Showing papers by "Nagoya University published in 1999"

The kinase TAK1 can activate the NIK-IκB as well as the MAP kinase cascade in the IL-1 signalling pathway

Abstract: Interleukin-1 (IL-1) is a proinflammatory cytokine that has several effects in the inflammation process When it binds to its cell-surface receptor, IL-1 initiates a signalling cascade that leads to activation of the transcription factor NF-kappaB and is relayed through the protein TRAF6 and a succession of kinase enzymes, including NF-kappaB-inducing kinase (NIK) and I kappaB kinases (IKKs) However, the molecular mechanism by which NIK is activated is not understood Here we show that the MAPKK kinase TAK1 acts upstream of NIK in the IL-1-activated signalling pathway and that TAK1 associates with TRAF6 during IL-1 signalling Stimulation of TAK1 causes activation of NF-kappaB, which is blocked by dominant-negative mutants of NIK, and an inactive TAK1 mutant prevents activation of NF-kappaB that is mediated by IL-1 but not by NIK Activated TAK1 phosphorylates NIK, which stimulates IKK-alpha activity Our results indicate that TAK1 links TRAF6 to the NIK-IKK cascade in the IL-1 signalling pathway

A requirement for neuropilin-1 in embryonic vessel formation

Abstract: Neuropilin-1 is a membrane protein that is expressed in developing neurons and functions as a receptor or a component of the receptor complex for the class 3 semaphorins, which are inhibitory axon guidance signals. Targeted inactivation of the neuropilin-1 gene in mice induced disorganization of the pathway and projection of nerve fibers, suggesting that neuropilin-1 mediates semaphorin-elicited signals and regulates nerve fiber guidance in embryogenesis. Neuropilin-1 is also expressed in endothelial cells and shown to bind vascular endothelial growth factor (VEGF), a potent regulator for vasculogenesis and angiogenesis. However, the roles of neuropilin-1 in vascular formation have been unclear. This paper reported that the neuropilin-1 mutant mouse embryos exhibited various types of vascular defects, including impairment in neural vascularization, agenesis and transposition of great vessels, insufficient aorticopulmonary truncus (persistent truncus arteriosus), and disorganized and insufficient development of vascular networks in the yolk sac. The vascular defects induced by neuropilin-1 deficiency in mouse embryos suggest that neuropilin-1 plays roles in embryonic vessel formation, as well as nerve fiber guidance.

Homogeneous Catalysis in Supercritical Fluids.

Abstract: Supercritical fluids (SCFs), compounds heated and pressurized beyond the critical point, have many unusual properties. Homogeneous molecular catalysts, which have far greater control over selectivity than heterogeneous solid catalysts, are now being tested in SCFs, and early results show that high rates, improved selectivity, and elimination of masstransfer problems can be achieved. As industry moves away from toxic or environmentally damaging solvents, supercritical carbon dioxide may be an ideal replacement medium for nonpolar or weakly polar chemical processes. More than simply substitutes for nonpolar solvents, SCFs can radically change the observed chemistry. Supercritical carbon dioxide is also an excellent medium for its own fixation, as demonstrated by studies of its hydrogenation.

Mitochondrial Electron Transport Complex I Is a Potential Source of Oxygen Free Radicals in the Failing Myocardium

Abstract: Oxidative stress in the myocardium may play an important role in the pathogenesis of congestive heart failure (HF). However, the cellular sources and mechanisms for the enhanced generation of reactive oxygen species (ROS) in the failing myocardium remain unknown. The amount of thiobarbituric acid reactive substances increased in the canine HF hearts subjected to rapid ventricular pacing for 4 weeks, and immunohistochemical staining of 4-hydroxy-2-nonenal ROS-induced lipid peroxides was detected in cardiac myocytes but not in interstitial cells of HF animals. The generation of superoxide anion was directly assessed in the submitochondrial fractions by use of electron spin resonance spectroscopy with spin trapping agent, 5, 5'-dimethyl-1-pyrroline-N-oxide, in the presence of NADH and succinate as a substrate for NADH-ubiquinone oxidoreductase (complex I) and succinate-ubiquinone oxidoreductase (complex II), respectively. Superoxide production was increased 2.8-fold (P<0.01) in HF, which was due to the functional block of electron transport at complex I. The enzymatic activity of complex I decreased in HF (274+/-13 versus 136+/-9 nmol. min(-1). mg(-1) protein, P<0.01), which may thus have caused the functional uncoupling of the respiratory chain and the deleterious ROS production in HF mitochondria. The present study provided direct evidence for the involvement of ROS in the mitochondrial origin of HF myocytes, which might be responsible for both contractile dysfunction and structural damage to the myocardium.

Memory of macromolecular helicity assisted by interaction with achiral small molecules

Abstract: The helicity of biological macromolecules such as DNA and proteins is largely governed by the homochirality of their components (D-sugars and L-amino acids). In polymer and supramolecular chemistry, control of helicity is an attractive goal because of possible applications in materials science, chemical sensing and enantioselective catalysis1,2,3,4,5,6,7,8,9,10,11,12,13. We reported recently that macromolecular helicity can be induced in a polymer by an optically active amine14. Here we show that this helicity can be ‘memorized’ when the amine is replaced by various achiral amines. Although the maintenance of helicity in the polymer is not perfect, it can ‘repair’ itself over time. Small structural changes in the achiral amines influence the efficiency of helicity retention markedly.

The TAK1–NLK–MAPK-related pathway antagonizes signalling between β-catenin and transcription factor TCF

Abstract: The Wnt signalling pathway regulates many developmental processes through a complex of β-catenin and the T-cell factor/lymphoid enhancer factor (TCF/LEF) family of high-mobility-group transcription factors1,2,3,4,5,. Wnt stabilizes cytosolic β-catenin, which then binds to TCF and activates gene transcription. This signalling cascade is conserved in vertebrates, Drosophila and Caenorhabditis elegans. In C. elegans, the proteins MOM-4 and LIT-1 regulate Wnt signalling to polarize responding cells during embryogenesis6. MOM-4 and LIT-1 are homologous to TAK1 (a kinase activated by transforming growth factor-β) mitogen-activated protein-kinase-kinase kinase (MAP3K)7 and MAP kinase (MAPK)-related NEMO-like kinase (NLK)8,9, respectively, in mammalian cells. These results raise the possibility that TAK1 and NLK are also involved in Wnt signalling in mammalian cells. Here we show that TAK1 activation stimulates NLK activity and downregulates transcriptional activation mediated by β-catenin and TCF. Injection of NLK suppresses the induction of axis duplication by microinjected β-catenin in Xenopus embryos. NLK phosphorylates TCF/LEF factors and inhibits the interaction of the β-catenin–TCF complex with DNA. Thus, the TAK1–NLK–MAPK-like pathway negatively regulates the Wnt signalling pathway.

Quantitative Analysis of Epstein-Barr Virus Load by Using a Real-Time PCR Assay

Abstract: To measure the virus load in patients with symptomatic Epstein-Barr virus (EBV) infections, we used a real-time PCR assay to quantify the amount of EBV DNA in blood. The real-time PCR assay could detect from 2 to over 10(7) copies of EBV DNA with a wide linear range. We estimated the virus load in peripheral blood mononuclear cells (PBMNC) from patients with symptomatic EBV infections. The mean EBV-DNA copy number in the PBMNC was 10(3.7) copies/microg of DNA in patients with EBV-related lymphoproliferative disorders, 10(4.1) copies/microg of DNA in patients with chronic active EBV infections, and 10(2.2) copies/microg of DNA in patients with infectious mononucleosis. These numbers were significantly larger than those in either posttransplant patients or immunocompetent control patients without EBV-related diseases. In a patient with infectious mononucleosis, the virus load decreased as the symptoms resolved. The copy number of EBV DNA in PBMNC from symptomatic EBV infections was correlated with the EBV-positive cell number determined by the in situ hybridization assay (r = 0.842; P < 0.0001). These results indicate that the real-time PCR assay is useful for diagnosing symptomatic EBV infection and for monitoring the virus load.

Small-Scale Anisotropy of Cosmic Rays above 1019 eV Observed with the Akeno Giant Air Shower Array

Abstract: With the Akeno Giant Air Shower Array, 581 cosmic rays above 1019 eV, 47 above 4 ) 1019 eV, and seven above 1020 eV were observed until 1998 August. The arrival direction distribution of these extremely high energy cosmic rays has been studied. While no signi—cant large-scale anisotropy is found on the celestial sphere, some interesting clusters of cosmic rays are observed. Above 4 ) 1019 eV, there are one triplet and three doublets within a separation angle of and the probability of observing 2i.5, these clusters by a chance coincidence under an isotropic distribution is smaller than 1%. The triplet is especially observed against expected 0.05 events. The distribution expected from the dark cos (h GC ) matter halo model —ts the data as well as an isotropic distribution above 2 ) 1019 and 4 ) 1019 eV, but the —t with the dark matter halo model is poorer than the isotropic distribution above 1019 eV. The arrival direction distribution of seven 1020 eV cosmic rays is consistent with that of lower energy cosmic rays and is uniform. Three of the seven are members of doublets above about 4 ) 1019 eV. Subject headings: cosmic raysgalaxies: generalGalaxy: halolarge-scale structure of universe

A slow thrust slip event following the two 1996 Hyuganada Earthquakes beneath the Bungo Channel, southwest Japan

Abstract: We report a “slow thrust slip event” that occurred beneath the Bungo Channel region, southwestern Japan. On Oct. and Dec., 1996, two Hyuganada earthquakes (both Mw = 6.7), followed by afterslips, occurred. In addition, a crustal movement characterized by an extremely slow rise was observed around the Bungo Channel, about 200 km north from the epicenters, and continued for about 300 days long. Assuming a slow slip on the plate boundary, we estimate its duration and surface displacements from GPS time series data by curve-fitting, and then, determine the fault slip distribution. We found that a slow slip without any earthquakes continued for nearly one year and released the seismic moment comparable to that of the Hyuganada earthquakes. Occurrence of the slow thrust slip event suggests that this kind of event may be a characteristic mode of stress release at a transition region of interplate coupling.

XIAP, a cellular member of the inhibitor of apoptosis protein family, links the receptors to TAB1–TAK1 in the BMP signaling pathway

Abstract: Signals elicited by transforming growth factor‐β (TGF‐β) superfamily ligands are generated following the formation of heteromeric receptor complexes consisting of type I and type II receptors. TAK1, a member of the MAP kinase kinase kinase family, and its activator, TAB1, participate in the bone morphogenetic protein (BMP) signaling pathway involved in mesoderm induction and patterning in early Xenopus embryos. However, the events leading from receptor activation to TAK1 activation remain to be identified. A yeast interaction screen was used to search for proteins that function in the pathway linking the receptors and TAB1–TAK1. The human X‐chromosome‐linked inhibitor of apoptosis protein (XIAP) was isolated as a TAB1‐binding protein. XIAP associated not only with TAB1 but also with the BMP receptors in mammalian cells. Injection of XIAP mRNA into dorsal blastomeres enhanced the ventralization of Xenopus embryos in a TAB1–TAK1‐dependent manner. Furthermore, a truncated form of XIAP lacking the TAB1‐binding domain partially blocked the expression of ventral mesodermal marker genes induced by a constitutively active BMP type I receptor. These results suggest that XIAP participates in the BMP signaling pathway as a positive regulator linking the BMP receptors and TAB1–TAK1.

High-level expression of maize phosphoenolpyruvate carboxylase in transgenic rice plants

Abstract: Using an Agrobacterium-mediated transformation system, we have introduced the intact gene of maize phosphoenolpyruvate carboxylase (PEPC), which catalyzes the initial fixation of atmospheric CO2 in C4 plants into the C3 crop rice. Most transgenic rice plants showed high-level expression of the maize gene; the activities of PEPC in leaves of some transgenic plants were two- to threefold higher than those in maize, and the enzyme accounted for up to 12% of the total leaf soluble protein. RNA gel blot and Southern blot analyses showed that the level of expression of the maize PEPC in transgenic rice plants correlated with the amount of transcript and the copy number of the inserted maize gene. Physiologically, the transgenic plants exhibited reduced O2 inhibition of photosynthesis and photosynthetic rates comparable to those of untransformed plants. The results demonstrate a successful strategy for installing the key biochemical component of the C4 pathway of photosynthesis in C3 plants.

Spontaneous assembly of ten components into two interlocked, identical coordination cages

Abstract: Supermolecules consisting of interlinked ring-like molecules (catenanes)1 are an interesting target for chemical synthesis both for their intrinsic interest as non-covalently bound but robust assemblies and because of the perspective they offer on materials chemistry2. Catenanes have been prepared by metal-ion templating3,4 and self-assembly through other non-covalent interactions5,6,7,8,9,10,11,12. Here we report the synthesis of a catenane composed not of two interlocking rings but of two cages. This structure is prepared by metal-mediated self-assembly13,14,15. The framework of each cage is assembled from five components: two tridentate ligands held together with three metal ions. Because each cage framework can bind an aromatic ring, two cage units will bind one another during their assembly process through the formation of a quadruple aromatic stack, giving rise to the ten-component interlocked supermolecule16,17.

A nanometre-sized hexahedral coordination capsule assembled from 24 components

Abstract: Molecular capsules consist of closed, hollow frameworks within which encapsulated molecules are isolated from interaction with external molecules1. In this environment, otherwise reactive molecules can be stabilized2,3,4,5. Although some molecular capsules have been prepared by conventional synthetic chemistry1, recent progress in non-covalent synthesis has allowed the creation of capsules held together by hydrogen bonds6,7,8,9. Here we report the use of transition-metal-based coordination chemistry10,11,12,13,14,15,16,17,18,19 to assemble a stable, nanometre-scale capsule from 24 small components: 18 metal ions and six triangular ligands. The capsule is roughly hexahedral and comprises six edge-sharing triangles with two metal ions on each edge. The internal space has a volume of 900 A3 and is fully closed to all but very small molecules.

Resistance of Crohn's disease T cells to multiple apoptotic signals is associated with a Bcl-2/Bax mucosal imbalance

Abstract: Crohn's disease (CD) is a condition characterized by excessive numbers of activated T cells in the mucosa. We investigated whether a defect in apoptosis could prolong T cell survival and contribute to their accumulation in the mucosa. Apoptotic, Bcl-2+, and Bax+ cells in tissue sections were detected by the TUNEL method and immunohistochemistry. T cell apoptosis was induced by IL-2 deprivation, Fas Ag ligation, and exposure to TNF-alpha and nitric oxide. TUNEL+ leukocytes were few in control, CD, and ulcerative colitis (UC) mucosa, with occasional CD68+ and myeloperoxidase+, but no CD45RO+, apoptotic cells. Compared with control and UC, CD T cells grew remarkably more in response to IL-2 and were significantly more resistant to IL-2 deprivation-induced apoptosis. CD T cells were also more resistant to Fas- and nitric oxide-mediated apoptosis, whereas TNF-alpha failed to induce cell death in all groups. Compared with control, CD mucosa contained similar numbers of Bcl-2+, but fewer Bax+, cells, while UC mucosa contained fewer Bcl-2+, but more Bax+, cells. Hence, the Bcl-2/Bax ratio was significantly higher in CD and lower in UC. These results indicate that CD may represent a disorder where the rate of T cell proliferation exceeds that of cell death. Insufficient T cell apoptosis may interfere with clonal deletion and maintenance of tolerance, and result in inappropriate T cell accumulation contributing to chronic inflammation.

Extended Framework for Modeling Choice Behavior

Abstract: We review the case against the standard model of rational behavior and discuss the consequences of various ‘anomalies’ of preference elicitation. A general theoretical framework that attempts to disentangle the various psychological elements in the decision-making process is presented. We then present a rigorous and general methodology to model the theoretical framework, explicitly incorporating psychological factors and their influences on choices. This theme has long been deemed necessary by behavioral researchers, but is often ignored in demand models. The methodology requires the estimation of an integrated multi-equation model consisting of a discrete choice model and the latent variable model system. We conclude with a research agenda to bring the theoretical framework into fruition.

Truncated CBP Protein Leads to Classical Rubinstein—Taybi Syndrome Phenotypes in Mice: Implications for a Dominant-Negative Mechanism

Abstract: A mouse model of Rubinstein-Taybi syndrome (RTS) was generated by an insertional mutation into the cyclic AMP response element-binding protein (CREB)-binding protein (CBP) gene. Heterozygous CBP-deficient mice, which had truncated CBP protein (residues 1-1084) containing the CREB-binding domain (residues 462-661), showed clinical features of RTS, such as growth retardation (100%), retarded osseous maturation (100%), hypoplastic maxilla with narrow palate (100%), cardiac anomalies (15%) and skeletal abnormalities (7%). Truncated CBP is considered to have been acting during development as a dominant-negative inhibitor to lead to the phenotypes of RTS in mice. Our studies with step-through-type passive avoidance tests and with fear conditioning test showed that mice were deficient in long-term memory (LTM). In contrast, short-term memory (STM) appeared to be normal. These results implicate a crucial role for CBP in mammalian LTM. Our CBP +/- mice would be an excellent model for the study of the role of CBP in development and memory storage mechanisms.

MAP kinase and Wnt pathways converge to downregulate an HMG-domain repressor in Caenorhabditis elegans

Abstract: The signalling protein Wnt regulates transcription factors containing high-mobility-group (HMG) domains to direct decisions on cell fate during animal development1 In Caenorhabditis elegans, the HMG-domain-containing repressor POP-1 distinguishes the fates of anterior daughter cells from their posterior sisters throughout development2,3, and Wnt signalling downregulates POP-1 activity in one posterior daughter cell called E (refs 2, 4, 5) Here we show that the genes mom-4 and lit-1 are also required to downregulate POP-1, not only in E but also in other posterior daughter cells Consistent with action in a common pathway, mom-4 and lit-1 exhibit similar mutant phenotypes and encode components of the mitogen-activated protein kinase (MAPK) pathway that are homologous to vertebrate transforming-growth-factor-β-activated kinase (TAK1) and NEMO-like kinase (NLK), respectively Furthermore, MOM-4 and TAK1 bind related proteins that promote their kinase activities We conclude that a MAPK-related pathway cooperates with Wnt signal transduction to downregulate POP-1 activity These functions are likely to be conserved in vertebrates, as TAK1 and NLK can downregulate HMG-domain-containing proteins related to POP-1 (ref 6)

Fluoroalkylsilane Monolayers Formed by Chemical Vapor Surface Modification on Hydroxylated Oxide Surfaces

Abstract: Water-repellent surfaces have been prepared by exposing Si substrates with a hydroxylated surface oxide to fluoroalkyl silane (FAS) vapor. Since this chemical vapor surface modification (CVSM) is based on the chemical reaction between organosilane molecules and hydroxyl groups at the oxide surface, prior to CVSM, the substrate surface was completely hydroxylated by irradiating in air with a 172-nm ultraviolet light until the water contact angle of the surface became almost 0°. Under atmospheric pressure, the substrate was then exposed to vapor of an FAS precursor, that is, one of three types of FAS having different perfluoroalkyl chain lengths [CF3(CF2)nCH2CH2Si(OCH3)3, where n = 0, 5, or 7, referred to as FAS-3, FAS-13, and FAS-17, respectively]. The FAS molecules chemically reacted with the hydroxyl groups on the substrate surface and adsorbed onto it, forming a thin layer of less than 2 nm in thickness. The water repellency of the substrate surface increased with an increase in perfluoroalkyl chain len...

Distortion of proximodistal information causes JNK-dependent apoptosis in Drosophila wing.

Abstract: Distinct and evolutionarily conserved signal-transduction cascades mediate the survival or death of cells during development. The c-Jun amino-terminal kinases (JNKs) of the mitogen-activated protein kinase superfamily are involved in apoptotic signalling in various cultured cells. However, the role of the JNK pathway in development is less well understood. In Drosophila, Decapentaplegic (Dpp; a homologue of transforming growth factor-beta) and Wingless (Wg; a Wnt homologue) proteins are secretory morphogens that act cooperatively to induce formation of the proximodistal axis of appendages. Here we show that either decreased Dpp signalling in the distal wing cells or increased Dpp signalling in the proximal wing cells causes apoptosis. Inappropriate levels of Dpp signalling lead to aberrant morphogenesis in the respective wing zones, and these apoptotic zones are also determined by the strength of the Wg signal. Our results indicate that distortion of the positional information determined by Dpp and Wg signalling gradients leads to activation of the JNK apoptotic pathway, and the consequent induction of cell death thereby maintains normal morphogenesis.