Publications

Disorders of consciousness (DoC) are severe neurological conditions in which consciousness is impaired to various degrees. They are caused by injury or malfunction of neural systems regulating arousal and awareness. Over the last decades, major efforts in improving and individualizing diagnostic and prognostic accuracy for patients affected by DoC have been made, mainly focusing on introducing multimodal assessments to complement behavioral examination. The present EU-funded multicentric research project "PerBrain" is aimed at developing an individualized diagnostic hierarchical pathway guided by both behavior and multimodal neurodiagnostics for DoC patients. In this project, each enrolled patient undergoes repetitive behavioral, clinical, and neurodiagnostic assessments according to a patient-tailored multi-layer workflow. Multimodal diagnostic acquisitions using state-of-the-art techniques at different stages of the patients' clinical evolution are performed. The techniques applied comprise well-established behavioral scales, innovative neurophysiological techniques (such as quantitative electroencephalography and transcranial magnetic stimulation combined with electroencephalography), structural and resting-state functional magnetic resonance imaging, and measurements of physiological activity (i.e. nasal airflow respiration). In addition, the well-being and treatment decision attitudes of patients' informal caregivers (primarily family members) are investigated. Patient and caregiver assessments are performed at multiple time points within one year after acquired brain injury, starting at the acute disease phase. Accurate classification and outcome prediction of DoC are of crucial importance for affected patients as well as their caregivers, as individual rehabilitation strategies and treatment decisions are critically dependent on the latter. The PerBrain project aims at optimizing individual DoC diagnosis and accuracy of outcome prediction by integrating data from the suggested multimodal examination methods into a personalized hierarchical diagnosis and prognosis procedure. Using the parallel tracking of both patients' neurological status and their caregivers' mental situation, well-being, and treatment decision attitudes from the acute to the chronic phase of the disease and across different countries, this project aims at significantly contributing to the current clinical routine of DoC patients and their family members.

Background, Key to curtailing the COVID-19 pandemic are wide-scale screening strategies. An ideal screen is one that would not rely on transporting, distributing, and collecting physical specimens. Given the olfactory impairment associated with COVID-19, we developed a perceptual measure of olfaction that relies on smelling household odorants and rating them online. Methods, Each participant was instructed to select 5 household items, and rate their perceived odor pleasantness and intensity using an online visual analogue scale. We used this data to assign an olfactory perceptual fingerprint, a value that reflects the perceived difference between odorants. We tested the performance of this real-time tool in a total of 13,484 participants (462 COVID-19 positive) from 134 countries who provided 178,820 perceptual ratings of 60 different household odorants. Results, We observe that olfactory ratings are indicative of COVID-19 status in a country, significantly correlating with national infection rates over time. More importantly, we observe indicative power at the individual level (79% sensitivity and 87% specificity). Critically, this olfactory screen remains effective in participants with COVID-19 but without symptoms, and in participants with symptoms but without COVID-19. Conclusions, The current odorant-based olfactory screen adds a component to online symptom-checkers, to potentially provide an added first line of defense that can help fight disease progression at the population level. The data derived from this tool may allow better understanding of the link between COVID-19 and olfaction.

Nonhuman terrestrial mammals sniff themselves and each other to decide who is friend or foe. Humans also sniff themselves and each other, but the function of this is unknown. Because humans seek friends who are similar to themselves, we hypothesized that humans may smell themselves and others to subconsciously estimate body odor similarity, which, in turn, may promote friendship. To test this, we recruited nonromantic same-sex friend dyads and harvested their body odor. We found that objective ratings obtained with an electronic nose, and subjective ratings obtained from independent human smellers converged to suggest that friends smell more similar to each other than random dyads. Last, we recruited complete strangers, smelled them with an electronic nose, and engaged them in nonverbal same-sex dyadic interactions. We observed that dyads who smelled more similar had more positive dyadic interactions. In other words, we could predict social bonding with an electronic nose. We conclude that there is indeed chemistry in social chemistry. An electronic nose can predict whether two strangers will have a positive social interaction.

Rapid diagnosis is key to curtailing the Covid-19 pandemic. One path to such rapid diagnosis may rely on identifying volatile organic compounds (VOCs) emitted by the infected body, or in other words, identifying the smell of the infection. Consistent with this rationale, dogs can use their nose to identify Covid-19 patients. Given the scale of the pandemic, however, animal deployment is a challenging solution. In contrast, electronic noses (eNoses) are machines aimed at mimicking animal olfaction, and these can be deployed at scale. To test the hypothesis that SARS CoV-2 infection is associated with a body-odor detectable by an eNose, we placed a generic eNose in-line at a drive-through testing station. We applied a deep learning classifier to the eNose measurements, and achieved real-time detection of SARS CoV-2 infection at a level significantly better than chance, for both symptomatic and non-symptomatic participants. This proof of concept with a generic eNose implies that an optimized eNose may allow effective real-time diagnosis, which would provide for extensive relief in the Covid-19 pandemic.

Functional magnetic resonance imaging (fMRI) has become the leading method for measuring the human brain response to sensory stimuli. However, olfaction fMRI lags behind vision and audition fMRI for two primary reasons: First, the olfactory brain areas are particularly susceptible to imaging artefacts, and second, the olfactory stimulus is particularly difficult to control in the fMRI environment. A component of the latter is related to the odorant-delivery human-machine interface, namely the point where odorants exit the dispensing apparatus to reach at the nose. Previous approaches relied on either nasal cannulas or nasal masks, each associated with particular drawbacks and discomforts. Here we provide detailed descriptions and instructions for transforming the MRI head-coil into an olfactory microenvironment, or odor canopy, where odorants can be switched on and off in less than 150 milliseconds without cannula or mask. In a proof-of-concept experiment we demonstrate that odor canopy provides for clearly dissociable odorant presence and absence, with no non-olfactory cues. Moreover, we find that odor canopy is rated more comfortable than nasal-mask, and we demonstrate that using odor canopy in the fMRI generates a typical olfactory brain-response. We conclude in recommending this approach for minimized discomfort in fMRI of olfaction.

In terrestrial mammals, body volatiles can effectively trigger or block conspecific aggression. Here, we tested whether hexadecanal (HEX), a human body volatile implicated as a mammalian-wide social chemosignal, affects human aggression. Using validated behavioral paradigms, we observed a marked dissociation: Sniffing HEX blocked aggression in men but triggered aggression in women. Next, using functional brain imaging, we uncovered a pattern of brain activity mirroring behavior: In both men and women, HEX increased activity in the left angular gyrus, an area implicated in perception of social cues. HEX then modulated functional connectivity between the angular gyrus and a brain network implicated in social appraisal (temporal pole) and aggressive execution (amygdala and orbitofrontal cortex) in a sex-dependent manner consistent with behavior: increasing connectivity in men but decreasing connectivity in women. These findings implicate sex-specific social chemosignaling at the mechanistic heart of human aggressive behavior.

Memory consolidation can be promoted via targeted memory reactivation (TMR) that re-presents training cues or context during sleep. Whether TMR acts locally or globally on cortical sleep oscillations remains unknown. Here, we exploit the unique functional neuroanatomy of olfaction with its ipsilateral stimulus processing to perform local TMR in one brain hemisphere. Participants learned associations between words and locations in left or right visual fields with contextual odor throughout. We found lateralized event-related potentials during task training that indicate unihemispheric memory processes. During post-learning naps, odors were presented to one nostril in non-rapid eye movement (NREM) sleep. Memory for specific words processed in the cued hemisphere (ipsilateral to stimulated nostril) was improved after local TMR during sleep. Unilateral odor cues locally modulated slow-wave (SW) power such that regional SW power increase was lower in the cued hemisphere relative to the uncued hemisphere and negatively correlated with select memories for cued words. Moreover, local TMR improved phase-amplitude coupling (PAC) between slow oscillations and sleep spindles specifically in the cued hemisphere. The effects on memory performance and cortical sleep oscillations were not observed when unilateral olfactory stimulation during sleep followed learning without contextual odor. Thus, TMR in human sleep transcends global action by selectively promoting specific memories associated with local sleep oscillations.

All primates, including humans, engage in self-face-touching at very high frequency. The functional purpose or antecedents of this behaviour remain unclear. In this hybrid review, we put forth the hypothesis that self-face-touching subserves self-smelling. We first review data implying that humans touch their faces at very high frequency. We then detail evidence from the one study that implicated an olfactory origin for this behaviour: This evidence consists of significantly increased nasal inhalation concurrent with self-face-touching, and predictable increases or decreases in self-face-touching as a function of subliminal odourant tainting. Although we speculate that self-smelling through self-face-touching is largely an unconscious act, we note that in addition, humans also consciously smell themselves at high frequency. To verify this added statement, we administered an online self-report questionnaire. Upon being asked, approximately 94% of approximately 400 respondents acknowledged engaging in smelling themselves. Paradoxically, we observe that although this very prevalent behaviour of self-smelling is of concern to individuals, especially to parents of children overtly exhibiting self-smelling, the behaviour has nearly no traction in the medical or psychological literature. We suggest psychological and cultural explanations for this paradox, and end in suggesting that human self-smelling become a formal topic of investigation in the study of human social olfaction. This article is part of the Theo Murphy meeting issue 'Olfactory communication in humans'.

Mammalian olfaction and reproduction are tightly linked, a link less explored in humans. Here, we asked whether human unexplained repeated pregnancy loss (uRPL) is associated with altered olfaction, and particularly altered olfactory responses to body-odor. We found that whereas most women with uRPL could identify the body-odor of their spouse, most control women could not. Moreover, women with uRPL rated the perceptual attributes of men’s body-odor differently from controls. These pronounced differences were accompanied by an only modest albeit significant advantage in ordinary, non-body-odor-related olfaction in uRPL. Next, using structural and functional brain imaging, we found that in comparison to controls, most women with uRPL had smaller olfactory bulbs, yet increased hypothalamic response in association with men’s body-odor. These findings combine to suggest altered olfactory perceptual and brain responses in women experiencing uRPL, particularly in relation to men’s body-odor. Whether this link has any causal aspects to it remains to be explored.

The olfactory bulbs (OBs) are the first site of odor representation in the mammalian brain, and their unique ultrastructure is considered a necessary substrate for spatiotemporal coding of smell. Given this, we were struck by the serendipitous observation at MRI of two otherwise healthy young left-handed women, yet with no apparent OBs. Standardized tests revealed normal odor awareness, detection, discrimination, identification, and representation. Functional MRI of these women's brains revealed that odorant-induced activity in piriform cortex, the primary OB target, was similar in its extent to that of intact controls. Finally, review of a public brain-MRI database with 1,113 participants (606 women) also tested for olfactory performance, uncovered olfaction without anatomically defined OBs in similar to 0.6% of women and similar to 4.25% of left-handed women. Thus, humans can perform the basic facets of olfaction without canonical OBs, implying extreme plasticity in the functional neuroanatomy of this sensory system.

In response to the coronavirus disease 2019 (COVID-19) pandemic, countries have implemented various strategies to reduce and slow the spread of the disease in the general population. For countries that have implemented restrictions on its population in a stepwise manner, monitoring of COVID-19 prevalence is of importance to guide the decision on when to impose new, or when to abolish old, restrictions. We are here determining whether measures of odor intensity in a large sample can serve as one such measure. Online measures of how intense common household odors are perceived and symptoms of COVID-19 were collected from 2440 Swedes. Average odor intensity ratings were then compared to predicted COVID-19 population prevalence over time in the Swedish population and were found to closely track each other (r = -0.83). Moreover, we found that there was a large difference in rated intensity between individuals with and without COVID-19 symptoms and the number of symptoms was related to odor intensity ratings. Finally, we found that individuals progressing from reporting no symptoms to subsequently reporting COVID-19 symptoms demonstrated a large drop in olfactory performance. These data suggest that measures of odor intensity, if obtained in a large and representative sample, can be used as an indicator of COVID-19 disease in the general population. Importantly, this simple measure could easily be implemented in countries without widespread access to COVID-19 testing or implemented as a fast early response before widespread testing can be facilitated.

After severe brain injury, it can be difficult to determine the state of consciousness of a patient, to determine whether the patient is unresponsive or perhaps minimally conscious(1), and to predict whether they will recover. These diagnoses and prognoses are crucial, as they determine therapeutic strategies such as pain management, and can underlie end-of-life decisions(2,3). Nevertheless, there is an error rate of up to 40% in determining the state of consciousness in patients with brain injuries(4,5). Olfaction relies on brain structures that are involved in the basic mechanisms of arousal(6), and we therefore hypothesized that it may serve as a biomarker for consciousness(7). Here we use a non-verbal non-task-dependent measure known as the sniff response(8-11) to determine consciousness in patients with brain injuries. By measuring odorant-dependent sniffing, we gain a sensitive measure of olfactory function(10-15). We measured the sniff response repeatedly over time in patients with severe brain injuries and found that sniff responses significantly discriminated between unresponsive and minimally conscious states at the group level. Notably, at the single-patient level, if an unresponsive patient had a sniff response, this assured future regaining of consciousness. In addition, olfactory sniff responses were associated with long-term survival rates. These results highlight the importance of olfaction in human brain function, and provide an accessible tool that signals consciousness and recovery in patients with brain injuries.

Wavelength is a physical measure of light, and the intricate understanding of its link to perceived colour enables the creation of perceptual entities such as metamers—non-overlapping spectral compositions that generate identical colour percepts1. By contrast, scientists have been unable to develop a physical measure linked to perceived smell, even one that merely reflects the extent of perceptual similarity between odorants2. Here, to generate such a measure, we collected perceptual similarity estimates of 49,788 pairwise odorants from 199 participants who smelled 242 different multicomponent odorants and used these data to refine a predictive model that links odorant structure to odorant perception3. The resulting measure combines 21 physicochemical features of the odorants into a single number—expressed in radians—that accurately predicts the extent of perceptual similarity between multicomponent odorant pairs. To assess the usefulness of this measure, we investigated whether we could use it to create olfactory metamers. To this end, we first identified a cut-off in the measure: pairs of multicomponent odorants that were within 0.05 radians of each other or less were very difficult to discriminate. Using this cut-off, we were able to design olfactory metamers—pairs of non-overlapping molecular compositions that generated identical odour percepts. The accurate predictions of perceptual similarity, and the ensuing creation of olfactory metamers, suggest that we have obtained a valid olfactory measure, one that may enable the digitization of smell.

Introduction Women's olfactory perception varies across the menstrual cycle. The influence of oral contraceptives on this variability remains unclear. Methods To further estimate this, we assessed discrimination performance for both body odors and ordinary odorants in 36 women, 18 naturally ovulating, and 18 using oral contraceptives. Each participant was tested once a week over the course of a month, and data was then parsed into menstrual phases. Results In naturally ovulating women, at the transition from follicular to luteal phases, there was a decline of 19% (p = 0.003) in olfactory discrimination of body odors but not ordinary odorants. In turn, in women using oral contraceptives, only at a later time of the month, at a point corresponding to the late luteal phase and shift from post-ovulation to pre-menstruation, was there a decline of 20% (p = 0.002) in olfactory discrimination performance. Moreover, when we reorganized the data from women using oral contraceptives in order to separately assess the contraceptive withdrawal period (the few days off pills), we observed a 23% reduction (p = 0.01) in discrimination accuracy of body odors but not ordinary odorants during this time alone. Conclusions Women have reduced ability to discriminate body odors during the withdrawal period of oral contraception. Implications If women indeed consider men’s body odor in their mate selections, then the oral contraception withdrawal period may not be the best time to make such decisions.

Olfactory stimulus acquisition is perfectly synchronized with inhalation, which tunes neuronal ensembles for incoming information. Because olfaction is an ancient sensory system that provided a template for brain evolution, we hypothesized that this link persisted, and therefore nasal inhalations may also tune the brain for acquisition of non-olfactory information. To test this, we measured nasal airflow and electroencephalography during various non-olfactory cognitive tasks. We observed that participants spontaneously inhale at non-olfactory cognitive task onset and that such inhalations shift brain functional network architecture. Concentrating on visuospatial perception, we observed that nasal inhalation drove increased task-related brain activity in specific task-related brain regions and resulted in improved performance accuracy in the visuospatial task. Thus, mental processes with no link to olfaction are nevertheless phase-locked with nasal inhalation, consistent with the notion of an olfaction-based template in the evolution of human brain function.

A common goal in olfaction research is modeling the link between odorant structure and odor perception. Such modeling efforts require large data sets on olfactory perception, yet only a few of these are publicly and freely available. Given that individual odor perception may be informative on personal makeup and interpersonal relationships, we hypothesized that people would gladly provide olfactory perceptual estimates in the context of an odor-based social network. We developed a web-based infrastructure for such a network we called SmellSpace and distributed 10 000 scratch-and-sniff registration booklets each containing a subset of 12 out of 35 microencapsulated odorants. Within similar to 100 days, we obtained data from similar to 1000 participants who rated the odorants along 13 verbal descriptors. To verify that these estimates are comparable to lab-collected estimates we tested 26 participants in a controlled lab setting using the same odorants and descriptors. We observed remarkably high overall group correlations between lab and SmellSpace data, implying that this method provides for credible group-representations of odorants. We further estimated the usability of the data by applying to it two previously published models that used odorant structure alone to predict either odorant pleasantness or pairwise odorant perceptual similarity. We observed statistically significant predictions in both cases, thus further implying that the current data may be helpful toward future efforts of modeling olfactory perception from structure. We conclude that an odor-based social network is a potentially useful instrument for collecting extensive data on olfactory perception and here post the complete raw data set from the first similar to 1000 participants.

Autism spectrum disorder (ASD) is characterized by impaired social communication, often attributed to misreading of emotional cues. Why individuals with ASD misread emotions remains unclear. Given that terrestrial mammals rely on their sense of smell to read conspecific emotions, we hypothesized that misreading of emotional cues in ASD partially reflects altered social chemosignaling. We found no difference between typically developed (TD) and cognitively able adults with ASD at explicit detection and perception of social chemosignals. Nevertheless, TD and ASD participants dissociated in their responses to subliminal presentation of these same compounds: the undetected 'smell of fear' (skydiver sweat) increased physiological arousal and reduced explicit and implicit measures of trust in TD but acted opposite in ASD participants. Moreover, two different undetected synthetic putative social chemosignals increased or decreased arousal in TD but acted opposite in ASD participants. These results implicate social chemosignaling as a sensory substrate of social impairment in ASD.

Improved easy-to-use diagnostic tools for infections are in strong demand worldwide. Yet, despite dramatic advances in diagnostic technologies, the gold-standard remains culturing. Here we offer an alternative tool demonstrating that a bacterial biosensor can efficiently detect Pseudomonas aeruginosa infections in patients suffering from otitis externa. Detection was based on specific binding between the biosensor and 2-aminoacetophenone (2-AA), a volatile produced by P. aeruginosa in high amounts. We collected pus samples from ears of 26 subjects exhibiting symptoms of otitis externa. Detection of P. aeruginosa using the biosensor was compared to detection using gold-standard culturing assay and to gas-chromatograph-mass-spectrometry (GC-MS) analyses of 2-AA. The biosensor strain test matched the culture assay in 24 samples (92%) and the GC-MS analyses in 25 samples (96%). With this result in hand, we designed a device containing a whole-cell luminescent biosensor combined with a photo-multiplier tube. This device allowed detection of 2-AA at levels as low as 2 nmol, on par with detection level of GC-MS. The results of the described study demonstrate that the volatile 2-AA serves as an effective biomarker for P. aeruginosa in ear infections, and that activation of the biosensor strain by 2-AA provides a unique opportunity to design an easy-to-use device that can specifically detect P. aeruginosa infections.

Individuals with cervical spinal cord lesions (SCLs) typically depend on caregivers to manually assist in coughing by pressing against their abdominal wall. Coughing can also be assisted by functional electric stimulation (FES) applied to abdominal muscles via surface electrodes. Efficacy of FES, however, depends on precise temporal synchronization. The sniff controller is a trigger that enables paralyzed individuals to precisely control external devices through alterations in nasal airflow. We hypothesized that FES self-triggering by sniff controller may allow for effective cough timing. After optimizing parameters in 16 able-bodied subjects, we measured peak expiratory flow (PEF) in 14 subjects with SCL who coughed with or without assistance. Assistance was either manual assistance of a caregiver, caregiver activated FES, button self-activated FES (for SCL participants who could press a button), or sniff-controlled self-activated FES. We found that all assisted methods provided equally effective improvements, increasing PEF on average by 25 +/- 27% (F[4, 52] = 7.99, p = 0.00004). There was no difference in efficacy between methods of assistance (F[3, 39] = 0.41, p = 0.75). Notably, sniff-controlled FES was the only method of those tested that can be activated by all paralyzed patients alone. This provides for added independence that is a critical factor in quality of life following SCL.

As far as I know, the only reason we need to sleep that is really, really solid is because we get sleepy. Coming from William C. Dement, one of the pioneers of contemporary sleep research, this statement depicts sleep as a neurobiological black box. One of the best ways to probe such black boxes is through exceptions, and olfaction stands out as such an exceptional sensory system during sleep. Specifically, whereas sensory stimuli presented during sleep typically wake, this is not the case with odors. In fact, odors may promote sleep. In turn, they remain processed by the sleeping brain, and provide a telling window onto sleeping brain capabilities. Here, we briefly review the foundations of sleep, and then extensively detail the literature on olfaction in sleep, concentrating on studies in humans. We speculate that the unique interplay of sleep and smell whereby odors are processed in sleep without causing wake reflects unique aspects of olfactory neurophysiology, particularly the direct projections from periphery to cortex without a thalamic relay. Finally, although the mechanisms allowing odor processing during sleep without arousal remain unclear, this phenomenon lends itself to using olfaction as a window onto sleep mentation. This approach has uncovered several aspects of learning and memory during sleep. We review these efforts, and conclude with detailing their potential application in the treatment of disease.

In a study by Gelstein etal., we found that human emotional tears act as a social chemosignal. In the first of three different experiments in that study we observed that sniffing women's emotional tears reduced the sexual attractiveness attributed by men to pictures of women's faces. In a study partly titled Chemosignaling effects of human tears revisited, Graanin etal. claim failed replication of this effect in a series of experiments, one they described as exactly the same procedure as Gelstein. Given that Graanin etal. refused our extended offer to jointly replicate the experiment at our expense, we can merely comment on their effort. We find that Graanin, who are not a chemosignaling laboratory, used methodology that falls short of standards typically applied in chemosignaling research. Thus, their experiments were profoundly different from Gelstein. Finally, we found that in reanalysing their raw data we could in fact replicate the effect from Gelstein. Thus, we conclude that the failed replication in Graanin is neither a replication nor failed.

Nasal airflow is greater in one nostril than in the other because of transient asymmetric nasal passage obstruction by erectile tissue. The extent of obstruction alternates across nostrils with periodicity referred to as the nasal cycle. The nasal cycle is related to autonomic arousal and is indicative of asymmetry in brain function. Moreover, alterations in nasal cycle periodicity have been linked to various diseases. There is therefore need for a tool allowing continuous accurate measurement and recording of airflow in each nostril separately. Here we provide detailed instructions for constructing such a tool at minimal cost and effort. We demonstrate application of the tool in 33 right-handed healthy subjects, and derive several statistical measures for nasal cycle characterization. Using these measures applied to 24-hour recordings we observed that: 1: subjects spent slightly longer in left over right nostril dominance (left = 2.63 +/- 0.89 hours, right = 2.17 +/- 0.89 hours, t(32) = 2.07, p

Rules linking patterns of olfactory receptor neuron activation in the nose to activity patterns in the brain and ensuing odor perception remain poorly understood. Artificially stimulating olfactory neurons with electrical currents and measuring ensuing perception may uncover these rules. We therefore inserted an electrode into the nose of 50 human volunteers and applied various currents for about an hour in each case. This induced assorted non-olfactory sensations but never once the perception of odor. To validate contact with the olfactory path, we used functional magnetic resonance imaging to measure resting-state brain activity in 18 subjects before and after un-sensed stimulation. We observed stimulation-induced neural decorrelation specifically in primary olfactory cortex, implying contact with the olfactory path. These results suggest that indiscriminate olfactory activation does not equate with odor perception. Moreover, this effort serendipitously uncovered a novel path for minimally invasive brain stimulation through the nose.

Most forms of suprathreshold sensory stimulation perturb sleep. In contrast, presentation of pure olfactory or mild trigeminal odorants does not lead to behavioral or physiological arousal. In fact, some odors promote objective and subjective measures of sleep quality in humans and rodents. The brain mechanisms underlying these sleep-protective properties of olfaction remain unclear. Slow oscillations in the electroencephalogram (EEG) are a marker of deep sleep, and K complexes (KCs) are an EEG marker of cortical response to sensory interference. We therefore hypothesized that odorants presented during sleep will increase power in slow EEG oscillations. Moreover, given that odorants do not drive sleep interruption, we hypothesized that unlike other sensory stimuli odorants would not drive KCs. To test these hypotheses we used polysomnography to measure sleep in 34 healthy subjects (19 women, 15 men; mean age 26.5 2.5 yr) who were repeatedly presented with odor stimuli via a computer-controlled air-dilution olfactometer over the course of a single night. Each participant was exposed to one of four odorants, lavender oil (n = 13), vetiver oil (n = 5), vanillin (n = 12), or ammonium sulfide (n = 4), for durations of 5, 10, and 20 s every 9-15 min. Consistent with our hypotheses, we found that odor presentation during sleep enhanced the power of delta (0.5-4 Hz) and slow spindle (9-12 Hz) frequencies during non-rapid eye movement sleep. The increase was proportionate to odor duration. In addition, odor presentation did not modulate the occurrence of KCs. These findings imply a sleep-promoting olfactory mechanism that may deepen sleep through driving increased slow-frequency oscillations.

2015 The American Laryngological, Rhinological and Otological Society, Inc.OBJECTIVES/HYPOTHESIS: Electronic noses can identify diseases, including head and neck squamous cell carcinoma (SCC) by the fingerprint of volatile organic compounds (VOCs) in exhaled air. However, whether these VOCs originated from the malignant lesion itself remains unclear. The objective was to test for the presence and properties of VOCs directly over the vocal folds in malignant and benign lesions, as a potential tool for noninvasive screening.STUDY DESIGN: Prospective observational case control study.METHODS: Samples of mucus directly covering vocal fold lesions were analyzed using gas chromatography mass spectrometry for detection of VOCs, and evaluation of the properties and quantity of VOCs in the samples. Additionally, samples of oropharyngeal mucus were analyzed to exclude VOCs found also in the vicinity of the lesion. Benign and malignant lesion groups were compared using a nonparametric (Mann-Whitney) test.RESULTS: We studied 14 patients, six with SCC and eight with benign pathology. We found an increased number of discrete VOC types in patients with SCC both above the lesion (SCC = 4.333 2.5, benign = 0.875 0.6; Z=3, P

In this study we identify a volatile compound produced by Pseudomonas aeruginosa, which can specifically activate the LuxR quorum-sensing response regulator of Vibrio fischeri. Comparative gas-chromatography analysis between P. aeruginosa wild type and a Delta lasR mutant strain implied that the active volatile is 2-aminoacetophenone. The use of synthetic 2-aminoacetophenone and in silico docking analyses verified the activity of the molecule and provided putative interacting residues within the binding site.

Internal action models (IAMs) are brain templates for sensory-motor coordination underlying diverse behaviors [1]. An emerging theory suggests that impaired IAMs are a common theme in autism spectrum disorder (ASD) [2-4]. However, whether impaired IAMs occur across sensory systems and how they relate to the major phenotype of ASD, namely impaired social communication [5], remains unclear. Olfaction relies on an IAM known as the sniff response, where sniff magnitude is automatically modulated to account for odor valence [6-12]. To test the failed IAM theory in olfaction, we precisely measured the non-verbal non-task-dependent sniff response concurrent with pleasant and unpleasant odors in 36 children-18 with ASD and 18 matched typically developing (TD) controls. We found that whereas TD children generated a typical adult-like sniff response within 305 ms of odor onset, ASD children had a profoundly altered sniff response, sniffing equally regardless of odor valance. This difference persisted despite equal reported odor perception and allowed for 81% correct ASD classification based on the sniff response alone (binomial, p

Each person expresses a potentially unique subset of similar to 400 different olfactory receptor subtypes. Given that the receptors we express partially determine the odors we smell, it follows that each person may have a unique nose; to capture this, we devised a sensitive test of olfactory perception we termed the "olfactory fingerprint." Olfactory fingerprints relied on matrices of perceived odorant similarity derived from descriptors applied to the odorants. We initially fingerprinted 89 individuals using 28 odors and 54 descriptors. We found that each person had a unique olfactory fingerprint (P

[No abstract available]

We hypothesize that true human olfactory abilities are obscured by cortical inhibition. Alcohol reduces inhibition. We therefore tested the hypothesis that olfactory abilities will improve following alcohol consumption. We measured olfaction in 85 subjects, 45 in a between-subjects design, and 40 in a repeated-measures within-subjects design before and after consumption of alcoholic or non-alcoholic beverages. Subjects were also assessed using neurocognitive measures of inhibition. Following alcohol consumption, blood alcohol levels ranged from 0.005% to 0.11%. Across subjects, before any consumption of alcohol, we found that individuals who were less inhibited had lower (better) olfactory detection thresholds (r = 0.68, p

Mammals have 1000 different olfactory receptor subtypes, each responding to a number of different odorants, and each odorant activating a number of different receptor subtypes. These molecular and anatomical underpinnings of olfaction imply a perceptual structure of very high dimensionality that relies on combinatorial coding. In contrast to this expectation, the study of olfactory perception reveals a structure of much lower dimensionality. Moreover, a low-dimensionality approach to olfaction enabled derivation of perception-based structural metrics for smell. These metrics provided meaningful predictions of odorant-induced neural activity and perception from odorant structure alone. Based on this low functional dimensionality, we speculate that olfaction likely does not functionally rely on 1000 different receptor subtypes, and their persistence in evolution may imply that they have additional roles in non-olfactory functions such as in guidance of embryogenesis and development.

Background: Detecting signs of consciousness in patients in a vegetative state/unresponsive wakefulness syndrome (UWS/VS) or minimally conscious state (MCS) is known to be very challenging. Plotkin et al. (2010) recently showed the possibility of using a breathing-controlled communication device in patients with locked in syndrome. We here aim to test a breathing based "sniff controller" that could be used as an alternative diagnostic tool to evaluate response to command in severely brain damaged patients with chronic disorders of consciousness (DOC). Methods: Twenty-five DOG patients were included. Patients' resting breathing-amplitude was measured during a 5 min resting condition. Next, they were instructed to end the presentation of a music sequence by sniffing vigorously. An automated detection of changes in breathing amplitude (i.e., >1.5 SD of resting) ended the music and hence provided positive feedback to the patient. Results: None of the 11 UWS/VS patients showed a sniff based response to command. One out of 14 patients with MCS was able to willfully modulate his breathing pattern to answer the command on 16/19 trials (accuracy 84%). Interestingly, this patient failed to show any other motor response to command. Discussion: We here illustrate the possible interest of using breathing dependent response to command in the detection of residual cognition in patients with DOG after severe brain injury.

Ample evidence suggests that social chemosignaling plays a significant role in human behavior. Processing of odors and chemosignals depends on sniffing. Given this, we hypothesized that humans may have evolved an automatic mechanism driving sniffs in response to conspecific sniffing. To test this, we measured sniffing behavior of human subjects watching the movie Perfume, which contains many olfactory sniffing events. Despite the total absence of odor, observers sniffed when characters in the movie sniffed. Moreover, this effect was most pronounced in scenes where subjects heard the sniff but did not see the sniffed-at object. We liken this response to the orienting towards conspecific gaze in vision and argue that its robustness further highlights the significance of olfactory information processing in human behavior.

Recent findings suggest that novel associations can be learned during sleep. However, whether associative learning during sleep can alter later waking behavior and whether such behavioral changes last for minutes, hours, or days remain unknown. We tested the hypothesis that olfactory aversive conditioning during sleep will alter cigarette-smoking behavior during ensuing wakefulness. A total of 66 human subjects wishing to quit smoking participated in the study (23 females; mean age, 28.7 +/- 5.2 years). Subjects completed a daily smoking diary detailing the number of cigarettes smoked during 7 d before and following a 1 d or night protocol of conditioning between cigarette odor and profoundly unpleasant odors. We observed significant reductions in the number of cigarettes smoked following olfactory aversive conditioning during stage 2 and rapid eye movement (REM) sleep but not following aversive conditioning during wakefulness (p

While effectively doubling the number of known odorant-to-receptor pairings in human olfaction, researchers explain a portion of perceptual variability that stems from genetic variability.

Like all mammals, humans use chemosignals. Nevertheless, only few such chemosignals have been identified. Here we describe an experimental arrangement that casts a wide net for the possible chemosignaling functions of target molecules. This experimental arrangement can be used in concert with various methods for measuring human behavioral and brain responses, including psychophysiology and brain imaging. Moreover, many of the methodological issues we describe are relevant to any study with human chemosignals.

To understand the brain mechanisms of olfaction we must understand the rules that govern the link between odorant structure and odorant perception. Natural odors are in fact mixtures made of many molecules, and there is currently no method to look at the molecular structure of such odorant-mixtures and predict their smell. In three separate experiments, we asked 139 subjects to rate the pairwise perceptual similarity of 64 odorant-mixtures ranging in size from 4 to 43 mono-molecular components. We then tested alternative models to link odorant-mixture structure to odorant-mixture perceptual similarity. Whereas a model that considered each mono-molecular component of a mixture separately provided a poor prediction of mixture similarity, a model that represented the mixture as a single structural vector provided consistent correlations between predicted and actual perceptual similarity (r >= 0.49, p

Animal studies of discriminative fear conditioning traditionally use stimuli that are distant in physical features and thus easily distinguished perceptually. Independently, human studies have shown that training mostly improves discrimination thresholds. We found that aversive learning actually induced an increase in discrimination thresholds in humans and that subjective aversion during conditioning predicted the individual threshold change. This counterintuitive performance deterioration occurred when using odors or sounds as aversive reinforcers and was not a result of attentional distraction or decision bias. In contrast, positive reinforcement or mere exposure induced the typically reported decrease in thresholds. Our findings indicate that aversive outcomes induce wider stimulus generalization by modulating perceptual thresholds, suggesting the engagement of low-level mechanisms. We suggest that for risk- or loss-related stimuli, less specificity could be a benefit, as it invokes the same mechanisms that respond quickly and efficiently in the face of danger.

During sleep, humans can strengthen previously acquired memories, but whether they can acquire entirely new information remains unknown. The nonverbal nature of the olfactory sniff response, in which pleasant odors drive stronger sniffs and unpleasant odors drive weaker sniffs, allowed us to test learning in humans during sleep. Using partial-reinforcement trace conditioning, we paired pleasant and unpleasant odors with different tones during sleep and then measured the sniff response to tones alone during the same nights' sleep and during ensuing wake. We found that sleeping subjects learned novel associations between tones and odors such that they then sniffed in response to tones alone. Moreover, these newly learned tone-induced sniffs differed according to the odor pleasantness that was previously associated with the tone during sleep. This acquired behavior persisted throughout the night and into ensuing wake, without later awareness of the learning process. Thus, humans learned new information during sleep.

In vision, two mixtures, each containing an independent set of many different wavelengths, may produce a common color percept termed "white." In audition, two mixtures, each containing an independent set of many different frequencies, may produce a common perceptual hum termed "white noise." Visual and auditory whites emerge upon two conditions: when the mixture components span stimulus space, and when they are of equal intensity. We hypothesized that if we apply these same conditions to odorant mixtures, "whiteness" may emerge in olfaction as well. We selected 86 molecules that span olfactory stimulus space and individually diluted them to a point of about equal intensity. We then prepared various odorant mixtures, each containing various numbers of molecular components, and asked human participants to rate the perceptual similarity of such mixture pairs. We found that as we increased the number of nonoverlapping, equal-intensity components in odorant mixtures, the mixtures became more similar to each other, despite not having a single component in common. With similar to 30 components, most mixtures smelled alike. After participants were acquainted with a novel, arbitrarily named mixture of similar to 30 equal-intensity components, they later applied this name more readily to other novel mixtures of similar to 30 equal-intensity components spanning stimulus space, but not to mixtures containing fewer components or to mixtures that did not span stimulus space. We conclude that a common olfactory percept, "olfactory white," is associated with mixtures of similar to 30 or more equal-intensity components that span stimulus space, implying that olfactory representations are of features of molecules rather than of molecular identity.

Emotional tearing is a poorly understood behavior that is considered uniquely human. In mice, tears serve as a chemosignal. We therefore hypothesized that human tears may similarly serve a chemosignaling function. We found that merely sniffing negative-emotion-related odorless tears obtained from women donors induced reductions in sexual appeal attributed by men to pictures of women's faces. Moreover, after sniffing such tears, men experienced reduced self-rated sexual arousal, reduced physiological measures of arousal, and reduced levels of testosterone. Finally, functional magnetic resonance imaging revealed that sniffing women's tears selectively reduced activity in brain substrates of sexual arousal in men.

Organization of receptive surfaces reflects primary axes of perception. In vision, retinal coordinates reflect spatial coordinates. In audition, cochlear coordinates reflect tonal coordinates. However, the rules underlying the organization of the olfactory receptive surface are unknown. To test the hypothesis that organization of the olfactory epithelium reflects olfactory perception, we inserted an electrode into the human olfactory epithelium to directly measure odorant-induced evoked responses. We found that pairwise differences in odorant pleasantness predicted pairwise differences in response magnitude; that is, a location that responded maximally to a pleasant odorant was likely to respond strongly to other pleasant odorants, and a location that responded maximally to an unpleasant odorant was likely to respond strongly to other unpleasant odorants. Moreover, the extent of an individual's perceptual span predicted their span in evoked response. This suggests that, similarly to receptor surfaces for vision and audition, organization of the olfactory receptor surface reflects key axes of perception.

The mammalian brain commonly uses structural proximity to reflect proximity in stimulus and perceptual space. Objects or object features that are near each other in physical structure or perception are also near each other in the brain. This generates sensory maps. The topography of olfactory connectivity implies a rudimentary map in the olfactory epithelium, a more intricate map in the olfactory bulb, but no ordered topography is evident in piriform cortex. Currently, we are largely unable to link the ordered topography in epithelium and bulb to meaningful olfactory axes within a strong predictive framework. We argue that the path to uncovering such a predictive framework depends on systematically characterizing olfactory perception, and we describe initial efforts in this direction.

Animal studies of discriminative fear conditioning traditionally use stimuli that are distant in physical features and thus easily distinguished perceptually. Independently, human studies have shown that training mostly improves discrimination thresholds. We found that aversive learning actually induced an increase in discrimination thresholds in humans and that subjective aversion during conditioning predicted the individual threshold change. This counterintuitive performance deterioration occurred when using odors or sounds as aversive reinforcers and was not a result of attentional distraction or decision bias. In contrast, positive reinforcement or mere exposure induced the typically reported decrease in thresholds. Our findings indicate that aversive outcomes induce wider stimulus generalization by modulating perceptual thresholds, suggesting the engagement of low-level mechanisms. We suggest that for risk- or loss-related stimuli, less specificity could be a benefit, as it invokes the same mechanisms that respond quickly and efficiently in the face of danger.

Paradoxically, improvements in emergency medicine have increased survival albeit with severe disability ranging from quadriplegia to "locked-in syndrome." Locked-in syndrome is characterized by intact cognition yet complete paralysis, and hence these individuals are "locked-in" their own body, at best able to communicate using eye blinks alone. Sniffing is a precise sensory-motor acquisition entailing changes in nasal pressure. The fine control of sniffing depends on positioning the soft palate, which is innervated by multiple cranial nerves. This innervation pattern led us to hypothesize that sniffing may remain conserved following severe injury. To test this, we developed a device that measures nasal pressure and converts it into electrical signals. The device enabled sniffs to control an actuator with speed similar to that of a hand using a mouse or joystick. Functional magnetic resonance imaging of device usage revealed a widely distributed neural network, allowing for increased conservation following injury. Also, device usage shared neural substrates with language production, rendering sniffs a promising bypass mode of communication. Indeed, sniffing allowed completely paralyzed locked-in participants to write text and quadriplegic participants to write text and drive an electric wheelchair. We conclude that redirection of sniff motor programs toward alternative functions allows sniffing to provide a control interface that is fast, accurate, robust, and highly conserved following severe injury.

By demonstrating that inactivation of gustatory cortex influences olfactory recognition, a new study finds that the interaction between taste and smell is bidirectional.

Paradoxically, although humans have a superb sense of smell, they don't trust their nose. Furthermore, although human odorant detection thresholds are very low, only unusually high odorant concentrations spontaneously shift our attention to olfaction. Here we suggest that this lack of olfactory awareness reflects the nature of olfactory attention that is shaped by the spatial and temporal envelopes of olfaction. Regarding the spatial envelope, selective attention is allocated in space. Humans direct an attentional spotlight within spatial coordinates in both vision and audition. Human olfactory spatial abilities are minimal. Thus, with no olfactory space, there is no arena for olfactory selective attention. Regarding the temporal envelope, whereas vision and audition consist of nearly continuous input, olfactory input is discreet, made of sniffs widely separated in time. If similar temporal breaks are artificially introduced to vision and audition, they induce "change blindness", a loss of attentional capture that results in a lack of awareness to change. Whereas "change blindness" is an aberration of vision and audition, the long inter-sniff-interval renders "change anosmia" the norm in human olfaction. Therefore, attentional capture in olfaction is minimal, as is human olfactory awareness. All this, however, does not diminish the role of olfaction through sub-attentive mechanisms allowing subliminal smells a profound influence on human behavior and perception.

To assess the feasibility of using odors as a potential mechanism for treating sleep apnea, we set out to test the hypothesis that odorants delivered during sleep would modify respiratory patterns without inducing arousal or wake in healthy sleepers. We used 2 mildly trigeminal odorants: the pleasant lavender and unpleasant vetiver oil and 2 pure olfactory odorants: the pleasant vanillin and unpleasant ammonium sulfide. During sleep, an olfactometer delivered a transient odorant every 9,12, or 15 min (randomized), providing 21-37 odorant presentations per night. Each of 36 participants was studied for 1 night and with 1 of the 4 different odorants tested. In addition to standard overnight polysomnography, we employed highly accurate measurements of nasal and oral respiration. Odorants did not increase the frequency of arousals or wake but did influence respiration. Specifically, all 4 odorants transiently decreased inhalation and increased exhalation for up to 6 breaths following odor onset. This effect persisted regardless of odorant valence or stage of sleep. These results suggest that the olfactory system may provide a path to manipulate respiration in sleep.

Olfaction is often referred to as a multidimensional sense. It is multidimensional in that similar to 1000 different receptor types, each tuned to particular odor aspects, together contribute to the olfactory percept. In humans, however, this percept is nearly unidimensional. Humans can detect and discriminate countless odorants, but can identify few by name. The one thing humans can and do invariably say about an odor is whether it is pleasant or not. We argue that this hedonic determination is the key function of olfaction. Thus, the boundaries of an odor object are determined by its pleasantness, which-unlike something material and more like an emotion-remains poorly delineated with words.

A primary goal for artificial nose (eNose) technology is to report perceptual qualities of novel odors. Currently, however, eNoses primarily detect and discriminate between odorants they previously "learned''. We tuned an eNose to human odor pleasantness estimates. We then used the eNose to predict the pleasantness of novel odorants, and tested these predictions in naive subjects who had not participated in the tuning procedure. We found that our apparatus generated odorant pleasantness ratings with above 80% similarity to average human ratings, and with above 90% accuracy at discriminating between categorically pleasant or unpleasant odorants. Similar results were obtained in two cultures, native Israeli and native Ethiopian, without retuning of the apparatus. These findings suggest that unlike in vision and audition, in olfaction there is a systematic predictable link between stimulus structure and stimulus pleasantness. This goes in contrast to the popular notion that odorant pleasantness is completely subjective, and may provide a new method for odor screening and environmental monitoring, as well as a critical building block for digital transmission of smell.

Odor identity is coded in spatiotemporal patterns of neural activity in the olfactory bulb. Here we asked whether meaningful olfactory information could also be read from the global olfactory neural population response. We applied standard statistical methods of dimensionality-reduction to neural activity from 12 previously published studies using seven different species. Four studies reported olfactory receptor activity, seven reported glomerulus activity, and one reported the activity of projection-neurons. We found two linear axes of neural population activity that accounted for more than half of the variance in neural response across species. The first axis was correlated with the total sum of odor-induced neural activity, and reflected the behavior of approach or withdrawal in animals, and odorant pleasantness in humans. The second and orthogonal axis reflected odorant toxicity across species. We conclude that in parallel with spatiotemporal pattern coding, the olfactory system can use simple global computations to read vital olfactory information from the neural population response.

Authors [1], poets [2], and scientists [3-6] have been fascinated by the strength of childhood olfactory memories. Indeed, in long-term memory, the first odor-to-object association was stronger than subsequent associations of the same odor with other objects [7]. Here we tested the hypothesis that first odor associations enjoy a privileged brain representation. Because emotion impacts memory [8-10], we further asked whether the pleasantness of an odor would influence such a representation. On day 1, we associated the same visual objects initially with one, and subsequently with a second, set of pleasant and unpleasant olfactory and auditory stimuli. One week later, we presented the same visual objects and tested odor-associative memory concurrent with functional magnetic resonance brain imaging. We found that the power (% remembered) of early associations was enhanced when they were unpleasant, regardless of whether they were olfactory or auditory. Brain imaging, however, revealed a unique hippocampal activation for early olfactory but not auditory associations, regardless of whether they were pleasant or unpleasant. Activity within the hippocampus on day 1 predicted the olfactory but not auditory associations that would be remembered one week later. These findings confirmed the hypothesis of a privileged brain representation for first olfactory associations.

LElectroolfactograms (EOGs) are the summated generator potentials of olfactory receptor neurons measured directly from the olfactory epithelium. To validate the sensory origin of the human EOG, we set out to ask whether EOGs measured in humans were odorant concentration dependent. Each of 22 subjects (12 women, mean age = 23.3 yr) was tested with two odorants, either valeric acid and linalool (n = 12) or isovaleric acid and l-carvone (n = 10), each delivered at four concentrations diluted with warm (37°C) and humidified (80%) odorless air. In behavior, increased odorant concentration was associated with increased perceived intensity (all F > 5, all P < 0.001). In EOG, increased odorant concentration was associated with increased area under the EOG curve (all F > 8, all P < 0.001). These findings substantiate EOG as a tool for probing olfactory coding directly at the level of olfactory receptor neurons in humans.

Olfactory information reaches olfactory cortex without a thalamic relay. This neuroanatomical substrate has combined with functional findings to suggest that, in olfaction, the typical thalamic role in sensory processing has shifted to the olfactory bulb or olfactory cortex. With this in mind, we set out to ask whether the thalamus at all plays a significant functional role in human olfaction. We tested olfactory function in 17 patients with unilateral focal thalamic lesions and in age-matched healthy controls. We found that thalamic lesions did not significantly influence olfactory detection but significantly impaired olfactory identification, and only right lesions altered olfactory hedonics by reducing the pleasantness of pleasant odors. An auditory control revealed that this shift in pleasantness was olfactory specific. These olfactory impairments were evident in explicit measures of perception, as well as in patterns of sniffing. Whereas healthy subjects modulated their sniffs in accordance with odorant content, thalamic patients did not. We conclude that, although the thalamus is not in the path of olfactory information from periphery to cortex, it nevertheless plays a significant functional role in human olfaction.

Background: The neural substrates of olfactory working memory are unknown. We addressed the questions of whether olfactory working memory involves a verbal representation of the odor, or a sensory image of the odor, or both, and the location of the neural substrates of these processes. Methodology/Principal Findings: We used functional magnetic resonance imaging to measure activity in the brains of subjects who were remembering either nameable or unnameable odorants. We found a double dissociation whereby remembering nameable odorants was reflected in sustained activity in prefrontal language areas, and remembering unnameable odorants was reflected in sustained activity in primary olfactory cortex. Conclusions/Significance: These findings suggest a novel dedicated mechanism in primary olfactory cortex, where odor information is maintained in temporary storage to subserve ongoing tasks.

In studies of vision and audition, stimuli can be systematically varied by wavelength and frequency, respectively, but there is no equivalent metric for olfaction. Restricted odorant-feature metrics such as number of carbons and functional group do not account for response patterns to odorants varying along other structural dimensions. We generated a multidimensional odor metric, in which each odorant molecule was represented as a vector of 1,664 molecular descriptor values. Revisiting many studies, we found that this metric and a second optimized metric were always better at accounting for neural responses than the specific metric used in each study. These metrics were applicable across studies that differed in the animals studied, the type of olfactory neurons tested, the odorants applied and the recording methods used. We use this new metric to recommend sets of odorants that span the physicochemical space for use in olfaction experiments.

Although the family of genes encoding for olfactory receptors was identified more than 15 years ago, the difficulty of functionally expressing these receptors in an heterologous system has, with only some exceptions, rendered the receptive range of given olfactory receptors largely unknown. Furthermore, even when successfully expressed, the task of probing such a receptor with thousands of odors/ligands remains daunting. Here we provide proof of concept for a solution to this problem. Using computational methods, we tune an electronic nose to the receptive range of an olfactory receptor. We then use this electronic nose to predict the receptors' response to other odorants. Our method can be used to identify the receptive range of olfactory receptors, and can also be applied to other questions involving receptor-ligand interactions in non-olfactory settings.

Whereas the rules underlying the perceived intensity of binary mixtures have been investigated, minimal efforts have been directed at elucidating the rules underlying the perceived pleasantness of such mixtures. To address this, 84 subjects ranked the pleasantness and intensity of 5 distinct binary mixtures (15 pairs, inter-stimulus interval = 4 s, inter-trial interval = 30 s, flow = 6 l/min, pulse = 2 s) constructed from different ratios (0:100%, 25:75%, 50:50%, 75:25%, and 100:0%, olfactometer-generated vapor phase). We found that in the majority of cases, the pleasantness of the mixture fell between the pleasantness values of its separated constituents and that it was strongly influenced by the relative intensities of the constituents. Based on these results, we proposed a prediction paradigm for the pleasantness of binary mixtures from the pleasantness of their separated constituents weighted by their respective perceived intensities. The uniqueness of the proposed paradigm is that it neither requires presetting an interaction constant between the mixture components nor require any factorization of the pleasantness weights. It does, nonetheless, require solid psychophysical data on the separated components at their different concentrations, and currently it can only explain the behavior of intermediate pleasantness of mixtures.

Whether olfactory working memory involves verbal representations or neural images of odor per se remains unclear. This study investigated whether verbal representation influences performance in an olfactory delayed-match-to-sample task and used monorhinal presentation to generate hypotheses as to the underlying anatomy of this mechanism. The main findings were that (a) nameable odorants were easier to remember than hard-to-name odorants and (b) the nameability effect was more pronounced when the evaluation was done across nostrils. Considering these results within a proposed model implies dual representation in olfactory working memory: All odors, nameable and hard to name, are represented both perceptually and verbally.

Olfaction consists of a set of transforms from a physical space of odorant molecules, through a neural space of information processing, and into a perceptual space of smell. Elucidating the rules governing these transforms depends on establishing valid metrics for each of the three spaces. Here we first briefly review the perceptual and neural spaces, and then concentrate on the physical space of odorant molecules. We argue that the lack of an agreed-upon odor metric poses a significant obstacle toward understanding the neurobiology of olfaction, and suggest two alternative odor metrics as possible solutions.

Whether mammalian scent-tracking is aided by inter-nostril comparisons is unknown. We assessed this in humans and found that (i) humans can scent-track, (ii) they improve with practice, (iii) the human nostrils sample spatially distinct regions separated by similar to 3.5 cm and, critically, (iv) scent-tracking is aided by inter-nostril comparisons. These findings reveal fundamental mechanisms of scent-tracking and suggest that the poor reputation of human olfaction may reflect, in part, behavioral demands rather than ultimate abilities.

A recent paper published by Kimchi, Xu, and Dulac in Nature describes the emergence of male-type sexual behavior in female mice following incapacitation of the accessory olfactory system. The authors argue that this implies a default male-type behavioral pattern that is otherwise constantly inhibited in the female brain by chemical signals transduced in the accessory olfactory system. In addition to reviewing these findings, we suggest in this Preview how these findings in the mouse could have relevance for human behavior.

We provide detailed instructions and part selections for construction of a five-channel air dilution olfactometer capable of generating neat odorants and binary mixtures at a range of known concentrations. At the heart of the olfactometer is an odorant canister that is (1) cheap and readily available, (2) safe and durable, (3) has minimal odor adherence, (4) is easily incorporated into any olfactometer, and critically (5) produces a highly consistent stimulus. By flowing a given carrier gas at a given flowrate through a given odorant in this canister, the same end-vapor is achieved. Flow/concentration outcomes are provided for several odorants routinely used in olfactometry. This toot will enable researchers to generate known concentrations without expensive analytical machinery. 

Irritation and negative valence are closely associated in perception. However, these perceptual aspects can be dissociated in olfaction where irritation can accompany both pleasant and unpleasant odorants. Whereas the sensation of odor reflects transduction at olfactory receptors, irritation reflects concurrent transduction of the odorant at trigeminal receptors. Thus a stimulus can be either a pure olfactant activating the olfactory receptors only or a bimodal odorant activating both types of receptors. Using event-related functional magnetic resonance imaging and a 2 x 2 experimental design contrasting odorant valence ( pleasant/unpleasant) and odorant type ( pure olfactant/bimodal) we found activity in piriform cortex to be associated with valence, and not type, of odors. In contrast, activity in the olfactory tubercle was associated with type, and not valence, of odors. Importantly, this was found when perceived intensity was held equal across odorants. These findings suggest that dissociable neural substrates subserve the encoding of irritation and valence in olfaction.

Rodents use chemosignals to alter endocrine balance in conspecifics. Although responses to human sweat suggest a similar mechanism in humans, no particular component of human sweat capable of altering endocrine balance in conspecifics has yet been isolated and identified. Here, we measured salivary levels of the hormone cortisol in women after smelling pure androstadienone (4,16-androstadien-3-one), a molecule present in the sweat of men that has been suggested as a chemosignal in humans. We found that merely smelling androstadienone maintained significantly higher levels of the hormone cortisol in women. These results suggest that, like rodents, humans can influence the hormonal balance of conspecifics through chemosignals. Critically, this study identified a single component of sweat, androstadienone, as capable of exerting such influence. This result points to a potential role for synthetic human chemosignals in clinical applications.

Although it is agreed that physicochemical features of molecules determine their perceived odor, the rules governing this relationship remain unknown. A significant obstacle to such understanding is the high dimensionality of features describing both percepts and molecules. We applied a statistical method to reduce dimensionality in both odor percepts and physicochemical descriptors for a large set of molecules. We found that the primary axis of perception was odor pleasantness, and critically, that the primary axis of physicochemical properties reflected the primary axis of olfactory perception. This allowed us to predict the pleasantness of novel molecules by their physicochemical properties alone. Olfactory perception is strongly shaped by experience and learning. However, our findings suggest that olfactory pleasantness is also partially innate, corresponding to a natural axis of maximal discriminability among biologically relevant molecules.

Hedonic-specific activity in piriform cortex during odor imagery mimics that during odor perception. J Neurophysiol 98: 3254-3262, 2007. First published October 3, 2007; doi:10.1152/jn.00349.2007. Although it is known that visual imagery is accompanied by activity in visual cortical areas, including primary visual cortex, whether olfactory imagery exists remains controversial. Here we asked whether cue-dependent olfactory imagery was similarly accompanied by activity in olfactory cortex, and in particular whether hedonic-specific patterns of activity evident in olfactory perception would also be present during olfactory imagery. We used functional magnetic resonance imaging to measure activity in subjects who alternated between smelling and imagining pleasant and unpleasant odors. Activity induced by imagining odors mimicked that induced by perceiving real odorants, not only in the particular brain regions activated, but also in its hedonic-specific pattern. For both real and imagined odors, unpleasant stimuli induced greater activity than pleasant stimuli in the left frontal portion of piriform cortex and left insula. These findings combine with findings from other modalities to suggest activation of primary sensory cortical structures during mental imagery of sensory events.

There is a growing appreciation for the role of sniffing in the formation of the olfactory percept. With this in mind, monitoring and measurement of sniffing is an important aspect of olfactory experiments. There are several methods for measuring human sniffs concurrent with odor delivery in olfactory experiments. Here, we set out to compare the temporal sensitivity and power of these different methods by applying them all simultaneously with an olfactory task. We discuss the advantages and disadvantages of each method and conclude in recommending the use of a nasal cannula linked to a pressure sensor whenever possible.

In this review, we use data obtained primarily from humans to argue that sniffs are not merely a stimulus carrier but are rather a central component of the olfactory percept. We argue that sniffs 1) are necessary for the olfactory percept, 2) affect odorant intensity perception and identity perception, 3) drive activity in olfactory cortex, 4) are rapidly modulated in an odorant-dependent fashion by a dedicated olfactomotor system, and 5) are sufficient to generate an olfactory percept of some sort even in the absence of odor.

Forty years ago, von Bekesy demonstrated that the spatial source of an odorant is determined by comparing input across nostrils, but it is unknown how this comparison is effected in the brain. To address this, we delivered odorants to the left or right of the nose, and contrasted olfactory left versus right localization with olfactory identification during brain imaging. We found nostril-specific responses in primary olfactory cortex that were predictive of the accuracy of left versus right localization, thus providing a neural substrate for the behavior described by von Bdkdsy. Additionally, left versus right localization preferentially engaged a portion of the superior temporal gyrus previously implicated in visual and auditory localization, suggesting that localization information extracted from smell was then processed in a convergent brain system for spatial representation of multisensory inputs.

Central to the concept of attention is the fact that identical stimuli can be processed in different ways. In olfaction, attention may designate the identical flow of air through the nose as either respiration or olfactory exploration. Here we have used functional magnetic resonance imaging (fMRI) to probe this attentional mechanism in primary olfactory cortex (POC). We report a dissociation in POC that revealed attention-dependent and attention-independent subregions. Whereas a temporal subregion comprising temporal piriform cortex (PirT) responded equally across conditions, a frontal subregion comprising frontal piriform cortex (PirF) and the olfactory tubercle responded preferentially to attended sniffs as opposed to unattended sniffs. In addition, a task-specific anticipatory response occurred in the attention-dependent region only. This dissociation was consistent across two experimental designs: one focusing on sniffs of clean air, the other focusing on odor-laden sniffs. Our findings highlight the role of attention at the earliest cortical levels of olfactory processing.

The past 15 years have seen significant advances in the study of olfaction, with particular emphasis on elucidating the molecular building blocks of the sensory process. However, much of the systems-level organization of olfaction remains unexplored. Here, we provide an overview at this level, highlighting results obtained from studying humans, whom we think provide an underutilized, yet critical, animal model for olfaction.

Functional imaging studies of olfaction have consistently reported odorant-induced activation of the cerebellum. However, the cerebellar role in olfaction remains unknown. We examined the olfactory and olfactomotor abilities of patients with unilateral cerebellar lesions, comparing performance within subjects across nostrils, as well as between subjects with age-matched and young controls. Regarding olfactory performance, initial testing revealed that patients had a contralesional impairment in olfactory identification but not olfactory detection threshold. However, when tested under conditions that prevented compensatory sniffing strategies, the patients also exhibited a contralesional olfactory detection impairment. Regarding olfactomotor function, a healthy olfactomotor system generates sniffs that are (1) sufficiently vigorous and (2) inversely proportional to odorant concentration in sniff mean airflow velocity, maximum airflow velocity, volume, and duration. Patients' sniffs were lower in overall airflow velocity and volume in comparison with control participants. Furthermore, reduced sniff velocity predicted poorer detection thresholds in patients. Finally, whereas young controls used concentration-dependent sniffs, there was a trend in that direction only for age-matched controls. Patients used sniffs that were concentration invariant. In conclusion, cerebellar lesions impacted olfactory and olfactomotor performance. These findings strongly implicate an olfactocerebellar pathway prominent in odor identification and detection that functionally connects each nostril primarily to the contralateral cerebellum.

There are large individual differences in the self-reported ability to form vivid olfactory mental imagery. Based on such self-reports, subjects have been classified as 'bad' or 'good' imagers. The present study examined whether a differential strategy in re-enacting the olfactomotor response during imagery may explain the dissociation between 'bad' and 'good' olfactory imagers. As previously reported, odor imagery was accompanied by sniffing. Although 'bad' and 'good' olfactory imagers did not differ in their overall sniffing volume, they used different strategies when re-enacting the motor component of olfaction during imagery. Particularly, as in real perception, 'good' but not 'bad' imagers generated bigger sniffs when imagining a pleasant smell compared with an unpleasant smell (P

The effects of sniffing different concentrations of the human sex-steroid derived compound 4,16-androstadien-3-one (AND) on autonomic nervous system function and mood were measured in 60 subjects. The effects were sex-specific and concentration-dependent. Only high concentrations of AND (0.00625 M) increased positive mood (p

Olfaction is typically described as behaviorally slow, suggesting neural processes on the order of hundreds of milliseconds to seconds as candidate mechanisms in the creation of olfactory percepts. Whereas a recent study challenged this view in suggesting that a single sniff was sufficient for optimal olfactory discrimination, a study by Abraham et al. in this issue of Neuron sets out to negate the challenge by demonstrating increased processing time for discrimination of similar versus dissimilar stimuli. Here we reconcile both studies, which in our view together support the notion of a speed-accuracy tradeoff in olfactory discriminations that are made within about 200 ms. These findings are discussed in light of the challenges related to defining olfactory perceptual similarity in nonhuman animals.

We asked whether the effects of exposure to two human sex-steroid derived compounds were context dependent. The effects of sniffing 4,16-androstadien-3-one (AND) and 1,3,5(10),16-estratetraen-3-ol (EST) on mood, memory, and autonomic nervous system responses were explored in 72 participants. Subjects were tested with AND, EST, or a Control compound within four mood contexts: neutral, sexually aroused, sad and happy. These moods were successfully induced using selected film segments (P

It has been estimated that ∼30% of the population is unable to detect the odor of androstenone. These estimates, however, were made using tests and criteria optimized for identifying detection. Such criteria favor Type II over Type I errors—that is, they are excellent at identifying true detectors at the cost of erroneously labeling some detectors as non-detectors. Because these criteria were used to identify non-detectors, it is possible that the rate of non-detection may have been overestimated. To test this we screened 55 subjects for non-detection employing previously used methods. This screen yielded nine putative non-detectors, a 16.3% putative non-detection rate. We then retested these putative non-detectors using a forced choice (yes–no) paradigm to obtain a precise measure of their sensitivity. We found that this group of putative non-detectors was significantly above chance at detecting androstenone (P < 0.001), despite very low self-confidence in their performance. Based on the results of the signal detection analysis in this sample, we estimate the rate of actual androstenone non-detection in young healthy adults is between 1.8 and 5.96%, which is significantly lower than previously estimated. This finding is significant considering the implications of specific anosmias on the understanding of odor discrimination.

Affective experience has been described in terms of two primary dimensions: intensity and valence. In the human brain, it is intrinsically difficult to dissociate the neural coding of these affective dimensions for visual and auditory stimuli, but such dissociation is more readily achieved in olfaction, where intensity and valence can be manipulated independently. Using event-related functional magnetic resonance imaging (fMRI), we found amygdala activation to be associated with intensity, and not valence, of odors. Activity in regions of orbitofrontal cortex, in contrast, were associated with valence independent of intensity. These findings show that distinct olfactory regions subserve the analysis of the degree and quality of olfactory stimulation, suggesting that the affective representations of intensity and valence draw upon dissociable neural substrates.

Neural representations created in the absence of external sensory stimuli are referred to as imagery(1), and such representations may be augmented by reenactment of sensorimotor processes(2). We measured nasal airflow in human subjects while they imagined sights, sounds and smells, and only during olfactory imagery did subjects spontaneously enact the motor component of olfaction-that is, they sniffed. Moreover, as in perception(3,4), imagery of pleasant odors involved larger sniffs than imagery of unpleasant odors, suggesting that the act of sniffing has a functional role in creating of olfactory percepts.

Sniffs are modulated in response to odor content. Higher concentrations of odor induce lesser-volume sniffs. This phenomenon implicates a neural feedback mechanism that measures sensory input (odor concentration) and modulates motor output (sniffing) accordingly. Here we used air-dilution olfactometry to probe the time course of this olfactomotor mechanism. A stainless-steel computer-controlled olfactometer, equipped with mass flow controllers, temperature and humidity control, and on-line photo-ionization detection, was coupled to a highly sensitive pneumatotachograph that measured nasal flow. The olfactometer was used to generate four ascending concentrations of the odorants propionic acid and phenethyl alcohol. Sniff volume was inversely related to odor concentration (P

The physiological and psychological effects of 2 human sex-steroid derived compounds, 4,16-androstadien-3-one (AND) and 1,3,5(10),16-estratetraen-3-ol (EST) were measured in 24 subjects who participated in a within-subjects, double-blind experiment. A dissociation was evident in the physiological effects of AND, in that it increased physiological arousal in women but decreased it in men. EST did not significantly affect physiological arousal in women or men. Neither compound significantly affected mood. AND is an androgen derivative that is the most prevalent androstene in human male sweat, male axillary hair, and on the male axillary skin surface. The authors argue that AND's opposite effects on physiology in men and women further implicate this compound in chemical communication between humans.

In this issue of Neuron, Small and colleagues used fMRI to find evidence for a neural segregation of two dimensions underlying human gustatory experience: intensity and valence. These results join several recent reports that challenge long-held notions regarding amygdaloid representation of negatively valenced events.

About 30% of the adult human population does not perceive an odour when sniffing the steroid androstenone (5-α-androst-16-en-3-one), but will become sensitive to its smell after repeated exposure to the compound1,2,3. Here we investigate the origin of the plasticity that governs this acquired ability by repeatedly exposing one nostril of non-detecting subjects to androstenone and then testing the unexposed nostril. We find that the exposed nostril and the naive nostril can both learn to recognize the smell, effectively doubling detection accuracy. As the two olfactory epithelia are not connected at the peripheral level, our results indicate that learning occurs in the brain by a mechanism that shares information from both nostrils.

Using PET, Savic et al., in this issue of Neuron, found a sexually dimorphic neural response to two putative human pheromones. The specific regions activated combined with the pronounced sex difference depict a pheromonal-type brain response in humans. Here, we preview this finding and suggest that human pheromones exist.

Although the presence of an olfactory impairment in Parkinson's disease (PD) has been recognized for 25 years, its cause remains unclear. Here we suggest a contributing factor to this impairment, namely, that PD impairs active sniffing of odorants. We tested 10 men and 10 women with clinically typical PD, and 20 age- and gender-matched healthy controls, in four olfactory tasks: (i) the University of Pennsylvania smell identification test; (ii and iii) detection threshold tests for the odorants vanillin and propionic acid; and (iv) a two-alternative forced-choice detection paradigm during which sniff parameters (airflow peak rate, mean rate, volume, and duration) were recorded with a pneomatotachograph-coupled spirometer. An additional experiment tested the effect of intentionally increasing sniff vigor on olfactory performance in 20 additional patients. PD patients were significantly impaired in olfactory identification (P < 0.0001) and detection (P < 0.007). As predicted, PD patients were also significantly impaired at sniffing, demonstrating significantly reduced sniff airflow rate (P < 0.01) and volume (P < 0.002). Furthermore, a patient's ability to sniff predicted his or her performance on olfactory tasks, i.e., the more poorly patients sniffed, the worse their performance on olfaction tests (P < 0.009). Finally, increasing sniff vigor improved olfactory performance in those patients whose baseline performance had been poorest (P < 0.05). These findings implicate a sniffing impairment as a component of the olfactory impairment in PD and further depict sniffing as an important component of human olfaction.

Paradoxically, attempts to visualize odorant-induced functional magnetic resonance imaging (fMRI) activation in the human have yielded activations in secondary olfactory regions but not in the primary olfactory cortex-piriform cortex. We show that odorant-induced activation in primary olfactory cortex was not previously made evident with fMRI because of the unique time course of activity in this region: in primary olfactory cortex, odorants induced a strong early transient increase in signal amplitude that then habituated within 30-40 s of odorant presence. This time course of activation seen here in the primary olfactory cortex of the human is almost identical to that recorded electrophysiologically in the piriform cortex of the rat. Mapping activation with analyses that are sensitive to both this transient increase in signal amplitude, and temporal-variance, enabled us to use fMRI to consistently visualize odorant-induced activation in the human primary olfactory cortex. The combination of continued accurate odorant detection at the behavioral level despite primary olfactory cortex habituation at the physiological level suggests that the functional neuroanatomy of the olfactory response may change throughout prolonged olfactory stimulation.

Air flow-rate is usually higher in one nostril in comparison to the other. Also, within bounds, higher nasal flow-rate improves odorant detection. It follows from the above that odorant detection should be better in the nostril with higher flow-rate in comparison to the nostril with lower flow-rate. Paradoxically, previous research has shown that odorant detection thresholds are equal for the high and low flow-rate nostrils. Here we resolve this apparent paradox by showing that when detecting through the nostril with lower air flow-rate, humans sniffed longer than when detecting through the nostril with higher air flow-rate, thus equalizing performance between the nostrils. When this compensatory mechanism was blocked, a pronounced advantage in odorant detection was seen for the nostril with higher air flow-rate over the nostril with lower air flow-rate. Finally, we show that normal birhinal sniff duration may enable only one nostril to reach optimal threshold. This finding implies that during each sniff, each nostril conveys to the brain a slightly different image of the olfactory world. It remains to be shown how the brain combines these images into a single olfactory percept.

The flow of air is greater into one nostril than into the other because there is a slight turbinate swelling in one1,2,3. The nostril that takes in more air switches from the left to the right one and back again every few hours4,5,6, but the effect of this switching on the sense of smell has been unclear7,8. Here we show that this difference in airflow between the nostrils causes each nostril to be optimally sensitized to different odorants, so that each nostril conveys a slightly different olfactory image to the brain.

EEG and behavioural evidence suggests that air-borne chemicals can affect the nervous system without being consciously detected. EEG and behaviour, however, do not specify which brain structures are involved in chemical sensing that occurs below a threshold of conscious detection, Here we used functional MRI to localize brain activation induced by high and low concentrations of the air-borne compound oestra-1,3,5(10),16-tetraen-3yl acetate, Following presentations of both concentrations, eight of eight subjects reported verbally that they could not detect any odour (P = 0.004), Forced choice detection performed during the presentations revealed above-chance detection of the high concentration, but no better than chance detection of the low concentration compound. Both concentrations induced significant brain activation, primarily in the anterior medial thalamus and inferior frontal gyrus, Activation in the inferior frontal gyrus during the high concentration condition was significantly greater in the right than in the left hemisphere (P = 0.03), A trend towards greater thalamic activation was observed for the high concentration than the low concentration compound (P = 0.08). These findings localize human brain activation that was induced by an undetectable air-borne chemical (the low concentration compound).

The sensation and perception of smell (olfaction) are largely dependent on snifting, which is an active stage of stimulus transport and therefore an integral component of mammalian olfaction(1,2). Electrophysiological data obtained from study of the hedgehog, rat, rabbit, dog and monkey indicate that sniffing (whether or not an odorant is present) induces an oscillation of activity in the olfactory bulb, driving the piriform cortex in the temporal lobe, in other words, the piriform is driven by the olfactory bulb at the frequency of sniffing(1-4). Here we use functional magnetic resonance imaging (fMRI) that is dependent on the level of oxygen in the blood to determine whether sniffing can induce activation in the piriform of humans, and whether this activation can be differentiated from activation induced by an odorant, We find that sniffing, whether odorant is present or absent, induces activation primarily in the piriform cortex of the temporal lobe and in the medial and posterior orbito-frontal gyri of the frontal lobe. The source of the sniff-induced activation is the somatosensory stimulation that is induced by air flow through the nostrils. In contrast, a smell, regardless of snifting, induces activation mainly in the lateral and anterior orbito-frontal gyri of the frontal lobe. The dissociation between regions activated by olfactory exploration (sniffing) and regions activated by olfactory content (smell) shows a distinction in brain organization in terms of human olfaction.

Functional magnetic resonance imaging was used to test whether odorants induce activation in the cerebellum of the human. The odorants vanillin and propionic acid both induced significant activation, primarily in the posterior lateral hemispheres. Activation was concentration-dependent, greater after stimulation with higher concentration odorants. By contrast, the action of sniffing nonodorized air induced significant activation in the anterior cerebellum, primarily in the central lobule. These findings demonstrate that the cerebellum plays a role in human olfaction. A hypothesis is proposed whereby the cerebellum maintains a feedback mechanism that regulates sniff volume in relation to odor concentration.

A method for generating olfactory stimuli for humans within a functional magnetic resonance imaging (fMRI) experimental design is described. The system incorporates a nasal-mask in which the change from odorant to no-odorant conditions occurs in less than 500 ms and is not accompanied by visual, auditory, tactile, or thermal cues. The mask provides an ordorant-free environment following prolonged ordorant presence. Specific imaging parameters that are conducive to the study of the human olfactory system are described. In a pilot study performed using these methods, the specific patterns of activation observed converged with published experimental and clinical findings.