. The formation of capillary tubes was promoted by α-isoproterenol administration. At low-frequency electrical stimulation, the formation of capillary tubes was inhibited. Both α-isoproterenol and electrical stimulation reduced the degeneration of skeletal muscles. This culture method of skeletal muscles may provide a useful model that can examine the effects of various drugs and physical stimulations

Adverse effect of cake collapse on the functional integrity of freeze-dried bull spermatozoa

Under optimal freeze-drying conditions, solutions exhibit a cake-like porous structure. However, if the solution temperature is higher than the glass transition temperature of the maximally freeze-concentrated phase (Tg′) during drying phase, the glassy matrix undergoes viscous flow, resulting in cake collapse. The purpose of the present study was to investigate the effect of cake collapse on the integrity of freeze-dried bull spermatozoa. In a preliminary experiment, factors affecting the Tg′ of conventional EGTA buffer (consisting of Tris–HCl, EGTA and NaCl) were investigated in order to establish the main experimental protocol because EGTA buffer Tg′ was too low (−45.0 °C) to suppress collapse. Modification of the EGTA buffer composition by complete removal of NaCl and addition of trehalose (mEGTA buffer) resulted in an increase of Tg′ up to −27.7 °C. In the main experiment, blastocyst yields after ooplasmic injection of freeze-dried sperm preserved in collapsed cakes (drying temperature: 0 or −15 °C) were significantly lower than those of sperm preserved in non-collapsed cake (drying temperature: −30 °C). In conclusion, freeze-dried cake collapse may be undesirable for maintaining sperm functions to support embryonic development, and can be inhibited by controlling both Tg′ of freeze-drying buffer and temperature during the drying phase.ArticleCRYOBIOLOGY. 68(3):354-360 (2014)journal articl

酢酸摂取と運動が脂肪代謝と運動耐久性に及ぼす影響

Previously, we found that acetic acid had effects on lipid metabolism in skeletal muscles and has functions that work against obesity and obesity-linked type 2 diabetes through the activation of AMPactivated protein kinase (AMPK). During exercise, AMPK is activated in skeletal muscle according to exercise intensity and it increases fatty acid oxidation. The purpose of this study was to investigate the interactive effects of chronic intake of acetic acid and exercise training on lipid metabolism and endurance performance. Six-week-old SD rats were randomly assigned to four groups: water-injected (rest-water), acetic acid-injected (rest-ace), exercise-trained after injection of water (water-ex), and exercise-trained after injection of acetic acid (ace-ex) for 4 weeks. Body weight (BW) in rest-ace and ace-ex groups was significantly lower than rest-water group. Exercise-training groups showed an increase of exercise capacity, by the addition of intake of acetic acid, lipid oxidation was promoted during exercise tolerance test. Skeletal muscle of rats treated with acetic acid and exercise training led to higher expressions of cytochrome c (cycs), and tended to stimulate expressions of peroxisome proliferator-activated receptor coactivator 1-α (PGC1-α ) and MHC1 genes than those of rest-water group. Those results indicate that treatments both of exercise training and intake of acetic acid contribute to enhancement of lipid metabolism and improvement of exercise capacity.これまで我々は、酢酸の摂取が骨格筋内のAMP活性化プロテインキナーゼ（AMPK）の活性化を介して脂質代謝と肥満、肥満に関連した2型糖尿病の予防に効果があることを示唆してきた。AMPKは運動によって骨格筋で活性化し、脂肪酸酸化を促進する。この研究は、4週間の継続的な酢酸摂取と運動トレーニングが運動中の脂肪代謝と運動耐久性に及ぼす影響について調べることを目的とした。　6週齢のSD系雄ラットを安静期に水を摂取するrest-water群、酢酸を摂取するrest-ace群、運動前に水を摂取するwater-ex群、運動前に酢酸を摂取するace-ex群に無作為に分け実験を行った。酢酸を継続的に摂取すると水摂取に比較して腹腔内脂肪量の減少と体重増加の抑制がみられた。また継続的な酢酸摂取および運動トレーニングにより、耐久性運動下でのグルコース利用の抑制および脂肪酸酸化の促進が見られた。酢酸摂取および運動トレーニング群の腓腹筋では、MHCIおよびcytochrome c等の遅筋線維マーカー遺伝子が増加していた。継続的な酢酸摂取と運動トレーニングにより、脂肪代謝と運動耐久性の向上が示唆された

Hemopoietic cell kinase (Hck) and p21-activated kinase 2 (PAK2) are involved in the down-regulation of CD1a lipid antigen presentation by HIV-1 Nef in dendritic cells

AbstractDendritic cells (DCs) play a major role in in vivo pathogenesis of HIV-1 infection. Therefore, DCs may provide a promising strategy to control and eventually overcome the fatal infection. Especially, immature DCs express all CD1s, the non-MHC lipid antigen -presenting molecules, and HIV-1 Nef down-regulates CD1 expression besides MHC. Moreover, CD1d-restricted CD4+ NKT cells are infected by HIV-1, reducing the number of these cells in HIV-1-infected individuals. To understand the exact role of DCs and CD1-mediated immune response during HIV-1 infection, Nef down-regulation of CD1a-restricted lipid/glycolipid Ag presentation in iDCs was analyzed. We demonstrated the involvement of the association of Nef with hemopoietic cell kinase (Hck) and p21-activated kinase 2 (PAK2), and that Hck, which is expressed strongly in iDCs, augmented this mutual interaction. Hck might be another therapeutic target to preserve the function of HIV-1 infected DCs, which are potential reservoirs of HIV-1 even after antiretroviral therapy

The Development and Assessment on the Qualities and Abilities Questionnaire in Elementary and Junior High School <Research Article>

本研究では，次期学習指導要領改訂の重要な観点である，主体性・多様性・協働性・学びに向かう力等の「育成すべき資質・能力」に着目して，小中学校の児童生徒を対象とした質問紙を開発し，教育現場での有効性を検討することを目的とした。質問紙開発に当たっては小学校校長や教諭等と協議を重ね，先行研究などから収集した項目を精選した。そして，資質・能力を多角的に見取るために，「社会性と情動」アンケート（田中・真井・津田・田中，2011）と共に，児童生徒約1600名に質問紙開発のための調査を実施した。その結果，育成すべき資質・能力に関して，「主体性・積極性」，「自尊心」，「協働する力」などの因子が得られた。続いて，各学校のニーズに応じて，児童一人一人の多様性の把握，学校全体，中学校区での分析を行ったところ，児童生徒の的確な見取りや，各校での取組の成果や課題の明確化につながった。以上の結果より，本資質・能力質問紙の有用性が示唆された。The present study aimed to develop a questionnaire concerning the "qualities and abilities that should be promoted," in terms of subjectivity, diversity, cooperation of labor, willingness to learn, and human nature, which will be important aspects of the next revision of the course of study; to conduct the questionnaire among children in elementary and junior high schools; and to examine its utility at these schools. When the questionnaire was developed, the author conferred repeatedly with the principal of the elementary school and with teachers, and the question items collected from previous works, etc., were selected. To identify the "qualities and abilities" multilaterally, the questionnaire was conducted with approximately 1600 children alongside a "sociality and affect" questionnaire (Tanaka, Sanai, Tsuda, and Tanaka, 2011 ). As a result, factors such as "subjectivity and activeness," "pride," and "cooperation of labor" could be related to "qualities and abilities that should be promoted." An analysis of findings in terms of the diversity of the children, the school as a whole, and the junior high school district was conducted by using the results in relation to the demands of each school, leading to an appropriate understanding of the children, clarification of the results of the teachers' approach, and the problems in each school. These findings suggest that this "qualities and abilities" questionnaire is useful

Oncofertility care in young women and the outcomes of pregnancy over the last 5 years

Aim: To ascertain the actual outcomes of oncofertility care in young women to provide more appropriate care. Materials & methods: We analyzed the data of 67 female patients under 43 years of age who underwent oncofertility care between January 2015 and September 2019. Results: There were 28 patients with breast cancer, 19 patients with hematologic cancer and 20 patients with other cancer diagnoses. Breast cancer patients tended to take longer than hematologic cancer patients to initiate oncofertility treatment. Despite undergoing oncofertility care, seven of nine pregnant patients did not choose assisted reproductive technology (ART). Conclusion: As spontaneous pregnancies were more common than ART pregnancies in our study, pregnancy by not only ART but also non-ART method is a viable option for young cancer survivors

Induction of tumor-specific acquired immunity against already established tumors by selective stimulation of innate DEC-205+ dendritic cells

Two major distinct subsets of dendritic cells (DCs) are arranged to regulate our immune responses in vivo; 33D1+ and DEC-205+ DCs. Using anti-33D1-specific monoclonal antibody, 33D1+ DCs were successfully depleted from C57BL/6 mice. When 33D1+ DC-depleted mice were stimulated with LPS, serum IL-12, but not IL-10 secretion that may be mediated by the remaining DEC-205+ DCs was markedly enhanced, which may induce Th1 dominancy upon TLR signaling. The 33D1+ DC-depleted mice, implanted with syngeneic Hepa1-6 hepatoma or B16-F10 melanoma cells into the dermis, showed apparent inhibition of already established tumor growth in vivo when they were subcutaneously (sc) injected once or twice with LPS after tumor implantation. Moreover, the development of lung metastasis of B16-F10 melanoma cells injected intravenously was also suppressed when 33D1+ DC-deleted mice were stimulated twice with LPS in a similar manner, in which the actual cell number of NK1.1+CD3− NK cells in lung tissues was markedly increased. Furthermore, intraperitoneal (ip) administration of a very small amount of melphalan (l-phenylalanine mustard; l-PAM) (0.25 mg/kg) in LPS-stimulated 33D1+ DC-deleted mice helped to induce H-2Kb-restricted epitope-specific CD8+ cytotoxic T lymphocytes (CTLs) among tumor-infiltrating lymphocytes against already established syngeneic E.G7-OVA lymphoma. These findings indicate the importance and effectiveness of selective targeting of a specific subset of DCs, such as DEC-205+ DCs alone or with a very small amount of anticancer drugs to activate both CD8+ CTLs and NK effectors without externally added tumor antigen stimulation in vivo and provide a new direction for tumor immunotherapy

Detection of deviance in Japanese kanji compound words

Reading fluency is based on the automatic visual recognition of words. As a manifestation of the automatic processing of words, an automatic deviance detection of visual word stimuli can be observed in the early stages of visual recognition. To clarify whether this phenomenon occurs with Japanese kanji compounds-since their lexicality is related to semantic association-we investigated the brain response by utilizing three types of deviants: differences in font type, lexically correct or incorrect Japanese kanji compound words and pseudo-kanji characters modified from correct and incorrect compounds. We employed magnetoencephalography (MEG) to evaluate the spatiotemporal profiles of the related brain regions. The study included 22 adult native Japanese speakers (16 females). The abovementioned three kinds of stimuli containing 20% deviants were presented during the MEG measurement. Activity in the occipital pole region of the brain was observed upon the detection of font-type deviance within 250 ms of stimulus onset. Although no significant activity upon detecting lexically correct/incorrect kanji compounds or pseudo-kanji character deviations was observed, the activity in the posterior transverse region of the collateral sulcus (pCoS)-which is a fusiform neighboring area-was larger when detecting lexically correct kanji compounds than when detecting pseudo-kanji characters. Taken together, these results support the notion that the automatic detection of deviance in kanji compounds may be limited to a low-level feature, such as the stimulus stroke thickness.Peer reviewe

The Role of Radiotherapy for Thymic Carcinoma

Objective: The aim of this study is to evaluate retrospectively the role of radiotherapy for thymic carcinoma. Methods: Between 1973 and 1998, 14 patients with thymic carcinoma were treated at Gunma Prefectural Cancer Center. Two patients who had hematogenous metastasis were excluded from this study, therefore 12 patients were analyzed. The Masaoka staging system was used; four patients were diagnosed with stage III disease and eight patients with stage IV disease. The pathological subtype according to the World Health Organization histological criteria for thymic tumors was squamous cell carcinoma (low-grade histology) in six cases and undifferentiated carcinoma (high-grade histology) in six. Ten patients underwent thoracotomy, and two patients underwent excisional biopsy without thoracotomy. Ten patients (83%) received radiotherapy as a curative intent, and the median dose was 60 Gy. Systemic chemotherapy was administered to four patients (33%), and the majority (75%) of the regimens contained cisplatin. Results: The 3-year overall survival rate was 25%. Histological subtype (low-grade versus highgrade), surgical resection (complete versus incomplete), radiotherapy and chemotherapy were evaluated as prognostic factors in a univariate analysis. Low-grade histology and complete resection were good prognostic factors, although these were not statistically significant. Patients who received radiotherapy had a better outcome than those who did not. The major sites of recurrence were the pleura and pericardium. Recurrence within the radiation field was observed in one of seven patients in whom failure patterns could be evaluated. Conclusion: Complete resection is mandatory if possible. Radiotherapy plays an important role in treating thymic carcinoma in terms of reducing local recurrence and prolonging survival time. Establishment of an innovative treatment protocol that includes chemotherapy is necessary to control intrathoracic relapse and distant metastasis