Background: Dopamine levels has been implicated in obesity, feeding behaviour, and hedonic control of appetite like olfactory cues and food palatability. Methylphenidate (MPH) is a dopamine reuptake inhibitor that increases brain dopamine levels and has been shown to reduce appetite and promote weight loss in patients with attention deficit hyperactivity disorder (ADHD). As such, the objectives of this study were to test the possible effect of MPH on appetite, olfaction, and food palatability as well as its effects on energy intake and body weight of healthy individuals with obesity. Methods: In a randomized, double-blind study, 12 participants (age 28.9±6.7 yrs) (BMI 36.1±4.5 kg/m2) were assigned to receive MPH (0.5mg/kg) (n=5) or placebo (n=7) twice daily for two months. Appetite and palatability (Visual Analog Scale (VAS)), odour threshold (Sniffin’ Sticks®), in-lab energy intake (ad libitum buffet), free-living energy intake (3-day food boxes) and body weight (DEXA scan) were measured at baseline (day 1) and final visit (day 60). Results: MPH intake caused significantly greater suppression of appetite sensations (desire to eat (p=0.001), hunger (p=0.008), and prospective food consumption (p=0.003)) and increase in fullness (p=0.028) over time compared to placebo. There was a significant increase in odour threshold scores in the MPH group (6.3±1.4 vs. 9.4±2.1) compared to placebo (7.9±2.3 vs. 7.8±1.9) (p=0.029). Both placebo and MPH groups showed decreases in their energy intake (p=0.021) and body weight (p=0.005) over time but with large effect sizes favouring greater reduction in the MPH group compared to placebo. Conclusions: Compared to placebo, MPH intake over 60 days suppressed appetite and improved olfactory sensitivity in individuals with obesity. These data provide novel findings into the possible efficacy of MPH to favourably impact appetite and therefore promoting weight loss in individuals living with obesity.

Sexually dimorphic behaviour of the house mouse (Mus musculus musculus) relies on various physical and chemical cues, however, chemical signals are the most essential cues for individual recognition and in causing various priming effects on reproductive behaviour of the receiver. House mice belong to macrosmatic mammals, and thus, their sense of smell is highly developed and is able to recognize a wide spectrum of ligands from other individuals and from their surrounding environment. Volatile signals belong to organic compounds that are produced by most tissues, and may have harmfull effects on cells, and thus they are transported out of the body with lipocalin transporters where some of them may function as signals. These volatile signals are able to stimulate chemosensory neuronal receptors, and thus, yield particular responses in neural circuits. The ligand sensing has a differential effect upon males and females, however, it has not been shown yet in wild mice whether these differences are also caused by the variation in receptors and neural processing, or rather by differential expression of signals typical for each sex. The aim of this thesis was to perform comparative analysis of orofacial mucosal tissues to determine the specificity of expression of particular lipocalins. For the first time...

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The development of an inexpensive, portable, stable, sensitive and selective biosensor for detection of odorants is a daunting task. Here, we hypothesized the development of a detector layer composed of the protein groups; the olfactory receptors (ORs) and the odorant binding proteins (OBPs), known to bind odorants in animal sensing. We report the design of 13 OR gene-vector constructs, and their subsequent transformation into Escherichia (E.) coli (BL21 (DE3)-STAR-pLysS) strain. Moreover, we report the expression of several ORs into an in vitro wheat germ extract using three separate detergent mixes for protein solubilization.   In addition, we describe the design of an odorant binding protein from the Aenopheles gambiae mosquito PEST strain (OBP-PEST) gene-vector construct under an IPTG (Isopropyl β-D-1-thiogalactopyranoside) inducible promoter. OBP-PEST was heterologously expressed in E.coli with an 8 amino acid long sequence (WSPQFEK) attached C-terminally, via a thrombin cleavage site and a flexible linker (GSSG). The WSPQFEK sequence, commonly referred to as a Strep-tag, enabled subsequent affinity chromatography purification of the protein, via binding to an engineered Streptavidin equivalent. Surprisingly, the OBP-PEST was found to contain a signal sequence leading to its truncation and secretion when expressed in E.coli.   Biophysical analyses were established using Circular Dichroism (CD) for the analysis of two lipocalins: Beta-lactoglobulin (BLG) and OBP-PEST. We studied the solubility, refoldability and the conformational transitions of BLG, as a result of change in solvent, pH and temperature. The secondary structure of OBP-PEST and its thermal stability was investigated.   In conclusion, this thesis work has enabled biophysical analyses of OBP-PEST and future analogs of interest to the development of a stable protein detector layer. Although further experiments are needed to fully characterize the biophysical properties, and to find odorant substrates of OBP-PEST, it was found to be a suitable alternative to ORs in a biosensor detector layer application. More importantly, an inherent OBP-PEST signal sequence was found to mediate protein secretion when expressed heterologously in E.coli. To the best of our knowledge this is the first lipocalin discovered to be secreted upon heterologous expression in E.coli.   We hypothesize that this signal peptide could be used as a means for targeted secretion and, hence, efficient protein purification.

Dementia is a growing burden for society, and it is of interest to discover it at an early stage. Both subjective cognitive decline (SCD) and subjective olfactory decline (SOD) has been associated with future cognitive decline and dementia. However, subjective measures have often been criticized and are still not fully understood. I aimed to examinate the frequency of SCD and SOD and whether they are likely to measure different things and what their longitudinal effects are. The baseline sample (N=784, 35-90 years, 51% female) were split into reported SCD, SOD, combined subjective olfactory and cognitive decline, and controls. Between-subjects and within-subjects statistical tests were conducted on a subset of participants (N=307, 45 to 90 years, 52% female) comparing SCD and SOD and their olfactory ability, cognitive performance, and demographics. In the baseline sample, a total of 21.1% reported a SOD whereas only 9.9% reported a SCD, only 2.7% reported both. SOD individuals had an emerged olfactory decline at follow up, their olfactory performance was associated with performance in several cognitive tests, this was not the case for the SCD individuals. The SOD and the SCD groups differ from each other, and they appear to be rather independent from each other. They might be complementary in understanding the aging brain.

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Le génome humain contient plus de huit cents gènes de récepteurs olfactifs et 25 gènes derécepteurs au goût amer, qui sont exprimés dans de nombreux tissus. Très peu de données existentsur leur présence au niveau des muqueuses nasosinusiennes humaines. Les récepteurs olfactifs ontprobablement des fonctions en dehors de l’olfaction. Il est donc nécessaire d’établir lesquels d’entreeux participent à l’olfaction en les recherchant au sein de la muqueuse olfactive. Par ailleurs, lesrécepteurs au goût amer participent potentiellement à d’autres processus physiologiquesindépendants de la gustation. Au niveau du nez et des sinus, ces récepteurs pourraient êtreimpliqués dans l’immunité innée nasale.La première partie de ce travail s’est intéressée à l’expression des gènes des récepteurs olfactifs dansla muqueuse olfactive humaine. Des prélèvements de muqueuse olfactive entière ont pu êtreréalisés lors de 26 autopsies, puis analysés par les techniques de RT-qPCR. Nous mettons en évidenceque 273 gènes sont exprimés en moyenne. Un groupe de 90 gènes est exprimé chez tous les sujets,un autre groupe de 140 gènes est exprimé chez plus de la moitié des sujets et enfin un derniergroupe de 125 gènes est exprimé chez moins de la moitié des sujets. Cette variabilitéinterindividuelle d’expression des gènes de récepteurs olfactifs pourrait donc intervenir dans lavariabilité des performances olfactives entre individus. Nous avons également réalisé une deuxièmesérie de six prélèvements dans le but de préciser si une différence d’expression existait entre lamuqueuse olfactive antérieure et postérieure. Nos résultats montrent une expression semblableentre ces deux parties.La deuxième partie de ce travail a été consacrée aux récepteurs au goût amer. Le premier objectifconsistait à déterminer parmi les gènes de cette famille de 25 récepteurs, lesquels étaient expriméset à quel niveau dans la sphère nasosinusienne. Nous avons eu recours à la même approche baséesur la RT-qPCR, pour identifier et quantifier les ARN messagers de ces récepteurs au sein de septrégions des muqueuses nasosinusiennes provenant de prélèvements obtenus lors de sept autopsies.Nos résultats montrent une expression similaire parmi les différentes régions étudiées.Enfin, partant de données de la littérature qui suggèrent un rôle du récepteur au goût amer T2R38dans l’immunité nasale innée, nous avons exploré in vitro la capacité de ce récepteur à reconnaîtredifférents métabolites bactériens. Nos résultats montrent 7 nouveaux agonistes, suggérant que lerécepteur T2R38 est capable de détecter des métabolites bactériens plus variés que ce qui étaitprécédemment connu. / Doctorat en Sciences médicales (Médecine) / info:eu-repo/semantics/nonPublished

Domestication is accompanied by marked changes in the phenotype of the domesticated form of a species relative to its wild ancestor. The domestic dog and its wild ancestor, the wolf, are a widely used model system to study the effects of domestication on cognition and behaviour. The aim of this study was therefore to compare the behavioural responses of wolves (Canis lupus) and domestic dogs (Canis lupus familiaris) to different odour stimuli. To this end, wooden logs were impregnated with one of four odours and repeatedly placed into the enclosure of a wolf or dog pack. The dogs (n=10) and wolves (n=9) both performed a significantly higher number of interactions with the wooden logs when these were impregnated with real blood compared to the blood odour component trans-4,5-epoxy-(E)-2-decenal, the fruity odour iso-pentyl acetate and the solvent diethyl phthalate. No significant difference in the frequency of interactions per animal was found between the wolves and dogs with any of the four odour stimuli. Sniffing was the most frequent behaviour in both wolves and dogs, followed by licking, biting, toying, and pawing. The only qualitative difference in behaviour was that the dogs, but not the wolves, displayed guarding behaviour when presented with the odorized wooden logs. The results of the present study suggest that domestication had little, if any, effect on olfactory behaviour in domesticated dogs. The results also indicate that odour-impregnated wooden logs are a suitable method of olfactory enrichment for captive wolves and domestic dogs.