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Biomedical subjects

H Amano

Publications and source records attributed to H Amano.

At least 55 records · Page 3Linked to original sources

Cultured human mast cells derived from umbilical cord blood cells in the presence of stem cell factor and interleukin-6 cannot be a model of human skin mast cells: fluorescence microscopic analysis of intracellular calcium ion mobilization.

To know whether cultured human mast cells raised from umbilical cord blood cells in the presence of stem cell factor (SCF) and interleukin-6 (IL-6) can be a model of human skin mast cells, the cells were stimulated, and intracellular calcium ion ([Ca(2+)](i)) mobilization was analyzed by fluorescence microscopic techniques in parallel with a measurement of histamine released from the cells. When IgE-sensitized mast cells were activated by anti-IgE, [Ca(2+)](i) elevation began at the periphery and subsequently proceeded toward the center of the cells. The increase in [Ca(2+)](i) in calcium ionophore A23187-stimulated mast cells began at the center and spread to the periphery of the cells. Significant histamine release was observed by each stimulation. However, either compound 48/80 or substance P failed to increase [Ca(2+)](i) with no appreciable histamine release. This study shows that there is heterogeneity of [Ca(2+)](i) mobilization in the activated human mast cells, and that cultured human mast cells derived from umbilical cord blood cells in the presence of SCF and IL-6 can not be a model of human skin mast cells.

Antibodies, Anti-Idiotypic↗

Process of egg formation in the female body cavity and fertilization in male eggs of Phytoseiulus persimilis (Acari: Phytoseiidae).

The process of egg formation in the body cavity of a phytoseiid mite, Phytoseiulus persimilis, was observed to examine fertilization of male eggs. After insemination, one of the ova at the periphery of the ovary began to expand, taking up yolk. Two pronuclei appeared in the expanded egg, located dorsally in the ovary, and yolk granules were formed gradually. After the egg became filled with yolk granules the two pronuclei fused. The egg moved via the narrow entrance at the ventral region into the oviduct, where the eggshell was formed. When the eggshell was complete, and while embryogenesis proceeded, the egg was deposited. In the meantime some ova began to expand sequentially and two joining pronuclei appeared in expanding eggs. The joining pronuclei in the first egg proved male diploidy. This is additional evidence of pseudo-arrhenotoky in this phytoseiid mite species, since the first eggs developed into males.

Animals↗

Walking speed as a good predictor for the onset of functional dependence in a Japanese rural community population.

OBJECTIVE: to investigate and compare the predictive values of four physical performance measures for the onset of functional dependence in older Japanese people living at home. DESIGN: a population-based prospective cohort study. SETTING: Nangai village, Akita Prefecture, Japan. METHODS: out of the population aged 65 years and older living in Nangai (n = 940) in 1992, we measured hand grip-strength, one-leg standing, and usual and maximum walking speeds in 736 subjects who were independent in the five basic activities of daily living. Their functional status was assessed each year for the subsequent 6 years. The outcome event was the onset of functional dependence, defined as a new disability in one or more of the five basic activities of daily living, or death of a subject who had shown no disability at the previous follow-up. RESULTS: even after controlling for age, sex and a number of chronic conditions, lower scores on each baseline performance measure showed increased risk for the onset of functional dependence. Maximum walking speed was most sensitive in predicting future dependence for those aged 65-74 years, while usual walking speed was most sensitive for people aged > or = 75 years. CONCLUSION: walking speed was the best physical performance measure for predicting the onset of functional dependence in a Japanese rural older population.

Activities of Daily Living↗

Impairment of endotoxin-induced macrophage inflammatory protein 2 gene expression in alveolar macrophages in streptozotocin-induced diabetes in mice.

To elucidate the mechanism of the high incidence of lower respiratory tract infections in patients with diabetes mellitus, we investigated the kinetics of production of macrophage inflammatory protein 2 (MIP-2), an important mediator of lung neutrophil recruitment, using mice with streptozotocin-induced diabetes. Intratracheal challenge with 1 mg of lipopolysaccharide (LPS), an endotoxin, per kg of body weight resulted in a time-dependent increase in the levels of MIP-2 protein in bronchoalveolar lavage (BAL) fluid, with the peak concentration (49.4 +/- 13 ng/ml) occurring at 3 h and significant neutrophil accumulation becoming apparent by 3 h in normal mice. In diabetic mice, the peak level of MIP-2 protein in BAL fluid did not occur until 6 h and was reduced to 21.9 +/- 10 ng/ml. Immunohistochemical studies using anti-MIP-2 antibody confirmed that the main cellular source of MIP-2 in the lung after LPS challenge was alveolar macrophages (AMs) in normal mice. The lungs in diabetic mice, however, showed no AMs staining for MIP-2 within 3 h after LPS challenge. PCR analysis using whole-lung RNA showed a time-dependent increase in MIP-2 mRNA levels after LPS instillation. The level of MIP-2 mRNA in diabetic mice was markedly decreased compared to that in normal mice. Our results indicate that impairment of MIP-2 mRNA expression in the AMs in diabetic mice resulted in delayed neutrophil recruitment in the lungs, and this may explain the development and progression of pulmonary infection in diabetes mellitus.

Animals↗

Effect of intravenous cyclophosphamide in systemic lupus erythematosus: relation to lymphocyte subsets and activation markers.

Since the mechanism mediating the beneficial effect of intravenous cyclophosphamide (IVCY) in systemic lupus erythematosus (SLE) is unknown, we investigated lymphocyte subsets and markers of activated lymphocytes in patients received IVCY, and compared the results with the effect of steroid pulse. In 55 patients with SLE, 34 patients receiving IVCY [21 cases (61.8%) were responsive] and 25 patients received steroid pulse [21 cases (84.0%) were responsive] (four patients who were resistant to steroid pulse therapy were transferred to IVCY). When the lymphocyte subsets and markers of activated lymphocytes were compared in the responsive and unresponsive group of IVCY, soluble CD4 levels and the ratio of HLA-DP-positive T cells were significantly higher in the unresponsive group. Further, the changes of these markers and costimulatory molecules [LFA-1 (CD11a), ICAM-1 (CD54), CD40 and CD40-ligand (CD154)] were also examined in the responsive patients. The ratio of HLA-DP-positive T cells did not change in the IVCY-responsive group, while it decreased in the steroid pulse therapy-responsive group. The ratio of CD11a on T cells increased and CD54 on B cells decreased in the IVCY-responsive group. The ratio of CD154 on T cells increased in the steroid pulse-responsive group, while it decreased in the IVCY-responsive group. These results suggest that the effect of IVCY is different to that of steroid pulse therapy and mainly related to B cell activation, and that these markers may contribute to predict the responsiveness of IVCY.

Adult↗

Importance of membrane- or matrix-associated forms of M-CSF and RANKL/ODF in osteoclastogenesis supported by SaOS-4/3 cells expressing recombinant PTH/PTHrP receptors.

SaOS-4/3, a subclone of the human osteosarcoma cell line SaOS-2, established by transfecting the human parathyroid hormone/parathyroid hormone-related protein (PTH/PTHrP) receptor complementary DNA (cDNA), supported osteoclast formation in response to PTH in coculture with mouse bone marrow cells. Osteoclast formation supported by SaOS-4/3 cells was completely inhibited by adding either osteoprotegerin (OPG) or antibodies against human macrophage colony-stimulating factor (M-CSF). Expression of messenger RNAs (mRNAs) for receptor activator of NF-kappaB ligand/osteoclast differentiation factor (RANKL/ODF) and both membrane-associated and secreted forms of M-CSF by SaOS-4/3 cells was up-regulated in response to PTH. SaOS-4/3 cells constitutively expressed OPG mRNA, expression of which was down-regulated by PTH. To elucidate the mechanism of PTH-induced osteoclastogenesis, SaOS-4/3 cells were spot-cultured for 2 h in the center of a culture well and then mouse bone marrow cells were uniformly plated over the well. When the spot coculture was treated for 6 days with both PTH and M-CSF, osteoclasts were induced exclusively inside the colony of SaOS-4/3 cells. Osteoclasts were formed both inside and outside the colony of SaOS-4/3 cells in coculture treated with a soluble form of RANKL/ODF (sRANKL/sODF) in the presence of M-CSF. When the spot coculture was treated with sRANKL/sODF, osteoclasts were formed only inside the colony of SaOS-4/3 cells. Adding M-CSF alone failed to support osteoclast formation in the spot coculture. PTH-induced osteoclast formation occurring inside the colony of SaOS-4/3 cells was not affected by the concentration of M-CSF in the culture medium. Mouse primary osteoblasts supported osteoclast formation in a similar fashion to SaOS-4/3 cells. These findings suggest that the up-regulation of RANKL/ODF expression is an essential step for PTH-induced osteoclastogenesis, and membrane- or matrix-associated forms of both M-CSF and RANKL/ ODF are essentially involved in osteoclast formation supported by osteoblasts/stromal cells.

Animals↗

[Alcohol consumption and change of activity of daily living among the elderly living in an urban community].

Alcohol consumption has both adverse and beneficial effects on survival. We examined the baseline status of alcohol consumption and the changes of instrumental activity of daily living (I-ADL) by a prospective survey for the elderly living in an urban community who were participants in the "Longitudinal and Interdisciplinary Study on Aging by the Tokyo Metropolitan Institute of Gerontology (TMIG-LISA)". Detailed information on alcohol intake was collected during a structured interview. At a baseline survey in 1991, 1) the frequency of current drinkers decreased lineally by age. 2) the most popular alcohol beverage was beer for both sexes and average ethanol intake per day were 41 (10-45) g in men and 24 (8-28) g in women. Current drinkers had significantly stronger grip strength, higher velocity in maximum walking and higher score on the TMIG Index of Competence, designed to measure the higher-level competence in elderly community residents. After 4 years follow-up, in 1995, 73.3 percent of the participants completed the examination. They were classified into five categories as "current drinkers", "lifetime abstainers", "beginning to drink during follow-up period", "quit drinking during follow-up period" and "quit drinking at baseline". The decline of I-ADL was significantly greater among the elderly who "quit drinking during follow-up period" than "current drinkers". Further "current drinkers" had significantly lower decline of I-ADL than "lifetime abstainers". Our findings show beneficial effects of moderate alcohol intake in the maintenance of a higher level of competence in the elderly living in a community.

Activities of Daily Living↗

[Proposal of a new lymph node mapping system for pancreatic head cancer].

The Japanese Pancreatic Society published the 4th edition of the general guidelines for the study of pancreatic cancer (JPS guidelines) in 1993. The current JPS guidelines differ from the UICC classification of lymph nodes. Thirty-five lymph node stations are divided into three groups in the JPS guidelines. Group 1 (n1) corresponds closely with the regional lymph nodes in the UICC classification. Group 2 (n2) and group 3 (n3) are distant metastases in the UICC classification. To clarify practical lymph node mapping, 2,449 resected tubular adenocarcinomas of the pancreas head registered with the JPS were analyzed. Lymph node involvement was observed most frequently in posterior pancreatoduodenal (PPD) and anterior pancreatoduodenal (APD) and then in paraaortic (n2) nodes. Fifty-six patients who achieved 5-year survival were registered, of whom 19 had lymph node involvement. The sites of lymph node involvement were limited to the PPD and/or APD nodes in 14 cases. PPD and APD nodes differ from the other n1 in terms of high incidence of metastasis and fair prognosis. In conclusion, we propose a new practical lymph node mapping system in which PPD and APD nodes are the first station and the other n1 are the second station.

Humans↗

Mechanisms of proton transport in isolated rat osteoclasts attached to bone.

Osteoclasts resorb bone by transporting protons (H+) into the space between the cell and the bone. To investigate the roles of a sodium-hydrogen exchanger (NHE) and a vacuolar type H+-ATPase (V-ATPase) in H+ extrusion of osteoclasts attached to bone, changes in intracellular pH (pHi) were monitored in osteoclasts isolated on glass coverslips and on bone slices using a fluorescent pHi indicator. Acid-loaded osteoclasts on glass coverslips recovered their pHi in the presence of Na+. The pHi recovery was inhibited by 5-(N, N-hexamethylene) amiloride (HMA), an inhibitor of NHE, or by removal of Na+. 7-chloro-4-nitrobenz-2-oxa-1, 3-diazol (NBD-CI), a blocker of V-ATPase, did not block the H+ transport in osteoclasts on glass coverslips. Acid-loaded osteoclasts on bone slices recovered their pHi in the absence of Na+, and this recovery was blocked by NBD-Cl. In addition, neither HMA nor NBD-Cl inhibited the pH, recovery in the presence of Na+. These results indicated that osteoclasts attached to bone extrude H+ by both V-ATPase and NHE and that osteoclasts attached to glass mainly extrude H+ by NHE.

4-Chloro-7-nitrobenzofurazan↗

Anthropogenic radionuclides in seawater of the Far Eastern Seas.

Large quantities of radioactive wastes have been dumped in the Far Eastern Seas by the former Soviet Union and the Russian Federation, and small amounts of radioactive wastes have been dumped by Japan and the Republic of Korea. In order to investigate the concentrations of anthropogenic radionuclides in the nine dumping areas, a second expedition was conducted in 1995 by Japan, the Republic of Korea, the Russian Federation and IAEA, following the first expedition in 1994. The results show that 137Cs, 90Sr and 239 + 240Pu concentrations in surface and bottom waters at dumping areas do not significantly differ from the values observed in background areas, and from historical values. There is no clear effect of possible contamination due to radioactive waste dumping. The concentrations and water column inventories of 137Cs, 90Sr and 239 + 240Pu in the Far Eastern seas are controlled by physical oceanic processes such as horizontal transport and biogeochemical processes such as scavenging.

Cesium Radioisotopes↗

Vascular endothelial growth factor can substitute for macrophage colony-stimulating factor in the support of osteoclastic bone resorption.

We demonstrated previously that a single injection of recombinant human macrophage colony-stimulating factor (rhM-CSF) is sufficient for osteoclast recruitment and survival in osteopetrotic (op/op) mice with a deficiency in osteoclasts resulting from a mutation in M-CSF gene. In this study, we show that a single injection of recombinant human vascular endothelial growth factor (rhVEGF) can similarly induce osteoclast recruitment in op/op mice. Osteoclasts predominantly expressed VEGF receptor 1 (VEGFR-1), and activity of recombinant human placenta growth factor 1 on osteoclast recruitment was comparable to that of rhVEGF, showing that the VEGF signal is mediated through VEGFR-1. The rhM-CSF-induced osteoclasts died after injections of VEGFR-1/Fc chimeric protein, and its effect was abrogated by concomitant injections of rhM-CSF. Osteoclasts supported by rhM-CSF or endogenous VEGF showed no significant difference in the bone-resorbing activity. op/op mice undergo an age-related resolution of osteopetrosis accompanied by an increase in osteoclast number. Most of the osteoclasts disappeared after injections of anti-VEGF antibody, demonstrating that endogenously produced VEGF is responsible for the appearance of osteoclasts in the mutant mice. In addition, rhVEGF replaced rhM-CSF in the support of in vitro osteoclast differentiation. These results demonstrate that M-CSF and VEGF have overlapping functions in the support of osteoclastic bone resorption.

Animals↗

Identification of the canarypox virus thymidine kinase gene and insertion of foreign genes.

We mapped the canarypox virus (CaPV) thymidine kinase (TK) gene within a 5.8-kbp XbaI fragment of the genome by Southern blotting using the fowlpox virus (FPV) TK gene as a probe. Nucleotide sequence analysis of the fragment revealed seven open reading frames (ORFs) showing gene organization similar to that of FPV. The TK gene contained in this region had an ORF of 179 amino acids encoding a polypeptide with a putative molecular mass of 20.0 kDa. An A/T-rich region and a transcription termination signal, TTTTTAT, were found upstream and at the end of the ORF, which is consistent with poxvirus early gene regulation. The consensus sequence of the late promoter TAAAT also overlapped with the initiation codon of the ORF. The amino acid sequence similarity between the TK genes of CaPV and FPV, avipoxviruses, was 64.2%, which was lower than the similarities between vaccinia and variola orthopoxviruses (97.2%) and between Shope fibroma and myxoma leporipoxviruses (82.6%). However, the monophyly of avian clades of CaPV and FPV was supported by phylogenetic analysis. We then inserted the genes encoding lacZ, luciferase (luci), and envelope of human T-lymphotropic virus type 1 (HTLV-1 env) into the TK gene of CaPV to evaluate its suitability as an expression vector. The recombinant viruses obtained were unstable, although the foreign genes were expressed efficiently in the mammalian cells infected with the viruses.

Amino Acid Sequence↗

Low p27 expression correlates with poor prognosis for patients with oral tongue squamous cell carcinoma.

BACKGROUND: p27, a cyclin-dependent kinase inhibitor, regulates progression from G1 to S phase. There have been a few clinical reports of low p27 expression associated with poor survival among patients with cancer; however, there have been no reports of such an association in cases of head and neck cancer. The authors investigated whether p27 expression in patients with oral tongue squamous cell carcinoma was associated with their prognosis. METHODS: Ninety-four patients with oral tongue squamous cell carcinoma were analyzed. The authors performed p27 immunohistochemistry on all patients and Western blot analysis on 19 available patients. Cox proportional hazards regression analysis that included gender, history of smoking and alcohol usage, presence of multiple primary cancers, stage, histologic grade, and p27 status was used to identify the multivariate predictive value of prognostic factors. RESULTS: Twenty-six patients had high p27 expression (> or =50% tumor cell nuclei positive), and 68 patients had low p27 expression (<50%) by immunohistochemistry. In those with low p27 expression, N(+) and advanced T (T3 or T4) were significantly higher than in those with high p27 expression (P = 0.02 and 0.04). The 5-year survival rate in the low p27 group was 44%, whereas that in the high p27 group was 68%, indicating a significant difference (P = 0.04). p27 expression was inferred from Western blot analysis, and an arbitrary quantity (<1, 1-5, or > or =5) from the ratio of tumor to normal tissue density was used to characterize, resulting in 8 (42%), 3 (16%), and 8 (42%) patients in the low (<1-fold), intermediate (1-5-fold), and high (> or =5-fold) groups, respectively. Results of immunohistochemical analysis for p27 were significantly correlated with those of Western blot analysis (P = 0.02). Multivariate analysis revealed that low intensity of p27 expression and advanced stage (Stage III or IV) were predictors of reduced survival (P = 0.02 and 0.001). CONCLUSIONS: Low p27 expression was associated with increasing lymph node metastasis and stage of tumor and resulted in a poor prognosis for patients with oral tongue squamous cell carcinoma. p27 is apparently a significant predictor of survival.

Actins↗

Cesium-137 and mercury contamination in lake sediments.

The contribution of fluvial discharge and global fallout of 137Cs and mercury to sedimentation fluxes in Lake Shinji and Lake Nakaumi, Japan, was studied. The fluvial flux through soil erosion accounted for 11 to 27% of accumulated 137Cs in the sediments in the 1950's and 1960's, which were the periods of the most extensive atmospheric input, and for 90 to 100% in the 1990's. The vertical profiles of mercury concentrations in the lake sediments studied showed a maximum between 1959 and 1963, which was originated mostly from the extensive use of mercury-agrochemicals in paddyfields of the watershed in the past. These findings are representative examples of long-term contamination of lake sediments caused by the contaminated ground soil erosion.

Cesium Radioisotopes↗

Response to cyclosporin and low-dose methylprednisolone in aggressive systemic mastocytosis.

BACKGROUND: There is no effective treatment for aggressive systemic mastocytosis. OBJECTIVE: The purpose of this study was to investigate the effect of cyclosporin and low-dose methylprednisolone in a 64-year-old man with aggressive systemic mastocytosis. METHODS: Immunohistochemical studies were done on biopsy specimens from the skin and other organs. Mast cells, predominantly containing tryptase, were derived from human umbilical cord blood cells cultured in the presence of stem-cell factor and IL-6. IgE-sensitized cultured human mast cells were activated by anti-IgE, and the effect of cyclosporin on histamine release was investigated. In addition, blood and urine levels of various mediators were measured in the patient before and after therapy. RESULTS: Biopsy specimens of the patient's skin lesions showed an increase of mast cells; cells containing tryptase (but not chymase) comprised 20% to 50% of the skin mast cells. Histamine release from activated cultured mast cells was inhibited by cyclosporin in a concentration-dependent manner. When the patient was treated with cyclosporin and low-dose methylprednisolone, he showed a good response. CONCLUSION: Cyclosporin combined with low-dose methylprednisolone may be a reasonable therapy for aggressive systemic mastocytosis.

Bone Marrow Cells↗

Nerve growth factor release with mast-cell-derived mediators in a patient with systemic mastocytosis after middle-wave ultraviolet irradiation.

BACKGROUND: The symptoms of a 56-year-old man with systemic mastocytosis became worse with exposure to sunlight. We evaluated mast-cell-derived mediators and cytokines before and after exposure to ultraviolet light in the patient. METHODS: The patient was irradiated with middle-wave ultraviolet light, so-called ultraviolet light B, and the levels of mediators and cytokines were measured serially. The point mutation Asp816Val in c-kit was investigated by analyzing polymerase chain reaction products from the complementary DNA of peripheral blood mononuclear cells. RESULTS: Before irradiation, the levels of mast-cell-derived mediators and metabolites were elevated. Among various cytokines measured, including soluble c-kit and stem cell factor, only the level of nerve growth factor was elevated. After irradiation, the nerve growth factor level was further increased along with the levels of mast-cell-derived mediators and metabolites. The point mutation Asp816Val in c-kit was not detected in peripheral blood mononuclear cells. CONCLUSIONS: Middle-wave ultraviolet light may activate mast cells to release nerve growth factor and mediators in systemic mastocytosis.

Cytokines↗

Levels of IL-12 in the sera of patients with systemic lupus erythematosus (SLE)--relation to Th1- and Th2-derived cytokines.

IL-12 is a cytokine that induces Th1-derived cytokines (interferon-gamma (IFN-gamma) and IL-2). The significance of IL-12 in human autoimmunity is no clear, and the serum levels of IL-12 in SLE are not clearly established. Therefore, we examined the levels of IL-12 in 39 patients with active SLE, with sandwich ELISA. The levels of IL-12 in patients were significantly higher than in normal subjects. Patients with high levels of IL-12 also had high levels of IFN-gamma, while their levels of IL-13 were significantly lower than in patients with normal levels of IL-12. Patients with pulmonary involvement had high levels of IL-12, and steroid therapy decreased the IL-12 level in three patients. In a retrospective study of seven patients, various changes of IL-12 and IL-13 were recognized before disease flare. Thus, in SLE patients, the level of IL-12 was increased and this increase was related to the change of Th1- or Th2-derived cytokines with some organ involvement.

Adult↗