Showing papers by "Tohoku University published in 1999"

Endoscopic assessment of oesophagitis: clinical and functional correlates and further validation of the Los Angeles classification.

Abstract: Background—Endoscopic oesophageal changes are diagnostically helpful and identify patients exposed to the risk of dis- ease chronicity. However, there is a seri- ous lack of agreement about how to describe and classify the appearance of reflux oesophagitis Aims—To examine the reliability of crite- ria that describe the circumferential ex- tent of mucosal breaks and to evaluate the functional and clinical correlates of pa- tients with reflux disease whose oesoph- agitis was graded according to the Los Angeles system. Methods—Forty six endoscopists from diVerent countries used a detailed work- sheet to evaluate endoscopic video record- ings from 22 patients with the full range of severity of reflux oesophagitis. In separate studies, Los Angeles system gradings were correlated with 24 hour oesophageal pH monitoring (178 patients), and with clini- cal trials of omeprazole treatment (277 patients). Results—Evaluation of circumferential extent of oesophagitis by the criterion of whether mucosal breaks extended be- tween the tops of mucosal folds, gave acceptable agreement (mean Œ value 0.4) among observers. This approach is used in the Los Angeles system. An alternative approach of grouping the circumferential extent of mucosal breaks as occupying 0-25%, 26-50%, 51-75%, 76-99%, or 100% of the oesophageal circumference, gave unacceptably high interobserver variation (mean Œ values 0-0.15) for all but the low- est category of extent (mean Œ value 0.4). Severity of oesophageal acid exposure was significantly (p<0.001) related to the se- verity grade of oesophagitis. Preteatment oesophagitis grades A-C were related to heartburn severity (p<0.01), outcomes of omeprazole (10 mg daily) treatment (p<0.01),and the risk for symptom relapse oV therapy over six months (p<0.05). Conclusions—Results add further support to previous studies for the clinical utility of the Los Angeles system for endoscopic grading of oesophagitis. (Gut 1999;45:172-180)

Signaling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase.

Abstract: The actin cytoskeleton undergoes extensive remodeling during cell morphogenesis and motility. The small guanosine triphosphatase Rho regulates such remodeling, but the underlying mechanisms of this regulation remain unclear. Cofilin exhibits actin-depolymerizing activity that is inhibited as a result of its phosphorylation by LIM-kinase. Cofilin was phosphorylated in N1E-115 neuroblastoma cells during lysophosphatidic acid–induced, Rho-mediated neurite retraction. This phosphorylation was sensitive to Y-27632, a specific inhibitor of the Rho-associated kinase ROCK. ROCK, which is a downstream effector of Rho, did not phosphorylate cofilin directly but phosphorylated LIM-kinase, which in turn was activated to phosphorylate cofilin. Overexpression of LIM-kinase in HeLa cells induced the formation of actin stress fibers in a Y-27632–sensitive manner. These results indicate that phosphorylation of LIM-kinase by ROCK and consequently increased phosphorylation of cofilin by LIM-kinase contribute to Rho-induced reorganization of the actin cytoskeleton.

Novel Mesoporous Materials with a Uniform Distribution of Organic Groups and Inorganic Oxide in Their Frameworks

Abstract: Novel organic−inorganic hybrid mesoporous materials have been synthesized, in which organic and inorganic oxide moieties are distributed homogeneously at the molecular level in the framework, forming a covalently bonded network. They are highly ordered at the mesoscale, with two- and three-dimensional hexagonal symmetries and well-defined external morphologies. Nitrogen adsorption measurements show a uniform pore-size distribution with pore diameters of 31 and 27 A, and high surface areas of 750 and 1170 m2/g. The synthetic procedure to polymerize the organosilane monomer containing two trialkoxysilyl groups in the presence of surfactant can be applied to the synthesis of a variety of highly ordered organic−inorganic hybrid mesoporous materials.

Light-driven monodirectional molecular rotor

Abstract: Attempts to fabricate mechanical devices on the molecular level1,2 have yielded analogues of rotors3, gears4, switches5, shuttles6,7, turnstiles8 and ratchets9. Molecular motors, however, have not yet been made, even though they are common in biological systems10. Rotary motion as such has been induced in interlocked systems11,12,13 and directly visualized for single molecules14, but the controlled conversion of energy into unidirectional rotary motion has remained difficult to achieve. Here we report repetitive, monodirectional rotation around a central carbon–carbon double bond in a chiral, helical alkene, with each 360° rotation involving four discrete isomerization steps activated by ultraviolet light or a change in the temperature of the system. We find that axial chirality and the presence of two chiral centres are essential for the observed monodirectional behaviour of the molecular motor. Two light-induced cis-trans isomerizations are each associated with a 180° rotation around the carbon–carbon double bond and are each followed by thermally controlled helicity inversions, which effectively block reverse rotation and thus ensure that the four individual steps add up to one full rotation in one direction only. As the energy barriers of the helicity inversion steps can be adjusted by structural modifications, chiral alkenes based on our system may find use as basic components for ‘molecular machinery’ driven by light.

Mammalian Cry1 and Cry2 are essential for maintenance of circadian rhythms.

Abstract: Many biochemical, physiological and behavioural processes show circadian rhythms which are generated by an internal time-keeping mechanism referred to as the biological clock. According to rapidly developing models, the core oscillator driving this clock is composed of an autoregulatory transcription-(post) translation-based feedback loop involving a set of 'dock' genes. Molecular clocks do not oscillate with an exact 24-hour rhythmicity but are entrained to solar day/night rhythms by light. The mammalian proteins Cryl and Cry2, which are members of the family of plant blue-light receptors (cryptochromes) and photolyases, have been proposed as candidate light receptors for photoentrainment of the biological clock. Here we show that mice lacking the Cryl or Cry2 protein display accelerated and delayed free-running periodicity of locomotor activity, respectively. Strikingly, in the absence of both proteins, an instantaneous and complete loss of free-running rhythmicity is observed. This suggests that, in addition to a possible photoreceptor and antagonistic clock-adjusting function, both proteins are essential for the maintenance of circadian rhythmicity.

Microstructural evolution of 6063 aluminum during friction-stir welding

Abstract: The microstructural distribution associated with a hardness profile in a friction-stir-welded, age-hardenable 6063 aluminum alloy has been characterized by transmission electron microscopy (TEM) and orientation imaging microscopy (OIM). The friction-stir process produces a softened region in the 6063 Al weld. Frictional heating and plastic flow during friction-stir welding create fine recrystallized grains in the weld zone and recovered grains in the thermomechanically affected zone. The hardness profile depends greatly on the precipitate distribution and only slightly on the grain size. The softened region is characterized by dissolution and growth of the precipitates during the welding. Simulated weld thermal cycles with different peak temperatures have shown that the precipitates are dissolved at temperatures higher than 675 K and that the density of the strengthening precipitate was reduced by thermal cycles lower than 675 K. A comparison between the thermal cycles and isothermal aging has suggested precipitation sequences in the softened region during friction-stir welding.

Molecular mechanisms of transcription activation by HLF and HIF1alpha in response to hypoxia: their stabilization and redox signal-induced interaction with CBP/p300.

Abstract: Hypoxia-inducible factor 1 alpha (HIF1alpha) and its related factor, HLF, activate expression of a group of genes such as erythropoietin in response to low oxygen. Transfection analysis using fusion genes of GAL4DBD with various fragments of the two factors delineated two transcription activation domains which are inducible in response to hypoxia and are localized in the C-terminal half. Their sequences are conserved between HLF and HIF1alpha. One is designated NAD (N-terminal activation domain), while the other is CAD (C-terminal activation domain). Immunoblot analysis revealed that NADs, which were rarely detectable at normoxia, became stabilized and accumulated at hypoxia, whereas CADs were constitutively expressed. In the mammalian two-hybrid system, CAD and NAD baits enhanced the luciferase expression from a reporter gene by co-transfection with CREB-binding protein (CBP) prey, whereas CAD, but not NAD, enhanced beta-galactosidase expression in yeast by CBP co-expression, suggesting that NAD and CAD interact with CBP/p300 by a different mechanism. Co-transfection experiments revealed that expression of Ref-1 and thioredoxin further enhanced the luciferase activity expressed by CAD, but not by NAD. Amino acid replacement in the sequences of CADs revealed a specific cysteine to be essential for their hypoxia-inducible interaction with CBP. Nuclear translocation of thioredoxin from cytoplasm was observed upon reducing O2 concentrations.

Glucose and fructose decomposition in subcritical and supercritical water: Detailed reaction pathway, mechanisms, and kinetics

Abstract: Experiments were performed on the products of glucose decomposition at short residence times to elucidate the reaction pathways and evaluate kinetics of glucose and fructose decomposition in sub- and supercritical water. The conditions were a temperature of 300−400 °C and pressure of 25−40 MPa for extremely short residence times between 0.02 and 2 s. The products of glucose decomposition were fructose, a product of isomerization, 1,6-anhydroglucose, a product of dehydration, and erythrose and glyceraldehyde, products of C−C bond cleavage. Fructose underwent reactions similar to glucose except that it did not form 1,6-anhydroglucose and isomerization to glucose is negligible. The mechanism for the products formed from C−C bond cleavage could be explained by reverse aldol condensation and the double-bond rule of the respective enediols formed during the Lobry de Bruyn Alberda van Ekenstein transformation. The differential equations resulting from the proposed pathways were fit to experimental results to obt...

Identification of a novel mechanism of regulation of Ah (dioxin) receptor function

Abstract: Ah receptor (AhR) is a ligand-activated transcription factor that mediates pleiotropic effects of environmental pollutants such as 2,3,7,8-tetrachlorodibenzo-p-dioxin on host animals. In addition to induction of drug-metabolizing enzymes, the liganded AhR complex was found to activate gene expression of a factor designated AhR repressor (AhRR), which inhibits AhR function by competing with AhR for dimerizing with Arnt and binding to the XRE sequence. Thus, AhR and AhRR form a regulatory circuit in the xenobiotic signal transduction pathway and provide a novel mechanism of regulation of AhR function that may determine tissue-specific sensitivity to environmental pollutants.

Presenilin-1 mutations downregulate the signalling pathway of the unfolded-protein response

Abstract: Presenilin-1 mutations downregulate the signalling pathway of the unfolded-protein response

Primary systemic carnitine deficiency is caused by mutations in a gene encoding sodium ion-dependent carnitine transporter

Abstract: Primary systemic carnitine deficiency (SCD; OMIM 212140) is an autosomal recessive disorder characterized by progressive cardiomyopathy, skeletal myopathy, hypoglycaemia and hyperammonaemia1,2,3. SCD has also been linked to sudden infant death syndrome4. Membrane-physiological studies have suggested a defect of the carnitine transport system in the plasma membrane in SCD patients5 and in the mouse model, juvenile visceral steatosis ( jvs; ref. 6 ). Although the responsible loci have been mapped in both human7 and mouse8, the underlying gene has not yet been identified. Recently, we cloned and analysed the function of a novel transporter protein termed OCTN2 ( ref. 9 ). Our observation that OCTN2 has the ability to transport carnitine in a sodium-dependent manner prompted us to search for mutations in the gene encoding OCTN2, SLC22A5 . Initially, we analysed the mouse gene and found a missense mutation in Slc22a5 in jvs mice. Biochemical analysis revealed that this mutation abrogates carnitine transport. Subsequent analysis of the human gene identified four mutations in three SCD pedigrees. Affected individuals in one family were homozygous for the deletion of a 113-bp region containing the start codon. In the second pedigree, the affected individual was shown to be a compound heterozygote for two mutations that cause a frameshift and a premature stop codon, respectively. In an affected individual belonging to a third family, we found a homozygous splice-site mutation also resulting in a premature stop codon. These mutations provide the first evidence that loss of OCTN2 function causes SCD.

Molecular Characterization of the Follicle Defects in the Growth Differentiation Factor 9-Deficient Ovary

Abstract: Growth differentiation factor-9 (GDF-9), a secreted member of the transforming growth factor-β superfamily, is expressed at high levels in the mammalian oocyte beginning at the type 3a primary follicle stage. We have previously demonstrated that GDF-9-deficient female mice are infertile because of an early block in folliculogenesis at the type 3b primary follicle stage. To address the molecular defects that result from the absence of GDF-9, we have analyzed the expression of several important ovarian marker genes. The major findings of our studies are as follows: 1) There are no detectable signals around GDF-9-deficient follicles for several theca cell layer markers [i.e. 17α-hydroxylase, LH receptor (LHR), and c-kit, the receptor for kit ligand]. This demonstrates that in the absence of GDF-9, the follicles are incompetent to emit a signal that recruits theca cell precursors to surround the follicle; 2) The primary follicles of GDF-9-deficient mice demonstrate an up-regulation of kit ligand and inhibin-α...

The human amygdala plays an important role in gaze monitoring. A PET study.

Abstract: Social contact often initially depends on ascertaining the direction of the other person's gaze. We determined the brain areas involved in gaze monitoring by a functional neuroimaging study. Discrimination between the direction of gaze significantly activated a region in the left amygdala during eye-contact and no eye-contact tasks to the same extent. However, a region in the right amygdala was specifically activated only during the eye-contact task. Results confirm that the left amygdala plays a general role in the interpretation of eye gaze direction, and that the activity of the right amygdala of the subject increases when another individual's gaze is directed towards him. This suggests that the human amygdala plays a role in reading social signals from the face.

Measurement of the Flux and Zenith-Angle Distribution of Upward Throughgoing Muons by Super-Kamiokande

Abstract: A total of 614 upward throughgoing muons of minimum energy 1.6thinspthinspGeV are observed by Super-Kamiokande during 537 detector live days. The measured muon flux is [1.74{plus_minus}0.07(stat){plus_minus} 0.02(sys)]{times}10{sup {minus}13} cm{sup {minus}2}thinsps{sup {minus}1}thinspsr{sup {minus}1} compared to an expected flux of [1.97{plus_minus}0.44(theor)]{times} 10{sup {minus}13} cm{sup {minus}2}thinsps{sup {minus}1}thinspsr{sup {minus}1} . The absolute measured flux is in agreement with the prediction within the errors. However, the zenith-angle dependence of the observed upward throughgoing muon flux does not agree with no-oscillation predictions. The observed distortion in shape is consistent with the {nu}{sub {mu}}{leftrightarrow}{nu}{sub {tau}} oscillation hypothesis with sin{sup 2}thinsp2{theta} {gt}0.4 and 1{times}10{sup {minus}3}{lt}{Delta}m{sup 2}{lt}1{times}1 0{sup {minus}1} eV{sup 2} at 90{percent} confidence level. {copyright} {ital 1999} {ital The American Physical Society}

Broadband antireflection gratings fabricated upon silicon substrates.

Abstract: We fabricated a two-dimensional subwavelength structured (SWS) surface upon a crystal silicon substrate. The SWS surface was patterned by electron beam lithography and etched by an SF(6) fast atom beam. The SWS grating had a conical profile, the period was 150 nm, and the groove was approximately 350 nm deep. The reflectivity was examined at 2002500-nm wavelengths. At 400 nm the reflectivity decreased to 0.5% from the 54.7% of the silicon substrate. We also used HeNe laser light to examine the reflectivity as a function of the incident angle.

Early evolution of disk galaxies: Formation of bulges in clumpy young galactic disks

Abstract: A new idea is proposed for the origin of bulges in spiral galaxies. Numerical simulations of protogalactic collapse suggest strongly that galactic bulges have been assembled from massive clumps formed in galactic disks in their early evolutionary phase. These clumps result from the gravitational instability of the gas-rich disks of young galaxies. Owing to dynamical frictions, those massive clumps, individual masses of which can be as large as ~109 M?, are able to spiral toward the galactic center within a few Gyr. Inward transport of disk matter by this process leads to the formation of a galactic bulge. A simple analytical model has been constructed in which the clumpy evolution of a disk galaxy is controlled by two parameters: the timescale with which the primordial gas in the halo accretes onto the disk plane (i.e., the collapse timescale) and the initial mass fraction of the gas relative to the galaxy total mass. Under plausible assumptions for the variation of these parameters among spiral galaxies, the clumpy evolution model can explain an observed trend in which the bulge-to-disk ratio increases as the total mass or the internal density of the galaxy increases. This success suggests that the clumpy evolution of the galactic disk constitutes an important ingredient of disk galaxy evolution. Star formation in primeval disk galaxies takes place mostly in the clumps. The resulting knotty appearance of these systems may explain the peculiar morphology observed in a number of high-redshift galaxies.

Germline-specific expression of the Oct-4/green fluorescent protein (GFP) transgene in mice.

Abstract: The Pic-1, Oct-1,2, Unc-86 (POU) transcription factor Oct-4 is specifically expressed in the germ cell line, and a previous study has indicated that the expression of the lacZ gene inserted into an 18 kb genomic fragment encompassing the Oct-4 gene can come close to mimicking the endogenous embryonic expression pattern of Oct-4 in transgenic mice. In the present study transgenic mice expressing green fluorescent protein (GFP) in the germ cell line were generated using the same Oct-4 genomic fragments and the expression pattern was analyzed in detail through all stages of germ cell development. The GFP expressing primordial germ cells were first detected as early as 8.0 days post-coitum (d.p.c.; early head fold stage) at the base of the allantois in living embryos. The GFP expression was thereafter found in both male and female germ cells at all developmental stages except in male germ cells after differentiating into type A spermatogonia in the postnatal testis. There was also a lower level of expression in female germ cells in the prophase of the first meiotic division. These transgenic mice therefore proved to be powerful tools for isolating living germ cells at various developmental stages to study their nature and to isolate new genes.

Structural and magnetic phase transitions in shape-memory alloys Ni 2 + x Mn 1 − x Ga

Abstract: The Heusler-type alloy ${\mathrm{Ni}}_{2+x}{\mathrm{Mn}}_{1\ensuremath{-}x}\mathrm{Ga}$ exhibits well defined shape memory properties in a ferromagnetic state, which means that the martensitic transition temperature is lower than the Curie point of this material. The change of composition makes these characteristic temperatures approach each other. To study this behavior, the measurements of specific heat, ac magnetic susceptibility, and dc resistivity were performed. The phase diagram of the cubic ferromagnet describing possible structural and magnetic transitions is obtained theoretically. This diagram is compared with experimental data on ${\mathrm{Ni}}_{2+x}{\mathrm{Mn}}_{1\ensuremath{-}x}\mathrm{Ga}.$ An estimate is given of the magnetic-field influence on the temperature of martensitic transformation in the studied alloys.

Selective expression of a novel surface molecule by human Th2 cells in vivo.

Abstract: The search for reliable marker molecules discriminating between human Th1 and Th2 cells identified a gene encoding a novel member of the G protein-coupled leukocyte chemoattractant receptor family, which is selectively expressed in Th2 but not Th1 lineage cells, thereby named CRTH2 (chemoattractant receptor-homologous molecule expressed on Th2 cells). Studies with anti-CRTH2 mAbs demonstrated that CRTH2 was expressed in a small population (0.4–6.5%) of CD4+ T cells in fresh PBMCs of healthy adults, but no remarkable expression was seen in B cells and NK cells. In some cases, CD8+ T cells (∼3.5%) expressed CRTH2. Phenotypes of CD4+ T cells expressing CRTH2 were CD45RA−, CD45RO+, and CD25+, similar to those of Ag-activated effector/memory T cells. Freshly isolated CRTH2+ CD4+ T cells produced Th2- but little or no Th1-type cytokines upon stimulation with PMA and ionomycin. In addition, an allergen-induced proliferative response in fresh PBMCs was significantly and selectively reduced by subtracting CRTH2+ cells. Together, these results indicate that CRTH2 is selectively expressed in vivo in an activated state of Th2 cells including allergen-responsive Th2 cells, suggesting its pivotal roles in ongoing Th2-type immune reactions.

Photic induction of mPer1 and mPer2 in cry-deficient mice lacking a biological clock.

Abstract: Mice lacking mCry1 and mCry2 are behaviorally arrhythmic. As shown here, cyclic expression of the clock genes mPer1 and mPer2 (mammalian Period genes 1 and 2) in the suprachiasmatic nucleus and peripheral tissues is abolished and mPer1 and mPer2 mRNA levels are constitutively high. These findings indicate that the biological clock is eliminated in the absence of both mCRY1 and mCRY2 (mammalian cryptochromes 1 and 2) and support the idea that mammalian CRY proteins act in the negative limb of the circadian feedback loop. The mCry double-mutant mice retain the ability to have mPer1 and mPer2 expression induced by a brief light stimulus known to phase-shift the biological clock in wild-type animals. Thus, mCRY1 and mCRY2 are dispensable for light-induced phase shifting of the biological clock.

Direct intestinal absorption of red fruit anthocyanins, cyanidin-3-glucoside and cyanidin-3,5-diglucoside, into rats and humans.

Abstract: We determined red fruit anthocyanins, cyanidin-3-glucoside (Cy-g) and cyanidin-3,5-diglucoside (Cy-dg), incorporated into plasma and liver of rats and human plasma by UV-HPLC. Fifteen minutes after an oral supplementation of a mixture of 320 mg of Cy-g and 40 mg of Cy-dg/kg of body weight, rats showed an increase to a maximum of 1563 μg (3490 nmol) of Cy-g/L and 195 μg (320 nmol) of Cy-dg/L in plasma and 0.067 μg (0.15 nmol) of Cy-g/g and a trace of Cy-dg together with methylated metabolites such as peonidin-3-glucoside in liver. In human plasma, 30 min after intake (2.7 mg of Cy-g and 0.25 mg of Cy-dg/kg of body weight), an average of 11 μg (24 nmol) of Cy-g/L and a trace of Cy-dg were found. Cyanidin as aglycone of Cy-g and Cy-dg was not found in such plasma samples, neither were conjugated and methylated anthocyanins. The results indicated that anthocyanins are incorporated keeping structurally intact glycoside forms, from the digestive tract into the blood circulation system in mammals. Keywords: Anth...

Deletion of Fcγ Receptor IIB Renders H-2b Mice Susceptible to Collagen-induced Arthritis

Abstract: Autoimmune diseases, like rheumatoid arthritis, result from a dysregulation of the immune response culminating in hyperactivation of effector cells leading to immune-mediated injury. To maintain an appropriate immune response and prevent the emergence of autoimmune disease, activation signals must be regulated by inhibitory pathways. Biochemical and genetic studies indicate that the type IIB low-affinity receptor for immunoglobulin (Ig)G (FcγRIIB) inhibits cellular activation triggered through antibody or immune complexes and may be an important component in preventing the emergence of autoimmunity. To investigate the role of FcγRIIB in the development of type II collagen (CII)-induced arthritis (CIA), a model for rheumatoid arthritis in humans, we have examined its contribution in determining the susceptibility to CIA in the nonpermissive H-2b haplotype. H-2b mice immunized with bovine CII do not develop appreciable disease. In contrast, immunization of the FcγRIIB-deficient, H-2b mice with bovine CII induced CIA at an incidence of 42.2%. The maximal arthritis index of the FcγRIIB-deficient mice developing CIA (6.9 ± 3.6) was comparable to that of DBA/1 mice (8.6 ± 1.9), an H-2q strain susceptible for CIA induction. IgG1, IgG2a, and IgG2b antibody responses against CII were elevated in the FcγRIIB-deficient animals, especially in those mice showing arthritis, but less pronounced than DBA/1 mice. Histological examinations of the arthritic paws from FcγRIIB-deficient mice revealed that cartilage was destroyed and bone was focally eroded in association with marked lymphocyte and monocyte/macrophage infiltration, very similar to the pathologic findings observed in DBA/1 mice. These results indicate that a nonpermissive H-2b haplotype can be rendered permissive to CIA induction through deletion of FcγRIIB, suggesting that FcγRIIB plays a critical role in suppressing the induction of CIA.