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Biomedical subjects

H Watanabe

Publications and source records attributed to H Watanabe.

At least 1,009 records · Page 56Linked to original sources

Serotyping of Streptococcus pyogenes isolated from common and severe invasive infections in Japan, 1990-5: implication of the T3 serotype strain-expansion in TSLS. The Working Group for Group A Streptococci in Japan.

To clarify the relationship between the epidemics of severe invasive group A streptococcal infections (streptococcal Toxic Shock-Like Syndrome: TSLS) and common group A streptococcal infections in Japan, we examined the T serotypes of S. pyogenes strains (group A streptococci) isolated from clinical specimens of the streptococcal infections (17999 cases) in the period 1990-5, including the severe infections (TSLS) (29 cases) in the period 1992-5. Characteristic points of the analyses were: (1) dominant serotypes of the infections in these periods were T12, T4, T1, T28 and TB3264, which were consistently isolated; (2) isolates of T3 rapidly increased through 1990 to 1994 while T6 decreased in the period 1990-3; (3) when Japanese area was divided into three parts, T3 serotype tended to spread out from the north-eastern to the south-western area; (4) strains of T3 and T1 serotypes were dominant in the TSLS. Dominant-serotype strains of streptococcal infections did not always induce severe infections and dominance of T3 serotype in the TSLS seemed to be correlated with the increase of T3 in streptococcal infections. These results may indicate that certain clones of S. pyogenes are involved in the pathogenesis of the TSLS.

Adolescent↗

Numerical and functional characteristics of lymphocyte subsets in centenarians.

The immune system in the aged is a very interesting subject for study. In this study, analysis was extended to extrathymic T cells as well as NK cells and "conventional" T cells (i.e., thymus-derived T cells) in terms of their constitution and function in both healthy and unhealthy centenarians. Middle-aged persons were used as controls. Healthy and unhealthy centenarians showed lower levels in the proportion and absolute number of lymphocytes. The major change in the constitution of lymphocyte subsets was increased levels in the proportion of NK cells (CD56+/CD57+) and extrathymic T cells (CD3+CD57+). Inversely, conventional T cells decreased in proportion and function (i.e., proliferative response to mitogen). Although NK cells increased in centenarians, NK activity by whole lymphocytes and the purified NK fraction decreased. The difference between healthy and unhealthy centenarians was small in all parameters, the only difference being a lower level of expression of CD56 antigens on CD57+ T cells in unhealthy centenarians. These results indicate that there is a major shift in lymphocyte population from conventional T cells to NK cells and extrathymic T cells with aging. Concerning the age-associated increases in CD56+ T and CD57+ T cells, these cells correspond to NK1+ T cells in mice.

Aged↗

A monoclonal antibody, DL10, which recognizes a sugar moiety of MHC class I antigens expressed on NK cells, NK+ T cells, and granulocytes in humans.

One mAb, DL10, was established from mice injected with dolphin lymphocytes. In addition to its reactivity against all dolphin lymphocytes, it reacted with some human leukocytes, including NK cells, NK+ T cells, and granulocytes. When its reactivity was examined in various animals, bovine, ovine, and equine leukocytes were DL10+. Murine, rat, and canine leukocytes were DL10-. Although the reactivity of DL10+ was similar to those of CD56 and CD57 antigens in humans, the actual molecules it recognized were different. Thus, all reactivity of DL10 disappeared after treatment of cells with glycopeptidase or after culture of cells with tunicamycin. Furthermore, the immunoprecipitation method revealed that DL10 indirectly recognized the heavy chain (45kD) of MHC class I antigen in humans and animals. Considering data from analysis of the N-terminal amino acid sequence of the DL10 molecule and the HLA typing of reactive cells, DL10 recognized a sugar moiety of some monomorphic MHC antigens and polymorphic MHC antigens such as HLA-B60 and -B61. If the donors are HLA-B60- and -B61 (> 80% in Japan and > 95% in the United States), DL10 would appear to be a very useful agent for the detection of pan-NK+ T cells.

Amino Acid Sequence↗

Self-reactive forbidden clones are confined to pathways of intermediate T-cell receptor cell differentiation even under immunosuppressive conditions.

It is believed that self-reactive forbidden T-cell clones are generated by 'failure' of the pathway of T-cell differentiation in the thymus, if it is disturbed. We examined how such forbidden clones are generated under immunosuppressive conditions. Mice were treated with an injection of deoxyspergualin, FK506, or cycloporin A. From day 3, the number of cells yielded by various organs decreased. Because of the resistance of intermediate (int) T-cell receptor (TCR) cells (i.e. TCRint cells), they became more prominent in proportion than TCRhigh cells. TCRhigh cells are conventional T cells generated through the mainstream in the thymus, whereas TCRint cells are primordial T cells generated by the extrathymic pathway or an alternative intrathymic pathway. Similar to untreated mice, forbidden V beta 3+ and V beta 11+ clones in C3H/He (Mls-1b2a) mice were confined to TCRint cells after treatment; there was no leakage of forbidden clones into TCRhigh cells in the thymus and periphery. In parallel with the increase in the proportion of TCRint cells, the proportion of forbidden clones also increased under immunosuppressive states, especially in the liver. Liver mononuclear cells isolated from treated mice still had the potential to mediate autologous killing. The present results suggest that the generation of self-reactive clones is highly restricted to the pathways of TCRint cell differentiation even under immunosuppressive conditions.

Animals↗

Autologous killing by a population of intermediate T-cell receptor cells and its NK1.1+ and NK1.1- subsets, using Fas ligand/Fas molecules.

Self-reactive clones, estimated by anti-V beta monoclonal antibodies (mAb) in conjunction with the Mls system, are confined to a population of intermediate (int) T-cell receptor (TCR) (or CD3) cells (i.e. TCRint cells), but are not found among TCRhigh cells. The next questions to be answered are whether autologous killing is confined to TCRint cells and how such killing is mediated. In this study, 51Cr-labelled thymocytes of syngeneic or allogeneic origin were used as target cells (4-hr assay). When liver and splenic mononuclear cells (MNC) obtained from B6 mice were used as effector cells, prominent autologous killing was seen in liver MNC, but not splenic MNC. Such killing was not seen when thymocytes from B6-lpr/lpr mice (i.e. Fas-) were used as target cells, nor when liver MNC from MRL-gld/gld mice (i.e. Fas ligand-) were used as effector cells (target thymocytes of MRL(-)+/+ mice). Cell separation experiments using a cell sorter revealed that autologous killing was mediated for the most part by CD3int cells, while allogeneic killing was mediated entirely by natural killer (NK) cells, TCRint cells and TCRhigh cells. Among CD3int cells, the NK1.1+ subset (i.e. NK1.1+ T cells) manifested a higher level of autologous killing than did the NK1.1- subset. Consistent with the results of a functional assay, it was found by reverse-transcription-polymerase chain reaction (RT-PCR) assay that CD3int cells among liver MNC showed the expression of Fas ligand mRNA, while thymocytes expressed Fas mRNA. When class I major histocompatibility complex (MHC)- thymocytes (from beta 2-microglobulin-deficient mice) were used as target cells, NK cells, but not CD3int cells, showed potent cytotoxicity. These results suggest that autologous killing is a major function of TCRint cells with self-reactivity, and that such killing is mediated by means of Fas ligand/Fas molecules.

Animals↗

Antimetastatic effect of NK1+ T cells on experimental haematogenous tumour metastases in the liver and lungs of mice.

Depletion of both natural killer 1.1+ (NK1+) intermediate alpha beta T-cell receptor (int T) cells and NK cells by in vivo treatment with anti-NK1 antibody greatly increased hepatic metastases of intravenously injected EL4 cells as well as pulmonary metastases of 3LL cells in C57BL/6 mice. However, depletion of NK cells alone by anti-asialo GM1 (AGM1) antibody treatment did not increase the metastases in either organ. Interleukin-12 (IL-12) administration into mice induced strong cytotoxicities of NK cell-depleted liver and lung mononuclear cells (MNC) comparable to those without NK-cell depletion and inhibited metastases in either organ. In contrast, in both NK cell- and NK1+ int T-cell-depleted mice, IL-12 could not induce cytotoxic activity of liver and lung MNC and metastases in both organs increased with or without IL-12 treatment. These results confirmed the fact that NK+ int T cells are more potent antitumour effectors than NK cells against experimental haematogenous tumour metastases.

Animals↗

Neonatal granulocytosis is a postpartum event which is seen in the liver as well as in the blood.

In a recent series of studies, we demonstrated that stress in humans and animals, with resultant sympathetic nerve strain, induces severe granulocytosis, because granulocytes carry adrenergic receptors on the surface. Because activated granulocytes produce free radicals and superoxides, they sometimes induce tissue damage if the stress is too strong or continuous. Human neonates are also known to show high levels of granulocytes in the peripheral blood. In this study, we investigated whether such neonatal granulocytosis are a stress-associated response at birth. Both human and mouse materials, before and after birth, were used. The number of leukocytes in the blood, as well as some other factors in the serum, were measured. Although levels of granulocytes were found to be low in fetal humans and mice, they increased sharply after birth. In parallel with this postpartal granulocytosis, transaminases in sera increased transiently. In reference to results of a transient elevation in the levels of catecholamines at birth in mice, all these phenomena resemble stress-associated responses. Indeed, fatty liver and hematopoietic destruction in the liver were also observed in mice and humans. At this time, the production of inducible nitric oxide synthase (iNOS) by granulocytes in the liver was evident. These results suggest that neonatal granulocytosis is a postpartum event which results from various stresses (e.g., oxygen stress) at birth. This event may be responsible for such well-known neonatal phenomena as the termination of fetal hematopoiesis in the liver and as neonatal jaundice.

Alanine Transaminase↗

The significance of tumor necrosis factor (TNF) levels for rejection of joint allograft.

This paper describes the efficacy of utilizing Tumor Necrosis Factor-alpha (TNF) as a detectable parameter of acute rejection after bone allografts. The authors used a bone-joint allograft model, transferring vascularized allograft knee joints across a major histocompatibility barrier, using three groups of rats divided into controls, non-immunosuppressed, and immunosuppressed with cyclosporine. The paper supplies preliminary information suggesting that TNF may be a marker for early bone-joint allograft rejection.

Animals↗

The omental flap with skin grafting for treatment of a difficult sternal closure.

A case report of difficult sternal closure is described. Chest closure following cardiac operation in critically ill patients can be a problem. Techniques have been described so that sternal closure can be delayed until the patient is hemodynamically stable and hemostasis has been achieved. However, in some instances, the sternal closure can not be performed because of prolonged cardiopulmonary instability. We describe the use of transposition of an omental flap with skin grafting in a 74-year-old patient with aortic valve stenosis, poor pulmonary function, and a thoracic deformity. This technique enabled incomplete sternal closure which maintained hemodynamic stability.

Aged↗

A competitive enzyme immunoassay for follicle-stimulating hormone in ovine plasma using biotin-streptavidin amplification.

To determine follicle-stimulating hormone (FSH) concentration in ovine plasma, a second-antibody enzyme immunoassay using biotin-streptavidin amplification (BS-EIA) was developed and was validated by comparison with a routine radioimmunoassay (RIA). Ovine FSH was labelled with D-biotinyl-epsilon-aminocaproic acid-N-hydroxysuccinimide ester (biotin-7-NHS). The minimum and maximum detection limits were 0.40 ng mL(-1) and 25.0 ng mL(-1) respectively in the BS-EIA. Plasma samples from three superovulated ewes were measured for FSH by both BS-EIA and RIA. Correlation between values obtained by BS-EIA and RIA was 0.86 (n = 106), and the linear regression had a slope of 0.88 and an intercept of -0.07 (R2 = 0.74). The baseline of BS-EIA was slightly lower than that of RIA (0.69 ng mL(-1) v. 1.10 ng mL(-1)). The intra-assay and inter-assay coefficients of variation in BS-EIA were 13.8% and 16.7% (n = 106) respectively. The recovery rates were 104.8+/-2.7% (mean+/-s.e.m., n = 14). Changes in plasma FSH concentration from superovulated ewes evaluated by BS-EIA corresponded well with the results of routine RIA.

Animals↗

Effects of heavy ions on the germination and survival of Arabidopsis thaliana.

Inhibition of germination and reduction in survival of Arabidopsis thaliana were investigated to study the effects of heavy ions on a multicellular system. Dry seeds of Col and Ler ecotypes were exposed to He, C, Ar and Ne ions with linear energy transfer (LET) in the range of 17-549 keV/micron and to electrons (LET = 0.2 keV/micron). The relative biological effectiveness (RBE) for the survival of both ecotypes showed the same pattern of variation with a maximum RBE of 11-12 at 252 keV/micron. For germination, RBE increased with increasing LET in Ler but not in Col, showing different sensitivities between the plant ecotypes. Inactivation cross sections of survival increased linearly in the range of 0.2-17 keV/micron and proceeded more steeply in the range of 113-252 keV/micron. At higher LET, cross sections appeared to reach a plateau at a little less than the size of the cell nucleus. When the value for survival was plotted against LET, it decreased steeply in the range about 113-252 keV/micron, indicating that heavy ions may have similar effects on both the shoulder and slope of the survival curve.

Arabidopsis↗

Absence of human papillomavirus-16 and -18 DNA and Epstein-Barr virus DNA in esophageal squamous cell carcinoma.

To elucidate the association of human papillomavirus (HPV) and Epstein-Barr virus (EBV) with carcinogenesis of the esophagus, 41 surgically resected specimens and 12 cell lines of esophageal squamous cell carcinoma were examined for the presence of HPV DNA and EBV DNA by polymerase chain reaction using primers for the E6 regions of HPV-16 and -18 and for the EBNA 1 region of EBV. We designed the reaction condition to amplify HPV and EBV DNA specifically and detected by gel electrophoresis. In ethidium bromide staining, no band was detected either for the HPV E6 region of for the EBV EBNA 1 gene in any of the surgically resected specimens and the cell lines, although the HPV sequence was detectable by Southern blot hybridization, which is a more sensitive detection method than staining; three out of 41 surgically resected specimens were positive for HPV-18 by Southern blot hybridization of polymerase chain reaction products. However, the number of viral genomes has been estimated as lower than 1 x 10(-3) copies per cell based on the intensity of the hybridization signals. Moreover, the DNA samples extracted from the corresponding non-cancerous esophageal mucosa were also positive for HPV-18, and the intensities of the hybridization signals were almost the same as those of the tumors. The results of our study indicate that HPV-16, HPV-18 and EBV are not generally associated with esophageal carcinogenesis.

Carcinoma, Squamous Cell↗

Metastatic esophageal tumors from distant primary lesions: report of three esophagectomies and study of 1835 autopsy cases.

Three cases of esophagectomy for secondary esophageal carcinoma metastasized from the ovary, breast and lung are reported. Long-term survival, 14 and 4 years, after esophagectomy was achieved in two patients. The intervals between surgery for primary cancer and dysphagia onset in these two patients were 16 and 7 years, respectively. An aggressive surgical approach appears to be the therapeutic procedure of choice for metastatic esophageal carcinoma when the primary tumor growth rate is suspected to be slow. Autopsy data on 1835 cases revealed 112 (6.1%) had metastasis to the esophagus. The lung was the most common primary tumor-bearing organ and the diffusely infiltrative type was the most common esophageal tumor observed macroscopically which corresponded to the findings in our three patients. When an esophageal stricture with normal mucosa is encountered, a metastatic tumor must be taken into consideration.

Adenocarcinoma↗

Characterization and regulation of taurine transport in Caco-2, human intestinal cells.

We characterized the taurine transport system in human intestinal Caco-2 cells and showed that it is subject to adaptive regulation. The activity of taurine transport in Caco-2 cells was evaluated by means of an Na+- and Cl(-)-dependent high-affinity transport system, the characteristics of which were similar to those of the beta-amino acid-specific taurine transport system previously described for various tissues. The activity of taurine transport was down-regulated on culturing in taurine-containing medium. This taurine-induced down-regulation was dependent on both the incubation time with taurine and the concentration of taurine. Hypotaurine and beta-alanine were also capable of inducing this adaptive regulation, whereas alpha-amino acids and gamma-aminoisobutyric acid were not. Kinetic analysis of control and taurine-treated cells suggested that the down-regulation was associated with a decrease in the maximal velocity of taurine transport and also with a decrease in the affinity of the taurine transporter. Cycloheximide treatment weakened the taurine-induced down-regulation. The mRNA level of the taurine transporter (HTAU type) in taurine-treated cells was markedly decreased compared with in control cells. These results indicate that a complex regulatory mechanism is involved in this down-regulation.

Adaptation, Physiological↗

L-ornithine decarboxylase from Hafnia alvei has a novel L-ornithine oxidase activity.

A novel activity producing gamma-aminobutyric acid (GABA) from L-ornithine in the presence of NAD(P)+ was found in the crude extract of L-ornithine-induced Hafnia alvei, in addition to L-ornithine decarboxylase (ODC) activity. The reaction system for the former activity consisted of two enzymes, L-ornithine oxidase (decarboxylating, OOD) and gamma-aminobutyraldehyde (GABL) dehydrogenase (GDH). OOD catalyzed the conversion of L-ornithine into GABL, CO2, NH3, and H2O2 in the presence of O2, and GDH dehydrogenated GABL to GABA in the presence of NAD(P)+. OOD, purified to homogeneity, had a high ODC activity and the activity ratio of ODC to OOD was almost constant throughout the purification (ODC/ OOD=160:1). The molecular mass of the OOD was about 230 kDa, probably consisting of three identical subunits of a 77 kDa peptide, and OOD had an absorption maximum at 420 nm as well as at 278 nm, the specific absorption for an enzyme containing pyridoxal phosphate (PLP). The content of PLP was estimated at about 1 mol per subunit. OOD was specific to L-ornithine, and other L-amino acids and polyamines including putrescine were inert. The enzyme was activated by PLP, but not by pyridoxamine 5'-phosphate, FAD, FMN, or pyrroloquinoline quinone, and it was inactivated by hydrazine, semicarbazide, and hydroxylamine. The holoenzyme can be resolved to the apoenzyme by incubation with hydroxylamine, and reconstituted with PLP. These properties of OOD were almost the same as those of ODC separately purified to homogeneity from H. alvei. Zn2+ and Cu2+, butanedione, and sodium borohydride inhibited both OOD and ODC in a similar manner. The OOD reaction required O2 and only the ODC reaction proceeded under anaerobic conditions. The substitution of air for oxygen in the reaction vessel and the addition of catalase-H2O, enhanced only the OOD reaction, resulting in an increase of the ratio of OOD/ODC to 1:30 and 1:4.1, respectively. These results suggested that OOD and ODC are identical and that the former is a side reaction of the latter in the presence of O2.

Aldehyde Oxidoreductases↗