Comparison of two stimulus-delivery systems for measurement of nasal pungency thresholds

Using representative members of each of three homologous series of chemicals-ketones, acetates and alcohols-we measured nasal pungency thresholds in anosmics via two stimulus-delivery systems. The first system consists of the fairly commonly used 270 ml, plastic 'squeeze bottles'. The second system consists of 1900 ml, glass vessels with Teflon tubing and nose-pieces. Although bulkier and more susceptible to mechanical breakage, the glass vessels possess advantages that can allow them to provide 'environmentally realistic' chemosensory thresholds, i.e. thresholds closer in absolute values to those that might be obtained under whole-body exposures. Such advantages include a larger volume of the vapor-source to accommodate whole sniffs, and a tight nose-nose-piece connection to avoid stimulus dilution. The outcome revealed that, for every chemical, the glass vessels provided nasal pungency thresholds significantly lower than those provided by the squeeze bottles. The difference amounted, on average, to a factor of 4.6, though the relative potency of the compounds remained the same under both systems. Additionally, when tested with the highest homologues used here, namely, octyl acetate and 1-octanol, anosmics using the glass vessels had little or no difficulty achieving the criterion for threshold whereas they did have difficulty when using the squeeze bottles.     

Determinants for nasal trigeminal detection of volatile organic compounds

We explored the influence of methodological and chemical parameters on the detection of nasal chemesthesis (i.e., trigeminal stimulation) evoked by volatile organic compounds (VOCs). To avoid odor biases, chemesthesis was probed via nasal pungency detection in anosmics and via nasal localization (i.e., lateralization) in normosmics, in both cases using forced-choice procedures. In the experiments with anosmics, 12 neat VOCs were selected based on previous reports of lack of chemesthetic response. Although none of the VOCs reached 100% detection, detectability and confidence of detection were higher when using a glass vessel system adapted with nosepieces to fit the nostrils tightly than when using wide-mouth glass jars. Half the stimuli were detected well above chance and half were not. When the latter were tested again after being heated to 37 degrees C, that is, body temperature (from room temperature, 23 degrees C), to increase their vapor concentration, only one, octane, significantly increased its detectability. Chemesthesis gauged with normosmics mirrored that with anosmics. Gas chromatography measurements showed that, even at 23 degrees C, the saturated vapor concentrations of the undetected stimuli, except vanillin, were well above the respective calculated nasal pungency threshold (NPT) from an equation that, in the past, had accurately described and predicted NPTs. We conclude that, except for octane and perhaps vanillin, the failure of the other four VOCs to precipitate nasal chemesthesis rests on a chemical-structural limitation, for example, the molecules lack a key property to fit a receptor pocket, rather than on a concentration limitation, for example, the vapor concentration is too low to reach a threshold value.     

Human responses to propionic acid. II. Quantification of breathing responses and their relationship to perception

In 20 normal and four anosmic participants, instantaneous inhalation and exhalation flow rates were recorded in response to 15 s stimulations with clean air or propionic acid concentrations (0.16, 1.14, 8.22 and 59.15 p.p.m., v/v) that ranged from peri-threshold for normals to clearly supra-threshold for anosmics. Each odorant/irritant delivery to the face-mask began with an exhalation. This allowed concentration to reach full value before stimulus onset, defined as the point where the participant began to bring the stimulus into the nose by inhalation. Two seconds after this stimulus onset, normals exhibited cumulative inhaled volume (CIV) declines of 39 and 14%, and latencies of 500 and 710 ms, with presentations of 59.15 and 8.22 p.p.m., respectively. With anosmics, 59.15 p.p.m. caused a 19% decline in CIV that began at 730 ms. Examination of the first inhalation after stimulus onset shows that the CIV declines in normals were achieved by a progressive decline in volume (InVol), beginning with a slight drop at 1.14 p.p.m., and a marked decline in duration (InDur) with only the highest concentration. Anosmics exhibited declines in InDur and InVol with only the 59.15 p.p.m. stimulus, and these declines were much more modest than the changes seen in normals. Comparison of these breathing results with perceptual responses from this same experiment demonstrates that: (i) in normals, odor perception rises slightly, but breathing does not change, with the lowest concentration; (ii) the higher breathing sensitivity (declines in InVol) of normals is paralleled by both the higher nasal irritation of these individuals and the presence of odor sensation; (iii) InDur declines in normals only with a stimulus concentration sufficient to cause marked nasal irritation in anosmics; and iv) in anosmics, modest but reliable declines in both InDur and InVol mirror the marked elevation in nasal irritation magnitude seen with only the highest concentration. In view of the failure of prior work to provide evidence that olfactory activation alone can cause any of the breathing changes we observed, we conclude that some breathing parameters are quite useful as rapid and sensitive measures of nasal irritation that arises from activation of nasal trigeminal afferents alone or in combination with the olfactory nerve.     

Computerized odor psychophysical testing in mice

An automated odor psychophysical procedure was developed andused to determine absolute sensitivity to n–amyl acetate.Mice were trained to initiate a trial by interrupting a photobeamat the rear of the test chamber, then sample an odor port andindicate the presence or absence of odorant by either quicklywithdrawing from the port or by continuing to sample the port.Once the air dilution olfactometer had been adjusted prior toa training or testing session, a microcomputer was used to recordall responses by the animal, to control the delivery of stimulito the odor port and to control all events in the test chamber.Correct reponses on both odor and control trials were reinforcedand incorrect responses on both types of trials were punishedwith a forced ‘time-out’ period. The odor sensitivityof all mice was estimated, using a tracking procedure, and wasthen studied in detail using schedules in which odor concentrationswere presented in ascending, descending and random order. Withall three schedules, thresholds to n-amyl acetate were between1x10^-12 and 1 x10^-13 M. Threshold estimates obtained from twoof these same animals more than 1 year later were within 0.25log units of the original values. This method should prove valuablein future studies of nasal chemoreception in mice.     

SPECIFIC ANOSMIA TO ISOBUTYRALDEHYDE: THE MALTY PRIMARY ODOR

About 36% of human subjects are specifically anosmic to theodor of isobutyraldehyde. Odor threshold measurements on 14aliphatic aldehydes and related compounds were made with panelsof specific anosmics and normal observers. The anosmia was mostpronounced with isobutyraldehyde, isovaleraldehyde and 2-methylbutanal.It is suggested that this specific anosmia corresponds withthe absence of a new olfactory primary, the ‘malty’odor. Isobutyraldehyde and its congeners occur in a wide varietyof foodstuffs, where they may serve as indicators for the availabilityof the indispensable amino acids valine, leucine and isoleucine. ^*Permanent address: Istituto di Industrie Agrarie della Universitàdegli St'udi, Pisa, Italy.     

Human responses to propionic acid. I. Quantification of within- and between-participant variation in perception by normosmics and anosmics

The objective of this study was to fully characterize normosmic perception of stimuli expected to cause widely varying degrees of olfactory and nasal trigeminal stimulation and to directly evaluate the possible role of olfactory nerve stimulation in nasal irritation sensitivity. During each of four identical test sessions, four anosmic and 31 normosmic participants were presented with a range of concentrations extending from peri-threshold for normosmics to supra- threshold for anosmics. For each session, odor (O) and nasal irritation (NI) sensitivities were summarized in terms of the concentrations required to produce four sensation levels ('iso-response' concentrations). Within-participant variation in these iso-response concentrations was < 10-fold for 95% of normosmics, for both O and NI. For O but not NI, these apparent fluctuations in sensitivity were largely accounted for by the uncertainty surrounding the iso-response concentrations calculated for each session. Anosmics exhibited minimal within- and between-participant variation in NI and required, for all but the highest perceptual level, a higher concentration than almost all normosmics. Between-participant variation, expressed in terms of 90% confidence interval widths, was approximately 0.5 log units for both O and NI for the highest perceptual level, but increased to approximately 0.8 and 1.8 log units, respectively, for the lowest (peri- threshold) level. Our findings suggest that: (i) most apparent variation over time in O sensitivity is actually a reflection of the uncertainty surrounding estimates of sensitivity obtained for each session; (ii) within- and between-participant variation in O sensitivity is far less than is commonly reported; and (iii) low to moderate levels of NI in normosmics are the result of relatively weak trigeminal stimulation combined with much greater olfactory activation.      

A quantitative structure activity analysis on the relative sensitivity of the olfactory and the nasal trigeminal chemosensory systems

We have applied a quantitative structure-activity relationship (QSAR) approach to analyze the chemical parameters that determine the relative sensitivity of olfaction and nasal chemesthesis to a common set of volatile organic compounds (VOCs). We used previously reported data on odor detection thresholds (ODTs) and nasal pungency thresholds (NPTs) from 64 VOCs belonging to 7 chemical series (acetate esters, carboxylic acids, alcohols, aliphatic aldehydes, alkylbenzenes, ketones, and terpenes). The analysis tested whether NPTs could be used to separate out "selective" chemosensory effects (i.e., those resting on the transfer of VOCs from the gas phase to the receptor phase) from "specific" chemosensory effects in ODTs. Previous work showed that selective effects overwhelmingly dominate chemesthetic potency whereas both selective and specific effects control olfactory potency. We conclude that it is indeed possible to use NPTs to separate out selective from specific effects in ODTs. Among the series studied, aldehydes and acids, except for formic acid, show clear specific effects in their olfactory potency. Furthermore, for VOCs whose odor potency rests mainly on selective effects, we have developed a QSAR equation that can predict their ODTs based on their NPTs.     

Odor and irritation thresholds for ammonia: a comparison between static and dynamic olfactometry

Odor and lateralization (irritation) thresholds (LTs) for ammonia vapor were measured using static and dynamic olfactometry. The purpose of the study was to explore the test-retest reliability and comparability of dynamic olfactometry methodology, generally used to determine odor thresholds following European Committee for Standardization guidelines in the context of odor regulations to outside emissions, with static olfactometry. Within a 2-week period, odor and LTs for ammonia were obtained twice for each method for 24 females. No significant differences between methods were found: mean odor detection thresholds (ODTs) were 2.6 parts per million (ppm) for either method (P = 0.96), and mean LTs were 31.7 and 60.9 ppm for the static and dynamic method, respectively (P = 0.07). Test-retest reliability was higher for the dynamic than for the static method (r = 0.61 vs. 0.14 for ODTs and r = 0.86 vs. 0.45 for LTs). The choice of optimal method for any application, however, depends not only on psychometric factors but also on practical factors such as physicochemical properties of the compound, availability of equipment and expertise, task efficiency, and costs.     

SPECIFIC ANOSMIAS TO 5{alpha}-ANDROST-16-EN-3-ONE AND {omega}-PENTADECALACTONE: THE URINOUS AND MUSKY PRIMARY ODORS

About 46% of human subjects are specifically anosmic to theodor of 5-androst-16-en-3- one, and about 9% are specificallyanosmic to the odor of -pentadecalactone. Odor threshold measurementson 33 representative steroids, synthetic musks and related compoundswere made with panels of normal observers and each of thesevarieties of specific anosmics. The androstenone anosmia wasmost pronounced with steroid ketones in the androstane seriesand some isosteric analogs. It is suggested that this specificanosmia corresponds with the absence of a recently-suspectedolfactory primary, the ‘urinous’ odor. The pentadecalactoneanosmia was associated with a number of diverse but approximatelyisosteric synthetic musks. This specific anosmia delineatesmore clearly the boundaries of the ‘musky’ odor,whose probably primacy has long been recognized. Threshold measurementswere made on mixtures of androst-16-en-3-one and pentadecalactone.The results depart little from the ‘rule of additivity’for odor ratios up to 100:1, and suggest a quantitative interpretationfor the anosmic defects measured with single compounds. ^* Permanent address: Istituto di Industrie Agrarie della Universitàdegli Studi, Pisa, Italy.     

Intranasal volume and olfactory function

The aim of this exploratory study was to identify the volume intranasal segments as they relate to parameters of olfactory function. Fifty healthy male volunteers (age range 22-59 years, mean age 28.5 years) were included. Olfactory function was measured by lateralized phenyl ethyl alcohol odor thresholds and odor discrimination, and by bilateral odor identification. Magnetic resonance imaging of the nasal cavity was performed immediately following olfactometry. To correlate the results of olfactometry with intranasal volume, each nasal cavity was divided into 11 segments. Significant correlations were found between the odor thresholds and volumes of the anterior part of the lower and upper meatus of the right nasal cavity. These results reveal that two nasal segments are important for inter-individual differences of odor thresholds in healthy subjects: (i) the segment in the upper meatus below the cribriform plate and (ii) the anterior segment of the inferior meatus. The latter finding is of special interest for nasal surgery, which allows modification of this volume through resection of the inferior turbinate and/or septoplasty.     

Individual differences in thresholds and quality reports of human subjects to various odors

Sixty-three subjects were selected on the basis of their relativeability (n = 30) or inability (n = 33) to detect the intenseurine–sweaty odor of the diastereoisomeric ketone, cis-4-(4'-t-butylcyclohexyl)-4-methyl-2-pentanone (pemenone). Absolute thresholds were determined,and quality reports and hedonic ratings were obtained for near-thresholdconcentrations of pemenone and five other odorous compounds.Several of these compounds were selected because large individualdifferences in sensitivity (specific anosmias) were known toexist. A principal compound analysis of threshold concentrationsindicated that three orthogonal factors well described the relationshipsbetween sensitivities to the compounds. Threshold concentrationsof pemenone and androstenone were highly correlated and thefirst factor was defined by relationships between those odorantsand isovaleric acid, all modally putrid-smelling compounds.Subjects relatively osmic for pemenone generally reported aputrid odor for pemenone, but anosmics reported mostly otherqualities. These relationships between relative sensitivityand the quality reports elicited by the different compoundsare consistent with a multiple-profile model of odor qualityperception. That model postulates that most odor molecules interactwith more than one perceptual channel (receptor process) andthat any individual alteration in the relative specificity ordeletion in the number of such receptor processes could alterthe pattern of interaction and thus should give rise to alterationsin the quality or intensity of the resulting odor perception.     

A STUDY OF OLFACTORY DISCRIMINATION IN THE RAT WITH THE AID OF A NEW ODOR DELIVERY TECHNIQUE

A technique for delivering odors at specific concentrationsand for precise periods into the nasal cavity of a freely movingrat is described. This is accomplished by linking the outletof an automatic micro-olfactometer to a cannula chronicallyimplanted into the nasal cavity of the rat. Details of the constructionand implantation of the cannula are given. Using this odor deliverytechnique rats were trained in a two-choice discrimination procedureto distinguish between the odors of benzaldehyde and benzonitrile(both are ‘almond like’), benzaldehyde and isobutylbutyrate(fruity), and benzonitrile-isobutylbutyrate. The rats foundthe olfactory properties of benzaldehyde and benzonitrile tobe similar, and each to be very different to the olfactory propertiesof isobutylbutyrate. It is proposed that the rat may use morethan a single olfactory property of each stimulus to discriminatebetween two odors. ^* This paper reports research undertaken at the School of BiologicalSciences, Macquarie University, and the CSIRO Division of FoodResearch.     

Genetic mapping of sulfur assimilation genes reveals a QTL for onion bulb pungency

Onion exhibits wide genetic and environmental variation in bioactive organosulfur compounds that impart pungency and health benefits. A PCR-based molecular marker map that included candidate genes for sulfur assimilation was used to identify genomic regions affecting pungency in the cross 'W202A' × 'Texas Grano 438'. Linkage mapping revealed that genes encoding plastidic ferredoxin-sulfite reductase (SiR) and plastidic ATP sulfurylase (ATPS) are closely linked (1–2 cM) on chromosome 3. Inbred F₃ families derived from the F₂ population used to construct the genetic map were grown in replicated trials in two environments and bulb pungency was evaluated as pyruvic acid or lachrymatory factor. Broad-sense heritability of pungency was estimated to be 0.78–0.80. QTL analysis revealed significant associations of both pungency and bulb soluble solids content with marker intervals on chromosomes 3 and 5, which have previously been reported to condition pleiotropic effects on bulb carbohydrate composition. Highly significant associations (LOD 3.7–8.7) were observed between ATPS and SiR Loci and bulb pungency but not with bulb solids content. This association was confirmed in two larger, independently derived F₂ families from the same cross. Single-locus models suggested that the partially dominant locus associated with these candidate genes controls 30–50% of genetic variation in pungency in these pedigrees. These markers may provide a practical means to select for lower pungency without correlated selection for lowered solids.     

Odor Perception Phenotypes: Multiple, Specific Hyperosmias to Musks

Olfactory detection thresholds for 11 structurally diverse muskodorants and one non-musk odorant were obtained from 32 subjects.Hierarchical cluster analysis produced four groups of subjects.One group (n = 12) was uniformly sensitive to all musks; another(n = 16) was uniformly insensitive. Two groups of subjects containedotherwise insensitive individuals who were exceptionally sensitiveto cyclopentadecanone and musk xylol (n = 2) and to delta9-hexadecenolactoneand tonalid (n = 2) respectively. We propose that the lattertwo groups are odor perception phenotypes (MSHM1 and MSHM2)that consist of multiple, specific hyperosmias to musk odorants.Chem. Senses 21: 411– 416, 1996. ¹Present address: Synesthetics, Inc., Montclair, NJ 07043, USA     

Chronobiology of Nasal Chemosensitivity: Do Odor or Trigeminal Pain Thresholds Follow a Circadian Rhythm?

Odor and trigeminal pain thresholds were studied four timeseach at 24:00, 04:00, 08:00, 12:00, 16:00 and 20:00 h in randomizedorder on different days in five healthy male volunteers. Nocircadian rhythm of olfactory or trigeminal thresholds wereobserved. However, the variability of odor, but not pain thresholds,increased from 04:00 h (thresholds between 0.4 and 1.2 p.p.m.)to 16:00 h (thresholds between 0.1 and 2 p.p.m.). It is hypothesizedthat environmental influences contribute to this increase invariance. Chem. Senses 22: 593–598, 1997.     

Handedness is a determining factor in lateralized olfactory discrimination

The study aimed to re-investigate differences in olfactory thresholds and odor discrimination between the left and right sides in relation to the handedness of healthy subjects. Twenty left- and 20 right-handers participated; all were in excellent health with no indication of any major nasal or health problems, and all were non-smokers. The two groups were comparable in terms of sex and age (left-handers: 11 women, 9 men, median age 25 years; right-handers: 9 women, 11 men, median age 26 years). Odor thresholds did not differ in relation to handedness. However, in the odor discrimination task the left-handers performed significantly better at the left side compared with the right nostril; this pattern was reversed in the right-handers. The data indicate that, similar to other sensory systems, higher olfactory functions exhibit a certain degree of lateralization.      

A simple and flexible device to odorize large stimulation areas

This paper describes a flow dilution olfactometer which allowsthe odorization of large stimulation areas and the easy manipulationof several odorants and/or concentrations. Generation of theodorized air is performed by mixing in two steps the odor vaporcontained in Tedlar bags with a pure air stream flowing continuouslyout of a nozzle. Discrete concentration values are obtainedby using pre-adjusted needle valves to change the vapor flowsampled in the bags. This kind of olfactometer was utilizedto study odor coding in the olfactory bulbs of rats and rabbits.Five Odorants were delivered at concentrations ranging from2 x 10^-4 to 1.5 x 10^-2 of the saturated vapor pressure. Measurementsshowed that lower concentrations can be obtained by fillingthe bags with a more diluted odor vapor. Furthermore, the numberof test odorants can be increased at low cost by increasingthe number of Tedlar bags.     

生姜的挥发性化学成分

用ＧＣ／ＭＳ方法,定性定量地分析了用３种不同方法提取的姜油的化学成分,分别鉴定了４６,５０,６１个化合物。蒸馏油主要含有单萜、倍半萜烯类化合物,未见生姜特有的辣味成分。冷榨油和超临界ＣＯ２萃取油除倍半萜类化合物外,主要含有辣味成分,含量分别为１８．６１％,２３．０９％。冷榨油及超临界ＣＯ２萃取油较好地保持了生姜的特征辛香气及辣味成分     

Modification of responses of newly hatched snails by exposure to odors during development

Predatory marine snail larvae and embryos were exposed to preyodors (oyster, mussel and barnacle) during development. Whenjuvenile snails hatched they were tested by bioassay to determineeffects of this prior odor exposure. Juvenile snails were testedwith oyster, mussel and barnacle odors and with partially purifiedbarnacle odor of known attractiveness. Independent of priorodor exposure, only solutions containing barnacle odor wereattractive. Snails exposed during development to prey odorsnot in themselves attractive were more responsive to barnacleodors than snails that developed in barnacle odors. Snails notexposed to prey odors during development were intermediate intheir responsiveness. Field bioassays showed detectable attractantlevels in an environment containing barnacles. Attractant activitywas not detected in an environment where barnacles were absent.Responses of snails that developed in field odor conditionswere similar to those of snails that developed in the presenceof barnacles. Odor exposure during development is importantin determining future responses of these predatory snails toprey odors. ¹Duke University Marine Laboratory, Pivers Island, Beaufort,NC 28516, USA ²University of Miami RSMAS/MAC 4600 Rickenbacker Causeway, Miami,FL 33149, USA ³Suffolk University, Department of Biology, Beacon Hill, Boston,MA 02114, USA     

Effect of topical prostaglandins on nasal patency in man

The effect of prostaglandin E₁ and 17 phenyl trinor PGE₂ on nasal patency has been studied in healthy volunteers and in patients with vasomotor and allergic rhinitis. Both drugs applied topically increased nasal patency. The effect of a single dose of either compound lasted for several hours. Prostaglandin E₁ produced nasal irritation and throbbing, lacrimation, headache and sore throat. Except for occasional brief nasal irritation, these side effects were not encountered with 17 phenyl trinor PGE₂.