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Biomedical subjects

M Kuwabara

Publications and source records attributed to M Kuwabara.

At least 91 records · Page 5Linked to original sources
PubMed1997

Relationship between radical production and natural killer cytotoxic factor (NKCF) in canine natural killer (NK) cell-mediated cytotoxicity.

The relationship between radical production and natural killer cytotoxic factor (NKCF) release via canine natural killer (NK)-mediated cytotoxic mechanism was examined. Radical production and NKCF release was induced in NK cells stimulated with either dead target cells, or their cytoplasmic membranes, as well as live target cells. Canine NKCF evoked target cell lysis but did not induce radical production. Radical production was inhibited by the addition of Tiron or n-propyl gallate, whereas NK-mediated cytotoxicity and NKCF release were only inhibited by the addition of n-propyl gallate. These results suggested that radical production and NKCF release may be induced by the contact and binding of NK cells to the target cell cytoplasmic membrane. Therefore, the release of NKCF from NK cells attached to the target cell cytoplasmic membrane may be associated with the production of radicals, especially hydroxyl radicals.

Animals↗
PubMed1997

Progressive effect of alpha-phenyl-N-tert-butyl nitrone (PBN) on rat embryo development in vitro.

In the present study we demonstrated the effects of the spin-trapping agent alpha-phenyl-N-tert-butylnitrone (PBN) on the in vitro development of rat embryos at the early stage. In rat embryos, PBN increased the speed of the first cleavage and had no toxicity during pregnancy after embryo culture. These results showed that reactive oxygen species (ROIs) that were formed by activating molecular oxygens through redox reactions regulated the speed of development for early-stage embryos. Thus, PBN caused a decrease in the level of ROIs and toxicity and an in increase in the level of the development of rat embryos. On the other hand, PBN could not decrease the 2-cell block in vitro nor increase the blastulation rate, in contrast to the fact that a scavenger of superoxide anions, SOD, is effective in doing so for mouse embryos. From these results it was concluded that free radicals play an important role in the in vitro development of rat embryos at the early stage, but play no role in the decrease of the 2-cell block or their blastulation rate. It should be noted that PBN had no toxicity for embryonic development at the 2-cell stage.

Animals↗
PubMed1997

Factors affecting M13 bacteriophage inactivation by methylene blue photosensitization.

We have investigated the factors that affect the virucidal activity of methylene blue (MB) photosensitization. The M13 bacteriophage was more rapidly inactivated at higher temperatures (6 degrees C < 24 degrees C < 38 degrees C). Rate constants for inactivation were 0.072, 0.139 and 0.260 (log10 inactivation)/ (J/cm2) at 6 degrees C, 24 degrees C and 38 degrees C, respectively. On the other hand, dye penetration into virus particles, which was monitored by the fluorescence of YOYO-1, was unchanged with incubation temperature. These data suggest that temperature dependency of M13 inactivation was due to factors other than dye permeability. The pH of the virus suspension also affected the rate of M13 inactivation by MB. The M13 bacteriophage was inactivated faster in basic suspensions and slower in acidic suspensions compared with neutral buffers. These results suggest that temperature and pH are factors that influence the extent of MB photosensitization, and hence, the control of these factors will be necessary for MB phototreatment of plasma products in transfusion medicine.

Antiviral Agents↗
PubMed1997

Potential involvement of both type I and type II mechanisms in M13 virus inactivation by methylene blue photosensitization.

We have investigated the mechanism of virus photoinactivation with methylene blue (MB) by conducting deuterium oxide (D2O), azide ion (N3-) and oxygen-dependent studies. Inactivation of M13 bacteriophage and singlet oxygen (1O2) generation by MB photosensitization were irradiation dose dependent. Inactivation of M13 was enhanced by D2O and inhibited by N3-, suggesting that 1O2 participates in M13 inactivation by MB photosensitization. However, N3- did not inhibit M13 inactivation completely. On the other hand, deoxygenating the reaction solution still caused 52-67% of M13 inactivation observed in the presence of oxygen. These results suggest that 1O2-mediated (Type II) and sensitizer-mediated (Type I) reactions may both play roles in M13 inactivation by MB photosensitization.

Antiviral Agents↗
PubMed1997

Reactions between hydroxyl-radical-induced 7,8-dihydro-8-oxo-2'-deoxyguanosine precursor and the spin trap alpha-phenyl-N-tert-butylnitrone.

An N2O-saturated aqueous solution containing 2'-dG and the spin trap agent PBN was examined by ESR and HPLC-ECD methods after X irradiation. ESR examination showed that the ESR spectrum obtained consisted of signals due to the PBN-OH and PBN-H adducts. The signal intensity of PBN-H adducts was larger in the presence of 2'-dG than in the absence of 2'dG, while that of PBN-OH adducts was smaller in the presence of 2'-dG than in the absence of 2'-dG. When the OH-radical-induced 8-oxodG was measured by HPLC-ECD, the yield of 8-oxodG was found to be enhanced about twofold in the presence of PBN. By contrast, usual OH-radical scavengers (DMSO, sodium formate and mannitol) inhibited the formation of 8-oxodG beyond expectation. The enhancement of the yield of PBN-H adducts by 2'-dG and the enhancement of the 8-oxodG formation by PBN were explained by the electron transfer and subsequent proton transfer reactions from OH-radical-induced guanine-N7 radicals to PBN to form 8-oxodG and PBN-H. The present study led us to conclude that PBN reacted with the precursor radical of 8-oxodG to accelerate the formation of 8-oxodG, while OH-radical scavengers reacted with it to diminish the formation of 8-oxodG.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed1997

Concentration-dependent modes of cell death in Chinese hamster V79 cells after treatments with H2O2.

Deterioration in the clonogenic ability of Chinese hamster V79 cells was observed after treatments with low concentrations of H2O2 (0.06-0.2 mM) for 1 h and subsequent incubation for 6-8 days, whereas no loss of ability of the cells to exclude trypan blue after treatments with H2O2 for 1 h and subsequent incubation for 6 h was observed in this concentration range. The loss of the dye-exclusion ability became observable when the cells were treated with concentrations greater than 0.9 mM H2O2 for 1 h and subsequently incubated for 6 h. Agarose gel electrophoresis of DNA extracted from cells treated with 1-5 mM H2O2 showed specific regular fragmentation of DNA, suggesting that apoptotic cell death was induced. The induction of apoptotic cell death was further confirmed by observing the protective effects of several modifiers, a protein synthesis inhibitor (cycloheximide), a Ca(2+)-chelator (BAPTA-AM) and an antioxidative compound PBN), against cell survival and DNA fragmentation. From these results, it was concluded that apoptotic cell death was induced by H2O2 in Chinese hamster cells at high concentrations, where their clonogenic ability completely disappeared.

Animals↗
PubMed1997

[A case of successful surgical treatment for active endocarditis 2 days after the onset of cerebral infarction].

We performed double valve replacement for a patient with active endocarditis 2 days after the onset of cerebral infarction because of intractable cardiac failure. The use of heparin and the hypotension brought by cardiopulmonary bypass can lead exacerbation of the cerebral symptoms after open heart surgery which is performed during acute phase of cerebral infarction. Perfusion pressure was maintained over 70 mmHg during cardiopulmonary bypass and activated clotting time was kept about 400 seconds to prevent aggravation of cerebral complications in this case. The patients recovered from surgery uneventfully. We described a case who was received double valve replacement 2 days after the onset of cerebral infarction successfully.

Aortic Valve↗
PubMed1997

Genetic profiles of newly inbred Dahl/Iwai salt-sensitive and salt-resistant rats.

Dahl-Iwai salt-sensitive (S) and salt-resistant (R) rat strains were established as inbred strains at Brookhaven National Laboratory, NY, and were introduced into Eisai Co., Ltd., Japan, and designated DIS/Eis and DIR/Eis. To examine whether there are different allele distributions among the substrains of inbred Dahl S and R rats, we determined biochemical and immunological alleles of DIS/Eis and DIR/Eis, and SS/Sea and SR/Sea, which were derived from SS/Jr and SR/Jr, which were developed by Rapp and Dene. Several differences of allele distribution were observed, indicating that the substrains have different genetic backgrounds. The phenotypic differences between the substrains, such as the severity of the hypertension induced, could be ascribed to the different genetic backgrounds.

Alleles↗
PubMed1997

Persistent sciatic artery aneurysm with ruptured internal iliac artery aneurysm.

A case of aneurysm of persistent sciatic artery with ruptured aneurysm of the internal iliac artery is reported. A 62-year-old man was admitted to our hospital in emergency because of abdominal severe pain. Abdominal CT and angiography showed aneurysm of the infrarenal aortic aorta and left internal iliac artery and left persistent sciatic artery. The persistent sciatic artery aneurysm was coursed down the left thigh. Left superficial femoral artery was supplying blood to the popliteal artery as normal connection. Emergency operation was performed. The left internal iliac aneurysm was ruptured to a retroperitoneal space. The aneurysm was replaced by a Y-shaped dacron double velour graft from the infrarenal abdominal aorta to right common iliac and left external iliac arteries, which was followed by a ligation at the origin of left persistent sciatic aneurysm. The pulsation of the left lower extremity was good and additional bypass procedure to the popliteal artery was not performed. The post-operative course of the patient was excellent and post-operative angiography showed normal anatomy of the left lower extremity. Optimal management of ruptured abdominal aneurysm thus requires not only an emergency operation but detailed observation of preoperative CT and angiography in order to scrutinize the associated vascular anomalies.

Aneurysm↗
PubMed1997

Magnetic resonance imaging of hepatocellular carcinoma in Long-Evans cinnamon rats under a magnetic field of 7.05 T.

Magnetic resonance (MR) images of livers in 3-, 12- and 29-month-old Long-Evans Cinnamon(LEC) rats (male) were taken under a magnetic field of 7.05 T. MR images of sagittal and transversal sections were obtained in 1-mm-thick slices by T1-weighted and two-dimensional Fourier transformation techniques. The data matrix size was 256 phase-encoded steps. Each image was obtained through four acquisitions. Three-month-old rats gave MR images with low signal intensity over the liver probably due to the shorting of its T1 and T2 relaxation times. However, 12-month-old rats gave hyperintense regions around hepatic veins in right hepatic lobe, which was assigned to hepatocellular carcinoma. In 29-month-old rats, MR images with hyperintensity throughout the hepatic lobe were observed. These MR images, therefore, suggested that hepatocellular carcinoma in LEC rats developed from the restricted regions surrounding hepatic veins. In the present study, T1-weighted MR imaging under a magnetic field of 7.05 T was shown to be applicable to the diagnosis of hepatic cancer in LEC rats.

Animals↗
PubMed1997

Effects of alpha-phenyl N-tert-butylnitrone, a spin trap reagent, on the proliferation of murine hematopoietic progenitor cells in vitro.

We investigated the effect of alpha-phenyl N-tert-butylnitrone (PBN), a spin trap reagent, on the proliferation of murine hematopoietic progenitor cells in vitro. During the addition of PBN to the liquid cultures of murine bone marrow cells containing a combination of interleukin-3, interleukin-6 and the c-kit ligand/stem cell factor, colony-forming cells in vitro (CFC) and the colony-forming unit in the spleen (CFU-S) increased about 1.6-fold and 2.0-fold, respectively, higher than the control culture. These effects were not observed when using dimethyl sulfoxide, which has the ability to scavenge radicals, and 5,5-dimethyl-1-pyrroline N-oxide, another spin trap reagent. Analysis of cultured cells from a 7-day liquid culture with PBN revealed that the ratio of the intracellular glutathione (GSH) and GSH/GSSG (oxidized GSH) content was higher than the control. Adding thiol N-acetylcysteine, a thiol reagent and a precursor of intracellular GSH, also showed similar effects on the liquid culture of murine hematopoietic progenitor cells and the level of intracellular GSH. In contrast, adding DL-buthionine-[S,R]-sulfoximine, a gamma-glutamylcysteine synthetase inhibitor, decreased the intracellular GSH level and did not increase the number of CFC and CFU-S. These results suggest that PBN regulates the content of intracellular thiol molecules, and the possibility of a relationship between the intracellular redox state and the proliferation and differentiation of hematopoietic stem cells.

Acetylcysteine↗
PubMed1996

Characterization of natural killer cytotoxic factor (NKCF) from canine NK cells.

We investigated the presence of canine natural killer cytotoxic factor (NKCF). Canine natural killer (NK) cell-mediated cytotoxicity measured by 51chromium (51Cr) release assay was found to be highest in the T-cell population, which was fractionated into the 35-40% Percoll fraction by discontinuous gradient centrifugation. The cytotoxicity of NKCF in the culture supernatant showed a similar tendency to NK activity. Release of NKCF was rapid after contact with target cells, and reached a plateau in 60 min. The cytotoxicity of NKCF could be detected within at least 15 min in coculture with CL-1 target cells, reaching a plateau in 60 min. We also characterized canine NKCF and found it to be a protein, which was stable against both heat and cold treatment. These findings suggest that canine NK cells release NKCF immediately after recognition and binding to the target cell, and that NKCF plays an important role in canine NK-mediated cytotoxicity.

Animals↗
PubMed1996

The effects of alpha-phenyl-tert-butyl nitrone (PBN) on copper-induced rat fulminant hepatitis with jaundice.

In the present study we demonstrated the protective effects of the spin-trapping agent alpha-phenyl-tert-butyl nitrone (PBN) against fulminant hepatitis with jaundice in LEC rats. In LEC rats an excess amount of copper is accumulated in the liver and causes hepatitis with severe jaundice. PBN was subcutaneously administered every 2 d at the concentration of 128 mg/kg, beginning with 13-week-old rats and continuing for 17 weeks. PBN prevented the loss of body weight, reduced death rate, and suppressed the increase in GTP and GOT values reflecting hepatic cell destruction. Ocular inspection also confirmed the suppressive effects of PBN on jaundice. In parallel with these phenomena, the amounts of thiobarbituric acid-reactive substances (TBARS) in livers of PBN-administered rats were found to be lower than those of non-PBN-administered rats. Little histological changes were observed in PBN-administered rats in comparison with non-PBN-administered rats. The protective effect of PBN on the formation of oxidative damage in liver DNA was observed but not so remarkable as that on lipid peroxidation. From these results, it was concluded that PBN had the liver-protective effects against fulminant hepatitis with jaundice. This suggested that free radicals play an important role in abnormally accumulated copper-induced liver injury and that PBN potentially has therapeutic value for the treatment of hepatitis.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed1996

Functional characteristics of enhanced Fc receptor expression of beta 2 integrin-deficient bovine mononuclear phagocytes.

Fc receptor expression, cytoplasmic Ca2+ signaling, chemiluminescent (CL) response, and electron spin resonance (ESR) combined with spin trapping of blood mononuclear phagocytes from control heifers and a heifer with leukocyte adhesion deficiency (LAD) were evaluated to elucidate the relationships between complement receptor type 3 (CR3) and Fc receptor expression and their functional responses. The mean fluorescence intensity of fluorescein isothiocyanate (FITC)-conjugated anti-bovine IgG bound to mononuclear phagocytes from the heifer with LAD was 1.8-fold higher than that of control heifers. The mean increments of cytoplasmic Ca2+ concentrations of mononuclear phagocytes from the heifer with LAD stimulated with OPZ, Agg-IgG, and PMA were 39.4 (P < 0.05), 118, and 71.6% compared with those of control heifers. A 1.27-fold increase in the CL response relative to control heifers was detected when mononuclear phagocytes from the heifer with LAD were stimulated with Agg-IgG. The OPZ-induced CL response of mononuclear phagocytes from the heifer with LAD was significantly (P < 0.05) decreased, whereas the PMA-induced CL response was similar to that of control heifers. The ESR spectrum of mononuclear phagocytes from the heifer with LAD was increased when stimulated with Agg-IgG, and was impaired when stimulated by OPZ compared with that of control heifers. The ESR spectrum of mononuclear phagocytes stimulated with PMA was similar in control heifers and the heifer with LAD. Fc receptors on mononuclear phagocytes from the heifer with LAD were enhanced, and their cytoplasmic Ca2+ signaling, CL response, and ESR-spin trapping when stimulated with Agg-IgG and OPZ appeared to be associated with enhanced Fc receptors.

Animals↗
PubMed1996

Adhesiveness for extracellular matrices and lysosomal enzyme release from normal and beta 2 integrin-deficient bovine neutrophils.

The adhesiveness of control and CD18-deficient bovine neutrophils on culture plates precoated with collagen I, collagen IV, fibronectin and laminin was measured to evaluate the possible factors for adherence to extracellular matrices. The release of N-acetyl-beta-D-glucosaminidase (NAGase) from control and CD18-deficient neutrophils stimulated with complement receptor type 3 (CR3) or Fc receptor dependent stimuli was also evaluated. The adhesive activities of CD18-deficient neutrophils to collagen I, collagen IV and fibronectin were significantly diminished (P < 0.05); however, similar adhesion to laminin was observed in CD18-deficient neutrophils and control neutrophils. The adhesive activity of control neutrophils on uncoated plates increased 2.5 times (P < 0.05) with the presence of PMA. The mean activities for NAGase release from CD18-deficient neutrophils stimulated with opsonized zymosan and aggregated bovine immunoglobulin G (Agg-IgG) were 46.7 and 82.7% that of the control neutrophils, respectively. The Agg-IgG-induced NAGase release from control and CD18-deficient neutrophils was eliminated by H7, a protein kinase C inhibitor. These results support that an association between CR3 and Fc receptors on neutrophils appears to play an essential role in neutrophil functions.

Acetylglucosaminidase↗
PubMed1996

T lymphocytes and silica-induced pulmonary inflammation and fibrosis in mice.

BACKGROUND: Silica-induced pulmonary inflammation and fibrosis in animals provides a good model for chronic pulmonary inflammation and fibrosis. Although lymphocytes are implicated in the pathogenesis of pulmonary fibrosis, experimental models using silica-treated athymic nude mice have not been successful in showing the fibrogenic mechanism regulated by T cells. The aim of this study was to re-evaluate the role of T lymphocytes in the development of silicosis by comparing the response to silica administration of nude athymic mutants with that of euthymic animals. METHODS: Suspensions of silica particles were transnasally administered to nude athymic mice (Balb/c nu/nu) as well as to their euthymic littermates (Balb/c nu/+). The degree of pulmonary inflammation and fibrosis was assessed on days 14, 28, and 56 based upon histological observation, analysis of collagen deposition in the lungs, and analysis of the cellular constituent, protein, and phospholipid content in the bronchoalveolar lavage fluid. RESULTS: Histologically, athymic mice developed less severe interstitial pneumonitis than euthymic mice. In euthymic mice the lung hydroxyproline content increased with time after silica administration from 6.48 (0.38) micrograms hydroxyproline/mg dry lung weight on day 0 to 8.87 (0.41) micrograms/mg on day 56. A gradual increase in lung hydroxyproline content was also observed in athymic mice but the increase was significantly smaller than in euthymic mice (6.63 (0.43) micrograms/mg on day 0, 7.90 (0.19) micrograms/mg on day 56). Administration of silica resulted in an increase in the number of macrophages and neutrophils and in the total protein and phospholipid content of the bronchoalveolar lavage (BAL) fluid in both mouse strains. No significant difference was detected between athymic and euthymic mice in the numbers of macrophages, but the increase in neutrophils in the BAL fluid of athymic mice was significantly smaller than in euthymic mice on days 14 and 56. The total protein and phospholipid content of the BAL fluid from athymic mice was lower than that from euthymic mice. CONCLUSIONS: T lymphocytes appear to be involved in the pathogenesis of silica-induced pneumonitis. Since pulmonary fibrosis develops even in nude athymic mice, T cells do not seem to play a primary part in fibrogenic response but they regulate, at least to some extent, the response of inflammatory cells and fibrogenesis of the lung.

Animals↗
PubMed1996

Damage induced by hydroxyl radicals generated in the hydration layer of gamma-irradiated frozen aqueous solution of DNA.

Aqueous DNA solutions with or without the spin trap alpha-phenyl-N-tert-butylnitrone (PBN) were exposed to gamma-rays at 77 K. After thawing the solutions, three experiments were carried out to confirm the generation of OH radicals in the hydration layer of DNA and to examine whether they act as an inducer of DNA strand breaks and base alterations. Observation with the ESR-spin trapping method showed ESR signals from PBN-OH adducts in the solution containing PBN and DNA, but there were few signals in the solution containing PBN alone, suggesting that reactive OH radicals were produced in the hydration layer of gamma-irradiated DNA and were effectively scavenged by PBN, and that unreactive OH radicals were produced in the free water layer of gamma-irradiated DNA. Agarose gel electrophoresis of DNA proved that PBN had no effect on the formation of strand breaks, whereas examination with the high-performance liquid chromatography-electrochemical detection (HPLC-ECD) method showed that PBN suppressed the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG). From these results it was concluded that OH radicals generated in the hydration layer of gamma-irradiated DNA did not induce DNA strand breaks but induced base alterations.

Animals↗