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Biomedical subjects

H Ushio

Publications and source records attributed to H Ushio.

At least 55 records · Page 3Linked to original sources

Protective immunity and mast cell and eosinophil responses in mice infested with larval Haemaphysalis longicornis ticks.

WBB6F1-+/+ mice were infested with larval Haemaphysalis longicornis ticks twice at an interval of 14 days: apparent resistance against ticks was expressed in the second infestation. The first infestation induced degranulation of a small number of mast cells at the feeding sites within 6 days, and resulted in two-fold increases of mast cell numbers on day 14 with a significant elevation of total immunoglobulin E (IgE) levels in sera and high proportion of IgE-bound mast cells. The second infestation resulted in the intensive degranulation of the increased mast cells at the feeding sites. Eosinophils infiltrated into the feeding sites of ticks: the second infestation led to a greater maximal level of the infiltrating eosinophils. These data suggest that the resistance against larval H. longicornis ticks in mice may be expressed as a result of immediate hypersensitivity and many eosinophils infiltrating from the blood to the feeding sites might contribute to the tick rejection.

Animals↗

Supernormal histamine release and normal cytotoxic activity of beige (Chédiak-Higashi syndrome) rat mast cells with giant granules.

The beige rat is an animal model of the Chédiak-Higashi syndrome. Since mast cells can be easily purified from the peritoneal cavity of rats, we investigated the function of beige rat mast cells with giant granules by using quantitative methods. Beige and normal rat mast cells were sensitized with anti-dinitrophenol (DNP) IgE antibodies and stimulated by DNP conjugated with human serum albumin. The proportion of histamine released to total histamine was significantly greater in beige rat mast cells than in normal rat mast cells. Since the supernormal histamine release of beige rat mast cells was observed after treatment with 12-O-tetradecanoylphorbol 13-acetate, calcium ionophore A23187, substance P or compound 48/80, it appeared to be attributable to the enlargement in granules in beige rat mast cells. Spontaneous cytotoxic activity of mast cells was assayed by incubating purified mast cells with 51Cr-labelled WEHI-164 cells. Both beige and normal rat mast cells showed significant cytotoxic activity, but no significant difference was detectable between beige and normal rat mast cells. Even after IgE-mediated stimulation, no significant difference in cytotoxic activity was detectable between beige and normal rat mast cells either. Giant granules of beige rat mast cells did not appear to influence the cytotoxic activity of mast cells.

Animals↗

Murine granulocyte-macrophage and mast cell colony formation promoted by nerve growth factor.

The nerve growth factor (NGF) is widely distributed in the target tissues of sympathetic neurons. Hemopoietic organs such as the bone marrow (BM) and spleens are known to be innervated by noradrenergic sympathetic neurons. Some of their constitutive cells express NGF receptors (lymphocytes and stroma cells) and its biologic effects have been extensively studied in the immune system and inflammatory responses. However, the effects of NGF on hemopoiesis have been little examined. Recently, we demonstrated that NGF promoted mast cell colony formation from murine BM cells (BMC) or BMC-derived cultured mast cells and induced the phenotypic changes in standard hemopoietic assays. Besides, we demonstrated NGF-enhanced murine neutrophil survival and functional properties. In this study, in order to investigate NGF activities on neutrophil differentiation, we examined granulocyte-macrophage (GM) colony formation from murine BMC or spleen cells (SC) in the methylcellulose culture. We also assessed mast cell colony formation. GM colonies were counted on day 5 and mast cell colonies were counted on day 20 in culture. Although NGF alone (50 ng/ml) neither supported GM nor mast cell colony formation, addition of various doses of NGF to the suboptimal dose of pokeweed mitogen-stimulated SC-conditioned medium (2.5%) or interleukin 3 (50 U/ml), well-known colony-stimulating factors, increased the number of GM and mast cell colonies from both BMC and SC in a dose-dependent manner. These colony formation-enhancing effects of NGF were inhibited by the addition of neutralizing sheep anti-NGF antibodies. The results suggest that NGF may act to develop granulopoiesis including neutrophil and mast cell differentiation in cooperation with hemopoietic factor(s) during inflammatory processes.

Animals↗

Nerve growth factor induces development of connective tissue-type mast cells in vitro from murine bone marrow cells.

The effect of nerve growth factor (NGF) on proliferation/differentiation of mast cells was investigated in vitro. Although NGF alone neither supported colony formation of bone marrow-derived cultured mast cells (BMCMC) nor induced development of mast cell colonies from nonadherent bone marrow cells (NBMC), addition of NGF to the suboptimal dose of interleukin 3 (IL-3) significantly increased the numbers of mast cell colonies produced by BMCMC or NBMC in methylcellulose. When stimulated by IL-3 alone, cells in mast cell colonies were not stained by berberine sulfate, a fluorescent dye. In contrast, mast cells developing in methylcellulose cultures obtaining both IL-3 and NGF were stained by berberine sulfate. The fluorescence was abolished by the treatment of heparinase but not of chondroitinase ABC, suggesting that mast cells stimulated by IL-3 and NGF produced and stored heparin proteoglycan. The histamine content of BMCMC maintained by IL-3 was also increased by addition of NGF. Since BMCMC showed mucosal mast cell-like phenotype, NGF appeared to induce the phenotypic change to connective tissue-type mast cells (CTMC). In the culture containing BMCMC, 3T3 fibroblasts, and IL-3, the phenotypic change of BMCMC to CTMC was observed as well. Since NGF was detected in this coculture and since addition of anti-NGF monoclonal antibody suppressed the phenotypic change, NGF produced by fibroblasts appeared to induce the phenotypic change. Neither BMCMC alone nor IL-3 alone increased the concentration of NGF. Therefore, there is a possibility that BMCMC stimulated by IL-3 may induce the production and/or release of NGF by fibroblasts.

Animals↗

2.5S nerve growth factor enhances survival, phagocytosis, and superoxide production of murine neutrophils.

The effect of 2.5S nerve growth factor (NGF) on survival, phagocytosis, and superoxide production of murine neutrophils was examined and compared with the effects of interleukin-3 (IL-3) and recombinant GM colony-stimulating factor (rGM-CSF). NGF enhanced the viability of neutrophils isolated from peripheral blood and peritoneal cavity in a dose-dependent way. IL-3 50 U/mL had no effect. rGM-CSF 50 U/mL had an effect similar to that of 50 ng/mL NGF. NGF also enhanced the phagocytosis of hydrophilic microspheres by peritoneal neutrophils, and the activity of NGF was greater than that of IL-3 or rGM-CSF. NGF enhanced the superoxide production induced by phorbol 12-myristate 13-acetate (PMA), which acts at postreceptor sites, and that induced by opsonized zymosan, a receptor-mediated ligand. The stimulating activity of NGF was largely comparable to that of rGM-CSF. The present data show that NGF bound to neutrophils enhances their survival, phagocytosis, and superoxide production. Thus, we postulate that NGF plays an important role in the inflammatory processes.

Animals↗

Purification and characterization of a calsequestrin-like calcium-binding protein from carp (Cyprinus carpio) sarcoplasmic reticulum.

1. A calsequestrin-like calcium-binding protein was purified from carp sarcoplasmic reticulum by column chromatographies using DEAE-cellulose and Butyl-Toyopearl 650S. 2. The mol. wt was estimated to be 50 kDa, which was larger than that of rabbit calsequestrin (42 kDa). 3. Carp calsequestrin-like protein bound Ca2+ with a higher affinity (apparent Kd = 400 microM) and lower capacity (25 mol/mol) compared with rabbit calsequestrin (1 mM and 40-50 mol/mol, respectively). 4. Anti-carp calsequestrin-like protein rabbit antiserum reacted with rabbit calsequestrin in immunoblotting analysis. 5. Carp calsequestrin-like protein was rich in acidic amino acids, as was rabbit calsequestrin.

Amino Acids↗

Necessity of IgE antibodies and mast cells for manifestation of resistance against larval Haemaphysalis longicornis ticks in mice.

Genetically mast cell-deficient WBB6F1-W/Wv mice showed an apparent defect in manifestation of the resistance against larval Haemaphysalis longicornis ticks, but their serum IgE levels increased more than 100-fold after the second tick infestation. Immune sera obtained from the WBB6F1-W/Wv mice were adoptively transferred to the other WBB6F1-W/Wv mice which had received intracutaneous injections of WBB6F1-+/+ mouse-derived cultured mast cells. Because the resistance against ticks was detectable only when both mast cells and IgE antibodies were available, immediate hypersensitivity reaction appeared to have a physiologic role in the manifestation of the resistance against H. longicornis ticks.

Animals↗

Study of cell kinetics in anaplastic thyroid carcinoma transplanted to nude mice.

The cell kinetics of the transplantable human anaplastic thyroid carcinoma were studied in athymic nude mice using bromodeoxyuridine (BrdU) pulse labeling. In order to estimate the cell cycle time in this study, the percent labeled mitosis curve was prepared with immunoperoxidase staining using monoclonal antibody to BrdU instead of 3[H]thymidine autoradiography. This technique was considered to be very useful because it can be done for a short term and can still be changed to the autoradiographic method. Comparing the results in this study with already reported data on various xenografts of human carcinomas in nude mice, the tumor doubling time (117.5 hr) and the cell cycle time (23.5 hr) were rather short, but the growth fraction (54%) and the cell loss factor (68%) were nearly equivalent to the median value. These findings suggest that the very rapid growth rate of this tumor was mainly due to the shortness of the cell cycle time.

Animals↗

A case of metastasis from gastric cancer to the thyroid gland.

Metastatic tumors in the thyroid gland are rarely seen in clinical practice. This report describes a case of metastasis from gastric cancer to the thyroid, found five years after removal of the primary gastric lesion. The patient had a large thyroid mass extending to the mediastinum, but there were no obvious metastatic lesions anywhere except in the thyroid. Subtotal thyroidectomy was performed and histological examination revealed the same findings a those of the original gastric cancer. Of additional interest are the findings that led us to believe this metastatic tumor produced alpha-fetoprotein. Seven months following the operation, the patient dies suddenly and, although it is difficult to say whether the patient's survival was prolonged, we believe that the thyroid surgery gene him considerable palliation.

Adenocarcinoma↗

Bilateral resection and reconstruction of internal jugular vein for thyroid carcinoma.

Two patients with thyroid carcinoma infiltrating bilateral internal jugular veins were treated. In reconstruction of the internal jugular vein by implantation of an autogenous venous segment or Gore-Tex artificial vessel, the repaired area soon became obstructed. The repaired area with the Impra-Flex artificial vessel became obstructed again one month after the implantation. In the reconstruction by ipsilateral end-to-end anastomosis between the internal and external jugular veins, good circulation was apparent even 2 years after the operation. The internal-external jugular vein anastomosis is expected to increase the safety of single-stage bilateral excision of the internal jugular vein, as an adjunct to total thyroidectomy, in the surgical treatment of thyroid carcinoma.

Adult↗

Histological reconstruction of a Von Meyenburg's complex on the liver surface.

We report here a case with four black nodules on a smooth liver surface. The cystically dilated part of the intralobular bile duct of the microhamartoma contained bile pigments. Neither liver cysts nor cystic kidney were found. Spicular processes from the dilated bile duct were attached directly to the liver cell cords, seemingly to receive the secreted bile juice from the bile canaliculi. A convoluted tubular bile duct communicated with an interlobular terminal bile duct in the neighbouring portal triad. Thus, the so-called microhamartoma had not developed separately in the liver acini but had arisen in connection with both the liver cell cords and the terminal bile duct as part of the still functioning bile duct.

Bile Duct Neoplasms↗

Application of flow cytometry for rapid detection of Lactococcus garvieae.

Flow cytometry (FCM) technique was applied to rapid determination of cell number of Lactococcus garvieae. An antiserum against L. garvieae was prepared and its immunological property was examined. The present antibody would recognize some epitopes of L. garvieae with a high specificity. The optimum conditions for the FCM assay were as follows: discriminate value, 60; dilution ratio of the antiserum, 1.0 x 10(4). Calibration curve for L. garvieae cells was linear, in the range of 2.4 x 10(4)-1.5 x 10(7) cells/mL. The detection of L. garvieae in cell suspensions contaminated with Escherichia coli was carried out. A good correlation was observed in the range of 20-90% for the mixing ratio of L. garvieae. One FCM assay could be completed within 2 min, and the total assay time, including the preparation of bacterial sample, was within 3 h.

Animals↗

A role for platelet release of serotonin in the initiation of contact sensitivity.

Finding contact sensitivity (CS) responses that were fairly normal in ear swelling, and in serotonin (5-HT) dependence in mast-cell-deficient mice, led to experiments to determine whether platelets supplemented mast cells as a source of 5-HT in CS. Severe depletion of platelets, and consequently blood 5-HT, with antiplatelet antibody, strongly inhibited CS, especially in mast-cell-deficient mice, suggesting that platelets supplemented mast cells. Furthermore, human platelets sensitized in vitro with anti-(tri-nitro-phenyl) IgE, and transferred intravenously together with isolated late-acting effector T cells, provided CS initiation due to local 5-HT release. Similar, IgE-dependent in vitro release of 5-HT was C dependent. These findings establish the importance of antigen-specific platelet release of 5-HT in CS initiation.

Amphetamine↗