Increased END/EPX in ongoing asthma.
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Adult↗
Biomedical subjects
J Chihara
Publications and source records attributed to J Chihara.
Serum markers of graft-versus-host disease after bone marrow transplantation.
BACKGROUND: Graft-versus-host disease is one of the major complications after allogenic bone marrow transplantation, but it is not easy to anticipate the onset. OBJECTIVES: The purpose of this study was to determine clinically useful markers of acute graft-versus-host disease. METHODS: We measured the serum levels of tumor necrosis factor-alpha, soluble tumor necrosis factor receptor 1, soluble c-kit, soluble Fas, soluble intercellular adhesion molecule-1, growth-related oncogene protein-alpha, thrombomodurin, and interleukin-16 in 13 patients at 1 to 7 weeks after allogenic bone marrow transplantation. RESULTS: The patients with acute graft-versus-host disease showed a significant increase of tumor necrosis factor, soluble tumor necrosis factor receptor 1, soluble Fas, soluble intercellular adhesion molecule-1, and growth-related oncogene protein-alpha, although there was a decrease of soluble c-kit. The increases of serum soluble tumor necrosis factor receptor 1, intercellular adhesion molecule-1, and growth-related oncogene protein-alpha were preceded by the elevation of soluble Fas. CONCLUSION: The patients with acute graft-versus-host disease had increased serum levels of tumor necrosis factor-alpha, soluble tumor necrosis factor receptor 1, soluble Fas, and soluble intercellular adhesion molecule 1 and a decreased soluble c-kit level. Tumor necrosis factor-alpha and soluble c-kit were shown to be sensitive and specific parameters for graft-versus-host disease after bone marrow transplantation, and soluble Fas was shown to be a predictor of acute graft-versus-host disease after bone marrow transplantation.
Adolescent↗
Delayed recovery of CDR3 complexity of the T-cell receptor-beta chain in recipients of allogeneic bone marrow transplants who had virus-associated interstitial pneumonia: monitor of T-cell function by CDR3 spectratyping.
BACKGROUND: In the T-cell receptor (TCR)-beta chain, complementary-determining region 3 (CDR3) contains specific peptide sequences essential for recognition. Diversity of this region is considered to contribute to immunocompetence in humans. OBJECTIVE: The purpose of this study was to define the process of reconstitution of CDR3 complexity of the TCR-beta chain after allogeneic bone marrow transplantation and to investigate the association between host immunocompetence and CDR3 complexity. METHODS: Diversity of the CDR3 region of the TCR-beta chain was examined by CDR3 size distribution analysis with the use of an automated DNA sequencer. RESULTS: Reconstitution of the alphabeta T-cell repertoire and CDR3 diversity was incomplete for at least 2 months after bone marrow transplantation. Delayed reconstitution of T-cell diversity was more marked in immunocompromised hosts. Unlike the situation in patients who received allogeneic bone marrow grafts, the recovery of CDR3 complexity was almost perfect by 2 months after transplantation in patients who received allogeneic blood stem cells. Clonal expansion of alphabeta T cells after allogeneic bone marrow transplantation was readily detected by CDR3 size spectratyping analysis. CONCLUSION: PCR-based CDR3 size spectratyping may be a useful tool for clinically monitoring immune reconstitution after allogeneic bone marrow transplantation.
Adolescent↗
Signaling through the beta2 integrin prolongs eosinophil survival.
BACKGROUND: Recently, adhesion molecules have been suggested to play an important role in allergic inflammatory diseases such as bronchial asthma. It is unclear whether eosinophil activation and paracrine or autocrine synthesis of eosinophilopoietic growth cytokines is mediated through signaling by intercellular adhesion molecule-1 (ICAM-1) and the beta2 integrin family. OBJECTIVE: We examined whether signaling by ICAM-1 and its ligands (beta2 integrins) could prolong eosinophil survival. METHODS: Eosinophils were isolated from patients with hypereosinophilia by modified CD16 negative selection. After culture with or without recombinant soluble ICAM-1, eosinophil viability was measured by trypan blue dye exclusion. RESULTS: Eosinophil survival was prolonged in cultures with recombinant soluble ICAM-1 compared with cultures without it (P <.01 on days 2, 4, and 6); this effect was dose-dependent. Eosinophil survival in cultures with recombinant soluble ICAM-1 was significantly inhibited by antibodies against ICAM-1 (P <.01), complement receptor 3 (P <.01), and lymphocyte function-associated antigen-1beta (P <.01). Anti-IL-3 showed no effect on eosinophil survival, whereas anti-IL-5 caused partial inhibition of survival. Interestingly, anti-granulocyte/macrophage colony-stimulating factor caused the complete inhibition of eosinophil survival in cultures with recombinant soluble ICAM-1. CONCLUSION: These results suggested the importance of the beta2 integrins in eosinophil-mediated allergic inflammation.
Cell Survival↗
Development of a novel enzyme-linked immunosorbent assay for blood and urinary eosinophil-derived neurotoxin: a preliminary study in patients with bronchial asthma.
BACKGROUND: Eosinophil-derived neurotoxin (EDN), also called eosinophil protein X (EPX), has been suggested to be a useful marker of eosinophilic inflammation. However, no commercial enzyme-linked immunosorbent assay (ELISA) kit for EDN is available yet. METHODS: EDN was purified from pooled urine from healthy male volunteers. Polyclonal and monoclonal anti-EDN antibodies were subsequently raised, and a sandwich ELISA for EDN was established. EDN levels in serum, plasma and urine from asymptomatic patients with bronchial asthma were measured by the ELISA method. Some of the blood samples were also measured by a commercial radioimmunoassay (RIA) kit. RESULTS: The ELISA method detected human EDN with a minimum detection limit of less than 0.62 ng/ml and did not cross-react with other eosinophil granule cationic proteins including eosinophil cationic protein. The intra- and interassay coefficients of variation of the ELISA method ranged from 2.6 to 3.6% and from 6.5 to 9.4%, respectively. Good linearity was observed with serially diluted different samples, and the recoveries of the purified EDN added to serum samples ranged from 85 to 110%. Median EDN concentrations in serum (36.9 vs. 19.1 ng/ml), plasma (23.0 vs. 14.5 ng/ml) and urine (118.2 vs. 72.1 microg/mmol Cr) were significantly raised in asymptomatic asthmatic patients compared with healthy control subjects. EDN levels in serum, plasma and urine from the patients significantly correlated with the number of peripheral blood eosinophils, but not total serum IgE levels. A significant relationship between EDN values measured by the EPX-RIA kit and the EDN-ELISA method was observed. CONCLUSIONS: We have developed a novel efficient ELISA method to specifically measure blood and urinary EDN, which may be useful to study the role of eosinophils in allergic diseases including bronchial asthma.
Asthma↗
An optimal condition of bronchial cell proliferation stimulated by insulin-like growth factor-I.
BACKGROUND: It is known that growth hormones such as insulin-like growth factor-I (IGF-I) and several kinds of cytokines are involved in the regeneration process of injured epithelial cells in chronic respiratory inflammatory diseases such as bronchial asthma. Repetitive degeneration/regeneration processes of the airway epithelial layer is supposed to be responsible for the remodeling and irreversible organic changes of the airway in bronchial asthma. The purpose of this study is to establish a simple and reliable in vitro method for studying airway epithelial cell growth and proliferation using IGF-I. METHOD: By altering the number of cultured epithelial cells (strain NCI-H(292)), culture duration before stimulation with IGF-I, concentration of IGF-I, and duration of IGF-I stimulation, the optimum conditions for epithelial cell growth was determined. The epithelial cell growth was evaluated using [methyl-(3)H]thymidine uptake. RESULT: Among various culture conditions, the epithelial cells cultured at 1 x 10(3) cells/well for 24 h followed by 24 h of stimulation by 10(-8) M of IGF-I showed the highest growth. CONCLUSION: The method for the evaluation of epithelial cell growth established in this study requires a small number of cells and has no complicated procedure. This simple model enables us to investigate the effect of various substances on bronchial epithelial cell growth in the presence of IGF-I.
Bronchi↗
[Advances in the clinical laboratory automation system of Akita University Hospital].
Abstract Clinical laboratory automation, which was widely discussed in the 1960's, was developed in late he 1970's. The computerized automatic analyzing system for clinical laboratories spread nationwide in the 1980's. Akita University Hospital was established in 1971. We have been making efforts to establish the automated clinical laboratory system. The hematological and clinical chemistry divisions were automated in 1981. In 1988, in order to establish a fully automated clinical laboratory, we introduced sample-conveying systems in each division including hematological, clinical chemistry, urinalysis, serological and immunological divisions. In 1998, we connected the sample-conveying systems which had been separately established in each division to create a fully automated clinical laboratory with an integrated sample-conveying system.
Automation↗
[Tests for bronchial asthma].
One characteristic feature of bronchial asthma is an allergic inflammation of the airways involving eosinophil activation. Since adhesion molecules, cytokines, and chemokines play a critical role in eosinophil infiltration into the tissues, it is of paramount importance to utilize these inflammation-related factors as clinical parameters to assess the status of asthma. For this reason, we measured the level of RANTES and soluble ICAM-1 in patients with asthma. The concentration of plasma RANTES was significantly elevated in asthmatic patients as compared with normal subjects. Patients with asthma attacks exhibited higher RANTES levels than those in remission. sICAM-1 concentration was also higher in serum and sputum in patients with asthma than in healthy subjects. In order to detect eosinophil activation directly, we studied intracellular EG2 expression in eosinophils using whole-blood flow-cytometric analysis. The number of EG2-positive eosinophils was significantly greater in patients with attacks than in asymptomatic subjects. We measured the temperature of expiratory flow and temperature flux as an alternative approach to assess airway inflammation. This study was based on a concept that inflammation would produce heat resulting in the higher temperature. The coefficient of temperature flux was significantly greater in asthmatic patients than in normal controls. Therefore, these new tests may be useful for the evaluation and treatment of allergic inflammation in asthma.
Asthma↗
[Clinical examinations on lung cancer].
The incidence of lung cancer is increasing in Japan and it is important to know whether a patient has a history of primary smoking/second-hand smoking because smoking is the main cause of lung cancer. Lung cancer is often found on chest roentgenography. In order to determine the strategy of the treatment, the stage of the disease can be studied by the examinations of morphology(chest computed tomography), pathology(biopsy under bronchoscopy) and biochemistry(tumor markers and genetic analysis). Most lung cancers are detected in the advanced stage with metastasis. Performance status should be useful for the estimation of the prognosis of patients. In the symposium, we reported two of our clinical cases, in which urinary deoxypyridinoline, a bone absorption marker, facilitated the discovery of the bone metastasis of lung cancers with negative bone scans. We were requested to verify the early diagnosis of lung cancers with a collection of data on morphology, pathology and biochemistry.
Amino Acids↗
Degranulation of eosinophils mediated by intercellular adhesion molecule-1 and its ligands is involved in adhesion molecule expression on endothelial cells-selective induction of VCAM-1.
BACKGROUND: Adhesion molecules and eosinophils may play an important role in the pathogenesis of allergic inflammatory reactions. OBJECTIVE: We attempted to clarify eosinophil activation, such as degranulation, by signaling through adhesion molecule and to determine whether degranulation is involved in adhesion molecule expression on endothelial cells. METHODS: Eosinophils were cultured with or without recombinant soluble intercellular adhesion molecule-1 (ICAM-1), and the levels of eosinophil cationic protein and eosinophil-derived neurotoxin were determined. The influence of these eosinophil granule proteins or supernatant from eosinophil cultured with ICAM-1 on the expression of ICAM-1 or vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells was also examined by flow-cytometric analysis. RESULTS: Supernatant levels of eosinophil granule protein were significantly increased by culture for 4 hourss or 16 hours with recombinant soluble ICAM-1, suggesting degranulation by adherence to ICAM-1. Both granule proteins and the supernatants of eosinophils cultured with recombinant soluble ICAM-1 induced expression of ICAM-1 and VCAM-1 on endothelial cells, with the latter showing a more prominant increase. CONCLUSION: Degranulation mediated through adherence to endothelial cells by ICAM-1 and its ligands may be involved in the expression of adhesion molecules, such as ICAM-1 or VCAM-1, on these cells. Our finding of the selective induction of VCAM-1 expression suggests that eosinophil adherence to endothelial cells, even if it is because of ICAM-1, may be involved in selective eosinophil recruitment and accumulation at sites of allergic inflammation.
Cell Adhesion Molecules↗
Eosinophil activation by eotaxin--eotaxin primes the production of reactive oxygen species from eosinophils.
BACKGROUND: The CC chemokine eotaxin has been shown to possess selective chemotactic activity for eosinophils, the major effector cells in allergic inflammation. Reactive oxygen species (ROS) from eosinophils may damage cells or tissue, such as the mucosal epithelium. In this study, we examined the effect of eotaxin on ROS from eosinophils and compared its activity with RANTES and interleukin (IL)-5. Moreover, we examined the signal transduction of eotaxin and the effect of dexamethasone on ROS from eosinophils. METHODS: Eosinophils were isolated by modified CD16-negative selection. ROS in luminol-dependent or lucigenin-dependent chemiluminescence reaction were examined. Calcium ionophore A23187 was added to the mixture of eosinophils with luminol or lucigenin, and then ROS were determined. RESULTS: Eotaxin primed the production of ROS in a dose-dependent manner. ROS from untreated eosinophils evoked with calcium ionophore A23187 in luminol-dependent chemiluminescence gave a maximal value of 4957+/-1035 intensity counts (IC) (mean+/-SE, n=7) and an integral value of 15.75+/-3.14 IC (x10(-4)), while eosinophils that were treated with eotaxin gave maximal values of 11 142+/-2300 IC (10 nM) and 29165+/-3718 IC (100 nM) and integral values of 41.07+5.44 IC (x10(-4)) (10 nM) and 152.90+/-22.38 IC (x10(-4))(100 nM). Moreover, eotaxin was less effective as a priming agent with lucigenin-sensitive pathways than luminol-sensitive pathways. Among several kinds of eosinophils activating cytokines and chemokines, the priming effect of eotaxin on RO5 was the most potent. Eotaxin-primed ROS were inhibited by pertussis toxin, which ADP-ribolysates G proteins; wortmannin, a phosphatidylinositol-3-kinase inhibitor; and genistein, a tyrosine kinase inhibitor, suggesting the involvement of pertussis toxin-sensitive G proteins, phosphatidylinositol-3-kinase, and tyrosine kinase in the signal transduction of eotaxin. Moreover, dexamethasone inhibited ROS from not only untreated eosinophils but also eosinophils treated with eotaxin. CONCLUSION: Eotaxin may play an important role in the pathogenesis of allergic inflammation through eosinophil activation by priming of eosinophil oxidative metabolism, as well as by involvement in selective eosinophil chemotaxis.
Acridines↗
Nucleus-electron model for states changing from a liquid metal to a plasma and the Saha equation.
We extend the quantal hypernetted-chain (QHNC) method, which has been proved to yield accurate results for liquid metals, to treat a partially ionized plasma. In a plasma, the electrons change from a quantum to a classical fluid gradually with increasing temperature; the QHNC method applied to the electron gas is in fact able to provide the electron-electron correlation at an arbitrary temperature. As an illustrating example of this approach, we investigate how liquid rubidium becomes a plasma by increasing the temperature from 0 to 30 eV at a fixed normal ion density 1.03x10(22)/cm(3). The electron-ion radial distribution function (RDF) in liquid Rb has distinct inner-core and outer-core parts. Even at a temperature of 1 eV, this clear distinction remains as a characteristic of a liquid metal. At a temperature of 3 eV, this distinction disappears, and rubidium becomes a plasma with the ionization 1.21. The temperature variations of bound levels in each ion and the average ionization are calculated in Rb plasmas at the same time. Using the density-functional theory, we also derive the Saha equation applicable even to a high-density plasma at low temperatures. The QHNC method provides a procedure to solve this Saha equation with ease by using a recursive formula; the charge population of differently ionized species are obtained in Rb plasmas at several temperatures. In this way, it is shown that, with the atomic number as the only input, the QHNC method produces the average ionization, the electron-ion and ion-ion RDF's, and the charge population that are consistent with the atomic structure of each ion for a partially ionized plasma.
Journal Article↗
Whole-blood flow-cytometric analysis of induction of adhesion molecules expression on eosinophils stimulated by phorbol-12-myristate-13-acetate/ionomycin.
BACKGROUND: Activation of eosinophils is closely associated with the pathology of allergic inflammatory disease, especially bronchial asthma. We recently investigated the activation of eosinophils by applying whole blood to a flow cytometer. We measured here beta1 and beta2 integrin on eosinophils stimulated by phorbol-12-myristate-13-acetate (PMA)/ionomycin to evaluate eosinophil activation in vitro using whole blood. METHODS: Heparinized whole blood was diluted with the same volume of RPMI 1640, then cells were incubated in the presence or absence of PMA and ionomycin for 45 min at 37 degrees C. After hemolyzation with lysing solution, flow-cytometric findings for CR3, LFA1-alpha, LFA1-beta and VLA-4 expression on eosinophils were examined. RESULTS: Mean fluorescent intensity (MFI) of CR3 and LFA1-beta stimulated by PMA and ionomycin was significantly higher than that of the unstimulated control. MFI of LFA1-alpha showed no significant difference from the unstimulated control. On the other hand, MFI of VLA-4 tended to decrease. CONCLUSIONS: Our method to distinguish eosinophils from various cell groups in whole blood is simple and time-saving, similar to conditions in vivo and may allow intensive investigation of eosinophils in clinical laboratories as well as in research laboratories. We are currently investigating the influence of different kinds of stimulations, regulation factors or agents on eosinophils using this method.
Carcinogens↗
Adhesion to fibronectin regulates expression of intercellular adhesion molecule-1 on eosinophilic cells.
BACKGROUND: Adhesion molecules may play an important role in eosinophilic activation as well as adherence of extracellular matrix (ECM) including fibronectin (FN) in the inflamed focus. Tissue eosinophils expressed inter-cellular adhesion molecule-1 (ICAM-1, CD54), whereas peripheral blood eosinophils did not. The molecular mechanisms of ICAM-1 expression on eosinophils are not clear. We immunologically examined the effect of adherence to FN on the expression of adhesion molecules in an eosinophilic cell line (EoL-1). METHODS: EoL-1 cells, suspended at a concentration of 2 x 10(6) cells/ml, were cultured at 37 degrees C in BSA or FN-coated 24-well plates. After 4 h, the cells were removed by pipetting, and the expression of adhesion molecules was evaluated by immunofluorescence. RESULTS: Adherence of EoL-1 to FN enhanced ICAM-1 (CD54) expression on EoL-1 (FN vs. BSA: 84.76 +/- 17.90 vs. 43.73 +/- 7.60, mean fluorescent intensity). Regarding other adhesion molecules on EoL-1 (CD11a, CD18, CD49d), the expression was not significantly augmented by adherence to FN. CONCLUSIONS: We concluded that the signal via adhesion of FN regulates the expression of ICAM-1 on eosinophilic cells. This study suggests that eosinophilic adhesion to ECM via adhesion molecules plays an important role in the pathogenesis of allergic inflammation through eosinophilic functional changes, including regulation of expressive adhesion molecules such as ICAM-1.
Cell Adhesion↗
CCR3 mRNA expression in bronchial epithelial cells and various cells in allergic inflammation.
BACKGROUND: RANTES and eotaxin are important chemokines involved in the activation and migration of eosinophils and are considered to play a major role in allergic inflammation. METHODS: In this study, we used RT-PCR to investigate the kinds of cells that express mRNA for CCR3, a common receptor of these chemokines, and eotaxin, a ligand for CCR3. RESULTS: CCR3 mRNA was expressed in eosinophils, peripheral mononuclear cells, an eosinophilic cell line (EoL-1), a bronchial epithelial cell line (NCI-H(292)), human endothelial cells and nasal washings from patients with allergic rhinitis. CONCLUSION: These results suggest that the CCR3-eotaxin system plays an important role in generating inflammation, since these substances are expressed not only in cells implicated in activation or migration of eosinophils but also in various other cells involved in allergic inflammation.
Bronchi↗
Effect of eotaxin on the generationof reactive oxygen species from eosinophil cell line, YY-1.
BACKGROUND: The CC chemokine eotaxin is a selective chemoattractant for eosinophils. Eosinophils have been considered to be the major effector cells in allergic inflammation, since not only eosinophil-specific granule proteins but also reactive oxygen species (ROS) from eosinophils may cause the damage to the cells or tissue of the mucosal epithelium. In this study, we examined the effect of eotaxin on ROS from an eosinophil cell line, YY-1. METHODS: ROS in luminol-dependent reaction were examined. Calcium ionophore A23187 were added to the mixture of YY-1 cells with luminol, and then ROS were determined. RESULTS: Eotaxin primed the production of ROS from YY-1 cells. ROS from untreated YY-1 cells evoked with calcium ionophore A23187 in luminol-dependent chemiluminescence gave a maximal value of 1,928 +/- 223 intensity counts (IC; mean +/- SE, n = 4) and an integral value of 17.04 +/- 1. 51 IC (x10(-4)), while eosinophils that were treated with eotaxin gave a maximal value of 2,264 +/- 86 IC (10 nM), 2,691 +/- 124 IC (100 nM) and an integral value of 21.22 +/- 0.67 IC (x10(-4); 10 nM), 26.20 +/- 1.41 IC (x10(-4); 100 nM). CONCLUSION: Eotaxin might play important roles in the pathogenesis of allergic inflammation through eosinophil activation by priming of eosinophil oxidative metabolism as well as involvement in selective eosinophil chemotaxis.
Cell Line↗
Overexpression of CD11b on eosinophils in atopic dermatitis: downregulation by cyclosporin A and upregulation by interleukin 5.
We examined the level of expression of CD11b on eosinophils in pripheral blood samples from patients with atopic dermatitis (AD) and non-AD volunteers. Eosinophils were defined using a new method employing CD14/CD45 and a backgate technique. Overexpression of CD11b was noted in eosinophils of AD patients. Treatment of AD with cyclosporin A resulted in clinical improvement as well as reduction in the expression of CD11b. Stimulation of eosinophils from patients with inactive AD by interleukin 5 upregulated the expression of CD11b on these cells. Our results suggest that the expression of CD11b surface molecule on eosinophils may play an important role in the activity of AD.
Cyclosporine↗
The position and size of radiological nephrogram in Japanese preschool children.
The early detection of retroperitoneal masses in children, such as neuroblastoma, Wilm's tumor, hydronephrosis and cystic renal diseases, has a great clinical importance for the improvement of their prognosis. The kidney is often affected in its size or position by these lesions, and occasionally allows clinicians to find a clue to reach the correct diagnosis before the patient become symptomatic. Since we had no clinically available nomogram on the position and the size of the kidney in Japanese children, we measured the size and position of the kidneys on plain abdominal x-rays in 347 Japanese children in preschool years with a special attention to their relationship with the spine. As a result, the nomogram showed age dependent growth of the kidneys keeping almost the same ratio with the spine, while the distance between the upper pole of the kidney and the spine remained less than 10 mm in all age groups. Our nomogram may be useful not only for picking up the malposition of the kidneys but also for the follow up of the patients with chronic renal diseases affecting the growth of the kidneys.
Aging↗