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Biomedical subjects

H Nishimura

Publications and source records attributed to H Nishimura.

At least 37 records · Page 2Linked to original sources

Isolation and characterization of a human thiamine pyrophosphokinase cDNA.

A human thiamine pyrophosphokinase cDNA clone (hTPK1) was isolated and sequenced. When the intact hTPK1 open reading frame was expressed as a histidine-tag fusion protein in Escherichia coli, marked enzyme activity was detected in the bacterial cells. The hTPK1 mRNA was widely expressed in various human tissues at a very low level, and the mRNA content in cultured fibroblasts was unaffected by the thiamine concentration of the medium. The chromosome localization of the hTPK1 gene was assigned to 7q34.

Amino Acid Sequence↗

Autoimmune dilated cardiomyopathy in PD-1 receptor-deficient mice.

Dilated cardiomyopathy is a severe pathology of the heart with poorly understood etiology. Disruption of the gene encoding the negative immunoregulatory receptor PD-1 in BALB/c mice, but not in BALB/c RAG-2-/- mice, caused dilated cardiomyopathy with severely impaired contraction and sudden death by congestive heart failure. Affected hearts showed diffuse deposition of immunoglobulin G (IgG) on the surface of cardiomyocytes. All of the affected PD-1-/- mice exhibited high-titer circulating IgG autoantibodies reactive to a 33-kilodalton protein expressed specifically on the surface of cardiomyocytes. These results indicate that PD-1 may be an important factor contributing to the prevention of autoimmune diseases.

Animals↗

Memory phenotype CD8(+) T cells in IL-15 transgenic mice are involved in early protection against a primary infection with Listeria monocytogenes.

We recently constructed IL-15 transgenic (Tg) mice using cDNA encoding a secretable isoform of the IL-15 precursor protein under the control of an MHC class I promoter. The IL-15 Tg mice exhibited resistance against a primary infection with Listeria monocytogenes. The numbers of memory CD8(+) T cells were markedly increased in the IL-15 Tg mice following Listeria infection accompanied by sustained IL-15 production. The increased CD44(+)CD8(+) T cells in the infected IL-15 Tg mice were not specialized to recognize Listeria-specific antigen but produced a large amount of IFN-gamma in response to bystander stimulation exogenous IL-15 in combination with IL-12. Furthermore, Listeria-specific Th1 response by CD4(+) T cells was significantly augmented in the IL-15 Tg mice compared with control mice following Listeria infection. In vivo depletion of the CD8(+) T cells by anti-CD8 monoclonal antibody and adoptive transfer of the T cells from naive IL-15 Tg mice indicated that the CD8(+) T cells functioned not only to eliminate bacteria at the early stage of infection but also to promote Th1 response to L. monocytogenes. Overexpression of IL-15 shed light on a novel role of memory CD8(+) T cells in early protection and promotion of Th1 response against a primary infection with L. monocytogenes.

Adoptive Transfer↗

Glutamate synthesis via photoreduction of NADP+ by photostable chlorophyllide coupled with polyethylene-glycol.

Chlorophyllide a was coupled with alpha-(3-aminopropyl)-omega-methoxypoly(oxyethylene) (PEG-NH2) to form a PEG-chlorophyllide conjugate through an acid-amide bond. The conjugate catalyzed the reduction of methylviologen in the presence of 2-mercaptoethanol. It also catalyzed the photoreduction of NADP+ or NAD+ in the presence of ascorbate as an electron donor and ferredoxin-NADP+ reductase as the coupling enzyme. Utilizing the reducing power of NADPH generated by PEG-chlorophyllide conjugate under illumination, glutamate was synthesized from 2-oxoglutarate and NH4+ in the presence of glutamate dehydrogenase. PEG-chlorophyllide conjugate was quite stable toward light illumination compared with chlorophyll a. The increase in the molecular weight of PEG in the PEG-chlorophyllide conjugates was accompanied by the enhancement of photostability of the conjugate and also by the increased solubility in the aqueous solution.

Carboxylic Ester Hydrolases↗

Relationship between bronchial hyperresponsiveness and development of asthma in children with chronic cough.

To evaluate the relationship between bronchial hyperresponsiveness (BHR) and the development of asthma in children with chronic cough, we performed methacholine inhalation challenges and transcutaneous oxygen pressure (tcPO2) measurements in 92 children with chronic cough aged from 1-13 years (55 boys and 37 girls; mean, 5.3 years) and followed them for > or = 10 years. Forty-four age-matched children with asthma (24 males and 20 females; mean, 6.5 years) and 44 age-matched children without cough or asthma served as controls (18 males and 26 females; mean, 4.6 years). Consecutive doubling doses of methacholine were inhaled until a 10% decrease in tcPO2 from baseline was observed. The cumulative dose of methacholine at the inflection point of the tcPO2 record (Dmin-PO2) was considered to represent hyperresponsiveness to inhaled methacholine. After 10 years or more of follow-up, 60 of the 92 subjects with cough answered our questionnaire, and 27/60 had been diagnosed with asthma. There was a statistical difference in Dmin-PO2 between the children who presented with chronic cough originally and who developed asthma (asthma-developed group) and those who did not develop asthma (asthma-free group). There was no difference in the value of Dmin-PO2 between the asthma-developed group and the asthma group, or between the asthma-free group and the age-matched control group. Among the children with chronic cough, there was no difference in Dmin-PO2 between girls and boys, either in the asthma-developed group or in the asthma-group. We conclude that 45% of the children with a chronic cough in early life developed asthma, and that BHR in children with chronic cough during the childhood period is a strong risk factor for the development of asthma.

Administration, Inhalation↗

Regional alterations of myocardial norepinephrine transporter density in streptozotocin-induced diabetic rats: implications for heterogeneous cardiac accumulation of MIBG in diabetes.

Cardiac scintigraphic studies using iodine-123 labeled metaiodobenzylguanidine ([123I]MIBG) have previously demonstrated the heterogeneous myocardial accumulation of radioactivity in diabetes. In this study, we investigated the myocardial regional distribution of [125I]MIBG and the effects of regional myocardial blood flow, myocardial norepinephrine (NE) content, and norepinephrine transporter (NET) function on regional [125I]MIBG accumulation in streptozotocin-induced diabetic (STZ-D) rats. Dual-isotope autoradiographic studies using [125I]MIBG and technetium-99m labeled hexakis (2-methoxy-2-isobutylisonitrile) (99mTc-MIBI), a tracer for the measurement of myocardial blood flow, were carried out to investigate the changes in regional myocardial blood flow in STZ-D rats. Uptake of [125I]MIBG was similar between the anterior wall and the inferior wall in control rats. On the other hand, in STZ-D rats, uptake of [125I]MIBG in the inferior wall was significantly less than that in the anterior wall. Uptake of 99mTc-MIBI was not significantly different between the anterior and inferior walls in control or STZ-D rats, indicating that myocardial blood flow did not change regionally in either control or STZ-D rats, and that the blood flow was not responsible for the heterogeneity of the distribution of [125I]MIBG in STZ-D rats. In STZ-D rats, cardiac NE concentrations determined using an HPLC-electrochemical detection (ECD) system were significantly increased in both the anterior and the inferior wall, although there was no significant difference in NE concentration between the anterior and inferior walls in control or STZ-D rats. Furthermore, the density and affinity of NET were investigated by studying the binding of [3H]desipramine to cardiac membranes. The Bmax values of the NET in the anterior wall were not significantly different between control and STZ-D rats, but the Bmax value of the NET in the inferior wall was significantly lower in STZ-D rats than in controls. In conclusion, myocardial MIBG uptake was reduced in the inferior wall of STZ-D rats compared with control rats; this decrease was correlated with the decrease in NET density, but was not dependent on the regional myocardial blood flow and NE concentration. These results suggest that regional fluctuations in NET levels in the inferior wall contribute to heterogeneous MIBG accumulation in diabetes.

3-Iodobenzylguanidine↗

Acute effects of prostaglandin D2 to induce airflow obstruction and airway microvascular leakage in guinea pigs: role of thromboxane A2 receptors.

BACKGROUND: Although prostaglandin D2 (PGD2), a mast cell-derived inflammatory mediator, may trigger allergic airway inflammation, its potency and the mechanism by which it induces airway microvascular leakage, a component of airway inflammation, is not clear. OBJECTIVE: We wanted to evaluate the relative potency of PGD2 to cause microvascular leakage as compared to airflow obstruction, because both responses were shown to occur simultaneously in allergic airway diseases such as asthma. The role of thromboxane A2 receptors (TP receptors) in inducing these airway responses was also examined. METHODS: Anesthetized and mechanically ventilated guinea pigs were given i.v. Evans blue dye (EB dye) and, 1 min later, PGD2 (30, 100, 300 or 1,000 nmol/kg). For comparison, the effect of 150 nmol/kg histamine or 2 nmol/kg leukotriene D4 (LTD4) was also examined. Lung resistance (R(L)) was measured for 6 min (or 25 min for selected animals) and the lungs were removed to calculate the amount of extravasated EB dye into the airways as a marker of leakage. In some of the animals, specific TP receptor antagonists, S-1452 (10 microg/kg) or ONO-3708 (10 mg/kg), or a thromboxane A2 synthase inhibitor, OKY-046 (30 mg/kg), was pretreated before giving PGD2. RESULTS: Injection of PGD2 produced an immediate and dose-dependent increase in RL (peaking within 1 min), which was significant at all doses studied. At 1,000 nmol/kg, PGD2 induced a later increase in R(L), starting at 3 min and reaching a second peak at 8 min. By contrast, only PGD2 at doses of 300 and 1,000 nmol/kg produced a significant increase in EB dye extravasation. The relative potency of 1,000 nmol/kg PGD2 to induce leakage as compared to airflow obstruction was comparable to histamine at most of airway levels, but less than LTD4. Both responses caused by PGD2 were completely abolished by S-1452 and ONO-3708, but not by OKY-046. CONCLUSION: PGD2 may induce airway microvascular leakage by directly stimulating TP receptors without generating TXA2 in guinea pigs. Microvascular leakage may play a role in the development of allergic airway inflammation caused by PGD2.

Airway Obstruction↗

Angiotensin receptors--evolutionary overview and perspectives.

The structure of the angiotensin molecule has been well preserved throughout the vertebrate scale with some amino acid variations. Specific angiotensin receptors (AT receptors) that mediate important physiological functions have been noted in a variety of tissues and species. Physiological and pharmacological characterization of AT receptors and, more recently, molecular cloning studies have elucidated the presence of AT receptor subtypes. Comparative studies suggest that an AT receptor subtype homologous to the mammalian type 1 receptor subtype (AT(1)), though pharmacologically distinct, is present in amphibians and birds, whereas AT receptors cloned from teleosts show low homology to both AT(1) and AT(2) receptor subtypes. Furthermore, receptors differing from both the AT(1)-homologue receptor and AT(2) receptor exist in some non-mammalian species. This may suggest that the prototype AT receptor evolved in primitive vertebrates and diverged to more than one type of AT receptor subtype during phylogeny. Furthermore, phenotypic modulation of AT receptors appears to occur during individual development/maturation.

Animals↗

Maturation-dependent neointima formation in fowl aorta.

Fowl show spontaneous elevation of blood pressure (BP) and neointimal plaque formation in the abdominal aorta at young ages. Maturation/age-dependent modulation of vascular lesions and a causal relationship between elevated BP and neointima formation, however, have not been clarified. We therefore intended to characterize, first, maturation/age-dependent neointimal plaque formation and vascular lesions and, second, their relationship to BP elevation. The BP measured in conscious domestic fowl, Gallus gallus, White Leghorn breed, DeKalb strain, via an indwelling catheter inserted into the ischiadic artery, increased with maturation in males; and at plateau level, BP (mmHg) was significantly (P<0.01) higher in males (194.0+/-4.6, n=11) than in females (169.3+/-3.1, n=10). Neointimal plaques consisting of neointimal cells and abundant extracellular matrix appeared initially in the distal segment of the abdominal aorta (lesion-prone area) of chicks as early as 6 weeks old. The area (size) of neointimal plaques right above the ischiadic bifurcation increased with maturation, whereas the plaque area became smaller with some degenerative changes in adult birds. In some birds, diffuse subendothelial hyperplasia and more extensive plaque formation at the branching points of the aorta were observed. The plaque area appears to be larger in birds, particularly in males that have higher BP (r=0.68). The width of aortic smooth muscle (SM) layers, measured in regions with no plaque, increased with age, whereas the number of cells per unit of area decreased, suggesting that hypertrophy of vascular SM occurs in response to exposure of the vascular wall to high BP. The number of cells was significantly (P<0.01) higher in the plaque than in underlying aortic SM layers or in layers with no plaque formation. Both neointimal plaques and underlying SM layers are immunohistochemically positive for alpha SM actin, suggesting that neointimal cells are modulated SM cells, whereas the staining with SM myosin heavy chain antibody is low in neointimal plaques. Furthermore, plasma arginine levels dropped in accordance with the time of neointimal plaque formation, whereas plasma cholesterol levels showed an age-dependent increase. The results suggest that spontaneous development of neointimal plaques may be a consequence of exposure to high BP and associated local hemodynamic changes.

Actins↗

PD-1: an inhibitory immunoreceptor involved in peripheral tolerance.

PD-1 deficiency causes a variety of autoimmune diseases. Inhibitory signaling through the PD-1 receptor might therefore be involved in the regulation of peripheral tolerance. The constitutive expression of PD-1 ligands (PD-L1 and PD-L2) on parenchymal cells of heart, lung and kidney suggests that the PD-1-PD-L system could provide unique negative signaling to help prevent autoimmune diseases.

Animals↗

Dichotomous effect of a traditional Japanese medicine, bu-zhong-yi-qi-tang on allergic asthma in mice.

To determine the potentiality of prophylactic and/or therapeutic approaches using a traditional herbal medicine, Bu-zhong-yi-qi-tang (Japanese name: Hochu-ekki-to, HOT), for the control of allergic disease, we examined the effects of oral administration of HOT on a murine model of asthma allergic responses. When oral administration of HOT was begun at the induction phase immediately after OVA sensitization, eosinophilia and Th2-type cytokine production in the airway were reduced in OVA-sensitized mice following OVA inhalation. The serum levels of OVA-specific immunoglobulin (Ig)E and IgG1 were significantly decreased, whereas the level of OVA-specific IgG2a was increased. Interleukin (IL)-4 production by spleen T cells in response to OVA was significantly suppressed, while Interferon (IFN)-gamma production was increased in mice treated with HOT in the induction phase. On the other hand, HOT given in the eliciting phase induced a predominant Th2 response with increased IgE production in OVA-sensitized mice following OVA inhalation. These results suggest that the oral administration of HOT dichotomously modulates allergic inflammation in a murine model for asthma, thus offering a different approach for the treatment of allergic disorders.

Administration, Oral↗

PD-L2 is a second ligand for PD-1 and inhibits T cell activation.

Programmed death I (PD-I)-deficient mice develop a variety of autoimmune-like diseases, which suggests that this immunoinhibitory receptor plays an important role in tolerance. We identify here PD-1 ligand 2 (PD-L2) as a second ligand for PD-1 and compare the function and expression of PD-L1 and PD-L2. Engagement of PD-1 by PD-L2 dramatically inhibits T cell receptor (TCR)-mediated proliferation and cytokine production by CD4+ T cells. At low antigen concentrations, PD-L2-PD-1 interactions inhibit strong B7-CD28 signals. In contrast, at high antigen concentrations, PD-L2-PD-1 interactions reduce cytokine production but do not inhibit T cell proliferation. PD-L-PD-1 interactions lead to cell cycle arrest in G0/G1 but do not increase cell death. In addition, ligation of PD-1 + TCR leads to rapid phosphorylation of SHP-2, as compared to TCR ligation alone. PD-L expression was up-regulated on antigen-presenting cells by interferon gamma treatment and was also present on some normal tissues and tumor cell lines. Taken together, these studies show overlapping functions of PD-L1 and PD-L2 and indicate a key role for the PD-L-PD-1 pathway in regulatingT cell responses.

Amino Acid Sequence↗

ORP150 protects against hypoxia/ischemia-induced neuronal death.

Oxygen-regulated protein 150 kD (ORP150) is a novel endoplasmic-reticulum-associated chaperone induced by hypoxia/ischemia. Although ORP150 was sparingly upregulated in neurons from human brain undergoing ischemic stress, there was robust induction in astrocytes. Cultured neurons overexpressing ORP150 were resistant to hypoxemic stress, whereas astrocytes with inhibited ORP150 expression were more vulnerable. Mice with targeted neuronal overexpression of ORP150 had smaller strokes compared with controls. Neurons with increased ORP150 demonstrated suppressed caspase-3-like activity and enhanced brain-derived neurotrophic factor (BDNF) under hypoxia signaling. These data indicate that ORP150 is an integral participant in ischemic cytoprotective pathways.

Animals↗

Thymus-dependent modulation of Ly49 inhibitory receptor expression on NK1.1+gamma/delta T cells.

Major histocompatibility complex (MHC) class I-specific inhibitory receptors are expressed not only on natural killer (NK) cells but also on some subsets of T cells. We here show Ly49 expression on gamma/delta T cells in the thymus and liver of beta2-microglobulin-deficient (beta2m-/-) and C57BL/6 (beta2m+/+) mice. Ly49C/I or Ly49A receptor was expressed on NK1.1+gamma/delta T cells but not on NK1.1-gamma/delta T cells. The numbers of NK1.1+gamma/delta T cells were significantly smaller in beta2m+/+ mice than in beta2m-/- mice with the same H-2b genetic background. Among NK1.1+gamma/delta T cells, the proportions of Ly49C/I+ cells but not of Ly49A+ cells, were decreased in beta2m+/+ mice, suggesting that cognate interaction between Ly49C/I and H-2Kb is involved in the reduction of the number of Ly49C/I+ gamma/delta T cells in beta2m+/+ mice. The frequency of Ly49C/I+ cells in NK1.1+gamma/delta T cells was lower in both lethally irradiated beta2m+/+ mice transplanted with bone marrow (BM) from beta2m-/- mice and lethally irradiated beta2m-/- mice transplanted with BM from beta2m+/+ mice than those in adult thymectomized BM-transplanted chimera mice. These results suggest that reduction of Ly49C/I+ NK1.1+gamma/delta T cells in beta2m+/+ mice is at least partly due to the down-modulation by MHC class I molecules on BM-derived haematopoietic cells or radioresistant cells in the thymus.

Animals↗

Bactericidal/permeability-increasing protein promotes complement activation for neutrophil-mediated phagocytosis on bacterial surface.

The neutrophil bactericidal/permeability-increasing protein (BPI) has both bactericidal and lipopolysaccharide-neutralizing activities. The present study suggests that BPI also plays an important role in phagocytosis of Escherichia coli by neutrophils through promotion of complement activation on the bacterial surface. Flow cytometric analysis indicated that fluorescein-labelled E. coli treated with BPI were phagocytosed in the presence of serum at two- to five-fold higher levels than phagocytosis of the bacteria without the treatment. In contrast, phagocytosis of the fluoresceined bacteria with or without treatment by BPI did not occur at all in the absence of serum. The phagocytosis stimulated by BPI and serum was dose-dependent. The effect of BPI on phagocytosis in the presence of serum was not observed on Gram-positive bacteria (Staphylococcus aureus). Interestingly, the complement C3b/iC3b fragments were deposited onto the bacterial surface also as a function of the BPI concentration under conditions similar to those for phagocytosis. Furthermore, the BPI-promoted phagocytosis was blocked completely by anti-C3 F(ab')(2) and partially by anti-complement receptor (CR) type 1 and/or anti-CR type 3. These findings suggest that BPI accelerates complement activation to opsonize bacteria with complement-derived fragments, leading to stimulation of phagocytosis by neutrophils via CR(s).

Antimicrobial Cationic Peptides↗

Therapeutic possibilities of long-term roxithromycin treatment for chronic diffuse sclerosing osteomyelitis of the mandible.

The clinical efficacy of long-term roxithromycin treatment was examined objectively in nine patients with chronic diffuse sclerosing osteomyelitis of the mandible. Roxithromycin was administered orally at a dose of 300 mg/day for between 68 days and 66 months. In seven of the nine cases (77.8%), the symptoms disappeared 1-12 months after the start of therapy. Radiography showed that osteolytic changes (evident from 'moth-eaten' appearance of bone) had improved but that osteosclerosis had persisted or become more predominant by the end of therapy. Therefore, the optimum duration of treatment should be decided according to the amelioration of symptoms along with the disappearance of osteolytic findings in radiographs. Diarrhoea and stomach discomfort occurred in one case, and liver dysfunction in another, but these adverse reactions were relatively mild. The mechanism of action of roxithromycin in this study is not yet fully understood, but our results indicate that long-term roxithromycin treatment may be useful for diffuse sclerosing osteomyelitis of the mandible and should be attempted before surgical treatment is considered.

Adult↗

Mice lacking interleukin-2 (IL-2)/IL-15 receptor beta chain are susceptible to infection with avirulent Salmonella enterica subsp. enterica serovar choleraesuis but mice lacking IL-2 are resistant.

Interleukin-2 (IL-2)/IL-15 receptor beta (IL-15R beta)(-/-) mice were susceptible to infection with avirulent Salmonella enterica subsp. enterica serovar Choleraesuis, whereas IL-2(-/-) mice were resistant. A natural killer cell response was not evident for both types of deficient mice. A Th1 response was detected in IL-2(-/-) but not in IL-2/IL-15R beta(-/-) mice infected with Salmonella, suggesting that IL-2/IL-15R beta signaling is important for the generation of protective Th1 cells.

Animals↗