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Biomedical subjects

M Okano

Publications and source records attributed to M Okano.

At least 19 recordsLinked to original sources

Simple assay for evaluation of Epstein-Barr virus specific cytotoxic T lymphocytes.

Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes (CTL) are considered to be one of major defenses against the activation of EBV infection. We have developed a simple assay for evaluation of EBV-CTL by means of culturing peripheral blood mononuclear cells at the concentration of 2 x 10(6) cells/ml infected by B95-8 EBV, in the presence or absence of cyclosporin A (CSA). No lymphoblastoid cell line was established from ten EBV-seropositive healthy individuals in the absence of CSA. In contrast, cell lines were established from ten EBV-seronegative healthy individuals in the absence of CSA, indicating lack of EBV-CTL in these individuals. Methods described herein are simple and correlate well with EBV-CTL activity when compared to conventional methods, such as outgrowth regression or chromium-51 release assays.

Antigen Presentation

Inhibitory effects of quassinoids on Epstein-Barr virus activation.

Short-term in vitro assays for tumor promoters and anti-tumor promoters (Epstein-Barr virus activation test) were carried out for 45 quassinoids. As a result, some quassinoids showed potent activity, more than 50% inhibition at a molar ratio of 1:1 (TPA/quassinoids). These results led to the following structure-activity relationships: (1) a methyleneoxy bridge and side chain enhance the activity and (2) a sugar moiety reduces the activity.

Antiviral Agents

Hemodynamic-force-induced difference of interendothelial junctional complexes.

1. The flow divider of the brachiocephalic branching of the rabbit aorta had both high and low shear stress regions, each of which was covered by endothelial cells with low and high permeability respectively, even in normolipidemic intact rabbits. When rabbits were placed on an atherogenic diet, low shear regions were the most vulnerable for lipid deposition, but the high shear regions were spared from deposition. 2. A freeze fracture study revealed that high shear regions both at the brachiocephalic branching and in the surgically coarctated abdominal aorta of rabbits had a more common appearance of zonular type tight junctions. Mean low shear regions had more macular and less zonular type. 3. Cultured porcine aortic endothelial cells exposed to laminar 30 dyn/cm2 shear stress in a flow chamber developed ridges of membranous protein particles at the cell-cell contact. 4. Increases of magnitude and duration of exposure to shear stress enhanced the structure of the protein ridge of the tight junction and immunohistochemical expression of proteins associated with both tight and adherens junctions.

Animals

Decreased cytoplasmic immunoglobulin A production during Epstein-Barr virus immortalization on lymphocytes from patients with ataxia-telangiectasia.

Previously, we have reported significantly lower immunoglobulin (Ig) A production in supernatants of cultured lymphoblastoid cells using enzyme-linked immunosorbent assay from patients with ataxia-telangiectasia (AT) when compared to that of age- and sex-matched healthy individuals. Here, we further assess the degree of cytoplasmic Ig production in these cells and also analyze it during the early phase of Epstein-Barr virus immortalization. All classes of cytoplasmic IgM, IgG, and IgA productions were demonstrated in cells from healthy controls. In contrast, cells from patients with AT showed only cytoplasmic IgM and IgG with low or nondetectable levels of IgA during and after the immortalizing process. These results suggest B lymphocytes bearing IgA are functionally immature and/or defective in patients with AT.

Adolescent

[Multiple eccrine spiradenoma. Histological association with dermal cylindroma].

A case of multiple eccrine spiradenomata is reported. One of the tumours was histopathologically associated with dermal cylindroma, and immunohistochemical studies showed that the cylindroma cells differentiate towards the secreting portion of the eccrine sweat gland. The relationship among multiple eccrine spiradenomata, dermal cylindroma and multiple trichoepitheliomata is discussed.

Adenoma

Bruceosides D, E, and F, three new cytotoxic quassinoid glucosides from Brucea javanica.

Three new quassinoid glucosides, bruceosides D [1], E [2], and F [3], were isolated from Brucea javanica, and their structures were elucidated by spectral evidence and chemical transformation to known compounds. Compounds 1-3 show selective cytotoxicity in the leukemia and non-small cell lung, colon, CNS, melanoma, and ovarian cancer cell lines with log GI50 values in the range of -4.14 to -5.72.

Antineoplastic Agents, Phytogenic

The reliability of a video-enhanced Hirschberg test under clinical conditions.

PURPOSE: To examine the reliability and usefulness of a video-based Hirschberg test under clinical conditions. METHODS: The authors estimated ocular deviation in 87 patients with strabismus through automated analysis of corneal reflex displacement using a video refractor. The reproducibility of measurement, the comparison with the prism and alternate cover test (PACT), and the distribution of the Hirschberg ratio were investigated. RESULTS: The 95% limits of agreement of the video-based Hirschberg test evaluated by repeated measurements were +/- 0.18 mm (equivalent to +/- 2.2 degrees or +/- 3.8 prism diopters [PD] of calculated strabismic deviation) for the horizontal deviation and +/- 0.28 mm (equivalent to +/- 3.4 degrees or +/- 5.9 PD) for the vertical deviation. The 95% limits of agreement between the Hirschberg measures and the PACT were within +/- 7.8 degrees or +/- 13.7 PD. The average (+/- SD) Hirschberg ratio was 12.3 +/- 1.2 degrees/mm or 21.8 +/- 2.1 PD/mm. CONCLUSIONS: The video-enhanced Hirschberg measurement shows good reproducibility and ease of application, even in the testing of infants. In quantitative analysis, however, systematic measurement error resulting from intersubject variance of the Hirschberg ratio should be taken into consideration.

Adolescent

Population analysis of cellular responses to synthetic peptides of Der p II, a major allergen molecule of Dermatophagoides pteronyssinus, in allergic and nonallergic subjects.

Responses of peripheral blood mononuclear cells to synthetic oligopeptides of Der p II, one of the major allergen molecules of Dermatophagoides pteronyssinus, were compared between allergic and nonallergic subjects. Healthy subjects showed positive responses to crude extracts of D. pteronyssinus, but only allergic subjects showed elevated cellular responses to Der p II. We synthesized three oligopeptides of Der p II in which motifs of a possible T-cell epitope were included. Of 14 subjects with positive response to Der p II, three responded to all three peptides, while five did not respond to any peptide tested. In 11 allergic patients who showed positive response to Der p II, responsiveness to the peptide K33-T47 was significantly higher than that to other peptides (P < 0.05). All the responding patients were also positive for scratch test to Der p II, suggesting that those epitopes induced IgE-promoting helper T-cell response in allergic persons. On the other hand, the in vitro cellular responses were not necessarily correlated to IgE production against Der p II in healthy subjects.

Adolescent

Processing and secretion of Alzheimer's disease amyloid precursor protein.

We studied in vivo expression and in vitro secretion of the Alzheimer's disease amyloid precursor protein (APP). The results indicate that secretion of APP is mediated by PKC and the initial step of the processing may occur in the acidic secretory granules of the glial cells. Our results suggest that a metabolic switch of APP in neural cells is critical in amyloid deposition.

Alzheimer Disease

Characterization of erythropoietin isolated from rat serum: biochemical comparison of rat and human erythropoietins.

We isolated erythropoietin (Epo) from anemic-rat serum with 1.3 x 10(6)-fold purification and 38% recovery using immunoaffinity chromatography. The isolated Epo migrated in SDS polyacrylamide gel with a molecular size of 37 kDa. Biological properties of rat Epo were compared with those of human Epo using target cells of primate and murine origins. When murine cells were used as target cells for assaying Epo, rat Epo stimulated proliferation of the cells with a 50% lower potency than did human Epo. The activity of rat Epo on human cells was only 25% of that of human Epo. Studies of Epo binding to the receptor indicated that rat and human Epos were not distinguishable in binding to murine cells; however, rat Epo bound to the receptor on human cells with an affinity much lower than that of human Epo. Rat Epo was digested with N-glycanase. Complete removal of N-linked sugars converted the native Epo to the deglycosylated form with 18 kDa. The in vitro activity of deglycosylated Epo was 2.5-fold higher than that of the native Epo.

Amino Acid Sequence

Nucleotide sequence of rat erythropoietin.

The cDNA for the rat erythropoietin (EPO) has been cloned and sequenced. The deduced amino acid sequence consists of 166 amino acid residues, which has a 79% and 95% homology with human and mouse EPOs, respectively. Many short stretches, highly conserved in primate and rodent EPOs, are found in the 3'-noncoding region when insertions and deletions are taken into consideration.

Amino Acid Sequence

Early activation of the proto-oncogene c-fgr during Epstein-Barr virus immortalization.

In an attempt to clarify the chronological relationships between Epstein-Barr virus (EBV) infection, B cell immortalization and c-fgr activation, we evaluated for the presence of EBV-determined nuclear antigen (EBNA), cellular DNA synthesis and expression of c-fgr-specific RNA following infection of human peripheral blood lymphocytes with B95-8 EBV. High expression of c-fgr was observed prior to EBNA detection and cellular DNA synthesis in EBV-infected cells. These results suggest that activation of c-fgr is an essential event during the early phase of EBV immortalization.

Antigens, Viral

Protein deficiency impairs erythropoiesis in rats by reducing serum erythropoietin concentration and the population size of erythroid precursor cells.

Erythropoietin (EPO), a glycoprotein produced mainly by the kidney, is the major physiological regulator of erythropoiesis. We developed a sensitive and rapid ELISA for measurement of rat serum EPO with two monoclonal antibodies that recognize different epitopes. To understand the mechanism by which erythropoiesis is impaired in rats deficient in dietary protein, we investigated the levels of the immunoreactive EPO (iEPO) in serums and erythroid precursor cells in hemopoietic tissues during protein deprivation. The iEPO level of 32-d-old rats fed a protein-free diet was lowered to one-third that of rats fed 20% casein at 6 h after protein deprivation began. Protein deprivation decreased the number of EPO-responsive cells in spleen. These results indicate that the impairment of erythropoiesis during protein deficiency is caused by the decrease in serum EPO and the subsequent reduction of the population size of erythroid precursor cells in spleen.

Animals

Analysis of antigenic determinants shared by two different allergens recognized by human T cells: house dust mite (Dermatophagoides pteronyssinus) and chironomid midge (Chironomus yoshimatsui).

To analyse the cross-reactivity of T-cell-mediated immunity between Dermatophagoides pteronyssinus (Dp) and Chironomus yoshimatsui (Cy), the most common allergens in Japan, we established antigen-specific human T-cell lines and clones. Some but not all of the Cy-induced T-cell lines showed a significant proliferative response not only to Cy, but also to Dp. No T-cell line responded to other unrelated antigens. When we stimulated the Dp-induced T-cell clones with Cy, 3 of the 40 clones (7.5%) showed a significant proliferation, and 2 of the 3 clones produced interleukin-4 and interferon-gamma, indicating their helper function. Cross-reactivity was diminished significantly after the absorption of Dp antigen in an anti-Cy affinity column. The cross-reactive epitopes were thought to be expressed on the Dp molecule of 45-53 kD. The presence of helper T cells reactive to both allergens suggests a possibility that this cross-reactivity might be involved in part in the high incidence of allergy to the 2 major allergens.

Allergens

Primary immunodeficiency diseases and Epstein-Barr virus-induced lymphoproliferative disorders.

Increased incidence of malignant disorders is noted in patients with both primary and acquired immunodeficiency diseases. The pathogenetic mechanism(s) for these disorders remain unclear. Defective immunosurveillance of these patients, however, is mainly postulated to be responsible for the increased risk of these malignant disorders. Of the malignant disorders, Epstein-Barr virus (EBV)-induced lymphoproliferative disorders (LPD) have been increasingly reported, possibly due to improved therapeutic management techniques such as bone marrow transplantation, which results in prolonged survival periods for the primary immunodeficiency; the dramatic development of immunosuppressive treatments for transplant recipients; and the growing numbers of acquired immunodeficiency syndrome (AIDS) patients. This review focuses on the primary immunodeficiency diseases and EBV-induced LPD, and discusses pathogenetic mechanism(s) for the increased incidence of these malignant disorders.

Adolescent

Endothelial cell morphometry of atherosclerotic lesions and flow profiles at aortic bifurcations in cholesterol fed rabbits.

Observations on shapes of endothelial cells both in sudanophilic and nonsudanophilic regions at bifurcations of the brachiocephalic (BC) and left subclavian (SA) arteries in hyperlipidemic rabbits were performed under a SEM. The stagnation point of flow and leading edges of flow dividers were nonsudanophilic and covered by round and long fusiform endothelial cells, respectively. The hips of flow dividers of both branchings, proven to be relatively low shear stress regions, by movement of microspheres in steady flow, were sudanophilic and covered by ellipsoidal cells. Similar studies were carried out in normolipidemic rabbits. It might be concluded that lipid deposition in hyperlipidemic rabbits occurs in relatively low shear stress regions, where endothelial cells are functionally activated, rather than in laminar high shear stress regions at the flow divider.

Animals