A modified formalin test in mice was investigated. The pain response curve induced by 0.5% formalin was biphasic, having 2 peaks, from 0 to 5 min (first phase) and from 15 to 20 min (second phase). A low concentration of formalin was used, allowing the effects of weak analgesics to be detected. Centrally acting drugs such as narcotics inhibited both phases equally. Peripherally acting drugs such as aspirin, oxyphenbutazone, hydrocortisone and dexamethasone only inhibited the second phase. Aminopyrine and mefenamic acid which acted on both central and peripheral sites inhibited both phases, but the second phase was inhibited by lower doses. Thus, this method enables one to easily distinguish the site of action of analgesics. Furthermore, pain response in the first phase was inhibited by capsaicin-treated desensitization and Des-Arg9-(Leu8)-bradykinin (bradykinin inhibitor). The second phase was inhibited by compound 48/80 pretreatment, indomethacin and bradykinin inhibitor. Therefore, it is suggested that substance P and bradykinin participate in the manifestation of the first phase response, and histamine, serotonin, prostaglandin and bradykinin are involved in the second phase. These results indicate that the first and second phase responses induced by formalin have distinct characteristic properties, and it is a very useful method for examining pain, nociception and its modulation by pharmacological or other means.

No abstract

The dental phenolic medicaments, eugenol and guaiacol, are partly similar in chemical structure to capsaicin, the pungent constituent of chili peppers, which selectively activates sensory neurons via a specific receptor. We have previously demonstrated that these phenolic compounds show capsaicin-like action. In the present study, an attempt was made to investigate the possibility that these compounds interact with the same cellular site as capsaicin, by using capsazepine, a selective and competitive anta-gonist of capsaicin. Intrathecal (i.t.) treatment with eugenol (12.5 to 50 micrograms), guaiacol (25 to 150 micrograms), or capsaicin (1 to 4 micrograms) for 24 h dose-dependently inhibited the formalin-induced nociceptive response. Capsazepine (5, 10 micrograms, i.t.) shifted these dose-response curves in parallel to the right. Similarly, capsazepine abolished antinociceptive effects of eugenol (50 micrograms), guaiacol (150 micrograms), or capsaicin (2 micrograms) in the acetic acid writhing test. These results suggest that eugenol and guaiacol may exert their antinociceptive effects via the capsaicin receptor located on sensory terminals in the spinal cord.

The activities of hydrogen ions in 20-200 μmol dm(-3) H(2)SO(4) solution were estimated by use of an ionic liquid salt bridge (ILSB), made of tributyl(2-methoxyethyl)phosphonium bis(pentafluoroethanesulfonyl)amide (TBMOEPC(2)C(2)N), sandwiched by two hydrogen electrodes. The experimental pH values (pH = -log a(H), where a(H) is the activity of hydrogen ions) were in good agreement, within 0.01 pH unit, with those calculated using the Pitzer model. The difference between the experimental and theoretical pH values at 50 μmol dm(-3) H(2)SO(4) solution was much smaller than that obtained by use of a glass electrode in combination with a reference electrode with a concentrated KCl salt bridge. The source of the small deviation can be explained by the residual diffusion potential due to the dissolution of TBMOEPC(2)C(2)N in the H(2)SO(4) solution (W) and the resultant increase in the ionic strength of W. The use of a reference electrode equipped with an ILSB opens the way to accurately estimate the pH in dilute aqueous solutions, for which we have not had effective means.