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Biomedical subjects

T Miyazawa

Publications and source records attributed to T Miyazawa.

At least 73 records · Page 4Linked to original sources
PubMed2000

Synthetically prepared Aamadori-glycated phosphatidylethanolaminecan trigger lipid peroxidation via free radical reactions.

This study for the first time confirmed the peroxidative role of the Amadori product derived from the glycation of phosphatidylethanolamine (PE), namely Amadori-PE. The product was synthesized from the reaction of dioleoyl PE with D-glucose, and then purified by a solid-phase extraction procedure, which was a key step in the next HPLC technique for the isolation of essentially pure Amadori-PE. When the synthetically prepared Amadori-PE was incubated with linoleic acid in the presence of Fe(3+) in micellar system, a remarkable formation of thiobarbituric acid reactive substances was observed together with increases in lipid hydroperoxides. In addition, the lipid peroxidation caused by Amadori-PE was effectively inhibited by superoxide dismutase, mannitol, catalase and metal chelator. These results indicated that Amadori-PE triggers oxidative modification of lipids via the generation of superoxide, and implied the involvement of 'lipid glycation' along with membrane lipid peroxidation in the pathogenesis of diabetes and aging.

Aging↗
PubMed2000

Fish oil diet affects on oxidative senescence of red blood cells linked to degeneration of spleen cells in mice.

The effect of dietary polyunsaturated fatty acids and alpha-tocopherol supplementation on erythrocyte lipid peroxidation and immunocompetent cells in mice was studied comparatively using seven dietary oils (15% oil/diet, w/w) including fish oil rich in eicosapentaenoic acid (EPA, 20:5, n-3) and docosahexaenoic acid (DHA, 22:6, n-3). A 43% increase in spleen weight, about twice as many spleen cells and no change in the subpopulations of spleen cells, as well as a significant depression of mitogen-induced blastogenesis of both T and B cells in the spleen were observed in mice fed fish oil for 30 days in comparison with soybean oil diet-fed mice. In the fish oil diet-fed mice, membranous lipid hydroperoxide (hydroperoxides of phosphatidylcholine and phosphatidylethanolamine) accumulation as a marker of oxidative senescence in red blood cells (RBC) was 2.7-3.5 times higher than that in mice fed soybean oil, although there was no difference in the plasma phosphatidylcholine hydroperoxide concentration. In spite of the supplementation of alpha-tocopherol to up to 10 times the level in the basal diet, the degeneration of spleen cells and the stimulated oxidative senescence of RBC found by the fish oil feeding could not be prevented. The results suggest that oral intake of excess polyunsaturated fatty acids, i.e. EPA and DHA, in a fish oil diet can lead to acceleration of membrane lipid peroxidation resulting in RBC senescence linked to the lowering of immune response of spleen cells, and that supplementation of alpha-tocopherol as antioxidant does not always effectively prevent such oxidative degeneration as observed in spleen cells and RBC in vivo.

Animals↗
PubMed2000

Do conjugated eicosapentaenoic acid and conjugated docosahexaenoic acid induce apoptosis via lipid peroxidation in cultured human tumor cells?

Conjugated eicosapentaenoic acid (CEPA) and conjugated docosahexaenoic acid (CDHA) with triene structure, isomerized by alkaline treatment, showed intensive cytotoxicity with LD(50) at 12 and 16 microM, respectively, in DLD-1 cells (colorectal adenocarcinoma), while they had no effect on normal human fibroblast cell lines such as MRC-5, TIG-103, and KMS-6 cells. Cytotoxic action of CEPA and CDHA was also demonstrated in other tumor cell lines including HepG2, A549, MCF-7, and MKN-7 cells. alpha-Tocopherol suppressed cytotoxicity of CEPA and CDHA in tumor cells, and the cytotoxicity involved membrane phospholipid peroxidation. CEPA and CDHA induced DNA condensation and fragmentation in DLD-1 cells, indicating the involvement of apoptosis in this cytotoxic mechanism. Furthermore, previous reports have shown that lipid peroxidation product induces cell death, including apoptotic cell death in different cell lines. CEPA and CDHA have been demonstrated in cultured cells to cause cell death via lipid peroxidation and apoptosis in the absence of alpha-tocopherol.

Antioxidants↗
PubMed2000

Analysis of experimental autoimmune encephalomyelitis induced in F344 rats by pertussis toxin administration.

To elucidate the factor(s) accelerating the autoimmune disease processes, we induced two types of experimental autoimmune encephalomyelitis (EAE), severe and very mild, in F344 rats by immunization with myelin basic protein (MBP) plus pertussis toxin (PT) (PT+) or with MBP alone (PT-) and compared the differences between the two. Immunohistochemical examinations showed that although the nature of inflammation was essentially the same between the two groups, the proportion of Vbeta8.2(+) T cells in the CNS lesion of PT (+) rats was larger than that of PT (-) rats. Cytokine analysis by competitive PCR revealed that IL-10 mRNA in the lymphoid organ was significantly suppressed in the PT(+) group, whereas levels of IFN-gamma,TNF-alpha and TGF-beta mRNA were insignificantly different after PT administration. In addition, T cells taken from PT (+) rats proliferated well in response to MBP, while those from PT (-) rats showed a marginal response to the same antigen. However, this finding does not indicate the switching of non-encephalitogenic to encephalitogenic T cells upon PT administration because PT (-) rats contained encephalitogenic T cells and/or their precursor cells as revealed by adoptive transfer experiments. Taken together, these findings suggest that suppression of IL-10 by PT administration is the major factor contributing to the exacerbation of EAE in PT(+) rats.

Animals↗
PubMed2000

Molecular cloning and sequencing of the cDNA encoding the feline FcgammaRIIIA (CD16) homologue.

We amplified the cDNA encoding the feline FcgammaRIIIA (CD16) homologue from peripheral blood mononuclear cells by polymerase chain reaction and cloned two forms of FCGR3A cDNA. Sequencing analysis revealed that the open reading frame of feline FCGR3A cDNA consists of 750 or 747 base pairs encoding 250 or 249 amino acid residues, respectively. Comparison of the predicted amino acid sequence of feline FCGR3A cDNA with those of other mammalians' homologues revealed that the extracellular domain has a relatively low homology. However, the cytoplasmic domain contained an 8-amino acid motif, Leu-Phe-Val-Val-Asp-Thr-Gly-Leu, which was considered to interact with an accessory molecule such as the gamma chain of Fc receptors for IgE to form heterodimeric complexes.

Amino Acid Sequence↗
PubMed2000

Newly recognized cytotoxic effect of conjugated trienoic fatty acids on cultured human tumor cells.

We investigated the cytotoxic effect of conjugated trienoic fatty acids on various human tumor cell lines: DLD- 1, colorectal; HepG2, hepatoma; A549, lung; MCF-7, breast; and MKN-7, stomach. Conjugated linoleic acid (CLA) and conjugated linolenic acid were prepared from linoleic acid (18:2, n-6) and alpha-linolenic acid (18:3, n-3), respectively, by treatment with 6.6% or 21% potassium hydroxide. Spectrophotometric readings at 235 nm for the conjugated diene formation, and at 268 nm for the conjugated triene, were confirmed for the respective conjugated fatty acids. In addition, tung oil (Aleurites fordii) fatty acids consisting principally of a conjugated triene (eleostearic acid, approximately 80% of total fatty acids) were prepared using an alkaline saponification procedure. All tumor cells were incubated for 24 h with 5-100 microM of the conjugated fatty acids, and MTT dye reduction was measured to verify the cell viability. Among the conjugated fatty acids examined, conjugated linolenic acid and tung oil fatty acids exhibited the most intense cytotoxic effects on DLD-1, HepG2, A549, MCF-7 and MKN-7 cells, while CLA was not cytotoxic to the tumor cells. These results demonstrate that conjugated trienoic fatty acids are more cytotoxic to human tumor cells than the conjugated dienoic fatty acid, CLA.

Antineoplastic Agents↗
PubMed2000

Absorption, metabolism and antioxidative effects of tea catechin in humans.

Green tea is consumed as a popular beverage in Japan and throughout the world. During the past decade, epidemiological studies have shown that tea catechin intake is associated with lower risk of cardiovascular disease. In vitro biochemical studies have reported that catechins, particularly epigallocatechin-3-gallate (EGCg), help to prevent oxidation of plasma low-density lipoprotein (LDL). LDL oxidation has been recognized to be an important step in the formation of atherosclerotic plaques and subsequent cardiovascular disease. Metabolic studies have shown that EGCg supplement is incorporated into human plasma at a maximum concentration of 4400 pmol/mL. Such concentrations would be enough to exert antioxidative activity in the blood stream. The potent antioxidant property of tea catechin may be beneficial in preventing the oxidation of LDL. It is of interest to examine the effect of green tea catechin supplementation on antioxidant capacity of plasma in humans by measuring plasma phosphatidylcholine hydroperoxide (PCOOH) as a marker of oxidized lipoproteins.

Antioxidants↗
PubMed2000

Real time monitoring of glutamate following fluid percussion brain injury with hypoxia in the rat.

In this study, extracellular glutamate (Glu) was monitored in real time using a biosensor following traumatic brain injury (TBI) either with or without inducing hypoxia in the rat Fluid-percussion model. We also measured the cortical contusion volume at 3 days after the insult. The animals were divided into 3 groups. Group 1 was subjected to TBI only, Group 2 to TBI followed by 20 min of moderate hypoxia (FiO2: 10%) and Group 3 to 20 min of moderate hypoxia without TBI. The surge increase in the extracellular Glu concentration occurred immediately after TBI in Groups 1 and 2. Group 2 showed a prolonged efflux of Glu during hypoxia. Group 3 Glu showed low continuous steady levels. The contusion volume in Group 2 was significantly larger than in Group 1. To test the possible involvement of apoptosis in Groups 1 and 2, rats were sacrificed at 1, 6, 24 and 72 h after TBI. Immunohistochemical studies showed an increased number of both CPP32 positive cells at 24 h and TUNEL cells at 72 h in Group 2. These results suggest that TBI with moderate hypoxia induced a prolonged efflux of Glu that resulted more cortical damage due to necrosis and apoptosis.

Animals↗
PubMed2000

Mn-SOD and Bcl-2 expression after repeated hyperbaric oxygenation.

To clarify the mechanism of ischemic tolerance induced by HBO, we investigated the effect of HBO on immunoreactivity to Bcl-2 and Bax, apoptosis-regulating protein, or Mn-SOD, a radical scavenging system, in the CA1 sector of the gerbil hippocampus. Pretreatment comprising, five sessions at 2 ATA (atmosphere absolute) every other day, but not that comprising, ten sessions at 3 ATA every day, caused significant increases in Bcl-2 and Mn-SOD immunoreactivity in the CA1 sector compared with in the sham pretreatment group. No significant differences in Bax immunoreactivity and neuronal density in the CA1 hippocampal neurons was observed between the groups. These results suggest that protection against mitochondrial alterations after ischemia through Mn-SOD and/or Bcl-2 expression is related to the ischemic tolerance induced by repeated HBO pre-treatment.

Animals↗
PubMed2000

Hepatocyte growth factor reduces infarct volume after transient focal cerebral ischemia in rats.

Hepatocyte growth factor (HGF) was originally discovered as a powerful mitogen for hepatocytes. HGF functions both as a neurotrophic factor as well as an angiogenetic factor. Furthermore, HGF has an anti-apoptotic effect on vascular endothelial cells. The present study examined the neuroprotective effect of HGF after transient focal cerebral ischemia in rats, in which an anti-apoptotic and an angiogenetic effect of HGF was assumed to contribute to the reduction of the infarct volume. The intraventricular administration of human recombinant HGF (90 micrograms) significantly reduced the infarct volume after 120 minutes occlusion of both the right middle cerebral artery (MCA) and the bilateral common carotid arteries (CCAs). In a separate series of experiments, we investigated both the anti-apoptotic effect on neurons and the angiogenetic effect of HGF histopathologically. The number of survival neurons and vascular lumina in the HGF group were significantly higher than those in the vehicle group. A large number of TUNEL positive neurons were observed in the inner boundary of the infarct area in the vehicle group, whereas only a few TUNEL positive neurons were observed in a corresponding area in the HGF group. In the HGF group, Bcl-2 protein was obviously represented in survival neurons as well as in vascular endothelial cells and in glial cells subjected to ischemia. These data suggest that HGF prevents apoptotic neuronal cell death by upregulating the production of Bcl-2 protein and by an angiogenetic effect in the central nervous system which affected transient focal cerebral ischemia.

Animals↗
PubMed2000

The role of lipid peroxidation on gastric mucosal lesions induced by water-immersion-restraint stress in rats.

This study was designed to determine the effect of water-immersion-restraint stress (WIRS) and pretreatment by reduced glutathione on both the production of gastric mucosal lesions and the content of gastric mucosal phosphatidylcholine hydroperoxide (PCOOH). Sprague-Dawley rats were divided into control and treatment groups (treated with reduced glutathione before stress). After graded durations of WIRS, the macroscopic ulcer index (UI) was measured and the PCOOH content was examined by a chemiluminescence-high performance liquid chromatography assay. The UI in the control group increased significantly in a time-dependent fashion. Elevation of the PCOOH level was observed in combination with the UI for up to 2h of WIRS, but then showed a declining tendency. Pretreatment with reduced glutathione significantly lowered both the UI and the PCOOH level. Lipid peroxidation is probably involved in the pathogenesis of gastric injury induced by WIRS at least in the early phases. However, in the late phases, other mechanisms causing gastric mucosal lesions may also be involved. We therefore believe this study to be the first to accurately and quantitatively assess the degree of lipid peroxidation in the gastric mucosa as a result of stress.

Animals↗
PubMed2000

Early experiences of haemostasis on brain tumour surgery with Argon Plasma Coagulation (APC).

OBJECTIVE: We first applied a novel haemostatic strategy involving Argon Plasma Coagulation (APC), an innovative no-touch electrocoagulation technique in which a high-frequency alternating current is delivered to the tissue through ionized argon gas, to brain tumour surgery, and report its usefulness and limitations. METHODS: The APC system we used comprised an APC 300 developed by ERBE Elektromedizin GmbH, Germany. We applied APC to 13 brain tumours in 12 patients (5 meningiomas, 4 sarcomas, 2 glioblastomas, and 2 pituitary adenomas). To avoid unnecessary thermal injury to the tissue as much as possible, power/gas flow settings of 20 and 40 W were used. The impact time was varied individually but was around several seconds per one impact. The argon jet (1.5-4.5 L/min) clears a field of pooled blood and evenly conducts electrical energy to the target tissue. A thin and flexible probe particularly increased the usefulness of APC for haemostasis on deep-seated skull base tumour operations under a microscope. CONCLUSION: All patients were successfully treated and satisfied with the surgical results without any complications due to APC. APC appears to be an excellent alternative strategy for achieving haemostasis on vascular-rich brain tumour surgery, and may be valuable for the management of patients with coagulation defects.

Adenoma↗
PubMed2000

Cystic schwannoma of the orbit: case report.

BACKGROUND: Cystic lesions of the orbit have diverse clinical findings and histopathological features. We report an orbital schwannoma composed of a single cyst. CASE DESCRIPTION: A 62-year-old woman with an intraconal cystic tumor in the left orbital cavity is described. The cystic tumor was totally removed and the pathological diagnosis of the cyst wall was schwannoma composed of both Antoni type A and B. CONCLUSIONS: A rare case of cystic schwannoma of the orbit is reported. This case may contribute to an improved preoperative differential diagnosis for cystic lesions of the orbit.

Cysts↗
PubMed2000

Efficient expression of the envelope protein of feline immunodeficiency virus in a recombinant feline herpesvirus type 1 (FHV-1) using the gC promoter of FHV-1.

We constructed two recombinant feline herpesvirus type 1 (FHV-1) expressing the envelope (Env) protein of feline immunodeficiency virus (FIV). One recombinant, designated dlTK-env, has the whole FIV env gene inserted at a thymidine kinase (TK) deletion site. The second recombinant, designated dlTK(gCp)-env, has a cassette containing a partial FIV env gene fused with the signal sequence of the gC protein of FHV-1 (under the control of the gC promoter) inserted at the same site. Growth kinetics of both the recombinants in Crandell feline kidney (CRFK) cells were similar to that of the parent strain of FHV-1. By indirect immunofluorescence assays and immunoblot analyses, we confirmed the expression of the FIV Env protein in CRFK cells infected with both recombinants. Enzyme-linked immunosorbent assays showed that the maximum Env expression level achieved by dlTK(gCp)-env was more than four times higher than that observed for dlTK-env. Flow cytometric analyses revealed that the Env protein produced by both recombinants was efficiently expressed on the cell surface. The dlTK(gCp)-env reported here may thus be a promising candidate for a live recombinant vaccine to protect against FIV infection.

Animals↗
PubMed2000

Comparison of the efficacy of two air-based contrast agents in dogs.

OBJECTIVE: The aim of this study was to evaluate the differences of the in vivo efficacies of Levovist(SH U 508 A) and Albunex, air-based contrast agents (CAs) with different shell materials, by direct comparison. METHODS: SH U 508 A, 300 mg/ml, and Albunex were injected intravenously at doses of 0.1 and 0.15 ml/kg, respectively, into the same dogs (n=6). Contrast enhancement in the left ventricle was evaluated visually and by off-line measurement of brightness. RESULTS: Both CAs yielded good peak contrast, while the duration of contrast enhancement with SH U 508 A was seven times that with Albunex. With Albunex, reduction of contrast enhancement at end-systole and in the late phase of diastole during one heartbeat was observed, and a positive correlation was observed between contrast enhancement and heart rate. Contrast enhancement was nearly constant during one heartbeat with SH U 508 A. CONCLUSION: SH U 508 A yielded consistent and longer contrast enhancement in the left ventricle than Albunex under the same conditions.

Air↗
PubMed2000

Digestion of plant monogalactosyldiacylglycerol and digalactosyldiacylglycerol in rat alimentary canal.

We investigated digestion of orally fed galactoglycerolipids such as monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) from wheat flour in the rat alimentary canal, especially focusing on the digestive fates of deacylated galactosylglycerol structures. After a single oral administration of MGDG (20 mg/rat), monogalactosylmonoacylglycerol and monogalactosylglycerol (MGG) were found to be major digestion products in the intestinal tract. Similarly, digalactosylmonoacylglycerol and digalactosylglycerol (DGG) were confirmed to be present in the intestinal tract after DGDG ingestion (20 mg/rat). In rats fed wheat flour glycolipids (42 mg MGDG and 81 mg DGDG per rat), completely deacylated galactosylglycerols (MGG and DGG) were not detected in portal plasma. Although the deacylated galactosylglycerols were not significantly decomposed by intestinal mucosa in vitro, they were hydrolyzed by cecal contents. The results demonstrated that orally ingested plant galactoglycerolipids in the rat alimentary canal are rapidly hydrolyzed into constituent fatty acids and that hydrophilic galactosylglycerols and the hydrophilic backbone galactosylglycerols are not absorbed from intestine or degraded into galactose and glycerol in the intestinal tract. Therefore, the presence of deacylated galactosylglycerols may affect the fermentative activity of enterobacteria in the cecum and colon.

Journal Article↗