Representation of odor habituation and timing in the Hippocampus

We performed simultaneous single-neuron recordings from the hippocampus and the olfactory bulb of anesthetized, freely breathing rats. Odor response properties of neurons in the olfactory bulb and hippocampus were characterized as firing rate changes or respiration-coupled changes. A panel of five odors was used. The rats had not been exposed to the odors on the panel before the experiment. The olfactory bulb and hippocampal neurons responded to repeated odor presentations in two ways: first, by changes in firing rate, and second, by respiratory tuning changes. Approximately 60% of bulbar neurons, 48% of hippocampal CA1 neurons, and 12% of hippocampal CA3 neurons showed statistically significant responses. None of the odor-responsive neurons in either the bulb or hippocampus responded to all of the odors on the panel. Repeated 10 sec odor stimuli presented at the intervals of 20, 30, 60, 110, and 160 sec were used to analyze the effect of the interval on odor response properties of the recorded neurons. Bulbar neurons were relatively nonselective for odor interval. Hippocampal neurons showed unexpected selectivity for the interval between repeated odor presentations. CA1 and CA3 neurons responded to only one to three of the intervals in the range. On the basis of these findings, we postulate that the hippocampus has the ability to keep track of the time elapsed between consecutive odor stimuli. This may act as a neuronal substrate for habituation and for complex tasks such as odor-guided navigation

Perception and representation of temporally patterned odour stimuli in the mammalian olfactory bulb

Sensory stimuli in natural environments are dynamic and complex. The neural circuits of sensory systems in the brain are therefore adapted to extract meaningful information from this dynamic input. An attractive model system for understanding how such sensory input is processed in neural circuits is the mammalian olfactory bulb (OB). The OB has a convenient dorsal anatomical location for e.g. probe implantation, viral delivery and a well-defined circuit architecture. Furthermore, olfaction in rodent models is extremely behaviourally salient and OB circuit function can therefore be efficiently investigated in the context of behavioural response. Historically, investigation of OB function has focused on encoding of odour quality, utilising static, square pulse stimuli to explore this problem. This is in stark contrast to odour transmission in natural environments, which is governed by the chaotic structure of air turbulence, creating odour plumes. There are a number of lines of evidence suggesting that temporal information in odour plumes – the fluctuations in odour concentration within this structure – can be behaviourally relevant for olfactory based navigation and odour scene segmentation. I here posit that temporal correlations in concentration for mixtures of odours transmitted in plumes are a potential mechanism by which animals identify odour objects: mixtures of odours emanating from a common source. Using neuronal imaging, high-throughput behavioural methods, high-speed odour delivery and physical recording of odour plume dynamics, I show that temporal correlations exist between pairs of odours emanating from the same source; that mice can perceive this correlation structure and that temporal correlation is represented in the output cells of the olfactory bulb. These results indicate that mammalian olfaction operates at a higher temporal bandwidth than previously thought, and that detection of temporal features in odour signals may represent a potential mechanism for olfactory scene segmenetation

Response and transcriptional regulation of rice SUMOylation system during development and stress conditions

Modification of proteins by the reversible covalent addition of the small ubiquitin like modifier (SUMO) protein has important consequences affecting target protein stability, sub-cellular localization, and protein-protein interactions. SUMOylation involves a cascade of enzymatic reactions, which resembles the process of ubiquitination. In this study, we characterized the SUMOylation system from an important crop plant, rice, and show that it responds to cold, salt and ABA stress conditions on a protein level via the accumulation of SUMOylated proteins. We also characterized the transcriptional regulation of individual SUMOylation cascade components during stress and development. During stress conditions, majority of the SUMO cascade components are transcriptionally down regulated. SUMO conjugate proteins and SUMO cascade component transcripts accumulated differentially in various tissues during plant development with highest levels in reproductive tissues. Taken together, these data suggest a role for SUMOylation in rice development and stress responses

Odors Pulsed at Wing Beat Frequencies are Tracked by Primary Olfactory Networks and Enhance Odor Detection

Each down stroke of an insect's wings accelerates axial airflow over the antennae. Modeling studies suggest that this can greatly enhance penetration of air and air-born odorants through the antennal sensilla thereby periodically increasing odorant-receptor interactions. Do these periodic changes result in entrainment of neural responses in the antenna and antennal lobe (AL)? Does this entrainment affect olfactory acuity? To address these questions, we monitored antennal and AL responses in the moth Manduca sexta while odorants were pulsed at frequencies from 10–72 Hz, encompassing the natural wingbeat frequency. Power spectral density (PSD) analysis was used to identify entrainment of neural activity. Statistical analysis of PSDs indicates that the antennal nerve tracked pulsed odor up to 30 Hz. Furthermore, at least 50% of AL local field potentials (LFPs) and between 7–25% of unitary spiking responses also tracked pulsed odor up to 30 Hz in a frequency-locked manner. Application of bicuculline (200 μM) abolished pulse tracking in both LFP and unitary responses suggesting that GABAA receptor activation is necessary for pulse tracking within the AL. Finally, psychophysical measures of odor detection establish that detection thresholds are lowered when odor is pulsed at 20 Hz. These results suggest that AL networks can respond to the oscillatory dynamics of stimuli such as those imposed by the wing beat in a manner analogous to mammalian sniffing

Novel measure of olfactory bulb function in health and disease

Present neuroimaging techniques are capable of recording the neural activity from all over the brain but the olfactory bulb (OB). The OB is the first olfactory processing stage of the central nervous system and the site of insult in several neurological disorders, particularly Parkinson’s disease (PD). It has been suggested that the OB has a pivotal role in the olfactory system anal-ogous to primary visual cortex (V1) and thalamus in the visual system. However, due to the existing technical limitations, there has not been any non-invasive technique that can reliably measure the OB function in humans, consequently limiting its functional recording to one in-tracranial study dating back to the 60s. Initially in Study I, a non-invasive method of measuring the function of human OB is devel-oped, so-called electrobulbogram (EBG). In line with previous animal literature as well as the only intracranial study in human OB, it was demonstrated that gamma oscillations on the EBG electrodes occurred shortly after the odor onset. Subsequently, applying source recon-struction analysis provided evidence that observed oscillations were localized to the OB. Ad-ditionally, the OB recording with the EBG method showed a test-retest reliability comparable with visual event related potentials. Notably, the detected gamma oscillations were demon-strated to be insensitive to habituation, the OB’s marked characteristic which has previously been demonstrated in rodents. Last, but not least, assessing the EBG response in an individual who did not have the bilateral OB indicated that the lack of OB results in disappearance of gamma oscillations in the EBG electrodes. Given that Study I determined the possibility of reliably measuring the function of the OB using the EBG, in Study II, I assessed the functional role of OB’s oscillations in the pro-cessing of the odor valence. Odor valence has been suggested to be linked to approach–avoidance responses and therefore, processing of odor valence is thought to be one of the core aspects of odor processing in the olfactory system. Consequently, using combined EBG and EEG recording, OB activity was reconstructed on the source level during processing of odors with different valences. Gamma and beta oscillations were found to be related to va-lence perception in the human OB. Moreover, the early beta oscillations were associated with negative but not positive odors, where these beta oscillations can be linked to preparatory neural responses in the motor cortex. Subsequently, in a separate experiment, negative odors were demonstrated to trigger a whole-body motor avoidance response in the time window overlapping with the valence processes in the OB. These negative odor-elicited motor re-sponses were measured by a force plate as a leaning backward motion. Altogether, the results from Study II indicated that the human OB processes odor valence sequentially in the gamma and beta frequency bands, where the early processing of negative odors in the OB might be facilitating rapid approach-avoidance behaviors. To further evaluate the functional role of the OB in odor processing, in Study III, OB’s communication with its immediate recipient, namely piriform cortex (PC), was assessed. These two areas are critical nodes of the olfactory system which communicate with each other through neural oscillations. The activity of the OB and the PC were reconstructed using a combination of EBG, EEG, and source reconstruction techniques. Subsequently, the cross spectrogram of the OB and the PC was assessed as a measure of functional connectivity where temporal evolution from fast to slow oscillations in the OB–PC connectivity was found during the one second odor processing. Furthermore, the spectrally resolved Granger causal-ity analysis suggested that the afferent connection form the OB to the PC occurred in the gamma and beta bands whereas the efferent connection from the PC to the OB was concen-trated in the theta and delta bands. Notably, odor identity could be deciphered from the low gamma oscillatory pattern in the OB–PC connectivity as early as 100ms after the odor onset. Hence, findings from this study elucidate on our understanding of the bidirectional infor-mation flow in the human olfactory system. Olfactory dysfunction, due to neurodegeneration in the OB, commonly appears several years earlier than the occurrence of the PD-related characteristic motor symptoms. Consequently, a functional measure of the OB may serve as a potential early biomarker of PD. In Study IV, OB function was assessed in PD to answer whether the EBG method can be used to dissociate individuals with a PD diagnosis from healthy age-matched controls. The spectrogram of the EBG signals indicated that there were different values in gamma, beta, and theta for PDs compared with healthy controls. Specifically, six components were found in the EBG re-sponse during early and late time points which together dissociate PDs from controls with a 90% sensitivity and a 100% specificity. Furthermore, these components were linked to med-ication, disease duration and severity, as well as clinical odor identification performance. Overall, these findings support the notion that EBG has a diagnostic value and can be further developed to serve as an early biomarker for PD. In the last study, Study V, the prevalence of COVID-19 was determined using odor intensity ratings as an indication of olfactory dysfunction. Using a large sample data (n = 2440) from a Swedish population, odor intensity ratings of common household items over time were found to be closely associated with prevalence prediction of COVID-19 in the Stockholm region over the same time-period (r = -.83). Impairment in odor intensity rating was further correlated with the number of reported COVID-19 symptoms. Relatedly, individuals who progressed from having no symptoms to having at least one symptom had a marked decline in their odor intensity ratings. The results from this study, given the relatively large sample size, provided a concrete basis for the future studies to further assess the potential association between the deficits in the OB function and olfactory dysfunction in COVID-19. In conclusion, our proposed method for non-invasive measurement of the OB function was shown to provide a reliable recording with a potential as a diagnostic tool for PD. Combining EBG and EEG allowed for reconstruction of the OB signal at the source level, where specific oscillations were found to be critical for odor valence processing and rapid avoidance re-sponse. Moreover, oscillations in different frequency bands were found to be critical for the OB reciprocal communications and transfer of odor identity information to higher order ol-factory subsystems. Finally, COVID-19 was found to be associated with a decline in olfactory acuity which might originate from damage to the patient’s OB. In conclusion, the results from the studies within this thesis provide a new perspective on the functional role of oscillations in the human OB

Correlating instrumental and sensory analyses of flavour

The relationship between in vivo captured data from an atmospheric pressure chemical ionisation mass spectrometer (APCI-MS) and sensory/psychophysical analyses was investigated. The stimuli used were mainly single volatiles under gas phase control or calibration by development of different olfactometry methods. Gas phase concentration retronasal (via the mouth to the nasal cavity) and orthonasal (via the nostrils) thresholds were determined for a trained panel of 13 individuals. Four volatiles were used with different sensory/physico-chemical properties and an adapted staircase method was employed to measure the individual thresholds. The data showed good repeatability over short durations of one week and also longer ones of eight months. It was used to test the hypothesis that thresholds varied between people due to differences in their in-nose concentration as measured or estimated by the APCI-MS. The analysis did not support this theory but relationships between orthonasal and retronasal thresholds were shown, in which the latter were -50 times lower than the former. Threshold determination of a larger group of 20 individuals revealed clusters of individuals. Methods of producing square edged pulses of aroma compound in the gas phase were developed using a modified chromatograph autosampler with a gas flow of 5 mL. min 1 and pulse rate of 0.6 secs. A trained panel of 23 individuals performed two types of sensory test using pulsed and constant olfactometer outputs of isoamyl acetate. The original intention was to reveal whether pulsed odorants were perceived as the same as or different to constant concentration. Initial experiments yielded results that were difficult to interpret, although the nature of the results was clarified when simultaneous breath by breath analysis techniques were employed. Here it was shown that each individual in different repetitions disrupted the olfactometer output pattern in unpredictable ways. This pattern disruption was measured in two instrumental configurations, as either volatiles in an exhalation or volatiles as they were inhaled together with two types of sensory test. In both sensory tests the pattern of aromas in an inhalation revealed a relationship with perception. In particular, the sensory response in the time intensity study was related to differences in the inhalation profiles between people, which in turn was related to an individual's breathing. This shows that physiological differences such as breathing and the structure of the nasal cavity have an impact on perception

Correlating instrumental and sensory analyses of flavour

The relationship between in vivo captured data from an atmospheric pressure chemical ionisation mass spectrometer (APCI-MS) and sensory/psychophysical analyses was investigated. The stimuli used were mainly single volatiles under gas phase control or calibration by development of different olfactometry methods. Gas phase concentration retronasal (via the mouth to the nasal cavity) and orthonasal (via the nostrils) thresholds were determined for a trained panel of 13 individuals. Four volatiles were used with different sensory/physico-chemical properties and an adapted staircase method was employed to measure the individual thresholds. The data showed good repeatability over short durations of one week and also longer ones of eight months. It was used to test the hypothesis that thresholds varied between people due to differences in their in-nose concentration as measured or estimated by the APCI-MS. The analysis did not support this theory but relationships between orthonasal and retronasal thresholds were shown, in which the latter were -50 times lower than the former. Threshold determination of a larger group of 20 individuals revealed clusters of individuals. Methods of producing square edged pulses of aroma compound in the gas phase were developed using a modified chromatograph autosampler with a gas flow of 5 mL. min 1 and pulse rate of 0.6 secs. A trained panel of 23 individuals performed two types of sensory test using pulsed and constant olfactometer outputs of isoamyl acetate. The original intention was to reveal whether pulsed odorants were perceived as the same as or different to constant concentration. Initial experiments yielded results that were difficult to interpret, although the nature of the results was clarified when simultaneous breath by breath analysis techniques were employed. Here it was shown that each individual in different repetitions disrupted the olfactometer output pattern in unpredictable ways. This pattern disruption was measured in two instrumental configurations, as either volatiles in an exhalation or volatiles as they were inhaled together with two types of sensory test. In both sensory tests the pattern of aromas in an inhalation revealed a relationship with perception. In particular, the sensory response in the time intensity study was related to differences in the inhalation profiles between people, which in turn was related to an individual's breathing. This shows that physiological differences such as breathing and the structure of the nasal cavity have an impact on perception

A Flight Sensory-Motor to Olfactory Histamine Circuit Mediates Olfactory Processing of Ecologically and Behaviorally Natural Stimuli

Environmental pressures have conferred species specific behavioral and morphological traits to optimize reproductive success. To optimally interact with their environment, nervous systems have evolved motor-to-sensory circuits that mediate the processing of its own reafference. Moth flight behavioral patterns to odor sources are stereotyped, presumably to optimize the likelihood of interacting with the odor source. In the moth Manduca sexta wing beating causes oscillatory flow of air over the antenna; because of this, odorant-antennal interactions are oscillatory in nature. Electroantennogram recordings on antennae show that the biophysical properties of their spiking activity can effectively track odors presented at the wing beat frequency. Psychophysical experiments using Manduca show that when odors are pulsed, as opposed to presented as a continuous stream, detection and discrimination thresholds are lowered. In this study, we characterized histamine immunoreactivity in the thoracic ganglia and brain of Manduca. We generated antibodies for and characterized the distribution of the histamine B receptor, the first known antibody for this receptor protein. Our results show an elaborate pair of neurons projecting from the mesothoracic ganglion to the brain, including axon innervation of the antennal lobe and antennal mechanosensory and motor centers. Additionally, histamine B receptor labeling overlapped with a subset of GABAergic and peptidergic local interneurons. Next, we characterized the response properties of these cells within the context of fictive flight behavior and found a tonic increase in activity. Furthermore, disrupting this circuit, with surgical ablation and pharmacology, disrupts antennal lobe projection neurons from entraining to odors presented at a natural 20 Hz frequency, as well as behavioral measures of detection and discrimination thresholds. Finally, we characterized the relationship between motor patterns/behaviors, and circuit structure of this pair of histamine immunoreactive neurons. Specifically, presence of MDHn axon collaterals entering the antennal lobe is correlated with olfactory-guided target approach behaviors in crepuscular and nocturnal moths who require stereotyped zigzagging and wing beating behaviors for locating an olfactory target have axonal ramifications in the antennal lobe. This study is the first characterization of a motor to olfactory corollary discharge circuit in invertebrates and may represent the first characterization of a higher order corollary discharge circuit in an invertebrate model

Synaptic depression and its relation to behavioral habituation within anterior piriform cortex.

The onset, time course, and extent of LOT synaptic depression during both in vitro electrical and in vivo odorant stimulation methods were similar. Similar to the odor specificity of cortical odor adaptation in vivo, there was no evidence of heterosynaptic depression between independent inputs in vitro. In vitro evidence suggests at least two mechanisms contribute to this activity-dependent synaptic depression: a rapidly recovering presynaptic depression during the initial 10--20 sec of the post-train recovery period and a longer lasting (120 sec) depression that can be blocked by the metabotropic glutamate receptor (mGluR) II/III antagonist (RS)-alpha-cyclopropyl-4-phosphonophenylglycine (CPPG) and by the beta-adrenergic receptor agonist isoproterenol. Importantly, in line with the in vitro findings, both adaptation of odor responses in the 15--35 Hz spectral range and the associated synaptic depression can also be blocked by intracortical infusion of CPPG in vivo. Additionally, blockade of group II/III mGluR activation in aPCX prevents habituation of the odor-evoked heart rate orienting response behavior in awake behaving rats.Short-term behavioral habituation is the response decrement observed in many behaviors that occurs during repeated presentation of non-reinforced stimuli. Within a number of invertebrate models of short-term behavioral habituation, depression of a defined synapse has been implicated as the mechanism. However, the synaptic mechanisms of short-term behavioral habituation have not been identified within mammals.Anterior piriform cortex (aPCX) neurons rapidly filter repetitive odor stimuli despite relatively maintained input from mitral/tufted cells. This cortical adaptation is correlated with short-term depression of afferent synapses, in vivo. The purpose of this study was to elucidate mechanisms underlying this non-associative neural plasticity using in vivo and in vitro preparations, determine its role in cortical odor adaptation and ultimately look at its relation to behavioral odor habituation. Lateral olfactory tract (LOT)-evoked responses were recorded in rat aPCX coronal slices. Extracellular and intracellular potentials were recorded before and after simulated odor stimulation of the LOT. Results were compared with in vivo intracellular recordings from aPCX layer II/III neurons and field recordings in urethane-anesthetized rats stimulated with odorants

Cross-modal interactions in complex food matrices

In the light of increasing rates of nutrition related diseases, such as obesity, diabetes, high blood pressure and stroke it is necessary to reduce sugar, salt and fat contents of industrial produced food. Reduction of those components generally leads to changes of the sensory properties of the products and rejection by the consumer. Traditional compensation strategies like the use of taste enhancers, artificial sweeteners or fat replacers often lead to off-flavours and consequently to consumer rejection. This thesis describes cross-modal interactions as an alternative strategy to reduce sugar, salt and fat. In the past cross-modal interactions have been describe to influence taste and texture perception, however up to now research focused on simple model systems lacking in perceptual, as well as chemical complexity. This thesis aimed on the application of cross modal interactions involving aroma modification to complex food systems, such as apple juice and cheese. It was investigated whether aromas are able to modify taste perception in complex food matrices and what the driving factors are to which extend aromas can enhance taste perception. Furthermore different cross modal approaches were combined in order to investigate possible additive or synergistic effects on taste enhancement. At first, the question whether individual aroma components can be identified that have the capacity to enhance taste perception in a complex beverage was addressed [chapter 2]. Apple juice was choosen to represent a complex matrix where interactions between taste and aroma are a natural product characteristic. Ethylhexanoate was identified to significantly enhance sweetness in apple juice, while three other esters selected on the same basis did not show sweetness enhancement. Ethylhexanoate induced sweetness enhancement appeared to be concentration dependent. Concentrations of 5 ppm were found to be most effective to enhance sweetness in this specific system. However, next to sweetness undesired attributes such as flowery and synthetic were also increased significantly. As a conclusion it has to be noticed that flavour balance in complex food is fragile and has to be carefully altered in order to use odour induced taste enhancement as a tool in sugar, salt and fat reduction. Chapter 3 describes the masking of off-flavours induced by ethylhexanoate as described in chapter 2. It was hypothesised that ethylhexanoate induced off-flavours can be masked by restoring flavour balance by equally adding combinations of all four esters. A combination of ethylhexanoate, ethylbutanoate and ethyl-2-methylbutanoate was found to be most effective to restore flavour quality while maintaining ethylhexanoate induced sweetness enhancement. It was concluded that all components that are naturally part of an aroma are needed to achieve a balanced product. Optimisation of odour presentation time in order to achieve maximum taste enhancement was subject to the study described in chapter 4. Swallowing is the key to aroma release during food consumption. It was demonstrated that aroma is most presented most effectively either 1.7 – 2.5 seconds before or 2.6 – 3.6 seconds after the moment of swallow. Aroma presented directly at the moment of swallow was found to be least effective to enhance taste. It is assumed that olfactory receptor neurons (ORN) do not transmit the activation signal to the brain at the exact moment of swallow in order to spare energy and function most effective, as during normal food consumption the aroma is released shortly after swallowing. This hypothesis was further tested in an fMRI study. Preliminary results support this theory, however at the moment of completion of this thesis data evaluation was still in progress. In chapter 5 further optimisation of odour/taste interactions was studied, by investigating the influence of temperature on odour induced taste enhancement. Consumption temperatures differ strongly among products. As aroma release strongly depends on the temperature of the food, it was hypothesised that the magnitude of aroma effects on taste perception changes over consumption temperature. Subjects consumed a sweet and a savoury system at four different temperatures (7, 25, 37 and 50°C). Stimuli temperature and odour presentation were fully controlled by temperature optimised gustometry and olfactometry. Both aroma/taste systems were known to have induced taste enhancement in the past (sweet: apple flavoured tea/ethylhexanoate; savoury: broth/sotolon). No significant effect of temperature was found for either of the stimulus pairs. Results indicate that the complexity of the experiment led to confusion by the panelists under fully randomised stimuli delivery conditions. It was therefore concluded, that temperature effects on the magnitude of odour induced taste enhancement need to be studied in a reduced experimental design. Temporal contrast as a strategy to enhance salty taste was studied in the experiments described in chapter 6. Salty solutions of different NaCl concentrations were presented in alternating sequence by a gustometer, creating a sensory contrast of low-in salt and high-in salt pulses. It was demonstrated that the sensory contrast induced by the pulsed delivery led to significant taste enhancement. It was shown that high concentrations of NaCl delivered in short pulses were most effective to enhance salty taste. It was concluded that pulsed stimulus delivery can be an additional tool to reduce salt and sugar concentrations in industrial produced food. Chapter 7 combines both cross-modal strategies to enhance taste in this thesis so far, odour induced taste enhancement and temporal contrast of stimulus delivery. Subjects were presented with taste and aroma pulses timed via a gustometer. The aroma was either presented in-phase or out-of-phase with the taste stimulus. A cumulative effect of aroma/taste interactions and temporal contrast of tastant delivery was found, resulting in higher taste enhancement than each of the strategies alone. Overall highest sweetness enhancement was observed when aroma and taste pulses were presented out-of-phase. Texture modification is a third cross-modal strategy to enhance taste perception. Combinatory effects of texture modification and odour induced taste enhancement are subject to the study reported in chapter 8. Apple juice containing gels were engineered differing in textural properties, aroma and sugar concentration. In contrast to the results presented in chapter 7, only an additive but no synergistic effect was found for those strategies. Texture modification was found to be more effective than aroma modification. Chapter 9 studies effects of aroma on the taste and the texture of cheese and dairy model gels. In the first study subjects consumed different types of cheese with and without a nose-clip. This way the contribution of the aroma phase on the flavour and texture perception of cheese was studied. Saltiness was influenced significantly by the aroma. It was concluded that the aroma of cheese strongly contributes to a cheeses salty taste. Furthermore it was observed that a decrease in cheese firmness strongly correlated with an increase in buttery aroma notes. This was further studied in fully controlled dairy model gels. For gels tasting of cream cheese a significant decrease in firmness was found upon increasing aroma concentration. No effect of butter aroma on firmness was found for yoghurt-like gels. This once more demonstrates the importance of congruency between stimuli. Only congruent sensory impressions can influence each other. In addition, an increase in creaminess was observed with increasing aroma concentration. The results described in chapter 9 clearly show that aromas cannot only modify taste perception, but also are a valid tool for texture modification. This thesis demonstrates for the first time, that flavour and texture modification and subsequently the reduction of sugar, salt and (possibly also) fat can be achieved by using cross modal interactions in complex food matrices. New methods have been developed and existing methods have been combined to study aroma/taste interactions in fully controlled settings. Its multidisciplinary approach combines chemical, physical and psychological knowledge in order to discuss and explain results. It furthermore shows that combinations of different strategies are most effective in order to achieve healthier products reduced in sugar, salt and fat. </p