Biomedical subjects

H Shiono

Publications and source records attributed to H Shiono.

At least 19 recordsLinked to original sources

PubMed1999

Nitric oxide production by bronchoalveolar cells during allograft rejection in the rat.

BACKGROUND: We reported the increased nitric oxide (NO) level in exhaled air of rat lung transplant recipients during acute rejection (AR). The aim of this study was to determine the site and level of NO production in the rejected graft. METHODS: Rat lung transplantation was performed in isografts and allografts. RESULTS: In isografts, no AR and no significant increase in NO production was identified. In allografts, grades I-II of AR was seen on postoperative day (POD) 3 and grade III on POD 5. NO produced by BAL cells increased on both POD 3 (11.8+/-2.0 parts per billion [ppb]) and POD 5 (115.3+/-66.9 ppb). There was a highly significant correlation between the level of NO and the severity of AR (p=0.862, P<0.005). BAL cells from allografts expressed iNOS mRNA. Among them, macrophages, lymphocytes and neutrophils were immunostained for iNOS. CONCLUSION: NO produced by BAL cells was detected in the early stages of rejection. Therefore, it may serve as a sensitive indicator of AR in lung transplantation.

Animals↗

PubMed1999

The polymorphisms of DYS388 and DYS392 on the Y chromosome in Japanese and German populations.

The analysis of a genotype survey in Japanese and German populations at the loci DYS388 and DYS392 located on the Y chromosome is reported. The gene diversities of DYS388 were 0.34 and 0.30 in the Japanese and German males, respectively, and six alleles were found in both groups. The gene diversities of DYS392 were 0.65 and 0.64 and the number of alleles was 8 and 9, respectively in the two populations. The distribution of DYS388 alleles in the Japanese population was different from the German population. The allele distribution of DYS392 showed significant differences among Asian populations.

Asian People↗

PubMed1999

T-cell development in human thymoma.

Human thymoma is derived from thymic epithelial cells and often associated with a large number of cortical thymocytes. Since thymic epithelial cells play key roles in T-cell development in the normal thymus, we hypothesized that the neoplastic epithelial cells of thymoma may support T-cell differentiation. We attempted to reconstitute the T-cell development in vitro by using neoplastic epithelial cells isolated from thymoma. CD34, a stem cell marker, was expressed on a proportion of CD4-CD8- cells in thymoma. These CD34+CD4-CD8- cells also expressed both IL-7R alpha-chain and common gamma-chain. Purified CD4-CD8- cells from thymomas were cultured with the neoplastic epithelial cells, and their differentiation into CD4+CD8+ cells via CD4 single positive intermediates was observed within 9 days' co-culture in the presence of recombinant IL-7. The CD34+CD4-CD8- cells purified from a normal thymus also differentiated to CD4+CD8+ cells in an allogeneic co-culture with the neoplastic epithelial cells of thymoma. In addition, a pleural dissemination from thymoma contained a large amount of cortical thymocytes. These results suggest that the neoplastic epithelial cells retain the function of thymic epithelium and can support T-cell development in thymomas.

Adult↗

PubMed1999

Impaired expression of MHC class II molecules in response to interferon-gamma (IFN-gamma) on human thymoma neoplastic epithelial cells.

A human thymoma is a neoplasm derived from the thymic epithelial cell, and is well known for its association with autoimmune diseases, especially myasthenia gravis. The neoplastic epithelial cells of thymoma clearly retain thymic epithelial functions, but the development of T cells in thymoma is somewhat impaired. In this study, we quantified by flow cytometry the in vitro expression of MHC molecules on neoplastic epithelial cells precultured with IFN-gamma. While MHC class I expression was comparable with that on normal thymic epithelial cells, the level of MHC class II molecules on neoplastic epithelial cells was lower than in controls, and also varied greatly from case to case. Additionally, there was a significant positive correlation between the expression level of MHC class II and the proportion of mature CD3+ cells in the CD4+CD8- subset. Thus, accumulation of CD3-CD4+CD8- cells in thymoma may result from impaired expression of the MHC class II molecules, suggesting that the function of the neoplastic epithelial cells might determine the maturation and the positively selected repertoire of T cells in thymomas.

Adult↗

PubMed1999

Preparation of chylomicrons and VLDL with monoacid-rich triacylglycerol and characterization of kinetic parameters in lipoprotein lipase-mediated hydrolysis in chickens.

To identify the substrate specificity of lipoprotein lipase (LPL) for triacylglycerol-rich lipoproteins with monoacid-rich triacylglycerols, monoacid-rich lipoproteins were prepared and kinetic parameters of LPL were characterized. Male broiler chickens were fed 8 g/100 g fat diets differing only in the fat source: palm oil (tripalmitin-rich), olive oil (triolein-rich), safflower oil (trilinolein-rich) and linseed oil (trilinolenin-rich). After diets were fed for 3 d, chickens were starved for 2 d and then force-fed emulsions containing one of the monoacid-triacylglycerols: tripalmitin, triolein, trilinolein or trilinolenin. The triacylglycerols in chylomicrons and very low density lipoprotein (VLDL) of chickens force-fed tripalmitin, triolein or trilinolein contained the corresponding acid at more than 70% of total acids. Linolenic acid was incorporated into chylomicrons and VLDL to a lower extent (51.2 and 57.2%, respectively) in chickens force-fed trilinolein. Major apolipoproteins and lipid compositions were not significantly different among all lipoproteins isolated from chickens fed the different fats. Vmax of LPL was significantly higher (P < 0.05) for palmitic acid-rich chylomicrons and VLDL and decreased with increasing chain length and unsaturation of monoacid: 16:0>18:1>18:2>18:3. The electron spin resonance analysis, order parameter (S), decreased with monoacid chain length and unsaturation. In addition, the Vmax of LPL increased linearly (P < 0.01, r = 0. 912) with an increase in the palmitic acid content of the lipoprotein triacylglycerols. These findings suggest that lipoprotein catalysis by LPL is modulated by the palmitic acid content of the lipoprotein triacylglycerol, which affects the fluidity of lipoproteins.

Animals↗

PubMed1999

Characterization of the carrier-mediated transport of levofloxacin, a fluoroquinolone antimicrobial agent, in rabbit cornea.

The cornea presents a formidable barrier to drug penetration. The fluoroquinolone levofloxacin, which is an effective antimicrobial agent, has the potential to be used in the topical treatment of ocular disease. Thus, we sought to characterize how levofloxacin penetrates the cornea. To perform this characterization, we measured the time dependent permeation of levofloxacin across the isolated rabbit cornea using a diffusion chamber, and compared it with antipyrine fluxes. Levofloxacin permeation into the receiver epithelial-side bathing solution (pH = 6.5) from the donor endothelial-side (pH = 7.4) reached 3.00 nmolcm(-2) cornea after 2h, whereas in the opposite direction permeation was 1.89 nmolcm(-2) cornea. Based on the temperature-dependent effects on permeation, the calculated energy of activation for permeation, Ea, was 31.3 kcal mol(-1), whereas Ea for antipyrine, a marker of diffusion, was 11.0 kcalmol(-1). The transport of levofloxacin from epithelium to endothelium was concentration-dependent and had both a linear and saturable component. Evaluation of the kinetic parameters, Jmax, apparent Km and k(d) showed that they were 38.78 pmol min(-1) cm(-2), 3.83 mM and 0.0135 microL min(-1) cm(-2), respectively. These results, coupled with the fact that levofloxacin permeation reached a maximum value at pH 6.5, suggest that levofloxacin transport across the cornea is carrier mediated. However, at present, it cannot be ascertained whether such a system is localized in either the corneal epithelial or the endothelial layer.

Animals↗

PubMed1998

Immaturity of lymphocytes in the metastatic lesions of thymoma.

Thymoma is a thymic epithelial tumor which often contains a large number of immature T cells. Although the metastatic lesions are also associated with abundant lymphocytes, their characteristics have not been assessed in detail. In this study, the phenotype was analyzed and compared with those in their primary lesions. Nine metastatic thymomas were obtained from seven patients. In the metastatic lesions, CD1a+ cells and CD4(+)CD8(+) cells accounted for 77.7 +/- 10.6 and 52.3 +/- 15.8% of all the lymphocytes, respectively. In five primary lesions and their metastatic lesions, CD3(-)CD4(+)CD8(-) cells accounted for 23.9 +/- 16.9 and 45.2 +/- 15. 5% of the CD4(+)CD8(-) cells, respectively. CD69 was expressed on 70. 9 +/- 9.5 and 53.1 +/- 11.8% of the CD4(+)CD8(-) cells, respectively. These results indicate that the metastatic lesions of thymoma are associated with abundant immature T cells which are phenotypically less mature than those in their primary lesions.

Adult↗

PubMed1998

Reduced dorsal hippocampal glutamate release significantly correlates with the spatial memory deficits produced by benzodiazepines and ethanol.

Memory deficits frequently occur after taking benzodiazepines and ethanol. We studied in vivo hippocampal presynaptic glutamate transmission in conjunction with memory deficits induced by benzodiazepines and ethanol in rats as an animal model of amnesia. These drugs potently impaired spatial memory formation as evaluated by the Morris water maze task, the rank order among tested treatments being the combination of triazolam (20 micrograms/kg) with ethanol (2 g/kg) > or = triazolam (100 micrograms/kg) > ethanol (2 g/kg) > or = triazolam (20 micrograms/kg) > rilmazafone (20 micrograms/kg). On the other hand, these drug treatments also reduced glutamate release in the dorsal hippocampus but not in the cerebellum measured by microdialysis: the combined administration of triazolam with ethanol potently inhibited glutamate release to 60% of basal output in the dorsal hippocampus. These decreases in hippocampal glutamate transmission closely correlated with the extent of impairment of spatial memory performance (r = 0.990). Thus, the present results strongly indicated that presynaptic dysfunction in dorsal hippocampal glutamatergic neurons would be critical for spatial memory deficits induced by benzodiazepines and ethanol.

Animals↗

PubMed1998

Neoplastic thymic epithelial cells of human thymoma support T cell development from CD4-CD8- cells to CD4+CD8+ cells in vitro.

Human thymoma is a thymic epithelial cell tumour which often contains a large number of immature T cells and is frequently associated with autoimmune diseases. Since thymic epithelial cells play key roles in the development and selection of T cells in the normal thymus, we hypothesized that the neoplastic thymic epithelial cells of thymoma may support T cell differentiation in the tumour. We characterized CD4-CD8- cells in thymoma and applied an in vitro reconstitution culture system using the CD4-CD8- cells and the neoplastic epithelial cells isolated from thymoma. CD34, a stem cell marker, was expressed on 29.9 +/- 12.2% of CD4-CD8- cells in thymoma. TCRgammadelta was expressed on 27.4 +/- 15.1% of CD4-CD8- cells and CD19, a B cell marker, was expressed on 14.1 +/- 23.1% of CD4-CD8- cells. CD4-CD8- cells expressed both IL-7R alpha-chain and common gamma-chain. Purified CD4-CD8- cells from thymomas were cultured with the neoplastic epithelial cells, and their differentiation into CD4+CD8+ cells via CD4 single-positive intermediates was observed within 9 days' co-culture in the presence of recombinant IL-7. Furthermore, we examined the reconstitution culture using CD34+CD4-CD8- cells purified from normal infant thymus. The CD34+CD4-CD8- cells in normal thymus also differentiated to CD4+CD8+ cells in the allogeneic co-culture with the neoplastic epithelial cells of thymoma. These results indicate that the tumour cells of thymoma retain the function of thymic epithelial cells and can induce differentiation of T cells in thymoma.

Adult↗

PubMed1998

Beta adrenergic antagonist permeation across cultured rabbit corneal epithelial cells grown on permeable supports.

PURPOSE: To determine whether cultured rabbit corneal epithelial cells (RCEC), grown on permeable supports, provide a suitable in vivo model for characterizing transcellular drug permeation and metabolism. METHODS: Primary rabbit corneal epithelial cells grown in DMEM-F12 were seeded on Transwell-COL inserts coated with fibronectin. The epithelial barrier integrity was evaluated, based on measurements of 14C-mannitol and 3H-PEG900, and their transepithelial electrical resistance (TEER). Ultrastructure evaluation was based on scanning electron microscopy and transmission electron microscopy, which were performed 8 days after seeding. Measurements of beta adrenergic antagonist permeability were performed to assess transcellular permeability. RESULTS: Eight days after seeding, the TEER reached a peak of 144 omega.cm2 and the 14C-mannitol and 3H-PEG900 permeabilities were 6.8 x 10(-6) and 2.9 x 10(-6) cm/sec, respectively. Ultrastructural analysis revealed a multilayered structure with numerous microplicae and typical cytoplasmic organelles along with desmosomes. The relationship between permeation of beta-blockers and lipophilicity resembled the intact isolated cornea. CONCLUSIONS: This is the first description of cultured RCEC grown on permeable support. Many of its properties mimic those described in the intact corneal epithelium. Even though its electrical tightness is less than that of the intact cornea, the transcellular permeability to lipophilic beta-antagonists is comparable to the isolated preparation. Therefore, this model will facilitate characterization of ocular permeation mechanisms of hydrophobic drugs whose route of permeation is transcellular.

Adrenergic beta-Antagonists↗

PubMed1998

[Use of a CO-oximeter for forensic diagnosis of hypothermia].

Blood samples were collected from the right and left atria separately in 9 cases of hypothermia and 18 cases of non-hypothermia during judicial and consented autopsy between March 1995 and December 1997. The oxyhemoglobin ratio (O2Hb), reduced hemoglobin ratio (RHb) and oxygen saturation (sO2) were measured by oximetric analysis. O2Hb, R-Hb and sO2 were significantly greater in the hypothermic group than the non-hypothermic group. We conclude that the diagnosis of hypothermia can be established in cases where O2Hb and sO2 in the left atrium exceed 80%, irrespective of values in the right atrium.

Adult↗

PubMed1998

Unnecessary surgical operation confirmed by individual identification by analyses of several short tandem repeat (STR) loci.

A 60-year-old man who was diagnosed as having hepatic cell carcinoma by needle punch biopsy from the liver, underwent right hepatic lobectomy. However, no malignant components were detected in the removed tissue. It was suspected that he was the wrong patient and had undergone the operation unnecessarily. Individual identification was performed using paraffin blocks of his liver tissue taken by needle punch biopsy, that of liver tissue taken at surgery and his peripheral blood by genotyping several STR loci. All of these specimens had the same genotypes. Histological re-examination revealed that the biopsied sample contained no malignant component. The end result was that the initial biopsied liver specimen was misdiagnosed as hepatic cancer and the patient underwent an unnecessary operation due to misdiagnosis.

Biopsy, Needle↗

PubMed1997

Human identification by genotyping of personal articles.

In cases of personal identification of unknown skeletal remains, forensic scientists normally analyze an individual's genotype or phenotype by comparison with those of other family members such as patients. However, such genotyping does not necessarily provide conclusive proof of identity. In this study, we attempted personal identification by comparison of several short tandem repeat (STR) loci in samples from three corpses in various stages of decomposition with samples from personal articles of the respective individuals. The first victim had been found dead in a burned car. Almost 100% of his body was charred. The genotypes of the iliac bone from the burned body with regard to several STR loci were compared with those from a single hair recovered from the vacuum cleaner from the candidate's home. The second victim was found in a forest after more than 5 years. Several STR loci genotypes of skeletal bone were compared with those of the candidate's mummified umbilical cord kept in his parent's house. In the last case the victim was found in the sea after more than 7 months. The genotypes of the loci in the pubic bone of the corpse were compared with those of a menstrual bloodstain on the candidate's underwear left in the washing machine at her home. We succeeded in identification of the three cases by comparison of several VNTR and STR loci in the corpses with those in personal articles. In conclusion, personal identification can sometimes be performed more precisely by genotyping of personal articles than by comparison with the genotypes of relatives.

Adult↗

PubMed1997

Expression of ubiquitin protein in each organ at death from hypothermia.

To examine the expression of heat shock protein (hsp) in each organ on autopsy in 11 deaths from hypothermia, we performed immunological staining of specimens from each organ using an antibody to hsp, ubiquitin (Ub). Staining of the liver, kidneys, lungs and pancreas revealed a high level of Ub expression in the cytoplasm and nuclei. However, staining of the same specimens of all controls revealed no Ub expression in the cytoplasm or nuclei. This finding suggests that cells are affected by stress even at a low temperature, and may be important in clarifying the cellular kinetics in death from hypothermia.

Adult↗

PubMed1997

Effect of sialic acid residues of human alpha 1-acid glycoprotein on stereoselectivity in basic drug-protein binding.

The function of sialic acid groups at the terminal of sugar chains of human alpha 1-acid glycoprotein (AGP) was investigated with respect to chiral discrimination between optical isomers of basic drugs, using high-performance capillary electrophoresis/frontal analysis (HPCE/FA), a novel analytical method developed for the determination of unbound drug concentration with ultramicrosample volume (100-200 nl). Native human AGP and desialylated AGP were used as test proteins, and propranolol (PRO) and verapamil (VER) were used as model drugs. The unbound concentration of (S)-VER was 1.31 times higher than that of (R)-VER in native AGP solution. This selectivity was not affected by desialylation. Further, enzymatic elimination of galactose residues, which neighbored sialic acid groups, did not change the binding of either isomer of VER. On the other hand, the unbound concentration of (R)-PRO was 1.27 times higher than that of (S)-PRO in native AGP solution. Desialylation caused the unbound concentration of (S)-PRO to rise to the same level of (R)-PRO, resulting in loss of enantioselectivity. Thus, it follows that sialic acid groups of AGP, as a whole, are not responsible for chiral recognition between enantiomers of VER but are involved in enantioselectivity toward the isomers of PRO.

Adrenergic beta-Antagonists↗

PubMed1997

Failure to down-regulate Bcl-2 protein in thymic germinal center B cells in myasthenia gravis.

The most unusual characteristic of myasthenia gravis (MG) is that the thymus has germinal centers (GC). Cultured thymic lymphocytes from MG patients spontaneously produce anti-acetylcholine receptor antibodies, indicating that autoreactive B cells have escaped negative selection. To investigate the underlying mechanism, we examined the expression of the apoptosis-related protein Bcl-2 in GC B cells (defined as CD19+ CD38+ cells) in the thymus in 14 MG patients using three-color flow cytometry. GC in MG patients did not show the normal down-regulation of Bcl-2 (the frequency of Bcl-2+ GC B cells in the MG thymus and in control tonsils 54.3 +/- 16.2% versus 20.6 +/- 8.0%; mean +/- SD. p < 0.0001). In contrast, Bcl-2 in GC in the mediastinal lymph nodes from four patients was down-regulated to a relatively normal level. Using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method to detect DNA fragmentation in situ, the frequency of TUNEL+ cells in GC in the MG thymus was lower than in control tonsils. These results suggest that autoreactive B cells which normally undergo apoptosis in GC may survive because of Bcl-2 up-regulation in this unusual location.

Adolescent↗

PubMed1997

Mature CD4 single positive thymocytes in human Thymoma: T cells may differentiate in the thymic epithelial cell tumor.

Human thymoma, which is occasionally associated with autoimmune disease, is a thymic epithelial cell tumor and often contains a large number of lymphocytes. In a previous study, we have shown that a proportion of CD4 single positive T cells in human thymomas lack CD3, suggesting immaturity. In this study, we focused on the rest of the CD4 single positive T cells in thymomas that expressed CD3/TcR alpha beta and investigated the maturity of single positive T cells by analyzing lymphocyte surface antigens and the cells' proliferative response to a mitogen. CD4 single positive cells that expressed CD3 or TcR alpha beta also expressed CD69 and had probably undergone positive selection in the tumor. Further, isolated CD4 or CD8 single positive cells from the thymomas responsed to a mitogen although at lower levels than the corresponding single positive cells in the peripheral blood. These results indicate that thymomas contain single positive T cells which have mature phenotype and proliferative ability, and suggest that T cells may differentiate in thymoma.

Adult↗

PubMed1997

Cellular contraction precedes membrane depolarization in Vorticella convallaria

Application of a mechanical stimulus to the cell body of the peritrich ciliate Vorticella convallaria evoked an all-or-nothing membrane depolarization, the large pulse. This was always accompanied by an all-or-nothing cellular contraction, and simultaneous recordings of the two events revealed that the large pulse was always preceded by the cellular contraction. A smaller graded membrane depolarization (the medium pulse) was sometimes produced in response to a weaker mechanical stimulus. The medium pulse was accompanied by a small, graded, localized contraction of the cell body and was occasionally followed by a large pulse. When a large pulse occurred during a medium pulse, it reached the same peak level as that of a large pulse evoked without a preceding medium pulse. When a medium pulse occurred during a medium pulse, summation of the two pulses was observed. Sustained contraction causes V. convallaria to become rounded, and in this state a mechanical stimulus stronger than that used to evoke the large pulse evoked a graded depolarizing mechanoreceptor potential in the cell. We conclude that both the large and medium pulses are caused by an inward receptor current that is activated mechanically following contraction of the cell body. A localized contraction evokes a small mechanoreceptor current, causing a medium pulse. An all-or-nothing contraction evokes a saturated, all-or-nothing mechanoreceptor current, causing a large pulse.

Journal Article↗