Development of a smell identification test using a novel stick-type odor presentation kit

The odor identification is strongly influenced by the social and cultural factors; therefore, the odorants used in a smell identification test should be familiar to the test population. In addition, the device used in the test is desired to be simply handled and retain odor quality over time. We developed a novel stick-type odor presentation kit that consists of microcapsules of odorant incorporated into stable cream and the smell identification test using it. Thirteen odorants were selected to be familiar to the test population. In the test, we used two identification methods: one was a modified forced-choice paradigm with "detectable but not recognizable" and "no smell detected" added as choices and the other was a two-step identification paradigm where the participant first selected one of eight odor categories and then chose the specific odor name from the selected category. We verified the performance of the odor stick and the test by stability, using a test-retest paradigm, comparing this test with another smell test, and testing Japanese people from a range of age groups. We conclude that this kit is a useful odor presentation device, and the test using it works effectively as a smell identification test.     

Odor similarity does not influence the time needed for odor processing

The brain's link between perception and action involves several steps, which include stimulus transduction, neuronal coding of the stimulus, comparison to a memory template and choice of an appropriate behavioral response. All of these need time, and many studies report that the time needed to compare two stimuli correlates inversely with the perceived distance between them. We developed a behavioral assay in which we tested the time that a honeybee needs to discriminate between odors consisting of mixtures of two components, and included both very similar and very different stimuli spanning four log-concentration ranges. Bees learned to discriminate all odors, including very similar odors and the same odor at different concentrations. Even though discriminating two very similar odors appears to be a more difficult task than discriminating two very distinct substances, we found that the time needed to make a choice for or against an odor was independent of odor similarity. Our data suggest that, irrespective of the nature of the olfactory code, the bee olfactory system evaluates odor quality after a constant interval. This may ensure that odors are only assessed after the olfactory network has optimized its representation.     

No influence of scopolamine hydrobromide on odor detection performance of rats

Despite speculation that the muscarinic cholinergic antagonist, scopolamine, may influence the olfactory sensitivity of rats, there have been no definitive studies on this point to date. In this study, we examined the influence of a range of doses of scopolamine hydrobromine (namely, 0.10, 0.125, 0.15 and 0.20 mg/kg i.p.) on the odor detection performance of 15 adult male Long-Evans rats to ethyl acetate. Air-dilution olfactometry and a go/no-go operant signal detection task were employed. The drug conditions and a saline control were administered to each animal in an order counterbalanced by Latin squares, with 2 day intervals interspersed between tests. Scopolamine had no significant influence on odor detection performance per se, as measured by percent correct S+ and S- responses and a non-parametric signal detection measure of sensitivity. This is in contrast to the relatively large effects previously observed in the same test paradigm for such drugs as the D-1 agonist SKF 38393 and the D-2 agonist quinpirole. These data suggest that scopolamine has no meaningful influence on a well-practiced odor detection task.     

Cross-cultural comparison of data using the odor stick identification test for Japanese (OSIT-J)

A new olfactory test, the odor stick identification test for Japanese (OSIT-J), has been developed in Japan. To determine if the OSIT-J would be effective cross-culturally, we administered the test to 52 US and 50 Japanese subjects reporting normal olfactory function. The average composite OSIT-J test score for US subjects was significantly lower (77%) than that for Japanese subjects (94%, P < 0.0001). Both US and Japanese subjects correctly identified eight of the 13 odorants included in the OSIT-J with scores of 80% or higher. However, for five odorants, the US subjects' scores fell below 80% and were consistently lower than Japanese subjects, presumably reflecting cultural differences in odor experience. Most of the US subjects found the OSIT-J to be easy, interesting, pleasant, and short in duration. Although the 13-odorant OSIT-J was found to be suitable for testing US populations, elimination of five test odorants that were unfamiliar to US subjects significantly enhanced the test's effectiveness. Findings from this study emphasize the importance of identifying test odorants that may have a cultural bias, a crucial issue when comparing data obtained from different smell tests used at smell and taste centers around the world.     

Activation of the anterior cingulate gyrus by 'Green Odor': a positron emission tomography study in the monkey

The equivalent mixture of cis-3-hexenol and trans-2-hexenal (hexenol/hexenal), 'green odor', is known to have a healing effect on the psychological damage caused by stress. Behavioral studies in humans and monkeys have revealed that hexenol/hexenal prevents the prolongation of reaction time caused by fatigue. In the present study, we investigated which brain regions are activated by the odor of hexenol/hexenal using positron emission tomography with alert monkeys. Regional cerebral blood flow (rCBF) in the prepyriform area (the primary olfactory cortex) was commonly increased by the passive application of odor: acetic acid, isoamylacetate or hexenol/hexenal. We observed rCBF increases in the orbitofrontal cortex (the secondary olfactory cortex) by these olfactory stimuli in two of three monkeys, and found no predominance of laterality of the activated hemisphere. Furthermore, rCBF increase in the cerebellum was observed in two of three monkeys, and the odor of acetic acid increased rCBF in the substantia innominata in all monkeys. In addition to these olfactory related regions, the anterior cingulate gyrus was activated by the odor of hexenol/hexenal. These findings suggest that the increase of rCBF in the anterior cingulate gyrus by the odor of hexenol/hexenal may contribute the healing effects of this mixture observed in the monkey.     

Effects of perceived and imagined odors on taste detection

We assessed the influence of different odors on detection of a sweet tastant, and the ability of imagined odors to elicit the same effects as perceived odors on taste perception. The tastant used was sucrose, and the two odorants were strawberry and ham. In the first experiment, participants either smelled or imagined one of two odors during taste detection tasks (between-subject design), whereas in the second one, subjects completed both the odor imagery and perception conditions with taste detection tasks (within-subject design). The effect was odorant-specific: detection of sucrose was significantly better when subjects smelled strawberry than when they smelled ham. Furthermore, imagined odors influenced taste perception in the same way as did perceived odors. We concluded that the odor-specific effect on taste perception is an authentic perceptual phenomenon. Our results also support the notion that odor-induced changes in taste perception are mediated centrally. Finally, our findings are in agreement with reports supporting the existence of odor imagery.     

The effect of meat consumption on body odor attractiveness

Axillary body odor is individually specific and potentially a rich source of information about its producer. Odor individuality partly results from genetic individuality, but the influence of ecological factors such as eating habits are another main source of odor variability. However, we know very little about how particular dietary components shape our body odor. Here we tested the effect of red meat consumption on body odor attractiveness. We used a balanced within-subject experimental design. Seventeen male odor donors were on "meat" or "nonmeat" diet for 2 weeks wearing axillary pads to collect body odor during the final 24 h of the diet. Fresh odor samples were assessed for their pleasantness, attractiveness, masculinity, and intensity by 30 women not using hormonal contraceptives. We repeated the same procedure a month later with the same odor donors, each on the opposite diet than before. Results of repeated measures analysis of variance showed that the odor of donors when on the nonmeat diet was judged as significantly more attractive, more pleasant, and less intense. This suggests that red meat consumption has a negative impact on perceived body odor hedonicity.     

Individual differences in sensitivity to the odor of 4,16-androstadien-3-one

Individual differences in sensitivity to the putative human pheromone androstadienone were investigated in three experiments. In experiment 1, the absolute detection threshold for androstadienone was determined to be 211 micro M using the method of constant stimuli. Detection for the related compound estratetraenol was also investigated but a threshold could not be determined. In experiment 2, using an adaptive threshold test on 100 participants, the sensitivity distribution for androstadienone, but not for the reference odor phenylethyl alcohol, was bimodal, with a smaller group of individuals with a high sensitivity to androstadienone (supersmellers). A lack of correlation between thresholds for androstadienone and phenylethyl alcohol further suggested that the bimodality for androstadienone was not due to individuals with a high general olfactory sensitivity. In line with an earlier observation, there was a statistical tendency for women to be more sensitive to androstadienone than men. Results of experiment 3 preclude the possibility that the bimodal sensitivity distribution for androstadienone would depend on individual differences in trigeminal activation. Altogether, the current study suggests that olfactory sensitivity to androstadienone is bimodally distributed in the population with a subgroup consisting of highly sensitive people.     

Human odor detectability: new methodology used to determine threshold and variation

Current ambiguity concerning the related issues of optimal means for measurement of odor sensitivity and the functional properties of the olfactory system hinders progress in basic and applied research on the human sense of smell. To address these needs, we selected n-amyl acetate (nAA) as a test odorant and developed a methodology in which participants (Ps) receive multiple presentations each session of several concentrations. Yes-no responses as to whether odor was detected are analyzed using binomial statistics, with the probability that a given proportion of yes responses (or greater) would occur by chance alone being treated as the inverse of detectability. Over the course of multiple sessions, this information is also used to maximize the collection of data in the peri-threshold region. Surprisingly, data collected over as many as 14 sessions were fit well by a single logistic regression model relating probability and concentration. Threshold concentrations, defined as those corresponding to a probability of 0.05, varied from 7.11 to 167.53 p.p.b. (v/v) for 11 Ps. Our approach and findings, if shown to be representative of other combinations of Ps and odorants, could accelerate the pace of research in human olfaction by providing a comprehensive operational definition of the limit of the olfactory system to detect odorant molecules.     

A simple and reliable method for clinical assessment of odor thresholds

We investigated whether presenting of dilutions of phenyl ethyl alcohol at random succession according to the method of constant stimuli can replace the standard procedure of presenting a various number of dilutions in a staircase paradigm. Forty-six men and 44 women, aged 19-76 years, participated in this study. Phenyl ethyl alcohol was diluted in a ratio of 1:2, starting from 4%. Presentation of the odorant followed a three-alternative, temporal forced-choice paradigm with two blanks in addition to the odorant. Twenty dilutions were administered in a randomized order. Odor threshold was obtained by logistic regression of the correct and incorrect identifications of the probe containing the odorant. Thresholds were also calculated on the basis of the first 16 dilution steps only. Results from these procedures were compared with 'gold-standard' threshold assessment employing a three-alternative, temporal forced-choice staircase paradigm with seven reversals using 16 dilutions of phenyl ethyl alcohol. The method of constant stimuli took a shorter and less variable testing time than the staircase technique. The use of 20 dilution steps provided no better results than the use of 16 steps. The method of constant stimuli exhibited a good test-retest reliability (r = 0.7; P < 0.001) comparable to that of the staircase method and provided unbiased results highly correlated (r = 0.8; P < 0.001) with those of the staircase technique with similar inter-test variability. Applying 16 dilutions (1:2 steps) of phenyl ethyl alcohol at random succession in a three-alternative, temporal forced-choice paradigm is thus a simple and reliable procedure for the reproducible assessment of odor thresholds that may be contemplated as an alternative to the 'gold-standard' staircase method of clinical odor threshold assessment.     

Primary culture of lobster (Homarus americanus) olfactory sensory neurons

Lobster olfactory sensory neurons have contributed to a number of advances in our understanding of olfactory physiology. To facilitate further study of their function, we have developed conditions allowing primary culture of the olfactory sensory neurons in a defined medium. The most common cells in the culture were round cell bodies with diameters of 10-15 micro m that often extended fine processes, features resembling olfactory sensory neurons. We discovered that acetylcholinesterase acted as a growth factor for these cells, improving their survival in culture. We also confirmed previous evidence from spiny lobsters that poly-D-lysine was a superior substrate for olfactory cells of this size and morphology. We then identified olfactory sensory neurons in the culture in two ways. Almost half the cells tested responded to application of a complex odorant with an inward current. An even more rigorous test was made possible by the development of an antiserum to OET-07, an ionotropic glutamate receptor homolog specifically expressed by Homarus americanus olfactory sensory neurons. It labeled a majority of the round cells in the culture, unequivocally identifying them as olfactory sensory neurons.     

Hemodynamic response of the frontal cortex elicited by intravenous thiamine propyldisulphide administration

The intravenous olfaction (IVO) test is a unique type of clinical olfactometry and is widely used in Japan. However, it is difficult to distinguish actual olfactory disturbance from feigned disturbance because the IVO test is a psychophysical test. To resolve this problem, we investigated the possibility of an objective IVO test assisted with near infrared spectroscopy (NIRS). IVO testing was performed according to the usual protocol with thiamine propyldisulphide (alinamin) administration. The relative oxy- and deoxyhemoglobin levels of the orbitofrontal area during olfactory stimulation by IVO test were measured by NIRS. Pairs of NIRS emitters and detectors were positioned on the bilateral frontal scalp. After administration of alinamin, oxyhemoglobin levels increased, though deoxyhemoglobin levels did not change. An increase in oxyhemoglobin levels was observed bilaterally. Administration of saline did not elicit any change in the oxy- or deoxyhemoglobin levels and concentration of the administered alinamin related increasing of the oxyhemoglobin level was observed. Oxyhemoglobin remained unchanged in anosmic subjects despite administration of alinamin. The latency of oxyhemoglobin increase on each side and smelling latency showed significant correlation. Latencies of oxyhemoglobin increases between the right and left sides also showed significant correlation. Oxyhemoglobin response appears to be linked to olfactory related response. NIRS is a useful technique for the development of an objective form of IVO testing.     

Metallic taste and retronasal smell

A series of experiments investigated the nature of metallic taste reports and whether they can be attributed to the development of a retronasal smell. Two studies showed that the metallic sensation reports following oral stimulation with solutions of FeSO4 were reduced to baseline when the nose was occluded. No such reduction was seen for CuSO4 or ZnSO4, which were more bitter and astringent, respectively, and less metallic. A discrimination test based on weak but equi-intense levels of FeSO4 and CuSO4 showed that FeSO4 could be discriminated from water with the nose open but not when occluded, but that discrimination of CuSO4 from water was not impaired by nasal occlusion. A discrimination test demonstrated that the headspace over solutions of FeSO4 was not different from water, although some subjects could discriminate FeSO4 solutions from water in the mouth when the nose was occluded, perhaps by tactile or astringent cues. These results confirm that metallic taste reports following oral stimulation with FeSO4 are likely due to development of a retronasal smell, possibly following a lipid oxidation reaction in the mouth. However, metallic taste reports may arise from different mechanisms with copper and zinc salts.     

The influence of age and sex on phagocyte chemiluminescence

The process of ageing is associated with increased susceptibility to infection. Phagocytes form the primary defence mechanism against infecting microorganisms, but the influence of ageing on phagocyte function remains controversial. In this study we have applied a microtitre plate phagocyte chemiluminescence (CL) assay suitable for clinical use to compare phagocyte oxidative metabolism in younger healthy subjects (age 20–60 years) and healthy older (60–70 years) subjects. Polymorphonuclear leukocytes (PMNL) and monocytes were stimulated using phorbol myristate acetate (PMA), serum opsonized zymosan (SOZ), and non-opsonized zymosan (ZYM) in the presence of both lucigenin and luminol. Monocytes showed a higher luminolenhanced CL response to PMA in males compared with females in the younger age group. No PMNL differences were observed between the sexes. Although no difference were found in relation to age when cells were stimulated with PMA and SOZ, significantly lower background (unstimulated) CL was obtained from PMNL with luminol. PMNL luminol-enhanced CL responses were also lower in response to ZYM. The findings suggest a reduced response of PMNL from older subjects to minimal stimulation. This could be related to abnormalities in the triggering of the respiratory burst or myeloperoxidase release due to ageing. The influence of age and sex should be taken into account in clinical studies of phagocyte CL.     

Effect of age and sex on copper-induced toxicity in theMacular mutant mouse

It was determined if the sensitivity inmacular mutant mouse to copper-induced toxicity was affected by sex or age. The sensitivity in 6–8-d-old or 3–4-wk-oldmacular mutant mouse to copper-induced toxicity was not affected by sex. However, 8–9-wk-old mutant females were more sensitive to copper-induced toxicity than mutant males. Furthermore, 6–8-d-old or 3–4-wk-old mutant males were more sensitive to copper-induced toxicity than 8–9-wk-old mutant males. However, age-related differences in sensitivity to copper-induced toxicity did not occur significantly in mutant females. On the other hand, in the case of normal mice, the sensitivity in 6–8-d-old or 3–4-wk-old mice to copper-induced toxicity was not also affected by sex. In contrast to mutant, however, 8–9-wk-old normal males were more sensitive to copper-induced toxicity than 8–9-wk-old normal females. Adult males were also more sensitive to copper-induced toxicity than 6–8-d-old or 3–4-wk-old males. However, age-related differences in sensitivity to copper-induced toxicity did not occur significantly in normal females. These results indicate that sex- and age-related differences in the copper-induced toxicity exist inmacular mutant mice.     

Sniff magnitude test: relationship to odor identification, detection, and memory tests in a clinic population

Recently a novel measure of olfactory function, the Sniff Magnitude Test (SMT), was developed that relies on changes in inhalation in response to an odor. The relationship of this unique test to that of other olfactory tests has received little investigation. In this study, we assessed, in 132 patients presenting to a chemosensory disorders clinic, the relationship of SMT scores to those from 3 standardized psychophysical tests: the University of Pennsylvania Smell Identification Test (UPSIT), a phenyl ethyl alcohol odor detection threshold test, and a short-term odor memory/discrimination test. SMT scores were roughly related to olfactory dysfunction categories defined for the UPSIT and correlated moderately with the other tests. Malodors (1% and 3% methylthiobutyrate [MTB], 1% ethyl 3-mercaptoproprionate) exhibited stronger correlations than nonmalodors (3% phenyl ethyl alcohol [PEA], 3% amyl acetate, 3% n-butanol) and elicited greater sniff suppression. In a principal component analysis, the SMT measures loaded on components different from those of the other tests, which loaded on a separate component. Anticipatory responses (i.e., smaller sniffs) occurred across trials for the first malodor (1% MTB), but not for the first nonmalodor (3% PEA), that was encountered. These results, along with those of an earlier factor analysis, suggest that sniff magnitude is influenced by odorant quality and intensity, as well as by cognitive factors.     

Odorant binding and conformational changes of a rat odorant-binding protein

Odorant-binding proteins (OBPs) are lipocalins secreted in the nasal mucus of vertebrates, which convey odorants to their neuronal receptors. We compared the binding properties of a recombinant rat OBP (OBP-1F) using a set of six odorants of various chemical structures. We examined the binding properties by both fluorescent probe competition and isothermal titration calorimetry. OBP-1F affinity constants, in the micromolar range, varied by more than one order of magnitude and were roughly correlated to the odorant size. The observed binding stoichiometry was found to be around one odorant per dimer. Using tyrosine differential spectroscopy, the binding of ligand was shown to induce local conformational changes. A three-dimensional structure of OBP-1F, modelled using the known structure of aphrodisin as template, allowed us to suggest the location of the observed structural changes outside of the binding pocket. These results are consistent with one binding site located in one of the two beta-barrels of the OBP-1F dimer and a subtle conformational change correlated with binding of an odorant molecule, which hampers uptake of a second odorant by the other hydrophobic pocket.     

Pheromone-binding proteins contribute to the activation of olfactory receptor neurons in the silkmoths antheraea polyphemus and Bombyx mori

The sensilla trichodea of the silkmoth Antheraea polyphemus are innervated by three types of receptor neurons each responding specifically to one of three pheromone components. The sensillum lymph of these sensilla surrounding the sensory dendrites contains three different types of pheromone-binding proteins (PBPs) in high concentrations. The sensilla trichodea of the silkmoth Bombyx mori are supplied by two receptor neurons each tuned specifically to one of the two pheromone components bombykol and bombykal, but only one type of PBP has been found so far in these sensilla. Recombinant PBPs of both silkmoth species in various combinations with pheromone components were applied to the receptor neurons via tip-opened sensilla during electrophysiological recordings. Over a fairly broad range of pheromone concentrations the responses of the receptor neurons depended on both, the pheromone component and the type of the PBP. Therefore, the PBPs appear to contribute to the excitation of the receptor neurons. Furthermore, bombykal in combination with the expressed PBP of B. mori failed to activate the corresponding receptor neuron of B. mori, but did so if combined with one of the PBPs of A. polyphemus. Therefore, a still unknown binding protein involved in bombykal transport might be present in B. mori.     

Different thresholds for detection and discrimination of odors in the honey bee (Apis mellifera)

Naturally occurring odors used by animals for mate recognition, food identification and other purposes must be detected at concentrations that vary across several orders of magnitude. Olfactory systems must therefore have the capacity to represent odors over a large range of concentrations regardless of dramatic changes in the salience, or perceived intensity, of a stimulus. The stability of the representation of an odor relative to other odors across concentration has not been extensively evaluated. We tested the ability of honey bees to discriminate pure odorants across a range of concentrations at and above their detection threshold. Our study showed that pure odorant compounds became progressively easier for honey bees to discriminate with increasing concentration. Discrimination is, therefore, a function of odorant concentration. We hypothesize that the recruitment of sensory cell populations across a range of concentrations may be important for odor coding, perhaps by changing its perceptual qualities or by increasing its salience against background stimuli, and that this mechanism is a general property of olfactory systems.     

The influence of female age and host size on the sex ratio of the parasitoid Opius concolor

The sex ratio of the progeny of single females parasitizing large hosts favoured the females (sex ratio=0.26); but on small hosts favoured the males (0.73). No differences in mortality of the sexes were detected. The sex ratio was independent of female age when large hosts were used. The percentage of males observed in the progeny of the first day of female oviposition was significantly greater than the mean, irrespective of the age at which female oviposition began. When females were exposed to small hosts, a greater percentage of females was observed in the progeny from the last days of oviposition.

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Résumé L'influence de la taille de l'hôteret de l'âge de la femelle sur le taux sexuel de la descendance a été étudiée sur le parasitoïde Opius concolor Szépl.Le taux sexuel de la descendance des femelles isolées est favorable aux femelles (t.s.=0,26) quand elles ont à leur disposition des hôtes de grande taille, tandis qu'avec des hôtes petits le taux sexuel est favorable aux mâles (t.s.=0.73). On n'a pas détecté de mortalité différentielle des sexes.Les pourcentage de mâles obtenu le premier jour de ponte des femelles sur les hôtes de grande taille est significativement différent de la moyenne, indépendamment de l'âge de la femelle à ce moment. Cependant, sur des hôtes petits, bien que restant favorable aux mâles dans l'ensemble, une plus grande proportion de femelles à partir des premiers jours de ponte a pu être observée.