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Naoki Harikai

Publications and source records attributed to Naoki Harikai.

11 recordsLinked to original sources

Acute heat stress induces jumping escape behavior in mice.

A new heat escape behavior was revealed in mice (ddY mice) under acute heat stress conditions. Mice in a fully covered cage were exposed to 24, 34, 37 and 38.5 degrees C for 60 min. Rectal temperature increased in conditions above 34 degrees C. Furthermore, serum osmolality and body weight loss also increased in conditions above 37 degrees C. At above 37 degrees C, a large number of mice attempted to escape from the partially covered cage, and so exhibited jumping behavior during a period of 60 min. However, mice exposed to 24 and 34 degrees C did not exhibit such behavior. These results indicated that acute heat stress above 37 degrees C induced evaporative water loss and jumping escape behavior in mice.

Analysis of Variance↗

[Inter-laboratory evaluation studies for development of notified ELISA methods for allergic substances (milk)].

Extracts of sausage, sauce, cookie, cereal and pasta sauce spiked with milk standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate in 10 laboratories. Coefficients of variation (CVs) of the three ELISA methods using a Milk Protein Casein ELISA Kit (Casein kit), a Milk Protein Beta-Lactoglobulin ELISA Kit (Beta-Lactoglobulin kit) and a FASTKIT Milk ELISA Kit (Milk ELISA kit) were mostly below 10%. Mean recoveries of the milk standard protein from the food extracts were over 40% in the three ELISA methods with a few excertions. The recoveries of milk standard protein from the sauce extract in Casein kit were improved by adjusting the extract to neutrality before the Casein kit assay. The recoveries of milk standard protein from cookie, cereal and pasta sauce were improved by the increasing the amount of antibody coated in the Milk ELISA kit. The detection limits of all the ELISA methods were 1 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of milk protein levels in extracts of sausage, sauce, cookie, cereal and pasta sauce.

Allergens↗

[Inter-laboratory evaluation studies for development of notified ELISA methods for allergic substances (wheat)].

Extracts of sausage, sauce, pasta sauce, fish paste and cereal spiked with wheat standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate in 10 laboratories. Coefficients of variation (CVs) of both ELISA methods using a Wheat Protein ELISA Kit (Gliadin kit) and a FASTKIT Wheat ELISA Kit (Wheat ELISA kit) were mostly below 10%. Mean recoveries of the wheat standard protein from the food extracts were over 40% in the two ELISA methods except those from cereal extract determined using the Wheat ELISA kit. Repeatability relative standard deviations of wheat standard protein in the five food extracts were in the ranges of 16-26.9% and 3.7-36.2% for the Gliadin kit and the Wheat ELISA kit, respectively. Reproducibility relative standard deviations of wheat standard protein in the five food extracts were 21.6-38.5%, 29.7-53.8% for the Gliadin kit and the Wheat ELISA kit, respectively. The recoveries of wheat standard protein from the cereal extract were improved by the increasing the amount of antibody coated on the plate in the Wheat ELISA kit. The detection limits of both ELISA methods were 1 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of wheat protein levels in extracts of sausage, sauce, pasta sauce, fish paste and cereal.

Allergens↗

[Inter-laboratory evaluation studies for establishment of notified ELISA methods for allergic substances (buckwheat)].

Inter-laboratory evaluation studies were conducted for the ELISA methods for allergic substances (buckwheat). Extracts of snack, bun and udon spiked with buckwheat standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate at 10 laboratories. Coefficients of variation (CVs) of the ELISA methods using a Buckwheat Protein ELISA Kit (Buckwheat kit) and a FASTKIT Buckwheat ELISA kit (Buckwheat ELISA kit) were mostly below 10%. Mean recoveries of the buckwheat standard protein from the food extracts were over 40% in the two ELISA methods. Repeatability relative standard deviations of buckwheat standard protein in three food extracts were in the ranges of 6.8-78.5% and 5.0-33.9% for the Buckwheat kit and the Buckwheat ELISA kit, respectively. Reproducibility relative standard deviations of buckwheat standard protein in three food extracts were 11.9-69.5% and 16.5-34.1% for the Buckwheat kit and the Buckwheat ELISA kit, respectively. The detection limits of both ELISA methods were 1 ng/mL in sample solutions. These results suggest that the notified ELISA methods are reliable and reproducible for the inspection of buckwheat protein levels in extracts of snack, bun and udon.

Allergens↗

[Inter-laboratory evaluation studies for establishment of notified ELISA methods for allergic substances (peanuts)].

Inter-laboratory evaluation studies were conducted for ELISA methods for allergic substances (peanuts). Extracts of biscuit, sauce, chocolate and butter spiked with peanut standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate in 10 laboratories. Coefficients of variation (CVs) of the ELISA methods using a Peanut Protein ELISA Kit (Peanut kit) and a FASTKIT Peanut ELISA kit (Peanut ELISA kit) were mostly below 10%. Mean recoveries of the peanut standard protein from the food extracts were over 40% in the two ELISA methods. Repeatability relative standard deviations of peanut standard protein in four food extracts were in the ranges of 15.2-49.7% and 3.0-28.3% for the Peanut kit and the Peanut ELISA kit, respectively. Reproducibility relative standard deviations of peanut standard protein in four food extracts were 23.5-44.4%, 9.6-28.4% for the Peanut kit and the Peanut ELISA kit, respectively. The detection limits of both ELISA methods were 2-2.5 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of peanut protein levels in extracts of biscuit, sauce, chocolate and butter.

Allergens↗

Differences in hypothalamic Fos expressions between two heat stress conditions in conscious mice.

Hyperthermia and dehydration were important physiological phenomena in heat stress. But, the degrees of these phenomena were changed by heat stress conditions, and the distinction between both phenomena is necessary for investigation of response for individual phenomenon. Heat stress at 34 degrees C for 60 min increased rectal temperature, and heat stress at 38.5 degrees C for 60 min further increased rectal temperature and increased osmolality in mice. We investigated the activated region in hypothalamus, which played a role in thermoregulation, fluid regulation and so on, using immunostaining for Fos protein under these conditions in conscious mice. At 34 degrees C, Fos-positive neurons increased in the median preoptic nucleus, lateral preoptic area and anterior hypothalamic area, which were known to be the thermoregulatory center, and the dorsomedial hypothalamic nucleus, which was known to control eating. At 38.5 degrees C, Fos-positive neurons further increased in the regions mentioned above and appeared in the lateral septal nucleus, medial preoptic area, lateral hypothalamic area and zona incerta, which were thought to be involved in thermoregulation and/or fluid regulation, and the paraventricular hypothalamic nucleus, supraoptic nucleus and supraoptic nucleus in the retrochiasmatic part, which were known to be involved in neuroendocrine effector systems. These results support that the activated regions in hypothalamus differed with heat stress conditions, which induced only hyperthermia and both hyperthermia and dehydration.

Animals↗

Oral sensitization of W/W(v) mice with ovalbumin and possible involvement of the decrease in gammadelta-T cells.

Mast-cell-deficient WBB6F1-W/W(v) mice (W/W(v)) and congenic wild-type (+/+) mice were sensitized by oral administration of 0.1 or 1.0 mg ovalbumin (OVA) in the form of gavage every day for 9 weeks, and active systemic anaphylaxis (ASA) was induced by intraperitoneal injection of OVA. Production of OVA-specific IgG1 in response to oral sensitization of the W/W(v) mice was very high, and the production of IL-4, IL-5 and IL-10 by splenocytes re-stimulated with OVA in vitro was increased. These findings suggest that Th2-dominant helper T-cell activation had occurred. By contrast, production of OVA-specific IgG1 was low in +/+ mice, and no significant increase in production of Th2-type cytokines by the splenocytes of +/+ mice was observed. Population analysis in Peyer's patches by flow cytometry revealed that the proportion of the CD11c(+) cell in the W/W(v) mice was slightly increased after antigen stimulation. Analysis of the cell surface markers of intraepithelial lymphocytes (IELs) by flow cytometry showed that the proportion of TCRgammadelta-T cells was extremely lower in the W/W(v) mice, especially in the antigen sensitized group. The proportion of TCRgammadelta-T cells in the splenocytes of W/W(v) mice was also lower than in +/+ mice. Taken together, the above findings indicate that W/W(v) mice seems to be a good model not only for studying the induction mechanism of food allergy but for examining the role of TCRgammadelta-T cells in food-induced hypersensitivity.

Anaphylaxis↗

Dynamic responses to acute heat stress between 34 degrees C and 38.5 degrees C, and characteristics of heat stress response in mice.

We focused on dynamic responses to acute heat stress between 34 degrees C and 38.5 degrees C. Physiological and neuroendocrinological changes between 34 degrees C and 38.5 degrees C were studied in mice. The influence of humid conditions, 85% relative humidity (RH), on these changes was also investigated. Rectal temperatures increased above 34 degrees C and hematocrit levels increased at 38.5 degrees C 85% RH for 60 min. Food consumption and body weight gains decreased after a daily 60 min exposure to 34, 37 and 38.5 degrees C for 2 weeks. The corticosterone and vasopressin levels in the blood, and catecholamine and serotonin metabolite levels in the hypothalamus were not changed at 34 degrees C, but increased when above 37 degrees C for 60 min. Above 37 degrees C, these physiological and neuroendocrinological changes were accelerated by humid conditions. These results indicated that food consumption and body weight gains decreased above 34 degrees C, and the neuroendocrinological changes, which were accelerated by humid conditions, were induced above 37 degrees C. In comparison with restraint and water immersion stress, heat stress at 37 degrees C 85% RH showed a slower increase in serum corticosterone levels, smaller changes in plasma dopamine and dihydroxyphenylacetic acid levels, and, after repeated exposure, larger decreases in food consumption and body weight gains. This study clarified the relationships between temperature and humidity conditions and physiological and neuroendocrinological changes, along with the characteristics of responses in acute heat stress.

Animals↗

[Inter-laboratory evaluation studies of notified ELISA methods for allergic substances (egg)].

Inter-laboratory evaluation studies were conducted for the notified ELISA methods for allergic substances (Egg). Standard extracts of egg spiked in extracts of sausage, sauce, cookie, bread and cereal at a level of 5-20 ng/mL as the sample solution were analyzed in replicate in 10 laboratories. Coefficients of variation (CVs) of all three ELISA methods using an Egg Protein ovalbumin ELISA Kit (ovalbumin kit), an Egg Protein ovomucoid ELISA Kit (ovomucoid kit) and a FASTKIT Egg ELISA kit (Egg ELISA kit) were mostly less than 10%. Mean recoveries of the standard extract of egg were over 40% in the three ELISA methods. Repeatability relative standard deviations of egg standard solution in five food extracts were in the ranges of 18.7-25.5%, 18.6-41.8%, 21.3-43.3% for the ovalbumin kit, the ovomucoid kit and the Egg ELISA kit, respectively. Reproducibility relative standard deviations of egg standard solution in five food extracts were 16.8-35.1%, 19.6-35.8%, 24.7-51.1% for the ovalbumin kit, the ovomucoid kit and the Egg ELISA kit, respectively. The detection limits of all the ELISA methods were 4-5 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of egg protein levels in extracts of sausage, sauce, cookie, bread and cereal.

Allergens↗

Effect of chondroitin sulfate on murine splenocytes sensitized with ovalbumin.

Chondroitin sulfate (CS) is a glycosaminoglycan that is widely present in animals organisms, and it has anti-inflammatory and chondroprotective properties. To examine the effects of CS on the immune system, splenocytes obtained from ovalbumin (OVA)-sensitized BALB/c mice were challenged with OVA in the presence of CS, and cytokine levels in the medium of the cultured cells were measured. CS induced secretion of Th1-type cytokines (IFN-gamma, IL-2, and IL-12) by OVA-sensitized splenocytes but suppressed secretion of Th2-type cytokines (IL-5 and IL-10). Flow cytometric assay showed a significantly higher percentage of helper T cells (CD4(+)CD8(-) cells) among the splenocytes cultured with OVA and CS than with OVA alone. Analysis of the IFN-gamma mRNA level of the splenocytes by the real-time quantitative RT-PCR technique revealed higher levels in the splenocytes cultured with OVA and CS than in the splenocytes cultured with OVA alone. This is the first demonstration that CS inhibits antigen-induced IgE production through induction of cytokine secretion by Th1 cells, and this finding suggests a potential use of CS in preventing IgE-mediated allergy.

Animals↗

Increases in plasma dihydroxyphenylacetic acid levels in decapitated mice after exposure to various stresses.

This study investigated changes in plasma levels of the dopamine metabolite dihydroxyphenylacetic acid (DOPAC) in decapitated mice in response to the variable stresses of restraint, restraint and water immersion, and foot shock. DOPAC levels, but not norepinephrine (NE) and epinephrine (EPI) levels, increased upon exposure to these stresses. Plasma DOPAC levels measured using the decapitation method in rats were then compared with those measured using the catheter method. The NE and EPI levels in plasma measured using the decapitation method were much higher than those using the catheter method under basal conditions. In contrast, differences in the levels of DOPAC in plasma were smaller than those of NE and EPI under basal conditions using in both methods; furthermore parallel changes in plasma DOPAC levels occurred during restraint and water immersion stresses. These results indicate that the plasma DOPAC levels measured in mice using the decapitation method were clearly increased by the different stresses. Furthermore, in rats there were correlations between the decapitation and catheter methods for plasma levels of DOPAC. Thus the change in plasma DOPAC levels measured using the decapitation method is a good indicator of stress responses involving sympathoneural activity.

3,4-Dihydroxyphenylacetic Acid↗