Biomedical subjects

H Horiuchi

Publications and source records attributed to H Horiuchi.

At least 55 records · Page 3Linked to original sources

PubMed2000

Cleavage of CD14 on human gingival fibroblasts cocultured with activated neutrophils is mediated by human leukocyte elastase resulting in down-regulation of lipopolysaccharide-induced IL-8 production.

Activated polymorphonuclear leukocytes (PMNs) release various types of proteases and express them on the cell surface. The proteases play important roles in PMN-mediated events. In the present study, flow cytometric analysis revealed that CD14 expression on human gingival fibroblasts (HGF) was markedly reduced by PMA-activated PMNs in a coculture system. We found that this reduction was caused by both secreted and cell surface proteases produced by activated PMNs. A protease responsible for the reduction was found to be human leukocyte elastase (HLE) secreted from the activated PMNs by use of various protease inhibitors, although HLE was only partially involved in CD14 reduction caused by cell-bound molecule(s) on fixed PMNs. Analysis with purified HLE revealed a time- and dose-dependent reduction of CD14 on HGF, and complete reduction was observed by 20 microg/ml HLE treatment for 30-60 min, but the other molecules such as CD26, CD59, CD157, and MHC class I on HGF were only slightly reduced. This reduction of CD14 resulted from direct proteolysis by HLE on the cell surface, because HLE reduced CD14 on fixed HGF and also on purified cell membranes. As a result of CD14 proteolysis, IL-8 production by HGF was suppressed when triggered by 10 ng/ml LPS, but not by IL-1alpha, indicating that HLE inhibited a CD14-dependent cell activation. These findings suggested that activated PMNs have a potential negative feedback mechanism for HGF function at the inflammatory site, particularly in periodontal tissues.

Adolescent↗

PubMed2000

Small GTPase Rab4 regulates Ca2+-induced alpha-granule secretion in platelets.

Upon activation, platelets release many active substances stored in alpha- and dense-core granules. However, the molecular mechanisms governing regulated exocytosis are not yet fully understood. Here, we have established an assay system using permeabilized platelets to analyze the Ca(2+)-induced exocytosis of both types of granules, focusing on RabGTPases. Incubation with Rab GDP dissociation inhibitor, an inhibitory regulator of RabGTPases, reduced membrane-bound RabGTPases extensively, and caused strong inhibition of the Ca(2+)-induced secretion of von Willebrand factor (vWF) stored in alpha-granules, but not that of [(3)H]5-hydroxytryptamine (5-HT) in dense-core granules. Specifically, Rab4 co-fractionated with vWF and P-selectin (an alpha-granule marker) upon separation of platelet organelles by density gradient centrifugation. Incubation of the permeabilized platelets with cell extracts expressing the dominant negative mutant of His-tagged Rab4S22N, but not with those of similar mutant His-Rab3BT36N, inhibited the vWF secretion, whereas neither of the cell extracts affected the [(3)H]5-HT secretion. Importantly, the inhibition of vWF secretion was rescued by depleting the cell extracts of the His-Rab4S22N with nickel beads. Thus, in platelets, the regulatory mechanisms governing alpha- and dense-core granule secretions are distinct, and Rab4 is an essential regulator of the Ca(2+)-induced exocytosis of alpha-granules.

Bacterial Proteins↗

PubMed2000

Junctional adhesion molecule (JAM) is phosphorylated by protein kinase C upon platelet activation.

Junctional adhesion molecule (JAM) is a member of the immunoglobulin superfamily (IgSF) expressed in tight junctions of epithelial cells and endothelial cells, and implicated in transendothelial migration of leukocytes. Recently, JAM is reported to be constitutively expressed on circulating monocytes, neutrophils, lymphocytes subsets, and platelets. However, the role of JAM is not known. Here, we examined how phosphorylaton of JAM is regulated upon platelet activation. Phosphorylation of JAM was induced by thrombin, collagen, but not by ADP. The phosphorylated amino acids were shown to be serine residues by phosphoamino acid analysis. Inhibition of JAM's phosphorylation by PKC inhibitors and Ca(++) chelator suggests the involvement of conventional types of PKCs. By in vitro kinase assays, we demonstrated that JAM could be directly phosphorylated by cPKCs. We also demonstrated phosphorylation of Ser 284, a putative PKC phosphorylation site, by immunoblotting with anti-phosphoserine-JAM antibody in thrombin-stimulated platelets. In addition to the phosphorylation, JAM seemed to form clusters at several sites of cell-cell contact in aggregated platelets by immunoelectron microscopic study. We speculate that JAM may be directly phosphorylated by cPKC(s)upon platelet activation and that the phosphorylationmight be involved in platelet activation.

Adenosine Diphosphate↗

PubMed2000

Poor bone quality or hip structure as risk factors affecting survival of total-hip arthroplasty.

BACKGROUND: The principal long-term complication after total hip arthroplasty (THA) has been aseptic fixation failure. Many hip prostheses and operative techniques have been developed to improve outcomes, but few measures have been taken to cope with poor bone quality or hip structure. We assessed risk factors for aseptic fixation failure after THA. METHODS: We assessed, by multivariate analysis, survival of 405 primary Charnley THAs to identify risk factors for aseptic fixation failures. We also investigated risk factors for development of rapid polyethylene wear (penetration depth of the femoral head into the socket polyethylene > or = 2 mm/year) FINDINGS: In the entire series of 405 THAs, with use of radiographic fixation failure or revision for a loose socket as the endpoint, development of rapid polyethylene wear and the preoperative diagnosis of atrophic osteoarthrosis (defined by scarce osteophyte formation) were identified as risk factors for socket loosening (p < or = 0.02). A medullary canal with an unfavourable geometry (a stovepipe canal, Noble's canal-flare index < 3.0) was the only risk factor for femoral fixation failure (p < or = 6.7x10(3)). The only variable related to development of rapid polyethylene wear was the type of steel used in the femoral prosthesis--Ortron 90 prostheses significantly lowered the rate of development of rapid wear from 12.7% to 0.4%. In the 248 THAs in which these femoral prostheses were used, socket survival was affected only by the preoperative diagnosis of atrophic osteoarthrosis (for radiographic fixation failure and revision, p=4.0x10(-5) and p=0.042, respectively). INTERPRETATION: In THA, the critical risk factors are poor bone quality, which manifests as atrophic osteoarthrosis, for socket survival and poor bone structure for femoral-prosthesis survival. To ensure longer durability of THAs, these factors should be assessed further and efforts, especially biological initiatives, should be made to resolve them.

Arthroplasty, Replacement, Hip↗

PubMed2000

Allopurinol induces renal toxicity by impairing pyrimidine metabolism in mice.

We investigated the relationship between the toxic effect of allopurinol and pyrimidine metabolism in mice. Allopurinol-induced increases in plasma transaminase levels in dinitrofluorobenzene (DNFB)-sensitized mice were not affected by uridine. In contrast, plasma creatinine and BUN tended to decrease 18 hr after the last injection of uridine. Both plasma and urinary orotidine (OD) were detected in DNFB-sensitized mice after administration of a single dose of allopurinol. In contrast, TEI-6720, a newly synthesized xanthine oxidase/xanthine dehydrogenase inhibitor, caused neither pyrimidine metabolism abnormality nor renal impairment in DNFB-sensitized mice. Also, normal mice administered high doses of allopurinol showed abnormal pyrimidine metabolism together with renal toxicity which could be ameliorated by uridine, indicating that allopurinol essentially causes pyrimidine metabolism abnormality leading to renal impairment. In DNFB-sensitized mice, allopurinol increased urinary OD excretion to an extent similar to that in normal mice administered the same dose of allopurinol. However, renal impairment by allopurinol was more striking in DNFB-sensitized mice than in normal mice. Histopathological observations showed that allopurinol induced calculus formation in the collecting tubules and papillary duct. Calculus formation was increased by DNFB and decreased by uridine. These observations indicate that the enhancement of the renal toxicity of allopurinol by DNFB-sensitization may be due to some biological interactions between DNFB and allopurinol. In humans, it is possible that there are some biological interactions which serve to enhance the toxicity of allopurinol, resulting in the development of allopurinol hypersensitivity syndrome (AHS). In contrast, TEI-6720, had no effect on pyrimidine metabolism and showed no toxic effect.

Allopurinol↗

PubMed2000

High-throughput pretreatment system in automated urinary sediment analyzer.

BACKGROUND: Urine contains microscopically observable particles that can indicate certain types of disease in the urinary tract system. Determining these various types of sediments by manual operation is a cumbersome and time-consuming task. To eliminate this labor, we developed an automated urinary sediment analyzer with high-throughput pretreatment system. METHODS: The pretreatment system mainly consists of four reaction vessels for dying samples (urine), a sheath flow chamber, and an unique sample carrier mechanism from the reaction vessel to the flow chamber, which enables overlapped processing, and rapid transfer of samples with small dispersion and a short buildup time. RESULTS: The buildup time was experimentally found to be 1.8 s, and the extra-sample volume beside that for measurement was only 4.9 microl (1/20 of the total sample volume). CONCLUSIONS: Short buildup time results in high throughput of 120 samples per hour, and relatively small extra-volume contributes to reduce carryover.

Automation↗

PubMed2000

Overexpression of low density lipoprotein receptor eliminates apolipoprotein B100-containing lipoproteins from circulation and markedly prevents early atherogenesis in apolipoprotein E-deficient mice.

Apolipoprotein E (ApoE) plays a pivotal role in the metabolism of apolipoprotein B (apoB)-containing lipoproteins. The defective apoE gene in humans can cause elevated plasma levels of apoB-containing lipoproteins such as chylomicron remnant and intermediate density lipoprotein (IDL). In this study, we examined whether liver-selective high-level expression of low-density lipoprotein receptor (LDLR) could affect the lipoprotein profile and atherogenesis in apoE-deficient (apoE-/-) mice. ApoE knockout mice expressing LDLR transgene in liver [apoE-/-;Tg(LDLR+/-)] were prepared after mating apoE-/- mice with the human LDLR transgenic mice. The apoE-/-;Tg(LDLR+/-) and littermate apoE-/- mice were fed a normal diet and sacrificed at 18 weeks of age. (1) The plasma levels of cholesterol and triglyceride in apoE-/-;Tg(LDLR+/-) mice were 51 and 33% lower than those of apoE-/- mice, respectively. (2) In the plasma of apoE-/-;Tg(LDLR+/-) mice, the levels of apoB-containing lipoprotein were reduced and apoB100-containg particles were totally eliminated. (3) By histochemical analysis, apoE-/-;Tg(LDLR+/-) mice showed drastic suppression of early atherogenesis; the lesion area of these mice was 1/70 of that in the littermate apoE-/- mice. These results indicate that, even in apoE-defective subjects, induction of hepatic LDLR expression could protect from early atherogenesis.

Animals↗

PubMed2000

Role of ras mutation in the progression of thyroid carcinoma of follicular epithelial origin.

The histological differentiation of thyroid carcinoma is known to correlate with prognosis. Ras oncogene mutations, which have been identified in various human cancers, have been suspected playing an important role in carcinogenesis and tumor progression. The purpose of this study was to clarify the mechanism of thyroid tumor progression, focusing on ras oncogenes. We examined ras mutations using nested polymerase chain reaction (PCR) and direct sequencing methods. The ras oncogene product was also examined immunohistochemically. Our results indicated that the incidence of ras mutations correlated with the histological differentiation of thyroid cancer. Three poorly differentiated carcinomas showed a higher rate of ras mutations than did 17 well-differentiated counterparts. Hot spots were not identified except for a relative accumulation of the N-ras gene at codon 61. There was a correlation between the immunoreactivity of the ras oncogene product and ras mutation, although the immunoreactivity of ras-p21 did not correlate with the histological differentiation. Mutation of the ras gene seemed to be one of the important events in the progression from well-differentiated carcinoma to poorly differentiated thyroid carcinoma.

Adenocarcinoma, Follicular↗

PubMed2000

Expression of heat shock proteins in osteosarcoma and its relationship to prognosis.

The prognosis of osteosarcoma has been improved by chemotherapy. Heat shock proteins (HSPs) assist in folding proteins at posttranslation and degeneration under stress. We investigated the effect of HSPs on survival in osteosarcoma. Conventional osteosarcomas of the extremities from 70 patients aged 30 years or younger were used. Preoperational chemotherapy was performed in all cases. Tissues at surgery and biopsy were immunohistochemically stained with anti-HSP27, HSP47, HSP60, HSP70, HSP90alpha, HSP90beta, and p53 antibodies. We classified the cases in which more than 10% of tumor cells were positive into the overexpressing group. Overall survival was compared between the groups either overexpressing HSPs or not using Wilcoxon's test and Cox's proportional hazard model. The overexpression rate at biopsy was 22% (HSP27), 88% (HSP47), 66% (HSP60), 48% (HSP70(, 47% (HSP90alpha), 31% (HSP90beta), and 17% (p53), respectively. The rate at surgery was 33% (HSP27), 94% (HSP47), 60% (HSP60), 49% (HSP70), 28% (HSP90alpha), 40% (HSP90beta), and 17% (p53), respectively. HSP27 and p53 overexpression at biopsy had a negative prognostic value. HSP27 showed the strongest negative prognostic value in osteosarcoma. It is therefore important to investigate further its function in cellular regulation and drug resistance.

Bone Neoplasms↗

PubMed2000

Phosphodiesterase inhibitors, pentoxifylline and rolipram, increase bone mass mainly by promoting bone formation in normal mice.

The administration of either Pentoxifylline (PTX), a methylxanthine derivative and an inhibitor of cyclic AMP (c-AMP) phosphodiesterases (PDEs), or Rolipram, an inhibitor specific to type-4 PDE (PDE4) in normal mice, significantly increased both cortical and cancellous bone mass. Vertebrae and tibiae from mice treated with PTX or Rolipram were analyzed by means of bone densitometry and histomorphometry. The results revealed that both PTX and Rolipram increased bone mass in normal mice mainly through the acceleration of bone formation. These findings suggest that both PTX and Rolipram can enhance physiological bone formation and thereby increase bone mass in normal mice. The possibility that these agents may be of value for the treatment of osteoporosis is discussed.

Absorptiometry, Photon↗

PubMed2000

Evidence that the Aspergillus nidulans class I and class II chitin synthase genes, chsC and chsA, share critical roles in hyphal wall integrity and conidiophore development.

Although many chitin synthase genes have been identified in a broad range of fungal species, there have been only a few reports about their role in fungal morphogenesis. In most cases, single gene disruption or replacement did not reveal their function, possibly because of functional redundancy among them. We obtained null mutants of Aspergillus nidulans chsA and chsC genes encoding non-essential class II and class I chitin synthases, respectively. The DeltachsA DeltachsC mutant exhibited growth defects on media supplemented with sodium dodecyl sulfate (SDS), high concentration of salts, chitin-binding dyes, or chitin synthase competitive inhibitors, suggesting loss of integrity of hyphal wall. Moreover, remarkable abnormalities of the double mutant were observed microscopically during its asexual development. The conidiophore population was drastically reduced. Interestingly, secondary conidiophores were occasionally produced from vesicles of the primary ones. The morphology of these conidiophores was similar to those of the A. nidulans developmental mutants, medusa (medA), abacus (abaA), and some kinds of bristle (brlA). In situ staining patterns suggested that chsA was mainly expressed in the metulae, phialides, and conidia, whereas chsC was expressed in hyphae as well as conidiophores. These results suggest that ChsA and ChsC share critical functions in hyphal wall integrity and differentiation.

Aspergillus nidulans↗

PubMed2000

Canal enlargement by Er:YAG laser using a cone-shaped irradiation tip.

To solve the problem of mechanical instrumentation, we developed a cone-shaped laser irradiation tip. This tip delivers 80% of the energy of the laser laterally and 20% of the energy forward. The tip was equipped with a water nozzle. The aim of this study was to examine the effect of Er:YAG laser irradiation using this tip on root canal enlargement and debridement. As a root canal model, a hole (0.5 x 5 mm) was drilled into a bovine dentin block. The tip was inserted into this hole and moved from the apex to the orifice (45 mm/min). The power of 10 mJ x 10 pps caused enlargement of the canal dimension by 106.5%. 20 and 40 mJ x 10 pps caused enlargements of 116.3 and 118.6%. 30 mJ x 10 pps caused the biggest change (129.8%). Scanning electron microscopic observations indicated that the dentin surface after laser preparation appeared cleaner than that obtained after preparation by drilling. This technique may have the advantage of decreasing the preparation time.

Animals↗

PubMed2000

Oxidized-LDL and atherosclerosis. Role of LOX-1.

The accumulation of substantial numbers of monocyte/macrophages and activated T lymphocytes in focal areas of the arterial intima appears to be a hallmark of atherosclerosis. Our report demonstrated that lysophosphatidylcholine (lyso-PC), a polar phospholipid component that is increased in atherosclerotic lipoproteins, such as oxidized LDL and remnant lipoproteins in diabetic and Type 3 hyperlipidemia, can upregulate adhesion molecules for monocytes and T lymphocytes, and growth factors, such as heparin-binding epidermal growth factor-like growth factor and PDGF A and B chains. Recently, we identified the novel receptor for oxidized LDL, named LOX-1. We summarize the importance of the interaction between oxidized LDL and its receptor, LOX-1, in terms of early stage atherogenesis.

Animals↗

PubMed2000

Cloning and characterization of EPD2, a gene required for efficient pseudohyphal formation of a dimorphic yeast, Candida maltosa.

Candida maltosa is a dimorphic fungus and its pseudohyphal growth is partly induced by additional copies of the centromeric DNA (CEN) region1) or n-alkane. In the course of analyzing the induction mechanism of pseudohyphal growth, we isolated EPD1, which is similar in sequence to PHR1 and PHR2 of Candida albicans. Epdlp could be involved in cell wall maintenance and is essential for pseudohyphal growth induced by CEN and n-hexadecane at pH 4 and by n-hexadecane at pH 7.) In this paper, we cloned EPD2 of C. maltosa, which is highly similar to EPD1, PHR1, and PHR2. The transcription of EPD2 is induced strongly when cells are grown in SD medium of higher pH (pH 7), but not in SD medium of lower pH (pH 4). This pattern of expression was an inverse of that of EPD1. This alternate expression is similar to that between PHR1 and PHR2. The expression of EPD2 was much higher when C. maltosa was grown on the n-hexadecane solid medium than grown in the n-hexadecane liquid medium. The efficiency of pseudohyphal formation of an epd2 null mutant on n-hexadecane medium at pH 7 or 7.5 was lower than that of the wild-type strain. These results suggest that Epd2p is required for efficient pseudohyphal formation induced by n-hexadecane in the medium at pH 7.

Amino Acid Sequence↗

PubMed2000

Bone mineral density of alveolar bone in rats during pregnancy and lactation.

BACKGROUND: The present study was designed to investigate the effect of pregnancy and lactation on the bone mineral density (BMD) of alveolar bone in rats fed diets containing different amounts of calcium (Ca). The effects of different levels of Ca intake by the mothers on the BMD of alveolar bone in their pups were also examined. METHODS: Ten-week-old female Wistar rats were housed with male rats for breeding and were divided into 3 groups fed diets containing 0.9, 0.3, and 0.02% Ca, respectively. They were further divided into 2 groups according to pregnancy or non-pregnancy. Animals in the pregnant group were raised with their own pups for lactation. After the experiment, all animals were sacrificed, their mandibles were dissected, and soft x-ray microradiographs were taken to determine BMD in the interdental area with an image analyzer. RESULTS: In both groups, BMD in alveolar bone decreased based on the amount of Ca in the diet, but the magnitude of this decrease was much greater in the pregnant group than in the non-pregnant group. There was no significant difference in BMD between 2 groups fed a 0.9% Ca diet. The BMD of alveolar bone in the pups also decreased depending on the amount of Ca in the diet. CONCLUSIONS: The above results suggest that pregnancy and subsequent lactation could be risk factors for alveolar bone loss, especially under conditions of Ca insufficiency or deficiency and that Ca insufficiency or deficiency in the mother caused decreases in the BMD of alveolar bone in the pups.

Alveolar Bone Loss↗

PubMed2000

Schwannoma in the sigmoid colon: report of a case.

Schwann cell tumor occurring in the intestines is rare. A 68-year-old female came to our hospital because of hematemesis. Barium enema and colonoscopic examination revealed submucosal tumor in the sigmoid colon. Laboratory data showed mild anemia. No other abnormal finding was found in the blood chemistry. Tumor marker levels of carcinoembryonic antigen (CEA), CA19-9, alpha feto protein (AFP) and neuron specific enolase (NSE) were within normal limits. The exploratory laparotomy confirmed a large sigmoid colon tumor. She received sigmoid colectomy. The resected specimen was a submucosal tumor with central depression, measuring 4.7 x 3.5 x 3.0 cm in size. The cut surface of the tumor was yellowish hue with necrosis. Histological examination showed spindle-shaped tumor cells with palisading comma-shaped nuclei and the nuclear pleomorphism. Immunohistochemical examination revealed that the tumor was positive for S-100 protein staining, and negative for Actin and for H.H.F. staining. These findings showed that this tumor was of Schwann cell origin. We report here the case in detail of a schwannoma in the sigmoid colon.

Aged↗

PubMed1999

Cutting edge: combined treatment of TNF-alpha and IFN-gamma causes redistribution of junctional adhesion molecule in human endothelial cells.

Proinflammatory cytokines such as TNF-alpha and IFN-gamma induce cell adhesion molecules in endothelial cells and promote transmigration of leukocytes across endothelial cells. However, when those two were administered together, leukocyte transmigration paradoxically decreased. We cloned a human and bovine homologue of the junctional adhesion molecule (JAM), a novel molecule at the tight junction, and examined the effects of proinflammatory cytokines on JAM in HUVECs. The combined treatment of TNF-alpha plus IFN-gamma caused a disappearance of JAM from intercellular junctions. However, flow cytometry, cell ELISA, and subcellular fractionation analysis demonstrated that the amount of JAM was not reduced. This suggested that JAM changed its distribution in response to proinflammatory cytokines. This redistribution of JAM might be involved in a decrease in transendothelial migration of leukocytes at inflammatory sites.

Amino Acid Sequence↗

PubMed1999

Molecular characterization of two endogenous double-stranded RNAs in rice and their inheritance by interspecific hybrids.

We completely sequenced 13,936 nucleotides (nt) of a double-stranded RNA (dsRNA) of wild rice (W-dsRNA). A single long open reading frame (13,719 nt) containing the conserved motifs of RNA-dependent RNA polymerase and RNA helicase was located in the coding strand. The identity between entire nucleotide sequence of W-dsRNA and that of the dsRNA of temperate japonica rice (J-dsRNA, 13,952 nt) was 75.5%. A site-specific discontinuity (nick) was identified at nt 1,197 from the 5' end of the coding strand of W-dsRNA. This nick is also located at nt 1,211 from the 5' end in the coding strand of J-dsRNA. The dsRNA copy number was increased more than 10-fold in pollen grains of both rice plants. This remarkable increase may be responsible for the highly efficient transmission of J-dsRNA via pollen that we already reported. J-dsRNA and W-dsRNA were also efficiently transmitted to interspecific F1 hybrids. Seed-mediated dsRNA transmission to F2 plants was also highly efficient when the maternal parent was wild rice. The efficiency of dsRNA transmission to F2 plants was reduced when the maternal parent was temperate japonica rice; however, the reduced rates in F2 plants were returned to high levels in F3 plants.

Base Sequence↗