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Biomedical subjects

M Ono

Publications and source records attributed to M Ono.

At least 541 records · Page 30Linked to original sources

Root surface caries of denture wearers in middle-aged and elderly people.

With an increasing number of elderly people retaining natural dentition, the number of exposed root surfaces and root surface caries is increasing. A total of 100 people, aged from 38 to 86, were examined for exposed root surfaces and root surface caries. Denture wearers and non-denture wearers were examined. The following results were obtained; 1. The percentages of exposed root surfaces were significantly higher (P < 0.05) in the 60's and 70's age groups. Root surface caries showed a tendency to increase until people were in their 60's. 2. Denture wearers had more exposed root surfaces (P < 0.05) and were more likely to have root surface caries than non-denture wearers. Denture wearers showed a tendency to have more severe grades of root surface caries than non-denture wearers. 3. Concerning denture wearers, there were not significant differences in the percentages of exposed root surfaces of buccal and proximal surfaces facing edentulous space. However, proximal surfaces facing edentulous space were more prone to root surface caries than buccal surfaces (P < 0.05).

Adult↗

Anti-angiogenic activity of arachidonic acid metabolism inhibitors in angiogenesis model systems involving human microvascular endothelial cells and neovascularization in mice.

We have established an in vitro angiogenesis model using human omental microvascular endothelial (HOME) cells, in which epidermal growth factor (EGF) or transforming growth factor-alpha (TGF-alpha) stimulated cell migration and tube formation. In this study, we examined whether alpha-guaiaconic acid (GR-12) and its synthetic 20 derivatives showed inhibition of cell migration and tubular formation of HOME cells. We found that GR-12 inhibits arachidonic acid metabolism, while GR-12 and one derivative, GS-01, inhibit tubular formation of endothelial cells in our model system. Confluent monolayers of HOME cells were damaged with a razor blade and incubated with or without TGF-alpha; HOME cell migration was stimulated about 1.5-fold over control values in the presence of TGF-alpha. Treatment of HOME cells with GR-12 or GS-01 inhibited both spontaneous and TGF-alpha-stimulated migration. GR-12 or GS-01 inhibited TGF-alpha-induced HOME-cell tube formation in type-1 collagen gels. We examined whether these compounds could modulate tubular formation of HOME cells induced by human cancer cells. Enhanced tube formation of HOME cells by co-cultured esophageal cancer cells was almost completely inhibited by co-administration of GR-12 or GS-01. Both compounds also inhibited formation of tubular networks of HOME cells on Matrigels. We also examined anti-angiogenic activity of these compounds in an in vivo model system of tumor angiogenesis in mice. In this system, GS-01 inhibited development of capillary networks at a rate comparable to that of a well-known anti-angiogenic compound, fumagillin, but GR-12 did not. The inhibitor of arachidonic acid metabolism is thus expected to modulate tumor angiogenesis.

Animals↗

[MR tractography--visualization of structure of nerve fiber system from diffusion weighted images with maximum intensity projection method].

We developed a new noninvasive technique to visualize the anatomical structure of the nerve fiber system in vivo, and named this technique magnetic resonance (MR) tractography and the acquired image an MR tractogram. MR tractography has two steps. One is to obtain diffusion-weighted images sensitized along axes appropriate for depicting the intended nerve fibers with anisotropic water diffusion MR imaging. The other is to extract the anatomical structure of the nerve fiber system from a series of diffusion-weighted images by the maximum intensity projection method. To examine the clinical usefulness of the proposed technique, many contiguous, thin (3 mm) coronal two-dimensional sections of the brain were acquired sequentially in normal volunteers and selected patients with paralyses, on a 1.5 Tesla MR system (Signa, GE) with an ECG-gated Stejskal-Tanner pulse sequence. The structure of the nerve fiber system of normal volunteers was almost the same as the anatomy. The tractograms of patients with paralyses clearly showed the degeneration of nerve fibers and were correlated with clinical symptoms. MR tractography showed great promise for the study of neuroanatomy and neuroradiology.

Aged↗

[Embolization of the internal maxillary artery for severe epistaxis--including an experience of the approach from the superficial temporal artery].

Six cases of severe epistaxis were treated with the super-selective intraarterial embolization of the internal maxillary artery. Tens of fragments of Gelfoam were delivered into the distal internal maxillary artery and, additionally, embolized by several pieces of the coils. In a case with cerebral infraction, catheter was inserted via the superficial temporal artery. There were no complications due to these procedures in all cases of this study. It is considered that therapeutic intraarterial embolization of the internal maxillary artery is an effective therapy for severe epistaxis and the superficial temporal artery approach is useful for selected cases.

Aged↗

Indispensable role of tissue-type plasminogen activator in growth factor-dependent tube formation of human microvascular endothelial cells in vitro.

Epidermal growth factor (EGF) stimulates the migration and proliferation of, and tissue-type plasminogen activator (tPA) synthesis in, human omental microvascular endothelial (HOME) cells in culture, as well as inducing the formation by these cells. In the present study, we examined the effects of various growth factors, i.e., transforming growth factor-alpha (TGF-alpha), insulin-like growth factor 1 (IGF-1), and hepatocyte growth factor (HGF) on HOME cells, and compared their effects with that of EGF. IGF-1 stimulated the proliferation and migration of these cells at a level comparable to EGF. EGF and TGF-alpha induced expression of tPA in HOME cells, while IGF-1 and HGF did not. EGF and TGF-alpha induced tube formation by HOME cells in type I collagen gel, while IGF-1 and HGF did not. The stimulatory effect of EGF on tube formation in the gel was blocked by anti-tPA antibody and by a serine protease inhibitor, aprotinin. When exogenous tPA and IGF-1 or HGF were added simultaneously to the culture, a marked induction of tube formation in the gel was observed. Exogenously added tPA alone, however, had no such inducible effect on tube formation. These results indicated an indispensable role of tPA in growth factor-dependent tube formation by HOME cells. Two subsets of growth factors appeared to modulate angiogenesis: One with fully active angiogenic activity which could induce PA (this included EGF and TGF-alpha), and the other, which could not induce PA and was not angiogenic, but could promote angiogenesis in the presence of PA. This subset included IGF-1 and HGF.

Cell Division↗

Morphological change and destabilization of beta-actin mRNA by tumor necrosis factor in human microvascular endothelial cells.

Tumor necrosis factor-alpha (TNF-alpha) produced morphological changes from a cobblestone-like shape into a spindle shape in human omental microvascular endothelial (HOME) cells and also a drastic rearrangement of actin filaments. Expression of beta-actin gene was diminished in HOME cells treated with TNF-alpha for 24 h. Northern blot analysis of the beta-actin gene demonstrated that the cellular level of beta-actin mRNA was decreased at 6-12 h after exposure to TNF-alpha. However, there appeared to be no changes in cellular mRNA levels of beta-tubulin, fibronectin, laminin B1, laminin B2, and laminin binding protein genes after treatment with TNF-alpha. Nuclear run-on assays showed increased transcription of the low-density lipoprotein receptor gene, but not of the beta-actin gene. These data suggested that the TNF-alpha-induced inhibition of beta-actin gene expression was not due to altered transcription activity. The degradation rates of beta-actin, plasminogen activator inhibitor-1, and epidermal growth factor receptor mRNAs were examined in the presence of actinomycin D. beta-Actin mRNA was found to be specifically destabilized in TNF-alpha-treated HOME cells, while other mRNA species were not. Coadministration of cycloheximide blocked the TNF-alpha-induced degradation of beta-actin mRNA. The TNF-alpha-induced destabilization of beta-actin mRNA and rearrangement of actin filaments are discussed in relation to the morphological changes in human microvascular endothelial cells.

Actin Cytoskeleton↗

Comparison of prostanoids and their precursor fatty acids in human hepatocellular carcinoma and noncancerous reference tissues.

Eicosanoid production and the compositions of precursor fatty acids were determined in human cancerous and reference liver tissues. Seventeen hepatectomized cases (12 cases of hepatocellular carcinoma (HCC) and 5 cases of metastatic liver cancer) were evaluated. The cancerous tissues and the noncancerous reference tissues were separated, lipids were extracted, and the fatty acids were determined as methyl esters by gas chromatography. Prostanoids (6-keto PGF1 alpha and TXB2) were measured by radioimmunoassay. In HCC, the levels of alpha-linolenic acid (omega-3) (0.41 +/- 0.38 x 100 micrograms/g) and docosahexaenoic acid (10.41 +/- 4.96 x 100 micrograms/g) in liver cancer tissue were significantly less than those in the reference tissues. In HCC, the levels of TXB2 reference (1.86 +/- 2.77 pg/mg wet weight) and 6-keto PGF1 alpha were 10-fold higher than those in reference tissues. We speculate that in HCC higher prostanoid levels in the liver are related in part to tumor metabolism.

6-Ketoprostaglandin F1 alpha↗

Inhibition of tumor metastasis by Arg-Gly-Asp-Ser (RGDS) peptide conjugated with sulfated chitin derivative, SCM-chitin-RGDS.

We have synthesized a new compound in which Arg-Gly-Asp-Ser (RGDS) was conjugated with 6-O-sulfated and 6-O-carboxymethyl-chitin (SCM-chitin), i.e. SCM-chitin-RGDS, and tested the inhibitory effect on lung and liver metastases of three different types of tumors in mice. SCM-chitin-RGDS was more effective for the inhibition of liver metastasis of L5178Y-ML25 lymphoma and lung metastases of colon 26 M3.1 cells than SCM-chitin, RGDS or their mixture. GRGDS peptide, however, required a higher dose (3000 micrograms) to obtain a sufficiently antimetastatic effect. Intermittent i.v. administration of SCM-chitin-RGDS before or after the i.v. inoculation of L5178Y-ML25 cells caused significant inhibition of liver metastasis as compared with the multiple administration of RGDS, SCM-chitin or untreated control. Co-injection of lymphoma cells with SCM-chitin-RGDS or multiple treatment of SCM-chitin-RGDS after tumor inoculation showed significantly enhanced survival rate. SCM-chitin-RGDS also showed the spontaneous lung metastasis produced by intrafootpad injection of B16-BL6 melanoma cells by the multiple i.v. administrations. These results demonstrate that the conjugation of RGDS peptide with SCM-chitin led to augmentation of therapeutic potential to cancer metastasis, thus implying an importance of the conjugation of cell-adhesive RGDS peptide with structurally heparin-like SCM-chitin, which possess binding ability to the heparin-binding domain of fibronectin or laminin and extremely low anticoagulant properties.

Amino Acid Sequence↗

Evidence that both cyclosporin and azathioprine prevent warm ischemia reperfusion injury to the rat liver.

The present work was undertaken to study whether the immunosuppressive agents cyclosporin (CyA) and azathioprine (AZA) ameliorate hepatic injury after warm ischemia. A temporary, normothermic liver ischemia was induced in female Sprague-Dawley rats. The rats were treated with CyA (10 mg/kg per day p.o.), AZA (8 mg/kg per day p.o.), or vehicles for 4 days before surgery. Seven-day survival rates after 60 min of ischemia improved significantly with CyA (76.2%, P < 0.005) and AZA (78.6%, P < 0.001) treatment, compared with 43.0% for the control group. The highest levels of serum aminotransferases in the treatment groups tended to be lower than those in the control group. The peak values for the percentage of liver necrosis, an indicator of the extent of hepatic necrosis, in the animals treated with CyA (26.1% +/- 7.2%, mean +/- SEM) and AZA (32.1% +/- 5.7%) were significantly lower than in the control group (47.4% +/- 3.7%). Lipid peroxidative damage after reperfusion, assessed as the hepatic malondialdehyde (MDA) concentration, was significantly suppressed by pretreatment with CyA and AZA. Histological findings coincided with other parameters. This study demonstrates that both AZA and CyA have beneficial effects on normothermic liver ischemia in rats. It is suggested that the diminished lipid peroxidative damage with these agents might be one of the mechanisms responsible for this.

Administration, Oral↗

Identification and nucleotide sequence of a gene in feline herpesvirus type 1 homologous to the herpes simplex virus gene encoding the glycoprotein H.

A gene encoding the glycoprotein H (gH) homologue of feline herpesvirus type 1 was identified and sequenced. It was located immediately downstream of the thymidine kinase gene within an EcoRI 6.6 kbp fragment. In addition, a partial UL21 homologous gene was located downstream of the gH homologous gene. The primary translation product of the gH homologous gene is predicted to consist of 821 amino acids with a molecular weight of 92.5 kDa. It possesses several characteristics typical of transmembrane glycoproteins, including a N-terminal hydrophobic signal sequence, C-terminal transmembrane domain, and putative N-linked glycosylation sites. Analysis of this protein revealed amino acid sequence homologies of 33.1% with equine herpesvirus type 1 (EHV-1) gH, 32.6% with EHV-4 gH, 29.1% with varicella-zoster virus gIII, 28.5% with pseudorabies virus gH, and 25.1% with herpes simplex virus type 1 gH. By Northern blot analysis, one of the transcripts specific for the gH homologous gene might be a mRNA of approximately 3.0 kb.

Amino Acid Sequence↗

Increase of kainate receptor mRNA in the hippocampal CA3 of amygdala-kindled rats detected by in situ hybridization.

To investigate whether lasting changes in excitatory amino acid (EAA) receptor subtypes occurred at their mRNA levels as a result of kindling, we carried out in situ hybridization of rat brain sections using synthetic oligonucleotide probes, which were complementary to cloned EAA receptor subunits, namely NMDAR1 for N-methyl-D-aspartate (NMDA), GluR-2 for alpha-amino-3-hydroxy-5-methylisoxazole 4-propionic acid (AMPA), KA-1 for kainate (KA) and mGluR1 for metabotropic EAA receptors. Rats in which left-amygdala-kindling had been established were decapitated 28 days after the last kindled seizure along with the matched controls, which had been subjected to electrode implantation but not to kindling, and the brain sections were hybridized with the probes. The amount of KA receptor mRNA detected with the KA-1 probe increased (25%) on both the left and right sides of the hippocampal CA3 region in the kindled rats, but in no other brain areas (hippocampal CA1, dentate gyrus, amygdala nuclei and pyriform cortex). There was no significant modification of NMDAR1, GluR-2 or mGluR1 receptor mRNAs in any brain area examined. The increase of KA receptor mRNA in the CA3 of amygdala-kindled rats may indicate that the excitability of the neural circuits mediated by KA receptors increased in the hippocampus as a consequence of kindling.

Animals↗

Reduction of the leukemogenic potential of malignant murine leukemic cells by in vivo treatment with recombinant human granulocyte colony-stimulating factor.

We have investigated the effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) administration on the leukemogenic potential of L-103 murine leukemic cells. Leukemogenic potential was assessed by comparing the regression lines drawn between the number of inoculated leukemic spleen cells and the mean survival time (MST) of the syngeneic recipients. rhG-CSF injected 2.5 micrograms daily for 14 days reduced the leukemogenic potential of spleen cells of the leukemic mice to 1/200 of the control. This phenomenon was not observed with the leukemic spleen cells treated with r-murine granulocyte-macrophage (rmGM)-CSF in vivo. Cytochemical study indicated that morphologically identifiable blast cells were fewer in the rhG-CSF-treated leukemic spleen. Furthermore, leukemic cells in the rhG-CSF-treated spleen were less proliferative than the control in spite of having more clonogenic cells in the leukemic cell preparation. Cytogenetical analysis showed that chromosome abnormalities found in the original leukemic cells were not altered by rhG-CSF administrations. It also showed that the frequency of the abnormal karyotype was reduced in rhG-CSF-treated leukemic spleen (4/17) as compared with the control (8/8), indicating that the mitotic fraction was smaller in the rhG-CSF-treated leukemic cells. These findings indicate that in addition to the reduced number of leukemic cells in the spleen cell preparation, a reduction of the proliferative capacity of the original leukemic cells is involved in the reduction of leukemogenic potential of leukemic cells treated with rhG-CSF in vivo.

Animals↗

Cloning and cDNA sequence analysis of nephritogenic monoclonal antibodies derived from an MRL/lpr lupus mouse.

Production of IgG3 in MRL/Mp-lpr/lpr (MRL/lpr) lupus mice is one of the major factors to develop glomerulonephritis (GN) in these mice. To examine molecular characteristics of IgG3 responsible for GN in these mice, hybridoma clones producing IgG3 antibodies were prepared from one unmanipulated MRL/lpr mouse. Two clones, 2B11.3 and 7B6.8, were nephritogenic; that is, they caused severe glomerular lesions when injected to normal mice, moreover with a different histopathological manifestation. The 2B11.3 clone generated diffuse cell-proliferative lesions, while those induced by the 7B6.8 clone resembled wire loop lesions in human lupus nephritis. The cDNA sequence analysis of 7B6.8 antibody and the other IgG3 antibody, 1G3, non-nephritogenic, revealed that the C regions of the heavy and light kappa chains were completely the same between them. Furthermore, they were identical in deduced amino acid sequences to those from non-autoimmune BALB/c mice, indicating no allelic difference of Igh-8 between these two strains. The V regions of 2B11.3 and 7B6.8 antibodies were composed of different sets of VH, D, JH, Vk and Jk. Although both of the VH belonged to the J558 family, they seemed to use a different VH germline gene. These findings suggest that GN in MRL/lpr mice is generated by the expansion of clonally different B cells producing particular antibodies possibly with a different pathogenetic potency.

Amino Acid Sequence↗

Increased levels of tumor necrosis factor and interleukin 1 in bronchoalveolar lavage fluids from pigs infected with Mycoplasma hyopneumoniae.

We examined the levels of tumor necrosis factor (TNF)-alpha and interleukin-1 (IL-1) in bronchoalveolar lavage fluids (BALF) from pigs experimentally infected with Mycoplasma hyopneumoniae using biological assays with WEHI-164 cells and D10.G4.1 cells, respectively. Increased TNF-alpha and IL-1 in BALF were found in infected pigs with gross and/or microscopic lesions. A time-course study suggested TNF-alpha and IL-1 to be persistently elevated in infected pigs. Their presence in BALF would thus appear to be associated with the development of pneumonic lesions in M. hyopneumoniae infected pigs.

Animals↗

Influence of gender and the endocrine environment on the distribution of androgen receptors in the lacrimal gland.

Androgens are known to regulate both the structure and function of lacrimal tissue in a variety of species. To explore the endocrine basis for this hormone action, the following study was designed to: (1) determine the cellular distribution of androgen receptors in the lacrimal gland; and (2) examine the influence of gender and the endocrine environment on the glandular content of these binding sites. Lacrimal glands were obtained from intact, castrated, hypophysectomized, diabetic or sham-operated male or female adult rats, mice or hamsters, as well as from orchiectomized rats exposed to placebo compounds or physiological levels of testosterone. The cellular location of androgen receptors was evaluated by utilizing an immunoperoxidase protocol, in which a purified rabbit polyclonal antibody to the rat androgen receptor was used as the first antibody. Our findings with lacrimal glands showed that: (1) androgen receptors are located almost exclusively in nuclei of epithelial cells; (2) the cellular distribution or intranuclear density of these binding sites is far more extensive in glands of males, as compared to females; (3) orchiectomy or hypophysectomy, but not sham-surgery or diabetes, lead to a dramatic reduction in the immunocytochemical expression of androgen receptors; and (4) testosterone administration to orchiectomized rats induces a marked increase in androgen receptor content, relative to that in placebo-exposed glands. Our results also reveal that a 10 kb androgen receptor mRNA exists in the rat lacrimal gland. Overall, these findings demonstrate that gender and the endocrine system may significantly influence the distribution of androgen binding sites in rat lacrimal tissue. Moreover, our results show that androgens up-regulate their own lacrimal gland receptors.

Animals↗

Reduction of drug accumulation in cisplatin-resistant variants of human prostatic cancer PC-3 cell line.

We have isolated cis-diamminedichloroplatinum (II) (CDDP)-resistant variants, P/CDP4 and P/CDP5, from human prostatic cancer PC-3 cells after a stepwise exposure to CDDP. P/CDP4 and P/CDP5 showed 11-fold and 23-fold higher resistance to CDDP than did PC-3. P/CDP5 was cross-resistant to carboplatin, mitomycin C, etoposide, m-AMSA, bleomycin and UV irradiation. Alkaline elution of DNA showed an increased amount of DNA interstrand cross-links in PC-3 but not in P/CDP5 when PC-3 and P/CDP5 were cultured with CDDP. Flameless atomic absorption spectrophotometry revealed that intracellular accumulation of CDDP in P/CDP4 and P/CDP5 was decreased to 18 to 34% and 9 to 18% of that of PC-3, respectively, when PC-3 and its CDDP-resistant counterparts were incubated with 5 and 10 micrograms./ml. of CDDP for 24 hours. These data suggest that decreased drug accumulation is involved in the development of CDDP-resistance in the PC-3 cell line.

Cisplatin↗

Association of the actin cytoskeleton with glass-adherent proteins in mouse peritoneal macrophages.

When mouse peritoneal macrophages adherent to glass surface were removed by treatment with triethanolamine and Nonidet P-40, fine thread structures of unique loops were left behind on glass at the sites of cell adhesion. To examine the ultrastructural relationship between such looped threads and cytoskeletal components in glass-adherent macrophages, we successfully used the 'zinc method' to remove most of the cytoplasm including nuclei and to expose the cytoskeleton associated with the ventral plasma membrane. The cytoskeleton was seen to be mainly composed of actin filaments forming dense networks. The network contained scattered star-like foci from which actin filaments radiated. When the ventral plasma membrane-cytoskeleton complex was further treated with Nonidet P-40, the membrane was dissolved to expose the glass surface with actin foci persisting on glass. When the complex was removed by further treatment with Nonidet P-40 and DNase I, the looped threads became visible. Confocal laser microscopy of glass-adherent macrophages stained with fluorescent phalloidin showed the preferential distribution of F-actin in the ventral cytoplasm along the plasma membrane, where intense fluorescent spots were also scattered. Confocal interference reflection microscopy revealed densely populated dark dots and striae of focal contact, which corresponded in overall distribution to actin foci and looped threads. These observations suggest that actin cytoskeleton is closely associated with looped threads to reinforce cell adhesion to glass.

Actins↗