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Responses to repeated oral irritation by capsaicin, cinnamaldehyde and ethanol in PROP tasters and non-tasters.

Both increases (sensitization) and decreases (desensitization) in oral irritation have been reported in response to repeated short-term stimulation by compounds such as capsaicin, zingerone and menthol. It is unclear why one irritant would show sensitization and another desensitization, and this is further complicated by substantial inter-individual variation in response patterns. These variations may be the result of individual differences such as that represented by sensitivity to 6-n-propylthiouracil (PROP), which has been associated with variation in the overall intensity of irritation. In addition, comparisons between irritants have almost always involved inter-study comparisons, entailing different subject groups and frequently different methods. In the studies reported here, responses to three irritants-capsaicin, cinnamaldehyde and ethanol-were examined as a function of PROP taster status. A common core of subjects also received all three irritants, allowing an assessment of the extent to which different response patterns between irritants seen previously were the result of different properties of the irritants themselves. Over a series of ten stimuli presented at 1 min intervals, PROP taster status differentiated subject responses on the basis of overall intensity, but not the pattern of responses over repeated stimulation. The group response to ethanol and cinnamaldehyde was desensitization, a pattern also shown by most of the individual subjects. In contrast, the group response to capsaicin was neither clear sensitization nor desensitization, reflecting much greater individual variability in response patterns. It is suggested that the time course to a single irritant stimulus largely determines between irritant response variations, while the inter-stimulus interval (ISI) used for a given irritant will have critical values for showing predominantly sensitization or desensitization.

Acrolein↗

Ibuprofen as a chemesthetic stimulus: evidence of a novel mechanism of throat irritation.

This paper reports a study of the oral and pharyngeal chemesthetic effects of the non-steroidal anti-inflammatory drug (NSAID) ibuprofen [2-(4-isobutylphenyl)propanoic acid], which pilot experiments had indicated produces an unusual sensory irritation of the throat. In experiment 1 subjects swallowed aqueous solutions of ibuprofen prepared with different buffering agents and gave ratings of irritation and taste in the mouth and throat. The results showed that ibuprofen irritates the throat much more than the mouth, and that its quality in the throat is characterized primarily as sting/prick, itch and tickle (often leading to cough). Based upon the results obtained with the different buffering agents, we hypothesized that the sting/prick/itch qualities of throat irritation were pH-dependent. Parametric manipulation of solution pH in experiment 2 confirmed this hypothesis. The same experiment revealed that, in contrast to other oral irritants (e.g. capsaicin and menthol), repeated stimulation caused neither sensitization nor desensitization of throat irritation. In the final experiment we found that ibuprofen's throat irritation could not be modulated by temperature, as it should be if stimulation occurred via capsaicin-sensitive receptors. We therefore conclude that ibuprofen has novel chemesthetic properties, which are not mediated by capsaicin-sensitive (vanilloid) receptors, and that a major component of the throat irritation it produces occurs via a pH-dependent receptor mechanism.

Administration, Oral↗

Autonomic nervous system responses to odours: the role of pleasantness and arousal.

Perception of odours can provoke explicit reactions such as judgements of intensity or pleasantness, and implicit output such as skin conductance or heart rate variations. The main purpose of the present experiment was to ascertain: (i) the correlation between odour ratings (intensity, arousal, pleasantness and familiarity) and activation of the autonomic nervous system, and (ii) the inter-correlation between self-report ratings on intensity, arousal, pleasantness and familiarity dimensions in odour perception. Twelve healthy volunteers were tested in two separate sessions. Firstly, subjects were instructed to smell six odorants (isovaleric acid, thiophenol, pyridine, L-menthol, isoamyl acetate, and 1-8 cineole), while skin conductance and heart rate variations were being measured. During this phase, participants were not asked to give any judgement about the odorants. Secondly, subjects were instructed to rate the odorants on dimensions of intensity, pleasantness, arousal and familiarity (self-report ratings), by giving a mark between 1 (not at all intense, arousing, pleasant or familiar) and 9 (extremely intense, arousing, pleasant or familiar). Results indicated: (i) a pleasantness factor correlated with heart rate variations, (ii) an arousal factor correlated with skin conductance variations, and (iii) a strong correlation between the arousal and intensity dimensions. In conclusion, given that these correlations are also found in other studies using visual and auditory stimuli, these findings provide preliminary information suggesting that autonomic variations in response to olfactory stimuli are probably not modality specific, and may be organized along two main dimensions of pleasantness and arousal, at least for the parameters considered (i.e. heart rate and skin conductance).

Adult↗

Oral irritation by mustard oil: self-desensitization and cross-desensitization with capsaicin.

We investigated the temporal pattern of oral irritation elicited by sequential application of mustard oil (allyl-isothiocyanate), and whether it exhibits self-desensitization and cross-desensitization with capsaicin. Mustard oil (0.125%, 40 micro l) was sequentially applied to one side of the tongue at 1 min intervals, and subjects rated the intensity of the irritant sensation elicited by each stimulus. Ratings successively declined across trials, indicating desensitization. In contrast, sequential application of capsaicin (10 ppm) elicited irritation that increased in intensity across trials (sensitization). To test for self-desensitization by mustard oil, a 10 min hiatus was imposed following the series of unilateral mustard oil stimuli, after which mustard oil was applied to both sides of the tongue. In a two-alternative forced-choice paradigm, subjects chose which side had stronger irritation and also independently rated the irritant intensity on each side. A significant majority of subjects chose the side not previously receiving mustard oil as more intense, and assigned significantly higher intensity ratings to that side, indicating self-desensitization. In two additional sessions, the same paradigm was used to show mustard oil cross-desensitization of irritation elicited by capsaicin, and capsaicin cross-desensitization of irritation from mustard oil. In a final session, sequential application of mustard oil at faster (20 s) intervals initially evoked a sensitizing pattern followed by desensitization. The temporal patterns of oral irritation exhibited by mustard oil, and its reciprocal cross-desensitization with capsaicin, are similar to those of menthol and nicotine.

Administration, Oral↗

The use of linear expressions of solute boiling point versus retention to indicate special interactions with the molecular rings of modified cyclodextrin phases in gas chromatography

The boiling points (degrees C, 1 x 10) of diverse C10 polar solutes from volatile oils are set against their relative retention times versus n-undecane to calculate linear equations for 12 commercial modified cyclodextrin (CD) capillary phases. Ten data points are considered for each CD, then solutes are rejected until 5 or more remain that give an expression with a correlation coefficient of at least 0.990 and a standard deviation of less than 5.5. Three phases give almost perfect correlation, and 3 other CDs have difficulty complying. Solutes involved in the equations (most frequently cuminal, linalol, and carvone) are presumed to have a 'standard' polar transient interaction with the molecular rings of the CDs concerned. Several remaining solutes (mostly citral, fenchone, and menthol) exhibit extra retention over the calculated standard (up to 772%), which is believed to indicate a firm 'host' CD or 'guest' solute molecular fit in some cases. Other solutes show less retention than calculated (mostly citronellal, citronellol, estragole, and pulegone). This suggests rejection by the CD, which behaves merely as a conventional stationary phase to them. The intercept constant in the equation for each phase is suggested to be a numerical relative polarity indicator. These b values indicate that 3 hydroxypropyl CDs show the most polarity with values from 28 to 43; and CDs that are fully substituted with inert groups fall in the range of 15 to 20.

Journal Article↗

Smoking and pregnancy outcome among African-American and white women in central North Carolina.

Despite extensive research on tobacco smoking during pregnancy, few studies address risks among African-American and white women, groups that differ in brand preference and smoking habits. The Pregnancy, Infection, and Nutrition Study is a prospective cohort study that included 2,418 women with detailed information on smoking during pregnancy, including brand, number of cigarettes per day, and changes during pregnancy. We analyzed risk of preterm birth (<37 and <34 weeks' gestation) and small-for-gestational-age deliveries in relation to tobacco use. Pregnant African-American smokers differed markedly from whites in brand preference (95% vs 26% smoked menthol cigarettes) and number of cigarettes per day (1% of African-Americans and 12% of whites smoked 20+ cigarettes per day). Smoking was not related to risk of preterm birth overall, but cotinine measured at the time of delivery was (adjusted odds ratio = 2.2, 95% confidence interval = 1.1-4.5). A clear association and dose-response gradient was present for risk of fetal growth restriction (risk ratio for 20+ cigarettes/day = 2.4, 95% confidence interval = 1.4-4.0). Associations of tobacco use with preterm premature rupture of amniotic membrane resulting in preterm birth were notably stronger than the associations with other types of preterm birth.

Black or African American↗

Cool (TRPM8) and hot (TRPV1) receptors in the bladder and male genital tract.

PURPOSE: Overactive bladder symptoms due to various etiologies have been successfully treated with capsaicin by desensitization of the temperature sensitive vanilloid receptor TRPV1. Recently another temperature sensitive receptor, TRPM8, activated by menthol and cool temperatures (8C to 28C) was described that may be the proposed cool receptor, at least in part mediating the bladder response in the diagnostic ice water test. We defined the sites of mRNA and protein expression of TRPM8 and TRPV1 in the rat and human genitourinary tract. MATERIALS AND METHODS: Prostate, testis, penis, bladder and dorsal root ganglion tissue was obtained from rats. Prostate, testicle, seminiferous tubules, corpus cavernosum, glans, overlying glans skin, scrotal skin and bladder were obtained from human patients. Reverse transcription-polymerase chain reaction was done using species specific primers for TRPM8 and TRPV1. Immunofluorescence staining for TRPM8 was performed in rat tissues as well as in cultured human urothelial cells. RESULTS: TRPM8 and TRPV1 mRNA were detected in all rat tissues. Human samples demonstrated TRPM8 mRNA in prostate, testicle, seminiferous tubules, scrotal skin and bladder. No TRPM8 mRNA was identified in human corpus cavernosum, glans or overlying glans skin. Separation of layers in human bladder demonstrated mRNA for TRPM8 only in the urothelium and not in the detrusor. Immunofluorescence location of TRPM8 was found in rat prostate, DRG and bladder, and in human urothelial cells in culture. TRPV1 mRNA was detected in all human genitourinary tract tissues. CONCLUSIONS: These results demonstrate that mRNA and protein for TRPM8 exist in multiple genitourinary organs in the rat and human, and it may be considered a possible new target, as is TRPV1, for the pharmacological treatment of detrusor overactivity or other urological disorders.

Animals↗

Effect of a topical herbal cream on the pain and stiffness of osteoarthritis: a randomized double-blind, placebo-controlled clinical trial.

This study compared the efficacy of an herbal ointment to a placebo ointment in relieving the pain and stiffness of osteoarthritis. The herbal preparation contained substances believed by alternative practitioners to be helpful in treating osteoarthritis. This study was a double-blind randomized placebo-controlled clinical trial. Subjects were solicited from patients attending an arthritis clinic in Dallas, Texas. Selection of subjects with osteoarthritis was based on the criteria developed by the American College of Rheumatology. Subjects were randomized to an active (n = 11) or a placebo (n = 8) group. Herbal ointment and control contained small amounts of capsaicin and menthol and were similar when applied. Subjects graded their levels of pain and stiffness for a 1-week no-treatment baseline. Subjects then applied the herbal or sham ointment to the involved joint(s) for 28 consecutive days and recorded their levels of pain and stiffness daily on visual analog scales. Significant differences between the active and placebo groups for pain (P < 0.05) and stiffness (P < 0.05) were found when the baseline phase was compared with the fourth week. An herbal ointment was shown to be effective in relieving the pain and stiffness of osteoarthritis without adverse effects. If future studies confirm effectiveness of this treatment, it may have a role as an adjunct to standard drug therapy.

Journal Article↗

Monoterpene metabolism. Cloning, expression, and characterization of (-)-isopiperitenol/(-)-carveol dehydrogenase of peppermint and spearmint.

The essential oils of peppermint (Mentha x piperita) and spearmint (Mentha spicata) are distinguished by the oxygenation position on the p-menthane ring of the constitutive monoterpenes that is conferred by two regiospecific cytochrome P450 limonene-3- and limonene-6-hydroxylases. Following hydroxylation of limonene, an apparently similar dehydrogenase oxidizes (-)-trans-isopiperitenol to (-)-isopiperitenone in peppermint and (-)-trans-carveol to (-)-carvone in spearmint. Random sequencing of a peppermint oil gland secretory cell cDNA library revealed a large number of clones that specified redox-type enzymes, including dehydrogenases. Full-length dehydrogenase clones were screened by functional expression in Escherichia coli using a recently developed in situ assay. A single full-length acquisition encoding (-)-trans-isopiperitenol dehydrogenase (ISPD) was isolated. The (-)-ISPD cDNA has an open reading frame of 795 bp that encodes a 265-residue enzyme with a calculated molecular mass of 27,191. Nondegenerate primers were designed based on the (-)-trans-ISPD cDNA sequence and employed to screen a spearmint oil gland secretory cell cDNA library from which a 5'-truncated cDNA encoding the spearmint homolog, (-)-trans-carveol-dehydrogenase, was isolated. Reverse transcription-PCR amplification and RACE were used to acquire the remaining 5'-sequence from RNA isolated from oil gland secretory cells of spearmint leaf. The full-length spearmint dehydrogenase shares >99% amino acid identity with its peppermint homolog and both dehydrogenases are capable of utilizing (-)-trans-isopiperitenol and (-)-trans-carveol. These isopiperitenol/carveol dehydrogenases are members of the short-chain dehydrogenase/reductase superfamily and are related to other plant short-chain dehydrogenases/reductases involved in secondary metabolism (lignan biosynthesis), stress responses, and phytosteroid biosynthesis, but they are quite dissimilar (approximately 13% identity) to the monoterpene reductases of mint involved in (-)-menthol biosynthesis. The isolation of the genes specifying redox enzymes of monoterpene biosynthesis in mint indicates that these genes arose from different ancestors and not by simple duplication and differentiation of a common progenitor, as might have been anticipated based on the common reaction chemistry and structural similarity of the substrate monoterpenes.

Alcohol Oxidoreductases↗

Biochemical characterization of a spearmint mutant that resembles peppermint in monoterpene content.

A radiation-induced mutant of Scotch spearmint (Mentha x gracilis) was shown to produce an essential oil containing principally C3-oxygenated p-menthane monoterpenes that are typical of peppermint, instead of the C6-oxygenated monoterpene family characteristic of spearmint. In vitro measurement of all of the enzymes responsible for the production of both the C3-oxygenated and C6-oxygenated families of monoterpenes from the common precursor (-)-limonene indicated that a virtually identical complement of enzymes was present in wild type and mutant, with the exception of the microsomal, cytochrome P-450-dependent (-)-limonene hydroxylase; the C6-hydroxylase producing (-)-trans-carveol in the wild type had been replaced by a C3-hydroxylase producing (-)-trans-isopiperitenol in the mutant. Additionally, the mutant, but not the wild type, could carry out the cytochrome P-450-dependent epoxidation of the alpha,beta-unsaturated bond of the ketones formed via C3-hydroxylation. Although present in the wild type, the enzymes of the C3-pathway that convert trans-isopiperitenol to menthol isomers are synthetically inactive because of the absence of the key C3-oxygenated intermediate generated by hydroxylation of limonene. These results, which clarify the origins of the C3- and C6-oxygenation patterns, also allow correction of a number of earlier biogenetic proposals for the formation of monoterpenes in Mentha.

Journal Article↗

Determination of the absolute configuration of a novel dipeptide isostere.

(1R-[1 alpha(1R*,2R*,3S*),2 beta,5 alpha])-2-(4-Morpholinocarbonyl)- 3-phenylcyclopropane 2-(1-methyl-ethyl)-5-methylcyclohexyl ester, C25H35NO4, Mr = 413 x 56, orthorhombic, P2(1)2(1)2(1), a = 9 x 359 (2), b = 10 x 1172 (14), c = 24 x 710 (4) A, V = 2339 x 8 (6) A3, Z = 4, Dx = 1 x 17 g cm-3, mu = 0 x 7332 cm-1, Mo K alpha radiation, lambda = 0 x 7107 A, F(000) = 896, T = 198 K, R = 0 x 0475 for 2460 reflections [Fo greater than or equal to 4 sigma(Fo)]. The assignment of the absolute configuration was based on internal comparison to the (-)-menthol moiety. The morpholenyl and cyclohexyl rings are in the chair conformation. The amide group is essentially planar [max. deviation 0.025 (2) A for N19] resulting in a close non-bonded contact between the amide oxygen, O18, and H24a of 2 x 24 (4) A.

Dipeptides↗

The complex problem of sensitive skin.

There exists within the population subsets of individuals who display heightened skin reactivity to materials the majority find tolerable. In a series of investigations, we have examined interrelationships between many of the endpoints associated with the term 'sensitive skin'. In the most recent work, 58 volunteers were treated with 10% lactic acid, 50% ethanol, 0.5% menthol and 1.0% capsaicin on the nasolabial fold, unoccluded, with sensory reactions recorded at 2.5 min, 5 min and 8 min after application. Urticant susceptibility was evaluated with 1 m benzoic acid and 125 mM trans-cinnamic acid applied to the volar forearm for 20 min. A 2 x 23-h patch test was also conducted using 0.1% and 0.3% sodium dodecyl sulfate, 0.3% and 0.6% cocamidopropyl betaine and 0.1% and 0.2% benzalkonium chloride to determine irritant susceptibility. As found in previous studies, increased susceptibility to one endpoint was not predictive of sensitivity to another. In our experience, nasolabial stinging was a poor predictor of general skin sensitivity. Nevertheless, it may be possible to identify in the normal population individuals who, coincidentally, are more generally sensitive to a range of non-immunologic adverse skin reactions. Whether such individuals are those who experience problems with skin care products remains to be addressed.

Administration, Topical↗

Development of a stable oral liquid dosage form of spironolactone.

A clear, stable, oral liquid dosage form of spironolactone has been developed. Solubility profiles of spironolactone were obtained in several co-solvent blends. Using this data, a co-solvent blend containing polyethylene glycol 400 (30% v/v), propylene glycol (10% v/v), glycerin (10% v/v) and ethyl alcohol (10% v/v) was used to solubilize spironolactone at a concentration of 2 mg/ml. The final formulation contained sweetening agents (sucrose, saccharin sodium), flavours (cherry, sweet), a desensitizing agent (menthol), a dye (FD&C Red #40) and a preservative (benzoic acid) to incorporate the desired organoleptic and preservative properties. A phosphate buffer was used to maintain a pH value of 4.5 (pH of maximum stability as reported earlier) to minimize hydrolysis. The final dosage form was stable for at least 93 days at 40 degrees C (loss of potency less than 4%). According to FDA guidelines, a tentative expiration date of 2 years at 25 degrees C is justifiable.

Administration, Oral↗

Comparative transcript and alkaloid profiling in Papaver species identifies a short chain dehydrogenase/reductase involved in morphine biosynthesis.

Plants of the order Ranunculales, especially members of the species Papaver, accumulate a large variety of benzylisoquinoline alkaloids with about 2500 structures, but only the opium poppy (Papaver somniferum) and Papaver setigerum are able to produce the analgesic and narcotic morphine and the antitussive codeine. In this study, we investigated the molecular basis for this exceptional biosynthetic capability by comparison of alkaloid profiles with gene expression profiles between 16 different Papaver species. Out of 2000 expressed sequence tags obtained from P. somniferum, 69 show increased expression in morphinan alkaloid-containing species. One of these cDNAs, exhibiting an expression pattern very similar to previously isolated cDNAs coding for enzymes in benzylisoquinoline biosynthesis, showed the highest amino acid identity to reductases in menthol biosynthesis. After overexpression, the protein encoded by this cDNA reduced the keto group of salutaridine yielding salutaridinol, an intermediate in morphine biosynthesis. The stereoisomer 7-epi-salutaridinol was not formed. Based on its similarities to a previously purified protein from P. somniferum with respect to the high substrate specificity, molecular mass and kinetic data, the recombinant protein was identified as salutaridine reductase (SalR; EC 1.1.1.248). Unlike codeinone reductase, an enzyme acting later in the pathway that catalyses the reduction of a keto group and which belongs to the family of the aldo-keto reductases, the cDNA identified in this study as SalR belongs to the family of short chain dehydrogenases/reductases and is related to reductases in monoterpene metabolism.

Alkaloids↗

Chewing-gum preservative induced toxidermic vasculitis.

This paper reports the case of a young female patient who presented with a cutaneous urticarial disseminated eruption. Drug-induced side effects were eliminated, and the only recent dietary change was the regular use of chewing-gums containing chlorophylla (E140), menthol and BHT (butylhydroxytoluene). Immunohistological analysis of a cutaneous lesion revealed signs of vasculitis. Within 1 week after stopping chewing gum, the eruption subsided. Oral provocation tests at 4-day intervals confirmed the responsibility of BHT by the reinduction of the cutaneous signs after a few hours.

Adult↗

Antimicrobial effects of essential oils in combination with chlorhexidine digluconate.

The aim of the present study was to compare antimicrobial effects of essential oils alone and in combination with chlorhexidine digluconate against planktonic and biofilm cultures of Streptococcus mutans and Lactobacillus plantarum. The essential oils included cinnamon, tea-tree (Melaleuca alternifola), manuka (Leptospermum scoparium), Leptospermum morrisonii, arnica, eucalyptus, grapefruit, the essential oil mouthrinse Cool Mint Listerine and two of its components, menthol and thymol. Cinnamon exhibited the greatest antimicrobial potency (1.25-2.5 mg/ml). Manuka, L. morrisonii, tea-tree oils, and thymol also showed antimicrobial potency but to a lesser extent. The combination effect of the essential oil-chlorhexidine was greater against biofilm cultures of both S. mutans and L. plantarum than against planktonic cultures. The amount of chlorhexidine required to achieve an equivalent growth inhibition against the biofilm cultures was reduced 4-10-fold in combination with cinnamon, manuka, L. morrisonii, thymol, and Listerine. We conclude that there may be a role for essential oils in the development of novel anticaries treatments.

Anti-Infective Agents, Local↗

Extemporaneous prescribing (1).

An outline of the pharmacology of important components of extemporaneous prescriptions will be undertaken over four issues of the journal. Substances such as dithranol, salicylic acid and coal tar will be examined in some depth with regard to their mode of action, uses and side effects. In addition, examples of extemporaneous formulations containing these substances will be provided along with advice regarding their use. Substances such as sulphur, calamine, podophyllin and menthol will also be reviewed using the same format but in less detail. The articles are designed to provide practical knowledge in the area of extemporaneous prescribing. Complicated areas of pharmaceutical chemistry, unless relevant to the clinical setting, will not be discussed.

Anthralin↗

The effects of 3,5,5-trimethylcyclohexanol on hepatic cholesterol synthesis, bile flow and biliary lipid secretion in the rat.

The chemical trimethylcyclohexanol (TMC) is closely related to menthol, the major component of a terpene preparation with known choleretic and cholelitholytic activity. Its effects on hepatic cholesterol synthesis and bile secretion were examined in the rat. In both acute and long-term dosing experiments TMC significantly inhibited hepatic S-3-hydroxy-3-methylglutaryl coenzyme A (HMGCoA) reductase. TMC was a potent choleretic, with detectable effects on bile flow at low doses, which also reduced coupling of cholesterol secretion to bile salt secretion. Single large doses tended to lower biliary cholesterol output and caused significant reduction in cholesterol saturation index after biliary diversion for 1 h. TMC and its widely prescribed ester cyclandelate, which is rapidly degraded to TMC after ingestion, should be investigated further as potential cholelitholytic treatments in man.

Animals↗