The CD8+ Dendritic Cell Subset Selectively Endocytoses Dying Cells in Culture and In Vivo

Dendritic cells (DCs) are able in tissue culture to phagocytose and present antigens derived from infected, malignant, and allogeneic cells. Here we show directly that DCs in situ take up these types of cells after fluorescent labeling with carboxyfluorescein succinimidyl ester (CFSE) and injection into mice. The injected cells include syngeneic splenocytes and tumor cell lines, induced to undergo apoptosis ex vivo by exposure to osmotic shock, and allogeneic B cells killed by NK cells in situ. The CFSE-labeled cells in each case are actively endocytosed by DCs in vivo, but only the CD8+ subset. After uptake, all of the phagocytic CD8+ DCs can form major histocompatibility complex class II–peptide complexes, as detected with a monoclonal antibody specific for these complexes. The CD8+ DCs also selectively present cell-associated antigens to both CD4+ and CD8+ T cells. Similar events take place with cultured DCs; CD8+ DCs again selectively take up and present dying cells. In contrast, both CD8+ and CD8− DCs phagocytose latex particles in culture, and both DC subsets present soluble ovalbumin captured in vivo. Therefore CD8+ DCs are specialized to capture dying cells, and this helps to explain their selective ability to cross present cellular antigens to both CD4+ and CD8+ T cells

The Possible Role of TASK Channels in Rank-Ordered Recruitment of Motoneurons in the Dorsolateral Part of the Trigeminal Motor Nucleus.

Because a rank-ordered recruitment of motor units occurs during isometric contraction of jaw-closing muscles, jaw-closing motoneurons (MNs) may be recruited in a manner dependent on their soma sizes or input resistances (IRs). In the dorsolateral part of the trigeminal motor nucleus (dl-TMN) in rats, MNs abundantly express TWIK (two-pore domain weak inwardly rectifying K channel)-related acid-sensitive-K(+) channel (TASK)-1 and TASK3 channels, which determine the IR and resting membrane potential. Here we examined how TASK channels are involved in IR-dependent activation/recruitment of MNs in the rat dl-TMN by using multiple methods. The real-time PCR study revealed that single large MNs (>35 μm) expressed TASK1 and TASK3 mRNAs more abundantly compared with single small MNs (15-20 μm). The immunohistochemistry revealed that TASK1 and TASK3 channels were complementarily distributed in somata and dendrites of MNs, respectively. The density of TASK1 channels seemed to increase with a decrease in soma diameter while there were inverse relationships between the soma size of MNs and IR, resting membrane potential, or spike threshold. Dual whole-cell recordings obtained from smaller and larger MNs revealed that the recruitment of MNs depends on their IRs in response to repetitive stimulation of the presumed Ia afferents. 8-Bromoguanosine-cGMP decreased IRs in small MNs, while it hardly changed those in large MNs, and subsequently decreased the difference in spike-onset latency between the smaller and larger MNs, causing a synchronous activation of MNs. These results suggest that TASK channels play critical roles in rank-ordered recruitment of MNs in the dl-TMN

Production and crystallization of lobster muscle tropomyosin expressed in Sf9 cells

AbstractA new form of muscle tropomyosin crystal has been obtained, by employing new strategies in protein preparation and crystallization. Non-polymerizable tropomyosin was prepared by removing 11 amino acids at the C-terminus. The truncated tropomyosin was expressed in Sf9 insect cells by use of the baculovirus-based expression system, to obtain highly homogeneous protein preparations. By routinely monitoring homogeneity by mass spectrometry, we found that the homogeneity played a key role in obtaining good crystals. The crystal quality was also dependent on isoforms; the crystals raised from a slow muscle-specific isoform diffracted to a higher resolution, compared with a fast muscle-specific counterpart. For crystallization, a high concentration of organic solvent was used as the precipitant; in the presence of 35% DMSO, tetragonal crystals were formed, which belong to space group P43(1)212 with cell constants of a = b = 105.6 Å, c = 506.9 Å. The crystals gave rise to reflections the intensities of which were characteristically determined by the transform of α-helical coiled-coil. Thus in the region of 10-5.5 Å resolution along the c∗-axis, the reflections were weak. For accurate measurement of these reflection intensities, beam-line ID2 in ESRF Grenoble was advantageous owing to the high brilliance and a low background. There the crystals diffracted to beyond 3.0 Å along the c∗-axis, whereas along the a∗–b∗-plane reflections were limited to 6.6 Å. Data analysis is under way on a data set from a PtCl4 derivative

Solution Structure of LC4 Transmembrane Segment of CCR5

CC-chemokine receptor 5 (CCR5) is a specific co-receptor allowing the entry of human immunodeficiency virus type 1 (HIV-1). The LC4 region in CCR5 is required for HIV-1 entry into the cells. In this study, the solution structure of LC4 in SDS micelles was elucidated by using standard 1H two-dimensional NMR spectroscopy, circular dichroism, and fluorescdence quenching. The LC4 structure adopts two helical structures, whereas the C-terminal part remains unstructured. The positions in which LC4 binds to the HIV-1 inhibitory peptide LC5 were determined by docking calculations in addition to NMR data. The poses showed the importance of the hydrophobic interface of the assembled structures. The solution structure of LC4 elucidated in the present work provides a structural basis for further studies on the HIV-1 inhibitory function of the LC4 region

Conditional deletion of Npt2b in phosphate transport

Background Hyperphosphatemia is common in chronic kidney disease and is associated with morbidity and mortality. The intestinal Na+-dependent phosphate transporter Npt2b is thought to be an important molecular target for the prevention of hyperphosphatemia. The role of Npt2b in the net absorption of inorganic phosphate (Pi), however, is controversial. Methods In the present study, we made tamoxifen-inducible Npt2b conditional knockout (CKO) mice to analyze systemic Pi metabolism, including intestinal Pi absorption. Results Although the Na+-dependent Pi transport in brush-border membrane vesicle uptake levels were significantly decreased in the distal intestine of Npt2b CKO mice compared with control mice, plasma Pi and fecal Pi excretion levels were not significantly different. Data obtained using the intestinal loop technique showed that Pi uptake in Npt2b CKO mice was not affected at a Pi concentration of 4 mM, which is considered the typical luminal Pi concentration after meals in mice. Claudin, which may be involved in paracellular pathways, as well as claudin-2, 12, and 15 protein levels were significantly decreased in the Npt2b CKO mice. Thus, Npt2b deficiency did not affect Pi absorption within the range of Pi concentrations that normally occurs after meals. Conclusion These findings indicate that abnormal Pi metabolism may also be involved in tight junction molecules such as Cldns that are affected by Npt2b deficiency

ダイガク キョウイク イインカイ ガッコウ ト レンケイ シタ キョウイク カイゼン ニ カンスル ジッセン キョウイク VII : ホンガク ト スズカシ キョウイク イインカイ ト ノ レンケイ ジギョウ ニ カンスル ガッコウ シエン ノ ケイカ

教職大学院を中心組織として，本学と鈴鹿市教育委員会は，平成２３年３月連携事業に関する協定書を交わした。事業内容は，以下の５点である。 ①本学教職大学院教員による公立学校を対象とした訪問指導 ②本学教職大学院教員による教育委員会及び校長会等への指導・助言の実施 ③鈴鹿市教育委員会による本学教職大学院への教員派遣の協力 ④鈴鹿市教育委員会による本学教職大学院が実施する教育実践研究に関する協力 ⑤その他，本学教職大学院と鈴鹿市教育委員会の協議結果に基づき実施する事業 この協定に基づき実施している連携事業の具体的方法を示すことを通して，大学，教育委員会，学校の三者の連携のあり方について考察した。本稿は，大学・教育委員会・学校と連携した教育改善に関する実践研究（I），（V）（VI）の継続論文である。On March 2011,Naruto University of Education, putting Advanced Practice of School Education as a center organization, signed an Agreement of Cooperative Project with Suzuka Board of Education. The five areas of the project are as follows: 1. On-site coaching to public schools by professors of Graduate School of Education. 2. Providing advices and instructions to the Board of Education and the principals by professors of Graduate School of Education. 3. Cooperation of teacher visits by Suzuka Board of Education to Graduate School of Education. 4. Suzuka Board of Education\u27s cooperation to practical educational research conducted by Graduate School of Education. 5. Other projects based on the discussion and agreement between Graduate School of Education and Suzuka Board of Education. Through concrete methods of this cooperative project based on this agreement, we studied the cooperation among universities, boards of education, and schools. This Study is a sequel of "Practical Study on Educational Improvement through Partnership among University, School and The Board of Education(I),(V)and(VI)

ダイガク キョウイク イインカイ ガッコウ ト レンケイ シタ キョウイク カイゼン ニ カンスル ジッセン ケンキュウ VI : ホンガク ト スズカシ キョウイク イインカイ トノ レンケイ ジギョウ ニ カンスル ガッコウ シエン ノ ケイカ

教職大学院を中心組織として，本学と鈴鹿市教育委員会は，平成２３年３月連携事業に関する協定書を交わした。事業内容は，以下の５点である。①本学教職大学院教員による公立学校を対象とした訪問指導②本学教職大学院教員による教育委員会及び校長会等への指導・助言の実施③鈴鹿市教育委員会による本学教職大学院への教員派遣の協力④鈴鹿市教育委員会による本学教職大学院が実施する教育実践研究に関する協力⑤その他，本学教職大学院と鈴鹿市教育委員会の協議結果に基づき実施する事業 この協定に基づき実施している連携事業の具体的方法を示すことを通して，大学，教育委員会，学校の三者の連携のあり方について考察した。本稿は，大学・教育委員会・学校と連携した教育改善に関する実践研究（I），（V）の継続論文である。On March 2011, Naruto University of Education, putting Advanced Practice of School Education as a center organization, signed an Agreement of Cooperative Project with Suzuka Board of Education. The five areas of the project are as follows: 1. On-site coaching to public schools by professors of Graduate School of Education. 2. Providing advices and instructions to the Board of Education and the principals by professors of Graduate School of Education. 3. Cooperation of teacher visits by Suzuka Board of Education to Graduate School of Education. 4. Suzuka Board of Education’s cooperation to practical educational research conducted by Graduate School of Education. 5. Other projects based on the discussion and agreement between Graduate School of Education and Suzuka Board of Education. Through concrete methods of this cooperative project based on this agreement, we studied the cooperation among universities, boards of education, and schools. This Study is a sequel of“ Practical Study on Educational Improvement through Partnership among University, School and The Board of Education (I) and (V).

ダイガク キョウイク イインカイ ガッコウ ト レンケイ シタ キョウイク カイゼン ニ カンスル ジッセン ケンキュウ VII : スズカシ チュウガッコウ ニオケル ガッコウ シンダン シツモンシ ノ コウセイ ニツイテ II

平成２３年度より，教職大学院を中心組織として，本学と鈴鹿市教育委員会は，平成２３年３月連携事業に関する協定書を交わした。この事業の一環として，平成２３・２４年度に鈴鹿市内の中学校において，生徒用，教職員用並びに保護者用の質問紙調査を実施した。本研究では，平成２３年度版の保護者用の調査項目の構成を検討して作成した平成２４年度版の学校診断質問紙の構成について検証した。平成２４年度版では汎用化のために，保護者用については３領域６下位領域の３３項目を設定した。平成２４年１１〜１２月に実施した調査結果に関する主成分分析の結果，保護者用では４因子が抽出された。設定領域との対照では，若干の項目については異なった関連性を示したが，多くの項目については設定領域の診断項目としての妥当性が示された。On March 2011, Naruto University of Education, putting Advanced Practice of School Education as a center organization, signed an Agreement of Cooperative Project with Suzuka Board of Education. We carried out the questionnaire survey of junior high school students, teachers and parents of the students in Suzuka City at 2011 and 2012 on this Project. The construction of the school assessment questionnaires for the parents of the 2012 version that examines those of the 2011 version are studied. In the 2012 version of the school assessment questionnaires, 33 items in the 3 categories / 6 subcategories at the parents were set up for generalized use. As a result of principal component analysis of survey data on November and December 2012, 4 factors were extracted in for the parents. Although a little item showed different relevancy from the setting, many items showed the validity as the assessment items of that