Cav2.3 (α1E) Ca2+ channel participates in the control of sperm function

AbstractTo know the function of the Ca2+ channel containing α12.3 (α1E) subunit (Cav2.3 channel) in spermatozoa, we analyzed Ca2+ transients and sperm motility using a mouse strain lacking Cav2.3 channel. The averaged rising rates of Ca2+ transients induced by α-D-mannose–bovine serum albumin in the head region of Cav2.3−/− sperm were significantly lower than those of Cav2.3+/+ sperm. A computer-assisted sperm motility assay revealed that straight-line velocity and linearity were greater in Cav2.3−/− sperm than those in Cav2.3+/+ sperm. These results suggest that the Cav2.3 channel plays some roles in Ca2+ transients and the control of flagellar movement

Interactive Rakuchu Rakugai-zu (Views in and Around Kyoto)

This book constitutes the refereed proceedings of the First International Conference on Culture and Computing, C&C 2020, held as part of the 22nd International Conference on Human-Computer Interaction, HCII 2020, in July 2020. The conference was planned to be held in Copenhagen, Denmark, but had to change to a virtual conference mode due to the COVID-19 pandemic.Rakuchu Rakugai-zu is a screen painting illustrating in and around Kyoto created from the beginning of 16th century until 17th century. As in the Funaki Version of Rakuchu Rakugai-zu everyday lives of over 2, 000 people in Kyoto are vividly painted, the painting is a good material to learn their customs in early 17th century in Kyoto. However, it is difficult to understand the details of the painting, if background information is not provided. How to document such information and how to provide people such information would be crucial for the future of museums. To cope with this, we are carrying out a project to develop the interactive Rakuchu Rakugai-zu, Funaki Version. Firstly, we digitize the painting with ultra-high resolution. Secondly, using historical books regarding the customs of that era., we identify clothes and behaviors of each person in the painting and linked them to the person in the painting. Thirdly, we develop the technology to provide such information interactively using text or voice. Fourthly, we develop present Funaki Version including various manga characters so that we can compare old and present customs of people. Integrating these contents and technologies we are now developing the interactive Rakuchu Rakugai-zu, Funaki Version

CAXII Is a Sero-Diagnostic Marker for Lung Cancer

To develop sero-diagnostic markers for lung cancer, we generated monoclonal antibodies using pulmonary adenocarcinoma (AD)-derived A549 cells as antigens by employing the random immunization method. Hybridoma supernatants were immunohistochemically screened for antibodies with AMeX-fixed and paraffin-embedded A549 cell preparations. Positive clones were monocloned twice through limiting dilutions. From the obtained monoclonal antibodies, we selected an antibody designated as KU-Lu-5 which showed intense membrane staining of A549 cells. Based on immunoprecipitation and MADLI TOF/TOF-MS analysis, this antibody was recognized as carbonic anhydrase XII (CAXII). To evaluate the utility of this antibody as a sero-diagnostic marker for lung cancer, we performed dot blot analysis with a training set consisting of sera from 70 lung cancer patients and 30 healthy controls. The CAXII expression levels were significantly higher in lung cancer patients than in healthy controls in the training set (P<0.0001), and the area under the curve of ROC was 0.794, with 70.0% specificity and 82.9% sensitivity. In lung cancers, expression levels of CAXII were significantly higher in patients with squamous cell carcinoma (SCC) than with AD (P = 0.035). Furthermore, CAXII was significantly higher in well- and moderately differentiated SCCs than in poorly differentiated ones (P = 0.027). To further confirm the utility of serum CAXII levels as a sero-diagnostic marker, an additional set consisting of sera from 26 lung cancer patients and 30 healthy controls was also investigated by dot blot analysis as a validation study. Serum CAXII levels were also significantly higher in lung cancer patients than in healthy controls in the validation set (P = 0.030). Thus, the serum CAXII levels should be applicable markers discriminating lung cancer patients from healthy controls. To our knowledge, this is the first report providing evidence that CAXII may be a novel sero-diagnostic marker for lung cancer

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

Aberrant nuclear beta-catenin expression in the spindle or corded cells in so-called corded and hyalinized endometrioid carcinomas. Another critical role of the unique morphological feature

Corded and hyalinized endometrioid carcinoma (CHEC), showing spindle and/or corded (SPICO) cells often in the background of hyalinized stroma, is a rare variant of uterine endometrioid carcinomas. The aim of our study was to explore the status of cell-adhesion molecules (beta-catenin, Ecadherin) in CHECs and to survey whether immunostains for beta-catenin and p53 can help to distinguish CHECs from their morphological mimics: malignant mixed mullerian tumors (MMMTs) and uterine tumors resembling ovarian sex-cord tumors (UTROSCTs). Immunohistochemistry was performed and scored for each element as follows: 0: negative, 1+: 30%. The SPICO patterns were classified as spindle/fusiform; 3, corded; 1, and both; 2. SPICO components consisted of <10%: 4, 10-30%: 1, >30%: 1. Five contained squamous components. In SPICO elements of all CHECs, nuclear beta-catenin expression (score: 1+; 1, 2+; 2, 3+; 3) and complete loss of membranous expression of E-cadherin was observed. In contrast, comparable components (sarcomatous ones for eight MMMTs or sex-cord-like ones for six UTROSCTs) showed no nuclear positivity for beta-catenin. p53 expression was observed in SPICO (64.7%), sarcomatous (87.5%), and sex-cord-like (50%) components, and sarcomatous areas of most MMMTs notably showed diffuse and intense staining. Sequence analysis of PCR amplification products of exon 3 of the beta-catenin gene revealed mutation in all cases, except two lacking SPICO components represented on microdissected areas. Our results suggest that alterations in beta-catenin/E-cadherin complex play a critical role in SPICO features. Immunostain for beta-catenin and p53 is a promising approach for distinguishing CHECs from MMMTs and UTROSCT