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Summary The possible mechanisms for reorganisation of outer LHCs of PSII (LHCII) upon state transitions in Chlamydomonas reinhardtii have been discussed for several decades [38, 43-54]. For a long time people adhered to the opinion that upon the transition from state 1 to state 2, 80% of LHCII detaches from PSII and attaches completely to PSI in Chlamydomonas reinhardtii [38, 45]. This thesis provides new insights for the mechanism of state transitions in Chlamydomonas reinhardtii. In the remainder of this thesis, the role of minor light-harvesting complexes in excitation energy transfer to reaction centers of photosystem II are discussed as well as multiexciton dynamics of the alloyed ZnCdTe quantum dots are studied in detail. In chapter 2, we demonstrate with picosecond-fluorescence spectroscopy on C. reinhardtii cells that although LHCs indeed detach from Photosystem II in state-2 conditions only a fraction attaches to Photosystem I. The detached antenna complexes become protected against photodamage via shortening of the excited-state lifetime. It is discussed how the transition from state 1 to state 2 can protect C. reinhardtii in high-light conditions and how this differs from the situation in plants. In chapter 3, we study the picosecond fluorescence properties of Chlamydomonas reinhardtti over a broad range of wavelengths at 77K. It is observed that upon going from state 1 (relatively high 680nm/720nm fluorescence ratio) to state 2 (low ratio), a large part of the fluorescence of LHC/PSII becomes substantially quenched, probably because of LHC detachment from PSII, whereas the fluorescence of PSI hardly changes. These results are in agreement with the proposal in chapter 2 that the amount of LHC moving from PSII to PSI upon going from state 1 to state 2 is very limited. In chapter 4, we used picosecond-fluorescence spectroscopy to study excitation-energy transfer (EET) in thylakoids membranes isolated from A. thaliana wild-type plants and knockout lines depleted of either two (koCP26/24 and koCP29/24) or all minor Lhcs (NoM). In the absence of all minor Lhcs, the functional connection of LHCII to the PSII cores appears to be seriously impaired whereas the “disconnected” LHCII is substantially quenched. For both double knock-out mutants, excitation trapping in PSII is faster than in NoM thylakoids but slower than in WT thylakoids. In NoM thylakoids, the loss of all minor Lhcs is accompanied by an over-accumulation of LHCII, suggesting a compensating response to the reduced trapping efficiency in limiting light, which leads to a photosynthetic phenotype resembling that of low-light-acclimated plants. Finally, fluorescence kinetics and biochemical results show that the missing minor complexes are not replaced by other Lhcs, implying that they are unique among the antenna subunits and crucial for the functioning and macro-organization of PSII. In chapter 5, we have performed picosecond fluorescence measurements on ZnCdTe ternary quantum dots at room temperature by using a streak-camera setup in order to investigate in detail the fluorescence kinetics for ZnCdTe quantum dots with different size and structure by using different excitation laser intensities. Our data show that the changes in fluorescence kinetics are mostly related to the changes in structure and size. In heterogeneous structured ZnCdTe quantum dots, the fluorescence kinetics become faster as compared to homogeneous structured ZnCdTe quantum dots. Also, in both homogeneous and heterogeneous ZnCdTe quantum dots, a new peak is observed in the high-energy region of the emission spectrum when using high excitation intensities, which shows that the radiative processes that occur from higher energy states become more favoured as the excitation intensity increases.

Oral coatings are residues of food and beverages that coat the oral mucosa after consumption. Several studies have reported on the lubrication properties in mouth, and the after-feel and after-taste impact of oral coatings. Further, oral coatings have been suggested to influence subsequent taste perception. Although it is well known that oral coatings can influence sensory perception, there was little information available on the chemical composition and physical properties of oral coatings. As such, the aim of this thesis was to understand which factors influence the composition of oral coatings and their sensory perception. This study started with the development of an appropriate calibration method for an already described methodology to quantify oil oral coatings: in vivo fluorescence. Further, the samples studied were shifted from pure oil (used on previous studies) to a more realistic food beverage: o/w emulsions. Pig´s tongues are known to be a good model of human tongue. As such, Chapter 2 used pig´s tongues on the calibration of the method, to mimic the fluorescence in mouth of oil coatings. On chapter 2, Confocal Scanning Laser Microscopy images showed that stable o/w emulsions (1-20% (w/w)) stabilised by Na-caseinate created individual oil droplets on the surface of the pigs tongue, as such a new descriptor for oil coatings was developed. Oil fraction, i.e. mass of oil per surface area of the tongue, was shown to be higher on the back compared to the front anterior part of the tongue. This is thought to be due to the morphology of the tongue and abrasion of the oil coating owed to the rubbing with the palate. Further, in vivo measurements showed that oil fraction deposited on the tongue increased linearly with oil content of o/w emulsions. Coating clearance from the tongue was a fast process with around 60% of the oil being removed on the first 45s. After-feel perception (Fatty Film and Flavour Intensity) was shown to be semi-logarithmic related to oil fraction on the tongue. Chapter 3, further investigated different properties of 10% (w/w) o/w emulsions that influence the oil fraction deposited on the tongue, its clearance and after-feel perception. Three different properties were studied: protein type, protein content and viscosity of the o/w emulsions. To study the influence of protein type, two different proteins which behave differently in-mouth were studied: Na-caseinate - creates emulsions which do not flocculate under in mouth conditions, and lysozyme – creates emulsions which flocculate under in mouth conditions. To study the influence of protein content, three concentrations of Na-caseinate and lysozyme were used (0.2, 3, 5.8% (w/w) all in excess to stabilize the water/oil interface). To study the influence of viscosity of o/w emulsions, three o/w emulsions stabilized with 3% (w/w) Na-caseinate were thickened with varying concentrations of xanthan gum (0-0.5%) (w/w). Generally, the irreversible flocculation of lysozyme stabilized emulsions with saliva did not create a significant difference on oil deposition compared to emulsions stabilized with Na-caseinate, immediately after expectoration of the emulsions. Nevertheless, lysozyme stabilised emulsions caused slower oil clearance from the tongue surface compared to emulsions stabilized with Na-caseinate. Protein content had a negative relation with oil fraction on the tongue for lysozyme stabilized emulsions and no relation for Na-caseinate stabilized emulsions. The presence of thickener decreased deposition of oil on tongue, although viscosity differences (i.e., thickener content) did not affect oil fraction. After-feel perception of creaminess and fatty-film was strongly influenced by the presence of thickener likely due to lubrication in-mouth, i.e., the higher the concentration of thickener in the emulsions the stronger was the perception. Oral coatings perception was further influenced by the protein used in the emulsions, with Na-caseinate stabilised emulsions creating coatings with higher perception on creaminess and fatty-film. Chapter 2 and chapter 3 provided knowledge on the deposition and clearance of oil coatings, but little was known on the formation of oil coatings. Chapter 4 focused on the formation of oil coatings formed by Na-caseinate stabilised o/w emulsions (1-20% (w/w)). The formation of oil coatings was a rapid process, where the maximum oil deposition was achieved at normal drinking behaviour (~3s). Further, in Chapter 4 we investigated the hypothesis often referred on literature, in which oil coatings form a physical barrier which prevents tastants to reach the taste buds, and thus create a reduction on taste perception. It was concluded that oil coatings formed by emulsions within one sip did not affect subsequent sweetness perception of sucrose solutions. We suggested that the oil droplets deposited on the tongue (as seen on chapter 2) did not form a hydrophobic barrier that is sufficient to reduce the accessibility of sucrose to the taste buds and consequently does not suppress taste perception. Previous chapters focused on oral coatings formed by liquid o/w emulsions, however studies describing oral coatings formed by semi-solids and solids are scarce. As such, chapter 5 focused on the formation, clearance and sensory perception of fat coatings from emulsion-filled gels. Four emulsion-filled gelatin gels varying in fat content and type of emulsifier (whey protein isolate - created fat droplets bound to matrix; tween 20 - created fat droplets unbound to matrix) were studied. As in for oil coatings formed by liquid o/w emulsions, fat coatings formed by emulsion-filled gels reach their maximum deposition in the first seconds of mastication. This suggests that the first bites are the most relevant for the formation of fat coatings on the tongue. Further, fat fraction deposited on tongue increased when oral processing time of the gels increased. This trend was clearer for gels with higher fat content (15%) compared to gels with lower fat content (5%). Fatty perception increased with increasing mastication time, and decreased after expectoration with increasing clearance time. Fat fraction deposited on tongue and fatty perception are higher in gels with unbound droplets compared to bound droplets, as well as in gels with 15% fat compared to 5% fat. To elucidate the role of protein on oral coatings, Chapter 6 focused on the development of a method to quantify protein in the oral coatings. Further, Chapter 6 studied the influence of protein content, in-mouth protein behaviour (lysozyme - protein which creates flocs with saliva vs. Na-Caseinate - protein which does not create flocs with saliva) and presence of thickener on the formation of protein oral coatings and sensory perception of protein coatings. Protein coatings were collected from the front and middle part of the anterior tongue using cotton swabs after subjects orally processed protein solutions for different time periods. Protein concentration of the coating (mass protein/mass coating) was quantified with the Lowry method. Similarly to oil/fat coatings, results show protein coatings are formed rapidly, reaching maximum deposition on the first seconds of the samples´ oral processing. Further, different protein in mouth-behaviour (Na-caseinate vs. lysozyme) did not create differences on protein deposition on the tongue. Presence of xanthan-gum in the processed samples decreased protein deposition on the tongue, compared to when samples without xanthan-gum were processed. The perception of protein coatings was strongly influenced by the viscosity and protein used in the samples. Higher viscosity of the samples lead to higher intensity on creaminess and thickness. Lysozyme samples created coatings with high sweetness and astringent intensity, which is related to the molecular structure of the protein. Changes in the viscosity of beverages can cause changes in thickness perception. The changes in thickness perception can be accompanied by differences in other sensory properties, such as sweetness and creaminess which might be undesirable when reformulating beverages or developing new products. Knowledge on the differences by which viscosity of beverages can be modified to create a difference in sensory perception is currently lacking. Chapter 7 focus on the determination of the Just Noticeable Difference (the minimal difference that can be detected between two stimuli) for thickness perception of beverages. Oral thickness sensitivity (K=0.26) was found to be comparable to literature values for kinesthetic food firmness and spreadability, creaminess, sourness and bitterness perception. The aim of this thesis was to determine and characterize factors influencing oral coatings and their sensory perception. For this purpose, reliable methods to quantify oil and protein deposited on the tongue had to be developed to later study the macronutrients deposition. Further, the influence of stimulus properties on the formation and clearance dynamics of oral coatings and their impact on sensory perception were investigated.