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Biomedical subjects

K Tonosaki

Publications and source records attributed to K Tonosaki.

At least 19 recordsLinked to original sources

(Meth)acrylate monomer-induced cytotoxicity and intracellular Ca(2+) mobilization in human salivary gland carcinoma cells and human gingival fibroblast cells related to monomer hydrophobicity.

To elucidate a possible link between the cytotoxicity and Ca(2+) mobilization by (meth)acrylates, we investigated the cell survival of and change in [Ca(2+)](i) in human salivary gland (HSG) cells (salivary gland carcinoma cell line) and human gingival fibroblasts (HGF) cells treated separately with 9 (meth)acrylate monomers used in dentistry. The cell survival was determined by the MTT method, and the [Ca(2+)](i) changes after the stimulation with the (meth)acrylate monomers were measured in floating indo-1/AM-loaded cells in Ca(2+)-free medium. For both HSG and HGF cells, the cytotoxicity of the monomers was approximately proportional to their hydrophobicity (logP). No increase in [Ca(2+)](i) was found with hydrophilic monomers, even with 10mm stimulation. [Ca(2+)](i) elevation by hydrophobic monomers occurred in a dose- and hydrophobic-dependent manner. The [Ca(2+)](i) change in HSG cells appeared as twin peaks, i.e., an initial sharp peak followed by a delayed broad one; whereas with the HGF cells only a single broad peak was seen, possibly dependent on their membrane quality. Pretreatment with n-butanol or methylmethacrylate enhanced the butylmethacrylate-induced [Ca(2+)](i) elevation, suggesting the [Ca(2+)](i) elevation by (meth)acrylate may be related to monomer hydrophobicity and cell type. The causal link between the cytotoxicity and [Ca(2+)](i) mobilization of monomers is discussed.

Butanols↗

A comparative study of the antioxidant/prooxidant activities of eugenol and isoeugenol with various concentrations and oxidation conditions.

Eugenol (compound in , 4-allyl-2-methyoxyphenol) and isoeugenol (compound in , 4-propenyl-2-methoxyphenol), both used as a flavor agent in cosmetic and food products, have both prooxidant and antioxidant activities. Their adverse effects such as allergic and inflammatory reaction may be due to their prooxidant activity. To clarify the mechanisms of their cytotoxicity and the factors affecting their antioxidant/prooxidant activities, we investigated the cytotoxicity, ROS production, and cellular glutathione (GSH) levels induced by eugenol and isoeugenol in a human submandibular cell line. The cytotoxicity (MTT method) of eugenol was 1 order of magnitude lower than that of isoeugenol (CC50: eugenol, 0.395 mM; isoeugenol, 0.0523 mM); and ROS production (CDF staining) was induced significantly by isoeugenol, but not by eugenol. Under treatment with H2O2 (100 microM) plus horseradish peroxidase (1 microg/ml) for 30 min or with visible light irradiation for 5 min, eugenol caused biphasic ROS production characterized by enhanced at lower eugenol concentrations (5-10 microM) and decreased at higher concentrations (500 microM). In contrast, isoeugenol enhanced ROS production over a wide range of concentrations (5-500 microM). Isoeugenol at 1000 microM significantly reduced GSH levels compared with eugenol at the same concentration. The high cytotoxicity of isoeugenol may be attributed to its induction of high ROS production and low GSH levels, possibly as a result of benzyl radical formation. In contrast, the cytotoxicity of eugenol is likely to be mediated by ROS-independent mechanisms, possibly involving phenoxyl radicals and/or eugenol quinone methide.

Antioxidants↗

Relationship between intracellular ROS production and membrane mobility in curcumin- and tetrahydrocurcumin-treated human gingival fibroblasts and human submandibular gland carcinoma cells.

OBJECTIVE: Curcumin is a well-known chemopreventive agent of oral cancers as well as stomach and intestinal cancers. The relationship between reactive oxygen species (ROS) production and cell membrane mobility was investigated to clarify the pro-oxidant mechanism of curcumin and tetrahydrocurcumin (TH-curcumin). METHODS: The intracellular ROS production and membrane mobility by curcumin or TH-curcumin were measured in human submandibular adenocarcinoma cells (HSGs) and human primary gingival fibroblasts (HGFs). ROS and mobility were measured by 5-(and -6)-carboxy-2',7'-dichlorofluorescein diacetate staining and fluorescence recovery after photo bleaching, respectively. RESULTS: Curcumin produced ROS dose-dependently. ROS appeared in the region surrounding the cell membrane. The membrane mobility coefficient of the curcumin-treated cells was significantly lower than that of control cells. The lowered membrane mobility induced by curcumin was reversed by the addition of glutathione, an antioxidant. In contrast, TH-curcumin did not affect the ROS production or the membrane mobility coefficient. The alternations induced by curcumin treated HSG cells were greater than those by HGF cells. CONCLUSION: The reduction in membrane mobility induced by curcumin was attributed to ROS production. The oxidative effects of curcumin may be related to the structure of the alpha, beta-unsaturated carbonyl moiety as well as the phenolic OH group of this compound.

Adenocarcinoma↗

Pleasant feeling from watching a comical video enhances free radical-scavenging capacity in human whole saliva.

OBJECTIVE: Free radicals have been implicated in aging, mutagenesis, inflammation and other pathological conditions. We conducted a study to clarify the relation between a pleasant feeling as a psychological eustress and free radical-scavenging capacity (FRSC) in saliva. METHODS: Saliva was collected from 27 healthy volunteers. FRSC before, during and after watching a cheerful comical video for 30 min was measured by using 1,1-diphenyl-2-picrylhydrazyl (DPPH). RESULTS: The median values of FRSC (micromol/ml) before, at 10 and 20 min during and after watching the video were 54.5, 66.8, 66.6 and 69.4, respectively. The FRSC values obtained after watching the video were significantly higher than those before watching it (P<.001). When the FRSC before watching was taken as 1, the value for the group that felt "Very good," "Good" or "Ordinary+Dull" while watching the video for 30 min was 1.38, 1.20 or 0.98, respectively (P<.01). CONCLUSION: Watching a comical video enhanced the FRSC in saliva; in addition, a pleasant feeling boosted it even more.

Adult↗

Acute anosmia in the mouse: behavioral discrimination among the four basic taste substances.

The importance of taste and smell in discrimination of tastes was examined in normal and anosmic mice. We studied the influence of olfaction on taste sensation using behavioral and electrophysiological methods in both normal animals and animals made anosmic mice by destroying their olfactory receptor cells with zinc sulfate (ZnSO(4)) solution. Electrophysiological responses from chorda tympani nerves showed that peripheral taste receptor cells transmitted taste signals normally to the central nervous system, even when the olfactory senses were abnormal. Behavioral observations showed that mice with abnormal olfaction could not differentiate tastes.

Animals↗

Evidence for involvement of dysfunctional teeth in the senile process in the hippocampus of SAMP8 mice.

In order to evaluate the involvement of dysfunctional teeth in age-related deficits in hippocampal function, we examined the effect of removal of molar teeth (molarless condition) on neuronal degeneration and glial fibrous acidic protein (GFAP) expression in the hippocampus and on learning ability in a water maze test in young, middle-aged, and aged accelerated senescence-prone mice (SAMP8). The molarless condition enhanced an age-dependent decrease in both learning ability and the number of neurons in the hippocampal CA1 subfield and the age-dependent increase in the number and hypertrophy of GFAP-labeled astrocytes in the same subfield. These observations suggest that the molarless condition may be involved in the senile process in the hippocampus in SAMP8 mice.

Aging, Premature↗

Effects of acids on neural activity elicited by other taste stimuli in the rat Chorda tympani.

The purpose of this study is whether the gustatory neural response of taste cell to a binary mixture with threshold concentration of acid becomes synergistic or antagonistic can be estimated from the whole chorda tympani (CT) nerve in the rat. The present data demonstrate that acids are synergistic enhancer for sugars, and suppressor for NaCl and QHCl, but no effect to glycine and alanine. These results suggest that the acid was modifying the interaction of the other stimulus with its transduction mechanism.

Acids↗

Electroencephalographic olfactometry (EEGO) analysis of odour responses in dogs.

Scientists in many fields have studied olfaction in dogs, but no simple method exists to study this function. The olfactory ability of dogs is used in many fields. However, the owners of the dogs have a frustrating experience because there is not an easy method to measure olfactory function. The purpose of this study was to examine the characteristics of olfactory function of the dog with electroencephalographic olfactometry (EEGO) recording. It was found that slow waves decreased and rapid waves increased in response to odours. From these results, it is suggested that the rapid waves of EEGO activity are important in determining a dog's olfactory ability. The EEGO recording is effective in diagnosing anosmia in the dog and is easier than a behavioural experiment. Additionally, this method may not cause distress to an animal.

Animals↗

Possible involvement of undissociated acid molecules in the acid response of the chorda tympani nerve of the rat.

To test whether undissociated acid is capable of exciting the chorda tympani nerves in rats, we have used buffered acid solutions as taste stimuli. These solutions were prepared by adding alkali to weak acids, such as acetic acid, so that the proportion of undissociated and dissociated acids was varied whereas keeping the total acid concentration constant. When acetic acid solutions, adjusted to wide ranges of pH by NaOH, were applied to the tongue, the response magnitude of the chorda tympani nerves was not varied systematically with pH changes. However, if the sodium effect was eliminated by amiloride or replacement of cation by potassium or Tris[hydroxymethyl]aminomethane; NH(2)C(CH(2)OH)(3) (Tris-base), the chorda tympani response was reduced systematically as pH increased. Similar results were obtained with citric acid and ascorbic acid. This pH-dependent change in taste nerve response to acid cannot be solely attributed to the proton gradient because the response magnitude induced by hydrogen itself, which was estimated from responses to strong acids, was much smaller than that by equi-pH acetic acid ( approximately 85%). Thus we cannot explain the pH-dependent responses of the chorda tympani nerves to weak acids unless effects of undissociated acid molecules are postulated. It is therefore concluded that undissociated acids in weak acid solutions can be a stimulant to taste receptor cells.

Acetic Acid↗

Low environmental temperature modulates gustatory nerve activity and behavioral responses to NaCl in rats.

We studied the effects of cold ambient temperature on chorda tympani nerve responses to taste stimuli such as sucrose, NaCl, quinine HCl (QHCl), and HCl in rats. The electrophysiological recordings of the whole chorda tympani nerves from control (22 degrees C) and cold-exposed (4 degrees C) rats revealed that the responses to sucrose, HCl, and QHCl were unaffected by cold exposure. In contrast, the nerve responses to NaCl were enhanced time dependently, reaching a maximum 7-14 days after cold exposure. Responses to sodium acetate were likewise elevated as they were to NaCl, whereas those to KCl were unchanged after cold exposure. In addition, the residual NaCl responses after lingual application of the sodium-channel blocker amiloride in cold-exposed rats were similar to those in control animals. It is thus most likely that cold exposure potentiates the chorda tympani nerve responses to Na(+), but not to Cl(-). Behavioral studies with the two-bottle preference test showed that the cold-exposed rats refused to drink NaCl solutions at 0.05 and 0.1 M, the concentrations being preferred by control animals. These results suggest that the ambient temperature influences taste cell function, and that the enhanced NaCl response of the chorda tympani nerve is related to the avoidance of NaCl intake under cold environment.

Amiloride↗

Role of hydrophobic amino acids in gurmarin, a sweetness-suppressing polypeptide.

The sweetness-suppressing polypeptide gurmarin isolated from Gymnema sylvestre consists of 35 amino acid residues and contains three intramolecular disulfide bonds. Nuclear magnetic resonance analysis showed that the hydrophobic side chains of Tyr-13, Tyr-14, Trp-28, and Trp-29 in gurmarin are oriented outwardly. Together with the hydrophobic side chains of Leu-9, Ile-11, and Pro-12, they form a hydrophobic cluster, and therefore these hydrophobic groups are assumed to act as the site for interaction with the receptor protein. To examine the roles of these hydrophobic amino acids, they were replaced by Gly. The resulting [Gly13,14,28,29] gurmarin and [Gly9,11,13,14,28,29]-gurmarin did not suppress the responses to sucrose, glucose, fructose, or Gly. This result strongly suggests that these hydrophobic amino acids are involved in the interaction with the receptor protein.

Amino Acid Sequence↗

Synthesis, characterization, and sweetness-suppressing activities of gurmarin analogues missing one disulfide bond.

The sweetness-suppressing polypeptide gurmarin isolated from Gymnema sylvestre consists of 35 amino acid residues and includes three intramolecular disulfide bonds. The roles of the three disulfide bonds were investigated by replacing each with two alanine residues by solid-phase synthesis. Nine analogues of [Ala3,18]gurmarin, [Ala10,23]gurmarin, and [Ala17,33]-gurmarin were obtained. Three analogues had native disulfide bonds, while the other six had non-native disulfide bonds. The three analogues with native disulfide bonds suppressed the response to sucrose, but not those to glucose, fructose, saccharin, or glycine in rats. In contrast, the six analogues with non-native disulfide bonds did not suppress the responses to any of these sweeteners. These results suggest that the native disulfide bonds of gurmarin are necessary for interaction with the receptor protein, and that the sucrose-specific receptor site is present in rats.

Amino Acid Sequence↗

Gustatory receptor cell responses to the sweeteners, monellin and thaumatin.

Mouse gustatory responses to sucrose, monellin and thaumatin were examined extracellularly from the chorda tympani nerve and intracellularly from individual taste receptor cells. Although monellin and thaumatin taste intensely sweet to humans and old-world monkeys, they do not appear to elicit chorda tympani nerve responses in rats and other mammals. However, there is considerable species variation in the taste responses of mammals, including differences in taste responses of different strains of mice. In the present study with Slc:ICR mice, we show that chorda tympani and taste receptor cell response profiles, and behavioral results for monellin and thaumatin, are similar to response profiles for sucrose.

Animals↗

Tastes activate different second messengers in taste cells.

Taste signal transduction occurs in the microvillous membrane of taste cells. Previously, we hypothesized that c-GMP may mediate sweet taste transduction. Some data indicated that IP3 may have a role in vertebrate bitter taste transduction. Here we report that the different second messengers are activated by different tastes. We used techniques designed for radioimmunoassay measurement. The results indicate that sucrose triggers an increase in c-GMP concentration and quinine increases the IP3 concentration in mouse taste cells. These results support the sweet and bitter taste transduction hypotheses.

Animals↗

Synthesis and characterization of the sweetness-suppressing polypeptide gurmarin and ent-gurmarin.

The sweetness-suppressing polypeptide gurmarin isolated from the leaves of Gymnema sylvestre consists of 35 amino acid residues including three intramolecular disulfide bonds. Herein, the total chemical synthesis of gurmarin was performed by the stepwise fluoren-9-ylmethoxy-carbonyl solid-phase method, the yield of reduced gurmarin being 1.9% based on the starting amino acid resin. Disulfide formation was carried out in the presence of a redox system of reduced glutathione/oxidized glutathione to give gurmarin in a yield of 35.5%. The product was identical to natural gurmarin by analytical reverse phase high performance liquid chromatography (RP-HPLC), mass spectroscopy (MS), and peptide mapping, and suppressed the responses to sucrose, D-glucose, and L-glucose in a rat. The D enantiomer (all D-amino acid gurmarin) was also synthesized, and was shown to be the mirror image of gurmarin. Interestingly, the D enantiomer (ent-gurmarin) also suppressed the responses to sucrose, D-glucose, and L-glucose in a rat.

Amino Acid Sequence↗

The effect of bilateral sectioning of the chorda tympani and glossopharyngeal nerves on the sweet taste in the mouse.

The chorda tympani nerve (CT) and the glossopharyngeal nerve (GL) have been considered important nerves for the sense of taste. We studied the effect of bilateral sectioning of the CT and/or GL on the sweet taste in the mouse. Before and after surgery we analyzed the daily drinking patterns, using the two-bottle preference test method. The normal mouse drank the low concentration sucrose solution (0.0125 M) more than distilled water. This report showed that the mouse who was bilateral sectioned, both CT and GL or bilateral sectioned CT, rejected drinking the low concentration of sucrose solution. In contrast, the mouse who was bilateral sectioned GL drank the low concentration sucrose solution like the normal mouse did. These phenomena suggested that the fungiform papilae play an important role to detect the low concentration of sucrose (0.0125 M) as a sweet favorable taste substance.

Animals↗

Cross-adaptation to odor stimulation of olfactory receptor cells in the box turtle, Terrapene carolina.

Electrical recording from small twigs of olfactory nerve and electro-olfactogram (EOG) from olfactory epithelium in a turtle shows that olfactory receptors in the nose are responsive to various odors. I have used the effects of cross-adaptation to odor stimulation on the olfactory receptors to investigate the stimulus-specific components of these responses and to provide information about the responsiveness of cells. The results of the cross-adaptation experiments strongly support the hypothesis that different categories of receptor cells exist in the olfactory epithelium.

Animals↗