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Biomedical subjects

M Umeda

Publications and source records attributed to M Umeda.

At least 37 records · Page 2Linked to original sources
PubMed2001

Downregulation of lipopolysaccharide-induced intercellular adhesion molecule-1 expression via EP2/EP4 receptors by prostaglandin E2 in human fibroblasts.

In the present study, the effect of prostaglandin E2 (PGE2) on intercellular adhesion molecule-1 (ICAM-1) expression in human gingival fibroblasts (HGF) stimulated with lipopolysaccharides (LPS) was investigated. LPS were isolated from periodontopathic bacteria, Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) and Porphyromonas gingivalis (P. gingivalis), by the phenol-water method and Escherichia coli (E. coli) LPS was used as a control. PGE2 significantly inhibited A. actinomycetemcomitans-, P. gingivalis- and E. coli-LPS-induced ICAM-1 expression. Next, of four PGE2 receptor subtypes (EP1, EP2, EP3 and EP4), we examined which subtype(s) was involved in inhibition of LPS-elicited ICAM-1 expression by PGE2. Eleven-deoxy-PGE1, a selective EP2/EP4 agonist, and butaprost, a selective EP2 agonist, attenuated A. actinomycetemcomitans-, P. gingivalis- and E. coli-LPS-elicited ICAM-1 expression, although butaprost was less potent than PGE2 and 11-deoxy-PGE1. Sulprostone, an EP1/EP3 agonist, and ONO-AP-324, an EP3 agonist, was inert to the LPS-elicited ICAM-1 expression. Furthermore, dibutyryl cAMP, a cAMP analogue, and forskolin, an adenylate cyclase activator, downregulated A. actinomycetemcomitans-, P. gingivalis- and E. coli-LPS-elicited ICAM-1 expression in HGF. Our data suggest that PGE2 downregulates A. actinomycetemcomitans- and P. gingivalis-LPS-induced ICAM-1 expression in HGF, via EP2/EP4 receptors by cAMP-dependent signaling pathways. The cAMP-elevating agents such as EP2/EP4 receptor activators may serve to control inflammatory and immune responses in periodontal disease.

Aggregatibacter actinomycetemcomitans↗
PubMed2001

Localization of sphingomyelin during the development of dorsal and tail epidermis of mice.

BACKGROUND: The water permeability barrier of the stratum corneum seems to be regulated primarily by lamellar bodies situated between the corneocytes; the lamellar bodies originate largely from polar lipid precursors, mainly sphingomyelin (SM), provided by the cells of the stratum granulosum via exocytosis of their lamellar body content. OBJECTIVES: The aim of our study was to evaluate the cellular distribution of SM during development of the epidermis. Methods In this study, we investigated the expression and localization of SM in both adult and fetal mouse skin by a cytochemical detection method, immunofluorescence microscopy and immunoelectron microscopy, using anti-SM antibody, a specific binding protein to SM (lysenin), and Nile red stain. In addition, we measured transepidermal water loss to estimate the barrier function of the fetal skin. RESULTS: We observed that SM was widely distributed from the basal layer to the granular layer in the adult mouse epidermis. An intense cytochemical reaction for SM was observed on embryonic day E14.5 of gestation just before the differentiation of the granular and squamous cells from the intermediate cells. The immunofluorescence indicating SM was detected in two regions, i.e. the most superficial zone of the granular layer and the upper spinous layer after the cell differentiation at the late gestational age. This distribution was not detected by conventional lipid staining, such as with Nile red stain. Immunoelectron microscopy revealed that SM was mainly localized in the intercellular spaces of the adult mouse epidermis and in the intracellular vesicles without a complete lamellar structure in the cytoplasm of epidermal cells of E14.5 fetuses. It is well known that the formation of the structurally mature cornified cell envelope occurs at E15.5 of development. The skin of fetuses at E16.5 showed a definite barrier function. CONCLUSIONS: These findings suggest that SM dynamics is related to the formation of the lipid envelope, cell differentiation, and epidermal barrier function during development.

Animals↗
PubMed2001

Experimental model of invasion and metastasis by orthotopic transplantation of oral squamous and adenoid cystic carcinomas into the tongue of nude mice.

Three squamous cell carcinoma (SCC) cell lines established from oral cancer, seven specimens of SCC and three of adenoid cystic carcinomas taken from the oral cavity during operations were transplanted into the tongues of nude mice. Metastases to the regional lymph nodes and the lungs were examined histologically. We were able to transplant every cell line or specimen of tissue into the tongue of nude mice, and found that cancer transplanted in the tongue invaded diffusely to the surrounding tissues without forming a capsule, and that the mode of invasion of the transplanted SCC was similar to that of the biopsy specimen of the patient from whom the material had been obtained. We also found that all three of the SCC cell lines, 3 of the 7 SCC specimens and 2 of the 3 adenoid cystic carcinoma tissues metastasized to the regional lymph node. SCC did not metastasize to the lung, but in two of the three adenoid cystic carcinomas we did see micrometastases to the lung. The study indicates that this method can be used as a model of metastasis in oral squamous cell carcinoma and adenoid cystic carcinoma to show the stages of metastasis in cancer.

Animals↗
PubMed2001

Functional reconstruction of mobile tongue and suprahyoid muscles after resection of cancer of the tongue.

The mobile tongue and the suprahyoid muscles were reconstructed in a man who had had a total resection of the mobile tongue with the oral floor and bilateral radical neck dissection (extensive resection of the bilateral suprahyoid muscles) for cancer of the tongue. A rectus abdominis musculocutaneous flap with the tenth intercostal nerve anastomosed to the remaining hypoglossal nerve was used for the reconstruction. The rectus abdominis muscle was inserted between the mandible and the hyoid bone, taking account of the anatomical positions of the tongue and suprahyoid muscles; the rectus sheath was then firmly sutured to the mandible and hyoid bone. This procedure created firm tendinous insertions between the mandible and hyoid bone. After the operation, the reconstructed tongue showed no sign of atrophy, and the reconstructed supra-hyoid muscles (rectus abdominis muscle) could be moved, and the hyoid bone could be raised.

Carcinoma, Squamous Cell↗
PubMed2001

Functional and aesthetic reconstruction of full-thickness cheek, oral commissure and vermilion.

PURPOSE: This paper presents surgical techniques for reconstruction of the cheek, oral commissure and vermillion in the repair of full-thickness cheek defects after resection of buccal-mucosal squamous cell carcinoma. PATIENTS AND METHODS: Four reconstructions in one-stage surgery with either a free radial forearm flap or a rectus abdominis musculocutaneous flap for cheek and oral commissure were carried out. There were combined with a new approach for vermillion advancement flaps. Most challenging was the need not only for morphological reconstruction of the orifice, but also for physiological reestablishment of sphincteric and sensory functions in the vermillion. RESULTS: Morphological and physiological reconstruction of the lip with sphincteric and sensory functions was attained. CONCLUSION: This valuable reconstruction technique was demonstrated in large, full thickness defects involving the cheek, oral commissure and vermilion.

Aged↗
PubMed2001

Rare variant of the intrasoleus musculocutaneous perforator: clinical considerations in raising a free peroneal osteocutaneous flap.

To improve the stability of the cutaneous portion of the vascularized free peroneal osteocutaneous flap (the so-called peroneal flap), various anatomic studies have been conducted, and a reliable notion of the course of cutaneous perforators has been provided; however, anatomic anomalies have occasionally been observed. The authors encountered a rare type of cutaneous perforator in the distal third of the lower leg. It was located under the inferior surface of the flexor hallucis longus muscle, after running in the soleus muscle parallel to the fibula, and then joined the posterior tibial artery. Since absolutely definite diagnostic methods have not been established for the course of the cutaneous perforator, surgery with the possibility of secondary anastomosis, always with a separate peroneal flap, should be a consideration.

Arteries↗
PubMed2001

Failure of the expression of phospholipid hydroperoxide glutathione peroxidase in the spermatozoa of human infertile males.

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) was intensely expressed in mitochondria in the midpiece of human spermatozoa by immunostaining with anti-PHGPx monoclonal antibodies. The PHGPx not only reduced phospholipid hydroperoxide but also scavenged hydrogen peroxide in human spermatozoa. We found a dramatic decrease in the level of expression of PHGPx in the spermatozoa of some infertile males by immunoblotting with anti-PHGPx monoclonal antibodies. These individuals accounted for about 10% of the group of 73 infertile males that we examined. All seven patients with PHGPx-defective spermatozoa were classified as suffering from oligoasthenozoospermia, a defect in which both the number and the motility of spermatozoa are significantly below normal. Males with PHGPx-defective spermatozoa accounted for 26% of the 27 infertile males with oligoasthenozoospermia. No defects in expression of PHGPx in spermatozoa were observed in 31 fertile volunteers. After a 3-h incubation, the relative number of motile spermatozoa with low-level expression of PHGPx was significantly lower than that of spermatozoa with normal expression of PHGPx. The PHGPx-defective spermatozoa failed to incorporate rhodamine 123, revealing a loss of mitochondrial membrane potential. Ultrastructual analysis of mitochondria by electron microscopy demonstrated that the morphology of mitochondria in PHGPx-defective spermatozoa was abnormal. The results suggest that failure of the expression of mitochondrial PHGPx in spermatozoa might be one of the causes of oligoasthenozoospermia in infertile men.

Antibodies, Monoclonal↗
PubMed2001

Rapid detection and quantification of five periodontopathic bacteria by real-time PCR.

The quantity of periodontopathic bacteria in plaque samples is an important determinant for understanding the etiologic role of bacteria. The real-time PCR method was used to detect and quantify periodontopathic bacteria, such as Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis, Treponema denticola, and Treponema socranskii, in saliva and subgingival plaque samples. There was good agreement between the results of conventional PCR and real-time PCR methods. Using the LightCycler system we were able to determine the amount of periodontopathic bacteria within an hour. The real-time PCR method was linear for samples containing from 10(3) to more than 10(8) cells (r2 = 0.999). The application of the real-time PCR method should be useful in the rapid detection and quantification of periodontopathic bacteria in clinical samples.

Actinobacillus↗
PubMed2001

Mutations in the novel membrane protein spinster interfere with programmed cell death and cause neural degeneration in Drosophila melanogaster.

Mutations in the spin gene are characterized by an extraordinarily strong rejection behavior of female flies in response to male courtship. They are also accompanied by decreases in the viability, adult life span, and oviposition rate of the flies. In spin mutants, some oocytes and adult neural cells undergo degeneration, which is preceded by reductions in programmed cell death of nurse cells in ovaries and of neurons in the pupal nervous system, respectively. The central nervous system (CNS) of spin mutant flies accumulates autofluorescent lipopigments with characteristics similar to those of lipofuscin. The spin locus generates at least five different transcripts, with only two of these being able to rescue the spin behavioral phenotype; each encodes a protein with multiple membrane-spanning domains that are expressed in both the surface glial cells in the CNS and the follicle cells in the ovaries. Orthologs of the spin gene have also been identified in a number of species from nematodes to humans. Analysis of the spin mutant will give us new insights into neurodegenerative diseases and aging.

Amino Acid Sequence↗
PubMed2001

Membrane lipid control of cytokinesis.

In the final stage of cell division, cytokinesis constricts and then seals the plasma membrane between the two daughter cells. The constriction is powered by a contractile ring of actin filaments, and scission involves rearrangement of the lipid bilayer of the cell membrane. We have shown that the lipid phosphatidylethanolamine (PE), which normally resides in the internal leaflet of the bilayer, is exposed on the external leaflet of the cleavage furrow as a result of enhanced transbilayer movement of the phospholipids during cytokinesis. To investigate the role of PE in cytokinesis, we employed two different approaches: manipulation of cell surface PE by a PE-binding peptide and establishment of a mutant cell line specifically defective in PE biosynthesis. Both approaches provide evidence that surface exposure of PE is essential for disassembly of the contractile ring at the final stage of cytokinesis. Based on these findings, we proposed that the transbilayer redistribution of PE plays a critical role in mediating coordinated movements between the contractile ring and the plasma membrane that are required for the proper progression of cytokinesis.

Actins↗
PubMed2001

Determination of D-sorbitol in human erythrocytes by an enzymatic fluorometric method with an improved deproteinization procedure.

We developed an improved enzymatic assay of D-sorbitol in human erythrocytes by employing highly specific D-sorbitol dehydrogenase from Pseudomonas sp. (EC 1.1.1.14) and replacing perchloric acid (HClO4) and potassium carbonate (K2CO3), generally used for deproteinization, with sodium hydroxide (NaOH) and zinc sulphate (ZnSO4). In this assay, erythrocytes were separated from plasma by centrifugation and washed once with physiological saline. Subsequently, the erythrocytes were lysed with distilled water and proteins precipitated with NaOH and ZnSO4. After centrifugation, the resulting colourless supernatant was mixed with a glycine buffer (pH 9.0) containing NAD+ and D-sorbitol dehydrogenase. After incubation for 30 min at 37 degrees C, the NADH produced was measured fluorimetrically. The fluorescence intensities were corrected for sample blanks, and the values of D-sorbitol were normalized for haemoglobin content. The method had an analytical range of 1-180 micromol/L. The intra- and inter-assay precisions were < 3.3% and < 5.8%, respectively. The detection limit was 0.65 micromol/L. In terms of the linearity, precision and sensitivity, the improved method using NaOH and ZnSO4 was superior to the conventional method using HClO4 and K2CO3.

Blood Chemical Analysis↗
PubMed2001

Treponema socranskii, Treponema denticola, and Porphyromonas gingivalis are associated with severity of periodontal tissue destruction.

BACKGROUND: The aim of the present study was to identify Treponema socranskii in addition to Treponema denticola and Porphyromonas gingivalis by polymerase chain reaction (PCR), and to clarify the relationship between the presence of these microorganisms and the severity of clinical periodontal parameters. METHODS: Saliva and subgingival plaque collected from 123 subjects (38 aggressive periodontitis patients, 65 chronic periodontitis patients, 20 healthy patients) were subjected to PCR to detect the aforementioned 3 microorganisms. RESULTS: Detection frequencies of T. socranskii, T. denticola, and P. gingivalis in plaque samples from aggressive periodontitis patients (71.1%, 73.7%, 84.2%, respectively) and chronic periodontitis patients (89.2%, 93.8%, 95.3%) were much higher than those from healthy subjects (30%, 5.0%, 10.0%). In aggressive and chronic periodontitis patients, these 3 species of bacteria were detected frequently at sites that showed deep periodontal pockets and severe attachment loss. The percentage of these bacteria-positive sites increased as the gingival index score of chronic periodontitis patients increased. T. socranskii was frequently detected together with T. denticola or P. gingivalis at the same sites, and coexistence of these microorganisms was frequently observed in deep periodontal pockets of aggressive periodontitis patients. CONCLUSIONS: T. socranskii, T. denticola, and P. gingivalis were frequently detected in periodontitis patients by PCR. The prevalence of these 3 microorganisms was correlated with various clinical parameters. Taken together, our findings suggest that T. socranskii, T. denticola, and P. gingivalis are associated with the severity of periodontal tissue destruction.

Adolescent↗
PubMed2001

Adaptive gastric cytoprotection is mediated by prostaglandin EP1 receptors: a study using rats and knockout mice.

Endogenous prostaglandins (PGs) play a central role in adaptive cytoprotection induced in the stomach by mild irritants. In the present study, we used taurocholate (TC) as a mild irritant in both rats and EP-receptor knockout mice, and examined which EP receptor is responsible for the adaptive gastric cytoprotection. Gastric lesions were induced by p.o. administration of HCl/ethanol (60% ethanol in 150 mM HCl). TC (5-20 mM) or PGE2 was administered p.o. 30 min before HCl/ethanol. HCl/ethanol-induced gastric lesions were dose dependently prevented by TC, and the effect at 20 mM was equivalent to that induced by PGE2 at 0.3 mg/kg. The protective effect of TC was significantly attenuated by indomethacin as well as ONO-AE-829, the EP1 antagonist, but not by either NS-398, the selective cyclooxygenase (COX)-2 inhibitor, or chemical ablation of capsaicin-sensitive sensory neurons. Likewise, the protective action of PGE2 was also antagonized by ONO-AE-829 but not chemical deafferentation. TC significantly increased PGE2 contents in the stomach, with or without chemical deafferentation, and this effect was blocked in the presence of indomethacin but not NS-398 or ONO-AE-829. TC increased the mucosal PGE2 contents similarly in both wild-type and knockout mice lacking EP1 or EP3 receptors, yet the protective action of TC against HCl/ethanol was observed in both wild-type and EP3 receptor knockout mice, but not in mice lacking EP1 receptors. The present findings confirmed a role for endogenous PGE2 produced by COX-1 in adaptive gastric cytoprotection and suggested that this action is mediated by activation of EP1-receptors but not associated with capsaicin-sensitive afferent neurons.

Adaptation, Physiological↗
PubMed2001

Anti-apolipoprotein A-I autoantibody: characterization of monoclonal autoantibodies from patients with systemic lupus erythematosus.

OBJECTIVE: The autoantibody to apolipoprotein A-I (apoA-I), a major constituent of high density lipoproteins (HDL), has been detected in sera of patients with systemic lupus erythematosus (SLE). We established a series of monoclonal anti-apoA-I antibodies (MAAI) from 2 patients with SLE and report the reactivities of MAAI with oxidized HDL, anionic substances, and blood coagulation factors. METHODS: Peripheral blood B cells from patients with SLE were immortalized by Epstein-Barr virus, and B cells secreting anti-apoA-I antibodies (AAI) were fused with mouse myeloma cells. Six MAAI reactive with human apoA-I in both ELISA and immunoblotting analysis were established. The reactivities of MAAI with HDL, ssDNA and dsDNA, phospholipids such as cardiolipin (CL), and coagulation factors were examined by ELISA. RESULTS: Although all MAAI bound effectively to apoA-I after the protein had been denatured and transferred to the filter membrane (in immunoblotting analyses), they bound less effectively to apoA-I present in HDL. Both oxidation of HDL in the presence of Mn2+ and an association of apoA-I with autoxidized trilinolein strongly enhanced the binding of MAAI to apoA-I, suggesting that MAAI recognize a defined region of apoA-I, which is exposed upon interacting with oxidatively modified lipids. MAAI showed a functional heterogeneity in their cross-reactivity with self-components: some MAAI were shown to cross-react with anionic substances such as CL and ssDNA, and one MAAI was shown to bind effectively to thrombin. CONCLUSION: We identified a novel family of AAI that shows preferential binding to apoA-I in oxidatively modified HDL. These AAI are composed of antibodies with heterogeneous cross-reactivities to various self-components such as anionic phospholipids, ssDNA, and thrombin.

Antibodies, Monoclonal↗
PubMed2000

Nadrin, a novel neuron-specific GTPase-activating protein involved in regulated exocytosis.

It has been proposed that the cortical actin filament networks act as a cortical barrier that must be reorganized to enable docking and fusion of the synaptic vesicles with the plasma membranes. We identified a novel neuron-associated developmentally regulated protein, designated as Nadrin. Expression of Nadrin is restricted to neurons and correlates well with the differentiation of neurons. Nadrin has a unique structure; it contains a GTPase-activating protein (GAP) domain for Rho family GTPases, a potential coiled-coil domain, and a succession of 29 glutamines. In vitro the GAP domain activates RhoA, Rac1, and Cdc42 GTPases. Expression of Nadrin in NIH3T3 cells markedly reduced the number of the actin stress fibers and the formation of the ruffled membranes, suggesting that Nadrin regulates actin filament reorganization. In PC12 cells, Nadrin colocalized with synaptotagmin in the neurite termini and also with cortical actin filaments in the subplasmalemmal regions. Expression of Nadrin or its mutant composed of the coiled-coil and GAP domain enhanced Ca(2+)-dependent exocytosis of PC12 cells, but a mutant lacking the GAP domain inhibited exocytosis. These results suggest that Nadrin plays a role in regulating Ca(2+)-dependent exocytosis, most likely by catalyzing GTPase activity of Rho family proteins and by inducing the reorganization of the cortical actin filaments.

Actins↗