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Biomedical subjects

K Komiyama

Publications and source records attributed to K Komiyama.

At least 19 recordsLinked to original sources

Co-production of vascular endothelial cadherin and inducible nitric oxide synthase by endothelial cells in periapical granuloma.

AIM: To clarify the mechanisms of inflammatory cell migration in human periapical granulomas by examining vascular endothelial (VE) cadherin and inducible nitric oxide synthase (iNOS)-producing cells. METHODOLOGY: Periapical tissues were obtained from patients during endodontic surgery and were divided into two portions. After fixing the tissues with acetone or 4% paraformaldehyde in phosphate-buffered saline, 5-microm-thick paraffin or cryostat sections were prepared, respectively. The paraffin sections of the inflamed tissues were evaluated histologically with haematoxylin-eosin stains. Cryostat sections of the tissue, diagnosed as periapical granulomas, were then examined by either immunohistochemistry using anti-human VE-cadherin or iNOS antibodies (Abs) for the characterization of infiltrating cells. In addition, co-localization of VE-cadherin and iNOS production was also analysed by two-colour immunofluorescence image analysis. RESULTS: Endothelial cells were strongly stained with iNOS Abs. Macrophages, lymphocytes, polymorphonuclear leucocytes and fibroblasts also exhibited iNOS production. These iNOS-positive cells accumulated around the blood vessels. On the other hand, VE-cadherin production was exhibited in only endothelial cells. Two-colour immunofluorescence image analysis using VE-cadherin and iNOS Abs demonstrated that iNOS-producing endothelial cells also showed VE-cadherin production. CONCLUSIONS: Vascular endothelial-cadherin produced by endothelial cells could be regulated by iNOS-producing cells in periapical granulomas and might play a pivotal role in vascular permeability.

Adult↗

Expression of type V secretory phospholipase A in myocardial remodelling after infarction.

AIMS: Secretory phospholipase A2 is associated with ischaemic injury in the human heart, but the distribution of type V secretory phospholipase A2 (sPLA2-V) remains unknown. The significance of sPLA2-V in myocardial infarction was investigated histopathologically. METHODS: Sequential changes in the localization of sPLA2-V and its mRNA in myocardial tissues obtained from 30 autopsied hearts were examined by immunohistochemistry and in situ hybridization and compared with those of fibronectin, vascular endothelial growth factor (VEGF), interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha, and cyclooxygenase-2 (COX-2). RESULTS: No expression of sPLA2-V was observed in normal heart, but it was promptly expressed in wavy myofibres positive for fibronectin just after the onset of infarction. sPLA2-V was subsequently expressed in ischaemic cardiomyocytes around the lesion. The expression decreased at the granulation tissue and disappeared at the chronic stage with scar formation. The distribution of the signal for sPLA2-V mRNA paralleled that of the protein. Ischaemic myocytes around the lesion expressed VEGF, IL-1beta, TNF-alpha and COX-2 at all stages. CONCLUSIONS: sPLA2-V production in myocardium is limited to the acute phase of infarction. sPLA2-V may play a dual role, acting both to remove degraded cell-membrane through cooperative activity with COX-2 in necrotic areas and to attack ischaemic myocytes around the lesion via degradation of membrane phospholipids.

Adult↗

Dendocarbins A--N, new drimane sesquiterpenes from the nudibranch Dendrodoris carbunculosa.

Fourteen new sesquiterpenes of the drimane series dendocarbins A--N (1--14) were obtained from ethanol extracts of the Japanese nudibranch Dendrodoris carbunculosa, together with two known compounds, isodrimeninol (15) and 11-epivaldiviolide (16). All structures were elucidated mainly from spectral data, and most of these sesquiterpenes were found to exhibit cytotoxicity. In addition, isodrimeninol (15), the major sesquiterpene of this animal, was found to have a sharp peppery taste.

Animals↗

New multiflorane-type triterpenoid acids from Sandoricum indicum.

Three new 12beta-hydroxymultiflorane triterpenoid acids, sandorinic acids A-C (1-3), were isolated from the stem bark of Sandoricum indicum together with five known triterpenes (4-8). The structures of 1-3 were elucidated by spectral data interpretation. Compounds 1-8 were evaluated for their inhibiting activity against several tumor cell lines and for their effects on lymphocyte proliferation.

Animals↗

Evidence for perforin-like activity associated with earthworm leukocytes.

Earthworm (Eisenia fetida) coelomic fluid contains several leukocytes (coelomocytes): basophils, acidophils and neutrophils as well as chloragocytes. Small coelomocytes and coelomocyte lysate are cytotoxic for the tumor cell target K562. The expression of a lytic factor was investigated by immunocytochemistry using light and transmission electron microscopy. A rat-anti-mouse-perforin-mAb labeled mainly small coelomocytes (nearly 20%) as visualized by light microscopy. TEM analysis using immunogold showed a homogenous labeling in the cytoplasm of small coelomocytes. The highest number of immunogold particles was estimated in coelomocytes with many small cytoplasmic granules. Coelomocytes with large lysosomal granules were also labeled but less intensely. No antibody binding was observed for chloragocytes either in light or electron microscopy. This suggests that the perforin-like activity is associated with only one cell type and that chloragocytes are responsible for other lytic activities. MALDI-MS revealed calreticulin usually associated with perforin in mammalian cells that mediate lysis (e.g. NK, CTL). Together, results strongly suggest the presence of putative perforin in earthworms. This in turn supports the hypothesis that perforin is a conserved component important in immune defense during evolution.

Journal Article↗

Histopathological and immunohistological analyses in IgA deficient lymphoplasia (aly/aly) mouse.

The aly/aly mouse has a severe immunodeficiency, because it lacks peripheral lymph nodes as well as IgA and IgG immunoglobulin synthesis. In the present study, we performed histopathological and immunohistological examinations to clarify histological disorders of various immune organs in these mice. Carbon CH40 injections into the apex of the tongue confirmed the absence of submandibular lymph nodes in aly/aly mice. The thymus had a poorly constructed cortex and medulla, and the number of lymphoid follicles was clearly decreased in the spleen. No IgG- or IgA- producing cells were found in any immune organs, including the mucosal immune sites, though several IgM -producing cells were identified. Other characteristic findings included perivascular lymphocytes accumulation in the salivary glands, lungs, liver and pancreas, which caused tissues damage. These results demonstrated that the various lymphoid tissues disorders and organ-specific lymphocyte infiltration cause immuno-deficiency in the aly/aly mouse.

Animals↗

Cloning and expression of the chicken immunoglobulin joining (J)-chain cDNA.

The J chain is a component of polymeric immunoglobulin (Ig) molecules and may play an important role in their polymerization and the transport of polymeric Ig across epithelial cells. In this study, the primary structure of the chicken J chain was determined by sequencing cDNA clones. The cDNA had an open reading frame of 476 nucleotides encoding a putative protein of 158 amino acid residues including the signal sequence. The 3' untranslated region consisted of 1216 nucleotides and a poly(A) tail. The deduced amino acid sequence of the chicken J chain had a high degree of homology to that of human, cow, rabbit, mouse, frog, and earthworm, with eight conserved Cys residues identical to the mammalian J chains. Northern blot hybridization performed with total RNA from various chicken tissues revealed high levels of J-chain mRNA expression in spleen, intestine, Harderian gland, and bursa of Fabricius, and low levels in the thymus. The J chain was expressed in the bursa as early as day 15 of embryogenesis. These data indicated that the chicken J-chain gene displays a high degree of homology with that of other species, and is expressed at an early stage of development of the chicken immune system.

Amino Acid Sequence↗

Immunoexpression of transforming growth factor beta in desmoplastic ameloblastoma.

Desmoplastic ameloblastoma (DA) is an unusual subtype of ameloblastoma histologically characterized by the pronounced collagenized stroma. In the present study, the immunolocalization of transforming growth factor beta (TGF-beta), one of the most potent local factors for modulating extracellular matrix formation, was observed in DA in order to study its participation in the stromal desmoplasia. Seven cases of DA, including a "hybrid" lesion, were studied together with ten cases of ordinary follicular and plexiform ameloblastomas as the control. In contrast to ordinary ameloblastomas, marked immunoexpression was observed in all DAs but one. In the "hybrid" lesion, TGF-beta was not expressed in the area of follicular ameloblastoma but in that of DA. These results show that TGF-beta produced by tumor cells of DA plays a part in the desmoplastic matrix formation.

Adolescent↗

Hepatitis G virus coinfection influences the liver histology of patients with chronic hepatitis C.

AIMS/BACKGROUND: The present investigation compared the histological features of the liver of chronic hepatitis C patients who are or are not coinfected with hepatitis G virus (HGV) to determine the histological and clinical characteristics of HGV infection. SUBJECTS AND METHODS: This study included 194 patients with chronic hepatitis C who visited our institution between 1993 and 1995. Detection of serum HGV RNA was performed by nested reverse transcription-polymerase chain reaction. Scores were assigned to indicate the severity of each of the following features on the liver biopsy of a patient: inflammatory cell infiltration in the periportal, parenchymal, and portal area; fibrous stage; lymphoid aggregates in the portal area; portal sclerotic change; perivenular fibrosis; pericellular fibrosis; bile duct damage; bridging necrosis; and irregular regeneration of hepatocytes (IR). RESULTS: HGV RNA was detected in the sera of 18 (9.3%) of the 194 patients. The histological features of the HGV RNA-positive patients show significantly more severe bile duct damage, perivenular fibrosis, pericellular fibrosis and IR than the liver of the HGV RNA-negative patients. The biochemical results in the two groups did not significantly differ. CONCLUSION: Our data suggest that chronic HGV coinfection worsens the histological features of liver disease.

Bile Ducts↗

Cytotoxic alkaloids of Pachysandra terminalis.

Four steroidal alkaloids, epipachysamines B (1) and E (2), pachystermine A (3) and pachysamine E (4), were isolated as cytotoxic principles from the MeOH extract of the stems of Pachysandra terminalis SIEB. et ZUCC. (Buxaceae). These alkaloids showed cytotoxic activity against P388 and P388/ADR leukemia cells in vitro. Three of the alkaloids (1-3) were previously isolated from this plant material, and this is the first report of their cytotoxic activity. Pachysamine E (4) is a new alkaloid.

Alkaloids↗

Helicobacter pylori in oral ulcerations.

Helicobacter pylori is an important pathogen involved in the development of gastrointestinal ulcers, but its involvement in oral ulcerous lesions is unclear. As culture is generally recognized as the gold standard for diagnosis of H. pylori infection, we employed this approach to assess the association of H. pylori with oral mucosal ulcerations. Samples were collected from patients with oral mucosal ulcerative disorders: 12 cases of recurrent aphthous stomatitis (RAS), 7 cases of herpes simplex virus (HSV) stomatitis, and 3 cases of erosive lichen planus (LP). Serum IgG antibodies against H. pylori were examined in all cases. All of the RAS and erosive LP cases were culture-negative for H. pylori, while two cases of HSV stomatitis were positive. The two culture-positive cases were also seropositve for the H. pylori antigen. It is suggested that H. pylori might not have a direct association with oral ulcerations. However, H. pylori in the oral cavity might exist in a non-culturable coccoid state without productive infection, and might form colonies only under special conditions such as HSV infection.

Adult↗

A histochemical study of inflammatory lesions of the maxillary sinus mucosa using biotinylated lectins.

The distribution of glycoconjugates in normal maxillary sinus tissues, in cases of maxillary sinusitis and in postoperative maxillary cysts (POMC), was examined using seven different lectins as probes. The results showed that wheatgerm agglutinin (WGA), peanut agglutinin (PNA), Ulex europaeus agglutinin-1 (UEA-1), Ricinus communis agglutinin-1 (RCA-1), and concanavalin A (ConA) strongly react with the cilia and goblet cells. The binding of WGA, PNA, UEA-1, and RCA-1 was increased in maxillary sinusitis and POMC compared with normal maxillary sinus epithelium, whereas that of ConA was decreased. The decreased binding of ConA suggested that there were fewer mannoside residues in the maxillary sinus epithelium in the inflammatory lesion. The PNA bound to the cilia, goblet cells and mucous glandular cells in maxillary sinusitis and POMC, but not in normal, uninflamed cells, indicating that D-galactose was produced by the inflammatory condition. Similar binding patterns of PNA and RCA-1 were found in the cilia and on the surface of the epithelium and in the goblet cells. It is assumed that the carbohydrate moiety in the sinus mucosa is altered in inflammatory conditions.

Acetylgalactosamine↗

Reversal of multidrug resistance by 7-O-benzoylpyripyropene A in multidrug-resistant tumor cells.

7-O-Benzoylpyripyropene A (7-O-BzP), a semi-synthetic analog of pyripyropene, was investigated for its reversing effect on multidrug-resistant (MDR) tumor cells. 7-O-BzP (6.25 microg/ml) completely reversed resistance against vincristine and adriamycin in vincristine-resistant KB cells (VJ-300) and adriamycin-resistant P388 cells (P388/ADR), respectively. 7-O-BzP alone had no effect on the growth of drug sensitive and drug-resistant cells. 7-O-BzP (6.25 microg/ml) significantly enhanced accumulation of [3H]vincristine in VJ-300 cells and completely inhibited the binding of [3H]azidopine to the P-glycoprotein in VJ-300 cells and P388/ADR cells. The result suggests that 7-O-BzP effectively reverses P-glycoprotein-related MDR by interacting directly with P-glycoprotein in drug resistant VJ-300 and P388/ADR cells.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Casein kinase II inhibitors isolated from two Brazilian plants Hymenaea parvifolia and Wulffia baccata.

Two dihydroflavonol rhamnosides (1 and 2) isolated from the bark of Hymenaea parvifolia and two pentacyclic triterpenoids (3 and 6) obtained from the leaves of Wulffia baccata have been found to exhibit inhibitory effects of casein kinase II (CK-II) dose-dependently, suggesting that at higher doses more than 10 microM, these four compounds may act as potent CK-II suppressors of the CK-II-mediated activation of 60S acidic ribosomal P proteins in vitro.

Brazil↗

Reversal of multidrug resistance by kopsiflorine isolated from Kopsia dasyrachis.

Kopsiflorine, an indole alkaloid of the aspidofractinine-type isolated from Kopsia dasyrachis, was examined for its effect in enhancing drug cytotoxicity in multidrug-resistant tumor cells. The cytotoxicity of vincristine was enhanced in a concentration-dependent manner by kopsiflorine in drug-resistant KB cells (VJ-300). Kopsiflorine alone had no effect on the growth of drug sensitive or resistant cells, but the intracellular accumulation of vincristine was enhanced by kopsiflorine in VJ-300 cells. Kopsiflorine (10 micrograms/ml) significantly inhibited the binding of [3H]azidopine to P-glycoprotein in VJ-300 cells. The results suggest that kopsiflorine interacts directly with P-glycoprotein and inhibits the efflux of antitumor agents in drug-resistant cells.

Drug Resistance, Multiple↗

Biochemical characterization of 60S acidic ribosomal P proteins associated with CK-II from bamboo shoots and potent inhibitors of their phosphorylation in vitro.

Three effective phosphate acceptors (35, 15 and 13 kDa polypeptides) for casein kinase II (CK-II) in the Superdex CK-II fraction prepared from a 0.5 M NaCl extract of bamboo shoots were selectively purified by glycyrrhizin (GL)-affinity column chromatography (HPLC). These three proteins (p35, p15 and p13) were identified as 60S acidic ribosomal P proteins by determination of their partial N-terminal sequences. CK-II was associated with p35 since the GL-affinity fraction was coprecipitated with an anti-serum against Drosophila CK-IIbeta. Moreover, a derivative (oGA) of glycyrrhetinic acid (GA) and several polyphenol-containing anti-oxidative compounds [quercetin, epigallocatechin gallate (EGCG) and two isoflavones, i.e., 3',4',7-trihydroxyisoflavone (3',4',7-THI) and 8-chloro-3',4',5,7-tetrahydroxyisoflavone (8C-3',4',5,7-THI)] inhibited the CK-II-mediated phosphorylation of 60S acidic ribosomal P proteins in vitro. Quercetin was found to be the most effective compound on CK-II activity since its ID50 was approx. 50 nM. These results suggest that (i) GL-affinity column chromatography is useful for the selective purification of 60S acidic ribosomal P proteins as a heterocomplex associated with CK-II from various cell sources; (ii) natural anti-oxidative compounds with polyphenols, but not GL and GA, act as potent CK-II suppressors; and (iii) CK-II mediates the regulation of the physiological functions of 60S acidic ribosomal P proteins in growing plant cells.

Casein Kinase II↗