Showing papers by "Hokkaido University published in 2000"

Immunologic Self-Tolerance Maintained by Cd25+Cd4+Regulatory T Cells Constitutively Expressing Cytotoxic T Lymphocyte–Associated Antigen 4

Abstract: This report shows that cytotoxic T lymphocyte–associated antigen 4 (CTLA-4) plays a key role in T cell–mediated dominant immunologic self-tolerance. In vivo blockade of CTLA-4 for a limited period in normal mice leads to spontaneous development of chronic organ-specific autoimmune diseases, which are immunopathologically similar to human counterparts. In normal naive mice, CTLA-4 is constitutively expressed on CD25+CD4+ T cells, which constitute 5–10% of peripheral CD4+ T cells. When the CD25+CD4+ T cells are stimulated via the T cell receptor in vitro, they potently suppress antigen-specific and polyclonal activation and proliferation of other T cells, including CTLA-4–deficient T cells, and blockade of CTLA-4 abrogates the suppression. CD28-deficient CD25+CD4+ T cells can also suppress normal T cells, indicating that CD28 is dispensable for activation of the regulatory T cells. Thus, the CD25+CD4+ regulatory T cell population engaged in dominant self-tolerance may require CTLA-4 but not CD28 as a costimulatory molecule for its functional activation. Furthermore, interference with this role of CTLA-4 suffices to elicit autoimmune disease in otherwise normal animals, presumably through affecting CD25+CD4+ T cell–mediated control of self-reactive T cells. This unique function of CTLA-4 could be exploited to potentiate T cell–mediated immunoregulation, and thereby to induce immunologic tolerance or to control autoimmunity.

The Vienna classification of gastrointestinal epithelial neoplasia

Abstract: Background—Use of the conventional Western and Japanese classification systems of gastrointestinal epithelial neoplasia results in large diVerences among pathologists in the diagnosis of oesophageal, gastric, and colorectal neoplastic lesions. Aim—To develop common worldwide terminology for gastrointestinal epithelial neoplasia. Methods—Thirty one pathologists from 12 countries reviewed 35 gastric, 20 colorectal, and 21 oesophageal biopsy and resection specimens. The extent of diagnostic agreement between those with Western and Japanese viewpoints was assessed by kappa statistics. The pathologists met in Vienna to discuss the results and to develop a new consensus terminology. Results—The large diVerences between the conventional Western and Japanese diagnoses were confirmed (percentage of specimens for which there was agreement and kappa values: 37% and 0.16 for gastric; 45% and 0.27 for colorectal; and 14% and 0.01 for oesophageal lesions). There was much better agreement among pathologists (71% and 0.55 for gastric; 65% and 0.47 for colorectal; and 62% and 0.31 for oesophageal lesions) when the original assessments of the specimens were regrouped into the categories of the proposed Vienna classification of gastrointestinal epithelial neoplasia: (1) negative for neoplasia/dysplasia, (2) indefinite for neoplasia/dysplasia, (3) non-invasive low grade neoplasia (low grade adenoma/ dysplasia), (4) non-invasive high grade neoplasia (high grade adenoma/ dysplasia, non-invasive carcinoma and suspicion of invasive carcinoma), and (5) invasive neoplasia (intramucosal carcinoma, submucosal carcinoma or beyond). Conclusion—The diVerences between Western and Japanese pathologists in the diagnostic classification of gastrointestinal epithelial neoplastic lesions can be resolved largely by adopting the proposed terminology, which is based on cytological and architectural severity and invasion status. (Gut 2000;47:251‐255)

Maze-solving by an amoeboid organism

Abstract: The plasmodium of the slime mould Physarum polycephalum is a large amoeba-like cell consisting of a dendritic network of tube-like structures (pseudopodia). It changes its shape as it crawls over a plain agar gel and, if food is placed at two different points, it will put out pseudopodia that connect the two food sources. Here we show that this simple organism has the ability to find the minimum-length solution between two points in a labyrinth.

Physical aspects of a real-time tumor-tracking system for gated radiotherapy

Abstract: Purpose: To reduce uncertainty due to setup error and organ motion during radiotherapy of tumors in or near the lung, by means of real-time tumor tracking and gating of a linear accelerator Methods and Materials: The real-time tumor-tracking system consists of four sets of diagnostic X-ray television systems (two of which offer an unobstructed view of the patient at any time), an image processor unit, a gating control unit, and an image display unit The system recognizes the position of a 20-mm gold marker in the human body 30 times per second using two X-ray television systems The marker is inserted in or near the tumor using image guided implantation The linear accelerator is gated to irradiate the tumor only when the marker is within a given tolerance from its planned coordinates relative to the isocenter The accuracy of the system and the additional dose due to the diagnostic X-ray were examined in a phantom, and the geometric performance of the system was evaluated in 4 patients Results: The phantom experiment demonstrated that the geometric accuracy of the tumor-tracking system is better than 15 mm for moving targets up to a speed of 40 mm/s The dose due to the diagnostic X-ray monitoring ranged from 001% to 1% of the target dose for a 20-Gy irradiation of a chest phantom In 4 patients with lung cancer, the range of the coordinates of the tumor marker during irradiation was 25–53 mm, which would have been 96–384 mm without tracking Conclusion: We successfully implemented and applied a tumor-tracking and gating system The system significantly improves the accuracy of irradiation of targets in motion at the expense of an acceptable amount of diagnostic X-ray exposure

In vivo Degradation of Resin-Dentin Bonds in Humans Over 1 to 3 Years

Abstract: The longevity of resin restorations is currently an area of great interest in adhesive dentistry. However, no work has been conducted to investigate the durability of resin-dentin bond structures using human substrate in vivo. The purpose of this study was to investigate the degradation of the resin-dentin bond structures aged in an oral environment for 1, 2, or 3 years. Cavities were prepared in primary molars, and an adhesive resin system (Scotchbond Multi-Purpose) was applied to the cavity. After 1 to 3 years, following the eruption of the succedaneous permanent teeth, the resin-restored teeth were extracted. Immediately after extraction, those teeth were sectioned perpendicular to the adhesive interface and trimmed to produce an hourglass-shaped specimen. Then, a micro-tensile test was performed at a crosshead speed of 1.0 mm/min. The mean bond strengths were statistically compared with one-way ANOVA and Fisher's PLSD test (p < 0.05). Further, all fractured surfaces were observed by SEM, and the area fraction of failure mode was calculated by means of a digital analyzer on SEM photomicrographs. There were significant differences in tensile-bond strength among all 3 groups (p < 0.05), with mean values ranging from 28.3 +/- 11.3 MPa (control), to 15.2 +/- 4.4 MPa (1 to 2 years), to 9.1 +/- 5.1 MPa (2 to 3 years). Moreover, under fractographic analysis, the proportion of demineralized dentin at the fractured surface in specimens aged in an oral environment was greater than that in control specimens. Furthermore, degradation of resin composite and the depletion of collagen fibrils was observed among the specimens aged in an oral environment. Analysis of the results of this study indicated that the degradation of resin-dentin bond structures occurs after aging in the oral cavity.

cAMP-GEFII is a direct target of cAMP in regulated exocytosis.

Abstract: Although cAMP is well known to regulate exocytosis in many secretory cells, its direct target in the exocytotic machinery is not known. Here we show that cAMP-GEFII, a cAMP sensor, binds to Rim (Rab3-interacting molecule, Rab3 being a small G protein) and to a new isoform, Rim2, both of which are putative regulators of fusion of vesicles to the plasma membrane. We also show that cAMP-GEFII, through its interaction with Rim2, mediates cAMP-induced, Ca2+-dependent secretion that is not blocked by an inhibitor of cAMP-dependent protein kinase (PKA). Accordingly, cAMP-GEFII is a direct target of cAMP in regulated exocytosis and is responsible for cAMP-dependent, PKA-independent exocytosis.

Four-dimensional treatment planning and fluoroscopic real-time tumor tracking radiotherapy for moving tumor.

Abstract: Purpose: To achieve precise three-dimensional (3D) conformal radiotherapy for mobile tumors, a new radiotherapy system and its treatment planning system were developed and used for clinical practice. Methods and Materials: We developed a linear accelerator synchronized with a fluoroscopic real-time tumor tracking system by which 3D coordinates of a 2.0-mm gold marker in the tumor can be determined every 0.03 second. The 3D relationships between the marker and the tumor at different respiratory phases are evaluated using CT image at each respiratory phase, whereby the optimum phase can be selected to synchronize with irradiation (4D treatment planning). The linac is triggered to irradiate the tumor only when the marker is located within the region of the planned coordinates relative to the isocenter. Results: The coordinates of the marker were detected with an accuracy of ± 1 mm during radiotherapy in the phantom experiment. The time delay between recognition of the marker position and the start or stop of megavoltage X-ray irradiation was 0.03 second. Fourteen patients with various tumors were treated by conformal radiotherapy with a “tight” planning target volume (PTV) margin. They were surviving without relapse or complications with a median follow-up of 6 months. Conclusion: Fluoroscopic real-time tumor tracking radiotherapy following 4D treatment planning was developed and shown to be feasible to improve the accuracy of the radiotherapy for mobile tumors.

Complications of pedicle screw fixation in reconstructive surgery of the cervical spine.

Abstract: Study Design. Retrospective evaluation of complications in 180 consecutive patients with cervical disorders who had been treated by using pedicle screw fixation systems.Objectives.To determine the risks associated with pedicle screw fixation in the cervical spine and to emphasize the importance of p

A defect in harmonin, a PDZ domain-containing protein expressed in the inner ear sensory hair cells, underlies Usher syndrome type 1C.

Abstract: Usher syndrome type 1 (USH1) is an autosomal recessive sensory defect involving congenital profound sensorineural deafness, vestibular dysfunction and blindness (due to progressive retinitis pigmentosa)1. Six different USH1 loci have been reported. So far, only MYO7A (USH1B), encoding myosin VIIA (ref. 2), has been identified as a gene whose mutation causes the disease. Here, we report a gene underlying USH1C (MIM 276904), a USH1 subtype described in a population of Acadian descendants from Louisiana3 and in a Lebanese family4. We identified this gene (USH1C), encoding a PDZ-domain–containing protein, harmonin, in a subtracted mouse cDNA library derived from inner ear sensory areas. In patients we found a splice-site mutation, a frameshift mutation and the expansion of an intronic variable number of tandem repeat (VNTR). We showed that, in the mouse inner ear, only the sensory hair cells express harmonin. The inner ear Ush1c transcripts predicted several harmonin isoforms, some containing an additional coiled-coil domain and a proline- and serine-rich region. As several of these transcripts were absent from the eye, we propose that USH1C also underlies the DFNB18 form of isolated deafness.

Maturation of Human Dendritic Cells by Cell Wall Skeleton of Mycobacterium bovis Bacillus Calmette-Guérin: Involvement of Toll-Like Receptors

Abstract: The constituents of mycobacteria are an effective immune adjuvant, as observed with complete Freund's adjuvant. In this study, we demonstrated that the cell wall skeleton of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS), a purified noninfectious material consisting of peptidoglycan, arabinogalactan, and mycolic acids, induces maturation of human dendritic cells (DC). Surface expression of CD40, CD80, CD83, and CD86 was increased by BCG-CWS on human immature DC, and the effect was similar to those of interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), heat-killed BCG, and viable BCG. BCG-CWS induced the secretion of TNF-alpha, IL-6, and IL-12 p40. CD83 expression was increased by a soluble factor secreted from BCG-CWS-treated DC and was completely inhibited by monoclonal antibodies against TNF-alpha. BCG-CWS-treated DC stimulated extensive allogeneic mixed lymphocyte reactions. The level of TNF-alpha secreted through BCG-CWS was partially suppressed in murine macrophages with no Toll-like receptor 2 (TLR 2) or TLR4 and was completely lost in TLR2 and TLR4 double-deficient macrophages. These results suggest that the BCG-CWS induces TNF-alpha secretion from DC via TLR2 and TLR4 and that the secreted TNF-alpha induces the maturation of DC per se.

The Group as the Container of Generalized Reciprocity

Abstract: The results of an experiment with 91 Japanese participants confirmed the hypothesis that players of a Prisoner's Dilemma game would cooperate more with an in-group member than with an out-group member in the simultaneous game but not in the sequential game. The game used in the experiment was constructed such that each player gave his or her partner a portion of his or her own endowment of 300 yen. The group membership was created on the basis of participant's preferences for Klee's or Kandinski's paintings; each participant played the game once with an in-group member and once with an out-group member. In the simultaneous game, the two players decided simultaneously how much to give to the partner without knowing what the partner would do. In the sequential game, the first player made the decision; then the second player followed with full information on the first player's behavior. All the participants in the sequential game in fact took the role of the first player. The differential effect of the partner's group membership had been predicted on the basis of Yamagishi and his colleagues' argument that expectations of generalized reciprocity from in-group members is the source of in-group favoritism in a minimal group. A group is clearly distinguished from a mere aggregate of people. People who sit in the waiting area of an airport, for example, hardly constitute a group, nor do all people who wear eyeglasses. They share certain characteristics such as sitting in the same place or wearing eyeglasses, but this does not make them a group. The defining feature

Global three-dimensional simulation of aerosol optical thickness distribution of various origins

Abstract: A global three-dimensional model that can treat transportation of various species of aerosols in the atmosphere is developed using a framework of an atmospheric general circulation model (AGCM). Main aerosols in the troposphere, i.e., soil dust, carbonaceous (organic and black carbon), sulfate, and sea-salt aerosols, are introduced into this model. Prior to the model calculations the meteorological parameters are calculated by the AGCM with the nudging technique using reanalysis data. To evaluate aerosol effects on the climate system and to compare simulated results with observations, the optical thickness and Angstrom exponent are also calculated taking into account the size distribution and composition. The model results are validated by both measured surface aerosol concentrations and retrieved aerosol optical parameters from National Oceanic and Atmospheric Administration/Advanced Very High Resolution Radiometer. A general agreement is found between the simulated result and the observation globally and seasonally. One of the significant results is that the simulated relative contribution of anthropogenic carbonaceous aerosols to the total optical thickness is comparable to that of sulfate aerosols at midlatitudes of the Northern Hemisphere, which agrees with recent observations. This result leads to a conclusion that the radiative effect evaluation of aerosols on the climate system is necessary to be modified because optical properties of carbonaceous aerosols are different from those of sulfate aerosols. The other finding is that the seasonal shift off the west coast of North Africa observed by satellites, i.e., the latitude of the maximum optical thickness moves seasonally, is also reproduced in consideration of a mixed state of soil dust and carbonaceous aerosols.

Water-Assisted Formation of Micrometer-Size Honeycomb Patterns of Polymers

Abstract: Micropatterned thin films have received increased interest in the past few years. Besides preparation by conventional lithography, self-assembly of materials around a template can be used for patterning. Recently a method that utilizes the condensation of micrometer-size water droplets on solutions of block copolymers was reported for the preparation of ordered micrometer-size honeycomb structures. Here we show that the formation of honeycomb-like porous films is a general method that can be used for patterning many materials, e.g., block copolymers, amphiphilic polyion complexes, organic/inorganic hybrids, and homopolymers, such as polystyrene. Stabilization of water droplets is indispensable for regular pattern formation.

Type I collagen‐induced osteoblastic differentiation of bone‐marrow cells mediated by collagen‐α2β1 integrin interaction

Abstract: Bone marrow cells are multipotent cells. When bone marrow cells were cultured with type I collagen matrix gels, they showed high alkaline phosphatase activity, collagen synthesis, and formed mineralized tissues. Furthermore, cells expressed osteocalcin and bone sialoprotein genes, which are osteoblast-specific genes. These findings indicate that type I collagen matrix gels induce osteoblastic differentiation of bone marrow cells. Type I collagen interacts with the α 2 β 1 integrin receptor on the cell membrane and mediates extracellular signals into cells. DGEA peptide is a cell-binding domain of type I collagen molecule. When collagen–integrin interaction was interrupted by the addition of Asp-Gly-Glu-Ala (DGEA) peptide to the culture, the expression of osteoblastic phenotypes of bone marrow cells was inhibited. Furthermore, anti-α 2 integrin antibody, which interacts with α subunit of integrin and blocks the binding of integrin with collagen, suppressed the expression of osteoblastic phenotypes. These findings imply that collagen-α 2 β 1 integrin interaction is an important signal for the osteoblastic differentiation of bone marrow cells. J. Cell. Physiol. 184:207–213, 2000. © 2000 Wiley-Liss, Inc.

The NPH4 locus encodes the auxin response factor ARF7, a conditional regulator of differential growth in aerial Arabidopsis tissue.

Abstract: Organ bending through differential growth represents a major mechanism by which plants are able to adaptively alter their morphology in response to local changes in the environment. Two plant hormones, auxin and ethylene, have been implicated as regulators of differential growth responses; however, the mechanisms by which they elicit their effects remain largely unknown. Here, we describe isolation of the NPH4 gene of Arabidopsis, which is conditionally required for differential growth responses of aerial tissues, and we report that NPH4 encodes the auxin-regulated transcriptional activator ARF7. The phenotypes of nph4 mutants, which include multiple differential growth defects associated with reduced auxin responsiveness, including impaired auxin-induced gene expression, are consistent with the predicted loss of function of a transcriptional activator, and these phenotypes indicate that auxin-dependent changes in gene transcription are prerequisite for proper organ bending responses. Although NPH4/ARF7 appears to be a major regulator of differential growth, it is not the sole regulator because phenotypes of nph4 null mutants were suppressed by application of ethylene. This latter finding illustrates the intimate connection between auxin and ethylene in the control of growth in higher plants.

A detailed linkage map of medaka, Oryzias latipes: comparative genomics and genome evolution.

Abstract: We mapped 633 markers (488 AFLPs, 28 RAPDs, 34 IRSs, 75 ESTs, 4 STSs, and 4 phenotypic markers) for the Medaka Oryzias latipes, a teleost fish of the order Beloniformes. Linkage was determined using a reference typing DNA panel from 39 cell lines derived from backcross progeny. This panel provided unlimited DNA for the accumulation of mapping data. The total map length of Medaka was 1354.5 cM and 24 linkage groups were detected, corresponding to the haploid chromosome number of the organism. Thirteen to 49 markers for each linkage group were obtained. Conserved synteny between Medaka and zebrafish was observed for 2 independent linkage groups. Unlike zebrafish, however, the Medaka linkage map showed obvious restriction of recombination on the linkage group containing the male-determining region (Y) locus compared to the autosomal chromosomes.

Band structure of acoustic waves in phononic lattices: Two-dimensional composites with large acoustic mismatch

Abstract: The finite-difference time-domain method is applied to the calculation of dispersion relations of acoustic waves in two-dimensional (2D) phononic lattices, i.e., periodic solid-solid, solid-liquid, and solid-vacuum composites, for which the conventional plane-wave-expansion method fails or converges very slowly. Numerical examples are developed for 2D structures with polyethylene, mercury, and vacuum cylinders forming a square lattice in an aluminum matrix. The implication of the calculated dispersion relations for ultrasound transmission experiments is discussed.

The complete nucleotide sequence of the mitochondrial genome of sugar beet (Beta vulgaris L.) reveals a novel gene for tRNACys(GCA)

Abstract: We determined the complete nucleotide sequence of the mitochondrial genome of an angiosperm, sugar beet (Beta vulgaris cv TK81-O). The 368 799 bp genome contains 29 protein, five rRNA and 25 tRNA genes, most of which are also shared by the mitochondrial genome of Arabidopsis thaliana, the only other completely sequenced angiosperm mitochondrial genome. However, four genes identified here (namely rps13, trnF-GAA, ccb577 and trnC2-GCA) are missing in Arabidopsis mitochondria. In addition, four genes found in Arabidopsis (ccb228, rpl2, rpl16 and trnY2-GUA) are entirely absent in sugar beet or present only in severely truncated form. Introns, duplicated sequences, additional reading frames and inserted foreign sequences (chloroplast, nuclear and plasmid DNA sequences) contribute significantly to the overall size of the sugar beet mitochondrial genome. Nevertheless, 55.6% of the genome has no obvious features of information. We identified a novel tRNA(Cys) gene (trnC2-GCA) which shows no sequence homology with any tRNA(Cys) genes reported so far in higher plants. Intriguingly, this tRNA gene is actually transcribed into a mature tRNA, whereas the native tRNA(Cys) gene (trnC1-GCA) is most likely a pseudogene.

Epstein-Barr virus and gastric carcinoma.

Abstract: The Epstein-Barr virus (EBV) is detected in the tissue of about 10% of gastric carcinoma cases throughout the world. In each case, 100% of carcinoma cells are infected with EBV. Analysis of EBV in carcinoma biopsies indicates that carcinoma is formed by the proliferation of a single EBV infected cell. These findings suggest that EBV plays an important role in the development of EBV positive gastric carcinomas. The EBV genes expressed are EBV determined nuclear antigen 1 (EBNA1), two small non-polyadenylated RNAs known as EBER1 and EBER2, and the transcripts from the BamHI-A region (BARF0); in addition, some cases also express a small amount of latent membrane protein 2A (LMP2A). Epithelial cells are refractory to EBV infection in vitro. This has hampered the study of the role of EBV in epithelial malignancies. The use of recombinant EBV carrying a selectable marker has enabled this difficulty to be overcome. EBV infected cell clones can be obtained from most carcinoma cell lines examined, and it was found that cell to cell contact was an efficient mode of EBV infection. Furthermore, it was possible to immortalise primary gastric epithelial cells by EBV infection. The cells expressed identical EBV genes to those typically seen in EBV positive gastric carcinoma, and showed accelerated malignant properties, including growth in soft agarose and tumorigenicity in severe combined immunodeficient (SCID) mice. These results suggest that EBV contributes to the maintenance of the malignant phenotype of EBV positive gastric carcinoma.

Association of autoantibodies against the phosphatidylserine-prothrombin complex with manifestations of the antiphospholipid syndrome and with the presence of lupus anticoagulant.

Abstract: Objective To clarify the association of autoantibodies against prothrombin with the clinical manifestations of the antiphospholipid syndrome (APS) and with the presence of lupus anticoagulant (LAC). Methods We examined 265 patients who visited our autoimmune disease clinic. IgG and IgM antiprothrombin antibodies were tested by enzyme-linked immunosorbent assay (ELISA) as either antiphosphatidylserine–prothrombin complex (aPS/PT) antibodies or as antibodies against prothrombin coated on irradiated ELISA plates (as antigen) (aPT). IgG, IgM, and IgA anticardiolipin (aCL) antibodies and their β2-glycoprotein I (β2GPI) dependency were also evaluated by ELISA. LAC was tested by 3 different methods. Results The presence of aPS/PT, but not of aPT, significantly correlated with the clinical manifestations of APS (odds ratio [OR] 4.39, 95% confidence interval [95% CI] 2.06–9.38), and aPS/PT antibodies were as specific as β2GPI-dependent aCL for APS (93.1% for both). IgG aPS/PT strongly correlated with the presence of LAC as detected using the dilute Russell viper venom time test (OR 38.2, 95% CI 13.4–109.1). Conclusion Antiprothrombin antibodies are heterogeneous and their clinical relevance depends on the method of detection applied. Positive results on the aPS/PT test can serve as a marker of thrombotic events in patients with autoimmune diseases.