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Biomedical subjects

J Chihara

Publications and source records attributed to J Chihara.

At least 55 records · Page 3Linked to original sources
PubMed1999

[Small increases of the serum alkaline phosphatase activity and malignant changes].

In order to see the clinico-pathological significance of the small increase of serum alkaline phosphatase (ALP) activity alone in routine laboratory examinations, we picked up 76 new out-patients of adult within the two-fold high ALP level in contrast to the reference range in the Akita University Hospital for one year, and then studied the relation between the histories and serum ALPs of 33 patients whose ALP had increased pathologically or decreased after intervention such as surgical operations. The ratio of the patients with malignant tumors to the tested all patients was 13 to 76 (17%). Immunochemical analysis of sera for the cancers of the lung, larynx and prostate showed that the small amount of the placental isozyme or intestinal-like isozyme was expressed selectively. It may be useful for earlier detection of malignancy to analyze the cancer-associated ALPs, when the small and sequential increase of the serum ALP activity is found in routine laboratory examinations.

Aged↗
PubMed1999

[A case-study in bed-side teaching of clinical laboratory medicine].

In Akita Medical School, a new curriculum on clinical undergraduate education has started since June 1997. The Department of Clinical and Laboratory Medicine had two-weeks practice by small groups of medical students. We introduced a case-study in the practice with a general instructional objective and specific behavioral objectives for students to understand more the clinico-pathological process of diseases. A patient for primary hypothyroidism with chronic renal failure was chosen as the case, which had been found based on the abnormality on essential laboratory tests. The reversed-CPC was carried out to make a diagnosis with the patient's history and clinical examinations, and then the clinico-pathological process for the patient was re-checked, discussed and explained by students for themselves. They felt harder to see how abnormal data were related to diseases, according to their own evaluation of the practice. The further education for the patho-physiology of diseases and the logic of the test combination will be required.

Clinical Chemistry Tests↗
PubMed1999

Evidence for a link between glycoxidation and lipoperoxidation in patients with chronic renal failure.

AIM: The purpose of this study is to examine whether or not there is a relationship between glycoxidation and lipid peroxidation in patients with chronic renal failure. SUBJECTS AND METHODS: Dermal samples from 26 living or autopsied subjects were sequentially extracted with NaCl, pepsin, collagenase, and NaOH to obtain four fractions (salt-soluble fraction: SSF; pepsin-soluble fraction: PSF; collagenase-soluble fraction: CSF; and insoluble fraction: ISF). The glycoxidation product was measured by pentosidine-linked fluorescence (ex: 335/em: 385) and the levels of lipid peroxide, malondialdehyde (MDA), were assessed by determining the MDA-linked fluorescence (ex: 390/em: 460) which was further confirmed by HPLC. RESULTS: In patients undergoing hemodialysis, MDA-linked fluorescence markedly increased in collagen-rich fractions, PSF, CSF, and ISF, while pentosidine-linked fluorescence increased in PSF and CSF, in comparison to the controls and the pre-dialysis patients with CRF. Interestingly, the increase in the lipid peroxides strongly correlated with the level of glycoxidation product in PSF, CSF, and ISF (p < 0.0001 in PSF, CSF; p < 0.01 in ISF). The HPLC data of MDA in the PSF was in good correlation with logistic levels of both MDA- (n = 9, r = 0.738, p = 0.023) and pentosidine-linked fluorescence (n = 9, r = 0.721, p = 0.028). In contrast, in SSF, the collagen-poor fraction (collagen content: less than 3% of the total extracted collagen), the data showed a significant increase in the MDA-linked fluorescence only in the pre-dialysis patients with CRF, but not in the HD patients with no correlation with the glycoxidation products. CONCLUSION: These findings suggest that both the lipid peroxidation and glycoxidation increased in close relation to each other in the matrix collagen and thus demonstrate a synergetic contribution to the tissue damage observed in patients with CRF

Female↗
PubMed1998

Inhibitory effect of ibudilast (KC-404) on the expression of the beta2 integrin family on an eosinophilic cell line (EoL-1).

BACKGROUND: Adhesion molecules are thought to play a key role in inflammatory processes in bronchial asthma. We previously observed an increased expression of the beta2-integrin family on an eosinophilic cell line (EoL-1) by platelet-activating factor (PAF). OBJECTIVE: In the present study, we examined the effect of ibudilast (KC-404), a novel anti-asthma agent, on beta2 integrin expression induced by PAF. MATERIAL AND METHODS: EoL-1 cells (1x10(6)/mL) were incubated in the presence or absence of 10(-6) M ibudilast (KC-404), then cells were cultured in the presence or absence of PAF (10(-7) M) for 45 minutes. Flow cytometric analysis for CD11a, CD11b, and CD18 expression was examined. RESULTS: Ibudilast had an inhibitory effect on beta2 integrin expression induced by PAF [CD11a: 84.8% versus 73.1% (preincubation with ibudilast), CD11b: 35.8% versus 26.2%, CD18 74.9% versus 65.6%]. CONCLUSIONS: Ibudilast (KC-404) has anti-inflammatory activities through its inhibitory effect on the expression of adhesion molecules on eosinophils.

Asthma↗
PubMed1998

Priming effect of RANTES on eosinophil oxidative metabolism.

BACKGROUND: RANTES has been shown to possess chemotactic activity for eosinophils, which have also been considered to play a role in allergic inflammation through reactive oxygen species. Thus, in this study, we examined the effect of RANTES on radical oxygen products from eosinophils. METHODS: Purified eosinophils by CD16-negative selection or an eosinophilic cell line (EoL-1) were incubated with or without RANTES (2.5 x 10(-6) M). To the mixture of eosinophils and luminol, calcium ionophore (A23187) or opsonized zymosan (OZ) was added, and radical oxygen products were determined by luminol-dependent chemiluminescence for 600 s. RESULTS: Eosinophil-mediated radical oxygen products of untreated eosinophils produced with A23187 gave a peak value of 14.09 +/- 2.40 (mean +/- SE, n = 12) relative light units (RLU) and an integrated value of 3232.20 +/- 513.09 RLU. However, with treatment with RANTES, a peak value of 18.66 +/- 2.40 RLU and an integrated value of 5301.05 +/- 561.02 RLU were obtained. Eosinophil oxidative metabolism-induced A23187 or OZ was apparently augmented by the preincubation with RANTES. In addition, the radical oxygen products of EoL-1 showed similar results. CONCLUSIONS: Thus, we concluded that RANTES may play an important role in the pathogenesis of allergic inflammation through its involvement in eosinophil activation, as evidenced by oxygen products, as well as in selective eosinophil infiltration as selective eosinophil chemoattractant.

Calcimycin↗
PubMed1998

RANTES augments eosinophil lucigenin-dependent chemiluminescence.

BACKGROUND: RANTES (regulated on activation, normal T expressed and secreted) has been shown to possess chemotactic activity for eosinophils. Eosinophils have been considered to play a key role in the allergic inflammation through the release of inflammatory molecules such as radical oxygen products. Thus, in this study, we examined the effect of RANTES on radical oxygen products from eosinophils. METHODS: Eosinophils were isolated from heparinized venous blood of patients with bronchial asthma by the modified CD16-negative depletion method. Radical oxygen products were examined in terms of lucigenin-dependent chemiluminescence. To a mixture of 50 microl of eosinophils (2x10(6)/ml) and 50 microl of lucigenin (5x10(-4)M), 50 microl of calcium ionophore A23187 (final concentration 10(-5)M) was added, and radical oxygen products were determined for 600 s. RESULTS: RANTES treatment resulted in the enhancement of peak value (0.64+/-0.23 RLU) and integrated value (119.08+/-20.52 RLU) as compared to untreated cells (0.15+/-0.03 RLU, 29.48+/-8.92 RLU, respectively). CONCLUSIONS We could conclude that RANTES might play an important role in the pathogenesis of allergic inflammation through involvement in selective eosinophil infiltration and eosinophil activation by augmentation of eosinophil oxidative metabolism.

Acridines↗
PubMed1998

Involvement of beta1 and beta2 integrin stimulation in RNA synthesis in an eosinophilic cell line (EoL-1).

In allergic inflammatory disease, especially in bronchial asthma, eosinophils play important roles as essential inflammatory cells. In the accumulation of eosinophils in airway inflammation, eosinophils receive very diverse stimuli. In this study, we investigated the influences of the signal between beta2 integrin/intercellular adhesion molecule-1 (ICAM-1) or beta1 integrin/fibronectin (FN) and RNA synthesis on proteins in eosinophilic cell line-1 (EoL-1), using recombinant soluble ICAM-1 (r-sICAM-1) as a global response of eosinophils. 3H-thymidine incorporation and 3H-uridine incorporation were used as indices for DNA synthesis and RNA synthesis, respectively. In a comparison of the RNA/DNA ratio with various durations of stimulation of 1 microg/ml of r-sICAM-1 and 100 microg/ml of FN, a time-dependent increase was observed, but the increase induced by FN rose slower than that induced by r-sICAM-1. From this result, a beta2 integrin/ICAM-1 signal induced an increase in the RNA/DNA ratio in EoL-1, implying that the signal promotes RNA synthesis, which suggests that various types of protein synthesis, such as the synthesis of various cytokines, are induced by the beta2 integrin/ICAM-1 signal. Similar results were obtained with a beta1 integrin signal using FN, but there was a difference in the time course between beta2 integrin/ICAM-1 and beta1 integrin/FN signals. This experimental method may be useful for understanding these manifestations as a global response of eosinophils.

Asthma↗
PubMed1998

Whole-blood flow-cytometric analysis of eosinophil EG2 expression as a marker of the pathological conditions of asthma.

UNLABELLED: Using a simple technique detecting the eosinophil fraction in whole-blood flow cytometry, we measured intracellular antigen EG2 (a monoclonal antibody to eosinophil cationic protein) in 56 asthmatic patients (26 during an attack and 30 during an asymptomatic period) and 22 healthy subjects to determine whether EG2 reflects the pathological stages of allergy. METHODS: In brief, preparations of the sample included the following procedures: (1) hemolyzation of heparinized or EDTA-mixed whole blood; (2) fixation of white blood cells with 0.4% parabenzoquinone (PBQ) or paraformaldehyde (PFA); (3) permeabilizing the cell membrane with n-octyl-beta-D-glucopyranoside, and (4) staining of intracellular EG2 antigen with monoclonal EG2 antibody and FITC-labeled secondary antibody. RESULTS: In PBQ-fixed samples, there was a clearer boundary of the eosinophil fraction with a higher yield and purity than in those fixed with PFA. The number of EG2-positive eosinophils was significantly greater in subjects during attacks than in asymptomatic patients. In addition, when compared with normal controls, asthmatic subjects had significantly greater numbers of EG2-positive eosinophils regardless of their current condition. CONCLUSION: Eosinophil intracellular EG2 may indicate the pathological stage of asthma. This simple technique for analysing the properties of eosinophils using whole-blood flow cytometry would save time and labor in laboratories.

Adult↗
PubMed1998

[Stress and immunoallergy].

We have previously reported that hyperresponsiveness in the restriction stress guinea pig group as compared to that in controls was observed. On the other hand, adhesion molecules, especially intercellular adhesion molecule-1 (ICAM-1), are considered to play an important role in inflammatory processes in allergic and immune reactions such as bronchial asthma. Therefore, in this study, our investigation was designed to clarify whether stress is involved in adhesion molecules expression on inflammatory cells and endothelial cells (EC). Results were as follows: 1) IL-1, which is considered to play an important role in psychoneuroimmunological phenomena, induced ICAM-1 expression on human EC. 2) Moreover, PAF and PF 4, which may be striking chemotactic agents and activating agents for eosinophils, also induced ICAM-1 expression on human EC. 3) The leukocyte adherence to plasma coated glass in the electric shock stress guinea pig model was augmented as compared to that of controls. 4) Eosinophil counts as well as eosinophil percentages in adherent leukocytes in the electric shock stress group were greater than those of controls. 5) Moreover, leukocyte adherence to autologous bronchial ciliary cells was also augmented in the stress group as compared to that of controls. In conclusion taken together, stress might be involved in adhesion molecules expression resulting in acceleration of allergic inflammatory reactions such as bronchial asthma.

Animals↗
PubMed1998

[The role of chemokines such as RANTES in allergic disease].

Much attention has recently been focused on the role of allergic inflammatory reaction in asthma. Eosinophils are considered to be the major type of inflammatory cell involved in bronchial asthma, since eosinophil-specific granule proteins can damage bronchial mucosal cells. Chemokines related to the beta subfamily, the so-called platelet factor 4 (PF4) superfamily have been shown to stimulate human eosinophils or basophils, and are considered to be important mediators of inflammation. RANTES may be released from activated platelets and is considered to play an important role in various immune and allergic disorders. RANTES is a potent chemoattractant for various inflammatory cells such as eosinophils, as well as for memory T cells and monocytes, thus potentially recruiting these cells from the circulation to an inflamed focus. Involvement of eosinophils and T cells in bronchial asthma has also been reported. To extend our understanding of the participation of eosinophils, T cells, and RANTES in the pathogenesis of allergic disease, we demonstrated the important roles of chemokines such as RANTES in allergic disease.

Asthma↗
PubMed1997

Elevation of the plasma level of RANTES during asthma attacks.

BACKGROUND: RANTES is considered to play an important role in various immune and allergic disorders since it is a potent chemoattractant for inflammatory cells such as eosinophils, memory T cells, and monocytes. OBJECTIVE: To investigate the possible role of RANTES in the pathogenesis of bronchial asthma. METHODS: The plasma level of RANTES was measured in 12 asthma patients and 15 normal controls by a sandwich enzyme-linked immunosorbent assay. RESULTS: In the patients with asthma, the plasma RANTES level was significantly elevated during acute attacks compared to that in the controls. In addition, it was higher than that during the asymptomatic state in the same patients. Plasma beta-thromboglobulin levels were also elevated in asthma patients during acute attacks and showed a correlation with the RANTES level. CONCLUSION: These findings suggest a role for RANTES in the pathogenesis of asthma and a possible role for platelets in RANTES release in asthma.

Adult↗
PubMed1997

Immunohistology of skin and oral biopsies in graft-versus-host disease after bone marrow transplantation and cytokine therapy.

BACKGROUND: Early diagnosis of graft-versus-host-disease (GVHD) after bone marrow transplantation is often difficult, particularly when the patients are immunosuppressed by chemotherapy or irradiation. OBJECTIVE: To investigate the influence of cytokines on skin lesions after bone marrow transplantation. METHODS: Biopsy specimens of skin and oral mucosa were obtained from bone marrow transplant patients with GVHD and were subjected to histologic and immunohistochemical examination. RESULTS: Administration of granulocyte-macrophage colony-stimulating factor caused atopic dermatitis-like lesions in two patients, who had infiltration of neutrophils, eosinophils, and lymphocytes around the hair follicles of the skin and no signs of GVHD in other organs. Only patients who were treated with cytokines developed acute GVHD. Immunohistochemical examination of skin biopsies from 18 patients with acute GVHD and 11 patients with chronic GVHD after cyclophosphamide administration or irradiation showed that the maculopapular skin lesions characteristic of acute GVHD contained infiltrates of CD4+ and CD8+ lymphocytes. There was also an increase in numbers of epidermal keratinocytes expressing intercellular adhesion molecule-I and HLA-DR antigens. CONCLUSION: These findings support the involvement of cytokines in GVHD and suggest that immunostaining of skin biopsies may be useful for the early diagnosis of this condition.

Acute Disease↗
PubMed1997

Soluble intercellular adhesion molecule-1 levels of patients with acute myeloid leukemia after allogeneic bone marrow transplantation.

Concentrations of serum-soluble intercellular adhesion molecule-1 (ICAM-1) were measured for 10 patients with or without chronic graft versus host disease after allogeneic bone marrow transplantation. Levels of soluble ICAM-1 were higher among patients with chronic graft versus host disease than among those without it, a statistically significant difference. The results indicated that measurement of serum-soluble ICAM-1 is useful for prediction of chronic graft versus host disease.

Acute Disease↗
PubMed1997

Coincident induction of K rev-1/rap 1A, rap 1B and H-ras mRNAs in the rat spinal cord by noxious stimulation.

Two cDNA fragments, K rev-1/rap 1A and rap 1B, were amplified from total cellular RNA of the rat spinal cord by reverse transcription-polymerase chain reaction with a set of oligonucleotide primers specific for the human rap 1A cDNA. We report here using Northern blot analysis with these cDNA probes that noxious stimulation causes a marked and coincident increase in rap 1A, rap 1B and H-ras mRNAs in the rat spinal cord. This suggests that Rap 1 participates in sensory processing in spinal neurons in parallel with Ras.

Amino Acid Sequence↗
PubMed1997

A monoclonal antibody modulates neutrophil adherence while enhancing cell motility.

A monoclonal antibody (MoAb) to human neutrophils, designated 3H9, was established by screening for the inhibition of neutrophil adherence to plastic plates containing a medium supplemented with fetal calf serum (FCS medium). The antigen recognized by 3H9 was shown to be present on human leukocytes and found at the highest levels on granulocytes. On Western blotting, 3H9 reacted with a molecule having a molecular weight of 80 kDa. When this MoAb was added at the same time as a neutrophil stimulant (fMLP), the inhibition of neutrophil adherence to plastic plates in the presence of FCS medium was observed after 60 min incubation. Furthermore, this MoAb enhanced not only fMLP-induced chemotaxis but random migration of neutrophils as well. The mechanisms of these phenomena are discussed.

Animals↗
PubMed1997

Enhanced production of RANTES, an eosinophil chemoattractant factor, by cytokine-stimulated epidermal keratinocytes.

In allergic skin diseases such as atopic dermatitis (AD), eosinophils migrate from the circulation to the skin. We investigated the mechanisms of eosinophil chemotaxis in atopic dermatitis by examining the effect of stimulation of epidermal keratinocytes (KC) by inflammatory cytokines, interferon-gamma (IFNgamma) and/or tumor necrosis factor-alpha (TNF alpha) on the production of eosinophil chemotactic factors. Simultaneous addition of IFNgamma and TNF alpha to culture KC synergistically increased eosinophil chemotaxis and the expression of RANTES mRNA and protein level on these cells. Anti-RANTES antibody blocked eosinophil chemotaxis by IFNgamma- and TNF alpha-stimulated KC. Our results indicate that the production of RANTES by KC may help to explain eosinophil infiltration into the skin in AD.

Chemokine CCL5↗