Biomedical subjects

Y Ikarashi

Publications and source records attributed to Y Ikarashi.

At least 109 records · Page 6Linked to original sources

PubMed1993

Rabbit eye irritation caused by wearing toxic contact lenses and their cytotoxicities: in vivo/in vitro correlation study using standard reference materials.

To clarify the relationship between eye irritancy and cytotoxic potential induced by irritant materials, we made lenses coated with standard reference materials (SRMs) prepared from various amounts of zinc diethyldithiocarbamate (ZDEC) and polyurethane (PU). Zinc diethyldithiocarbamate was classified as a mild irritant by the Draize eye irritation test. When ZDEC-SRM coated contact lenses were applied to rabbit eyes, Draize scores increased in proportion to both the ZDEC and PU concentrations used for coating. Furthermore, correlation with the cytotoxic potential (gamma = -0.93) was better than with lactate dehydrogenase (LDH) activities of tears from rabbit eyes wearing these coated lenses (gamma = 0.78). In conclusion, in vivo eye irritancy induced by wearing lenses could be estimated quantitatively with the cytotoxic potentials using a colony assay. Furthermore, we could compare different sensitivities caused by the same set of SRMs among three different sites of tissue. As a result, the order of sensitivity was eye > muscle >> skin.

3T3 Cells↗

PubMed1993

A sensitive mouse lymph node assay with two application phases for detection of contact allergens.

A predictive test using mice for the identification of contact sensitizing chemicals was developed. Contact sensitizing activity is measured as a function of draining lymph node activation following application of test chemical. Experimental conditions for assessment of induced lymph node cell (LNC) responses have been optimized. BALB/c mice were initially treated with intradermal injections of test chemical in Freund's complete adjuvant emulsion. Five days after intradermal injection, mice were exposed topically to chemical in vehicle on the ears daily for 3 consecutive days. Next day following the final exposure, changes in lymph node weight, total cell number in the draining lymph nodes and LNC proliferation for 24 h culture were assessed. The performance of the method was evaluated with ten sensitizing chemicals and a non-sensitizing irritant, sodium lauryl sulfate (SLS). The LNC proliferation induced by combination of intradermal injection and topical application of sensitizing chemicals was more clearly increased than that following only topical application. With the single exception of sulfanilic acid, the method developed was able to detect the sensitizing capacity of chemicals that failed to induce sensitization in the local lymph node assay. Under the conditions used, SLS did not induce measurable lymph node responses. These results suggest that the mouse lymph node assay can provide a sensitive screening test for weak to moderate sensitizers. In addition, the assay offers the advantages of objective and quantitative endpoints, and is suitable for the evaluation of colored or irritant chemicals.

Administration, Topical↗

PubMed1993

Compensation between sympathetic nerves and adrenal medullary activity:effects of adrenodemedullation and chemical sympathectomy on catecholamine turnover.

To clarify functional compensation between the sympathetic nerves and the adrenal medulla, the effects of adrenodemedullation and chemical sympathectomy on norepinephrine (NE) turnover in some sympathetically innervated tissues and on NE and epinephrine (E) turnover in the adrenal gland were studied in rats. The rate of NE turnover was significantly accelerated by bilateral adrenodemedullation in the pancreas and in interscapular brown adipose tissue (IBAT). Chemical sympathectomy accelerated E turnover in the adrenal gland. These data indicate that some adrenergic nerves show functional compensation under conditions of depression of the adrenal medulla, and that compensatory acceleration of the adrenal medullary function occurs under conditions of adrenergic dysfunction.

Adipose Tissue, Brown↗

PubMed1993

Dopamine output upon death reflects intraneuronal aspects while alive: accumulation of releasable dopamine during tetrodotoxin perfusion.

Intraneuronal and extracellular dopamine (DA) metabolism in rat striatum during inhibition of neurotransmission processes by tetrodotoxin (TTX) perfusion was examined by analysis of the striatal DA content and the amount of initial post-mortem DA output. In spite of there being an excess amount of DA already present in the striatum, the contents of DA and its metabolites were increased time-dependently by TTX perfusion through a microdialysis membrane. A massive increase in extracellular level of DA occurred immediately after death. The amount of the post-mortem DA output was further increased in proportion to the preceding TTX perfusion time. The ratio of the amount of post-mortem DA output to the striatal DA content was also increased by previous perfusion with TTX. Verapamil, a Ca2+ channel blocker, delayed the appearance of the peak of the DA output. These results suggest that releasable DA accumulates in neurons during TTX perfusion, and that the early part of the DA output upon death reflects intraneuronal aspects of DA metabolism while the animal is alive.

Animals↗

PubMed1993

Effects of choline-free plasma induced by choline oxidase on regional levels of choline and acetylcholine in rat brain.

Choline-free plasma (CFP) was induced in rats by intravenous (IV) injection of 56.0 x 10(2) units kg-1 of choline oxidase (ChO) which completely metabolized the free Ch circulating in the plasma for at least 15.0 h and caused subsequent significant decrease in the concentration of free Ch in the three brain regions examined, the striatum, hippocampus, and cortex. However, the treatment did not affect concentrations of acetylcholine (ACh) in these regions. By contrast, intraperitoneal (IP) injection of 1.0 mmol kg-1 Ch chloride resulted in a maximum concentration of free Ch in plasma in 5 min, after which tissue Ch in all regions examined increased (p < 0.001). Concomitant increases were observed in cortical and hippocampal ACh (p < 0.05) 20 min after the injection. It is thus suggested that the brain may possess compensative mechanisms to prevent the supply of free Ch from circulating to the brain during synthesis of ACh in the brain. It is also suggested that the CFP rat would be a useful and readily available animal model for future study.

Acetylcholine↗

PubMed1993

Acute neurochemical changes in mouse brain following cerebral ischemia.

Acute changes in neurochemical levels induced by ischemia were studied in the mouse brain. Contents of neurochemicals in the frontal, parietal and occipital cortices and hippocampus were determined immediately after 15 min of ischemia (0), and then at 15, 30, 90 and 180 min after recirculation following ischemia. These data were compared with those for sham-operated control mice. Choline (Ch) contents in ischemic animals increased by 530-630% from control levels immediately after ischemia, and returned to control levels by 90 min. Decreases in levels of norepinephrine (NE) and serotonin (5-HT) were observed during 30 min after recirculation. There were no significant changes in levels of acetylcholine (ACh) or dopamine (DA), throughout recirculation. On the other hand, DA and 5-HT metabolites (3,4-dihydroxyphenylacetic acid and 5-hydroxyindoleacetic acid) significantly increased. Thus, comprehensively investigating the various neurotransmitters will provide meaningful information regarding the disturbance of central nervous system induced by cerebrovascular disease.

Acetylcholine↗

PubMed1993

Rejection of cultured keratinocyte allografts in presensitized mice.

The fate of cultured keratinocyte (KC) allografts remains controversial. Although prolonged survival of cultured KC allografts in naive mice has been reported, the detailed mechanisms remain undetermined. Furthermore, it was also reported that in the human cultured KC allografts do not survive permanently, and they are rapidly replaced by recipient cells. In the present study, we have addressed this issue and obtained findings that cultured KC allografts survive for a prolonged period in naive mice under the conditions in which reepithelization by recipient cells is prevented. However, the same cultured KC allografts were rejected when they were grafted onto recipients primed with allogeneic spleen cells or full-thickness skin grafts. To clarify the mechanisms behind these findings, the allostimulatory ability of cultured KC and their susceptibility to alloreactive cytotoxic T cells were examined in vitro. In a mixed epidermal cell-lymphocyte reaction, cultured KC were unable to induce allospecific proliferative responses of naive T cells. On the other hand, primed T cells from presensitized mice showed weak but significant proliferative responses against allogeneic KC. It also was confirmed that cultured KC are susceptible to lysis by alloreactive cytotoxic T cells. These data indicate that prolonged survival of cultured KC allografts in naive mice is attributable to a defect in the afferent, but not the efferent, phase of the rejection process that is caused by the weak allostimulatory ability of cultured KC. This assumption was also supported by the finding that spleen cells from the recipient mice bearing long-surviving KC allografts retain in vitro responsiveness against stimulator cells syngeneic to the grafted KC. Taken together, these findings indicate that long-term survival of cultured KC allografts in naive mice may be due solely to the weak allostimulatory ability of cultured KC, but not to loss of susceptibility to alloreactive cytotoxic T cells after culture of KC or induction of tolerance in the recipient mice bearing KC allografts.

3T3 Cells↗

PubMed1993

The role of nitric oxide in cholinergic neurotransmission in rat trachea.

1. We have investigated the role of nitric oxide (NO) in cholinergic contraction in rat trachea. 2. Methylene blue (10 nM to 30 microM) potentiated cholinergic contraction induced by electrical field stimulation (EFS) at 5 Hz in a concentration-dependent fashion. At a concentration of 30 microM, methylene blue decreased responses to log EFS frequency, producing 50% of maximum contraction from a control value of 0.74 +/- 0.09 Hz to 0.30 +/- 0.05 Hz without a significant effect on concentration-response curves to acetylcholine (ACh). 3. NG-monomethyl-L-arginine (L-NMMA; 100 microM) also potentiated cholinergic contraction induced by EFS at 5 Hz (131.5 +/- 4.6% of control) without having any effect against ACh (3 microM)-induced contractions. Likewise, L-NMMA (100 microM) significantly increased EFS (5 Hz)-evoked release of ACh from tracheal segments into the bath solution (51.4 +/- 4.0 pmol ml-1 in the presence of L-NMMA and 35.0 +/- 1.8 pmol ml-1 in the absence of L-NMMA, respectively). 4. Administration of NO (present in acidified solution of NaNO2) (1 nM to 10 microM) and sodium nitroprusside (100 nM to 10 microM) concentration-dependently reduced EFS (5 Hz)-induced cholinergic contractions without having a significant effect on ACh (3 microM)-induced contractions. These results were unaffected by prior exposure of the tissues to L-NMMA (100 microM). 5. Dibutyryl cyclic GMP (3 mM) also reduced cholinergic contractions induced by EFS at 5 Hz (70.1 +/- 3.6% of control) without any significant effect on ACh (3 microM)-induced contractions. 6. Pretreatment of tissues with capsaicin (30 microM) or a-chymotrypsin (1 u ml-') failed to inhibit methylene blue (30 microM)-induced potentiation of responses to EFS at 5 Hz.7. These results suggest that an endogenous NO-like factor may mediate prejunctional inhibition of cholinergic contraction through a cyclic GMP-dependent mechanism in rat trachea.

Acetylcholine↗

PubMed1993

Evaluation of contact sensitivity of rubber chemicals using the murine local lymph node assay.

The sensitizing abilities of 4 rubber additives, tetramethylthiuram disulfide (TMTD). 2-mercaptobenzothiazole (MBT), N-isopropyl-N'-phenyl-p-phenylenediamine (IPPD) and zinc diethyldithiocarbamate (ZDEC), were evaluated using the murine local lymph node assay. Exposure to IPPD induced a significant increase of lymph node cell proliferation in the draining lymph nodes even at a low concentration. Exposure to TMTD and MBT induced moderate proliferative responses, while ZDEC induced a weak proliferation even at the higher concentrations. The sensitizing potency of each chemical was described in terms of the concentration that increased lymph node cell proliferation by a factor of 2 over that in the vehicle-treated control group. The concentrations of IPPD, TMTD, MBT and ZDEC were 0.14%, 0.35%, 4.2% and > 10%, respectively.

Animals↗

PubMed1993

Influence of irritants on lymph node cell proliferation and the detection of contact sensitivity to metal salts in the murine local lymph node assay.

Dimethyl sulfoxide (DMSO) and sodium lauryl sulfate (SLS) are known to cause irritation of the skin, and to enhance the penetration of chemicals into the epidermis. In the present study, the lymph node cell (LNC) proliferative response following exposure to irritants, such as SLS and DMSO, was examined in the murine local lymph node assay (LLNA). Exposure to DMSO or SLS aqueous solution induced a small increase in lymph node cell proliferation compared with aqueous solution alone. Exposure to SLS in DMSO caused a significant increase in LNC proliferation. Further, the effect of addition of the irritants in a vehicle on the detection of contact sensitivity to metal allergens was examined. Application of potassium dichromate and nickel sulfate in DMSO or SLS aqueous solution caused increases in LNC proliferation. Exposure to metal allergen with SLS in DMSO also induced a significant LNC proliferative response, but did not induce a significant increase in stimulation index (increase in 3H-thymidine incorporation relative to vehicle-treated control group). This was because of increased 3H-thymidine incorporation following exposure to SLS-DMSO in the control group. These results suggest that irritants enhance the LNC proliferative responses to metal allergens. The use of SLS in aqueous solution is effective for the detection of sensitivity to water-soluble allergens, such as metal allergens, in the LLNA, as well as the use of DMSO as an application vehicle.

Animals↗

PubMed1993

Immunosuppressive activity of macrophages in mice undergoing graft-versus-host reaction due to major histocompatibility complex class I plus II difference.

In a previous report, we showed that the injection of parental CD4+ T cells into major histocompatibility complex (MHC) class II disparate F1 hybrid mice induces autoimmune-like graft versus-host reaction (GVHR) resembling systemic lupus erythematosus (SLE) and the hepatic lesion of primary biliary cirrhosis (PBC). In the present study, we examined whether or not simultaneous or subsequent injection of CD8+ T cells changes the GVHR form. When parental CD8+ T cells together with CD4+ T cells were injected into MHC class I plus class II-disparate F1 mice, autoimmune phenomena did not develop and alternatively a profound immunosuppressive state was induced. Furthermore, ongoing autoimmune-like GVHR induced by CD4+ T cells was also suppressed by later injection of CD8+ T cells. In these mice, an increase of donor type CD8+ T cells and a marked decrease of host B and T cells in the spleen were observed. The spleen cells from these mice strongly inhibited the mitogenic response of normal spleen cells against lipopolysaccharide (LPS). In vitro studies demonstrated that this immunosuppression was not induced by CD8+ T cells themselves but by macrophages which produced suppressive factor(s) by LPS stimulation. These findings were discussed with reference to suppressive mechanisms of GVHR.

Animals↗

PubMed1993

Effect of the novel histamine H2-antagonist 5,6-dimethyl-2-[4-[3-(1- piperidinomethyl)phenoxy]-(z)-2-butenylamino]-4(1H)-pyrimidine dihydrochloride on histamine-induced gastric acid secretion in Heidenhain pouch dogs.

Effects of IGN-2098 (5,6-dimethyl-2-[4-[3-(1-piperidinomethyl)phenoxy]- (z)-2-butenylamino]-4(1H)-pyrimidone dihydrochloride, CAS 126869-04-3) a novel histamine H2-antagonist, on histamine-induced gastric acid secretion were investigated in Heidenhain pouch dogs in comparison with those of famotidine, roxatidine acetate HCl and cimetidine. Orally administered IGN-2098 (0.03-1.0 mg/kg), famotidine (0.01-0.3 mg/kg), roxatidine acetate HCl (0.1-1.0 mg/kg) and cimetidine (0.3-3.0 mg/kg) showed dose-dependent inhibition on histamine-induced gastric acid secretion, and ED50 values of IGN-2098, famotidine, roxatidine acetate HCl and cimetidine were 0.077, 0.024, 0.200 and 0.585 mg/kg, respectively. IGN-2098 was effective even at 6 h after administration and ED50 value was 0.315 mg/kg. IGN-2098 was effective also by intravenous route. The inhibitory effect of IGN-2098 on histamine-induced gastric secretion was not affected by the repeated administration of IGN-2098 (1 mg/kg b.i.d. for 14 days). These results show that IGN-2098 is a potent and long acting antisecretory agent and is a useful antisecretory drug for the treatment of peptic ulcer disease.

Administration, Oral↗

PubMed1992

Differences of draining lymph node cell proliferation among mice, rats and guinea pigs following exposure to metal allergens.

Contact sensitivities of three well known metal allergens (nickel sulfate, potassium dichromate and cobalt chloride) were examined using the local lymph node assay in CBA/N mice, F344 rats and Hartley guinea pigs. The effect of various species sera on lymph node cell (LNC) proliferation was also investigated. Exposure to potassium dichromate and cobalt chloride induced significant LNC proliferative responses in the three species. The LNC responses to potassium dichromate in the rats were higher than those in the mice and guinea pigs. Mice exhibited the highest response to cobalt chloride among the three species, whereas, exposure to nickel sulfate failed to induce a marked LNC proliferation. Increased draining lymph node weights and LNC numbers were also observed following exposure to the metal salts. However, these parameters were less sensitive compared with the LNC proliferative response. There was a large difference in the lymph node weight between individual guinea pigs. The [methyl-3H]thymidine incorporation into LNC of each species cultured in the presence of the homologous serum in vitro was lower than in the presence or absence of fetal calf serum. However, there was no significant difference in stimulation indices among the different culture conditions. The local lymph node assay may be performed in rats as well as in mice for the detection of metal allergens.

Allergens↗

PubMed1992

Mechanism of the rejection of major histocompatibility complex class I-disparate murine skin grafts: rejection can be mediated by CD4+ cells activated by allo-class I + II antigen in CD8+ cell-depleted hosts.

In the preceding article, we analyzed the immunohistochemical rejection mechanism of major histocompatibility complex (MHC) class I (H-2K)-disparate murine skin grafts, and showed that only CD8+ cells infiltrated at the site of the epithelial tissue of MHC class I-disparate graft. We also showed that perfect survival of MHC class I-disparate grafts were attained in thymectomized recipients treated with anti-Lyt-2 monoclonal antibody. In this report, we showed that these long-surviving allo-class I grafts were rejected in the absence of CD8+ cells by stimulation with allo-MHC class I + II-disparate graft as the second stimulation. Furthermore, it was immunohistochemically revealed that under that condition, a large number of CD4+ cells infiltrated into the epithelial tissue of these long-surviving class I grafts, which were going to be rejected 2-5 d after the transplantation of a second graft with MHC class I + II difference. This result directly shows that CD4+ cells are able to became effectors for the rejection of allo-MHC class I (H-2K) skin graft.

Animals↗

PubMed1992

Glassy carbon pre-column for direct determination of acetylcholine and choline in biological samples using liquid chromatography with electrochemical detection.

The determination of acetylcholine and choline has been quite successfully accomplished using liquid chromatography with electrochemical detection following the original reports of Potter et al. [J. Neurochem., 41 (1984) 188]. A post-column reactor containing acetylcholinesterase and choline oxidase allows conversion of the desired species into hydrogen peroxide, an electrochemically active substance. However, the direct injection of tissue homogenates and other biological samples into such a system exhibits quite large solvent fronts and unidentified peaks. Using a pre-column packed with glassy carbon particles, we were able to dramatically decrease the size of the solvent front for such injections and tentatively identify the unknown peaks to be caused, at least in part, by common catecholamines. The glassy carbon pre-column, in addition to increasing the selectivity of the results, allowed the required chromatographic time per sample to be decreased from 20 to 10 min.

Acetylcholine↗

PubMed1992

Details of an isolation method for hepatic lymphocytes in mice.

The liver comprises a unique lymphocyte population, i.e., extrathymic alpha beta T cells with TcR of intermediate intensity. In the present study, we attempted to determine what pretreatments were appropriate to isolate hepatic mononuclear cells (MNC) containing such intermediate alpha beta TcR cells in mice. Hepatic MNC were isolated from untreated mice and mice subjected to either bleeding or liver perfusion, and the intermediate alpha beta TcR cells in each preparation were identified. For reasons of simplicity, cell purity and cell yields, hepatic lymphocytes should be obtained from mice subjected to total bleeding. Additional information on extrathymic alpha beta T cells obtained by using the recommended method is also presented.

Animals↗

PubMed1992

Identification of O-methyldopa in the ventricular fluid of patients with Parkinson's disease.

A newly developed liquid chromatographic system with a multiple electrode detector was used for quantification of neurochemical substances in ventricular fluid of patients with Parkinson's disease. During the analysis, an unknown peak was observed at almost the same high-performance liquid chromatography retention time as 5-hydroxytryptophan. Through the use of ion suppression techniques, voltammographic analysis and fast atom bombardment mass spectrometry, the compound was identified as O-methyldopa, the major metabolite of L-dopa.

Cerebral Ventricles↗

PubMed1992

Comparative studies by cell culture and in vivo implantation test on the toxicity of natural rubber latex materials.

Colony assay using V79 cells, the agar diffusion assay with L929 cells, and the 7-day rabbit muscle implantation test were employed to evaluate the cytotoxicity and tissue toxicity of natural rubber latex (NRL) materials. The in vivo implantation test showed that, among 13 histological parameters, thickness of inflammatory layer was the most useful index to evaluate tissue responses quantitatively. A comparison of the in vivo and in vitro parameters revealed the following correlations between the thickness of the inflammatory layer and cytotoxicity indices: Colony assay of the extracts, IC50: r = 0.80; Agar diffusion assay, Zone index: r = 0.73; Lysis index: r = 0.61. From these results, it appears that the colony assay provides a more reliable prediction of the tissue response than the agar diffusion assay.

Animals↗