Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Achieving Olfactory Expertise: Training for Transfer in Odor Identification
Morquecho-Campos, Paulina ; Larsson, Maria ; Boesveldt, Sanne ; Olofsson, Jonas K. - \ 2019
Chemical Senses 44 (2019)3. - ISSN 0379-864X - p. 197 - 203.
Memory - mixtures - perceptual learning - smell - transfer effect

Human olfactory function requires the identification of everyday odors. A characteristic feature of olfaction is that most people find it hard to identify and name common odors, and when odors are presented simultaneously in mixtures, performance is even further compromised. Few studies have systematically assessed how training might enhance identification of single odors and mixtures. This study compared how odor identification training with either single odors or binary mixtures affected identification performance, as well as transfer effects to untrained tasks and odors. Twenty-seven healthy participants (22 F; 28.0 ± 4.7 years old) completed identification training of 8 odors using a list of 16 veridical names. The study included 8 training sessions, as well as pretest and posttest evaluations. Results suggest notable effects of learning, as well as transfer to novel tasks and odors. Overall, training with single odors led to slightly better results than the binary mixture condition, suggesting that in novices, odor identification may be facilitated via consolidation of single odor objects, before learning to dissociate binary mixtures. Overall, odor identification may be trained to generate transfer of learning, although transfer effects were observed in both training methods. Our work suggests that odor identification abilities, while often limited, are highly trainable.

Fibrillar structures in mixed systems
Peng, Jinfeng - \ 2016
Wageningen University. Promotor(en): Erik van der Linden, co-promotor(en): Paul Venema; K.P. Velikov. - Wageningen : Wageningen University - ISBN 9789462578265 - 284
cellulose - bacteria - fibres - protein isolates - whey - mixtures - emulsions - mechanical properties - cellulose - bacteriën - vezels - eiwitisolaten - wei - mengsels - emulsies - mechanische eigenschappen

Fibrillar structures are important structuring elements for food products. Understanding the behaviour of fibrillar structures in complex food systems is essential for successful industrial applications. This thesis presents the behaviour of two different fibrillar structures, i.e. whey protein isolate (WPI) fibrils and bacterial cellulose (BC) microfibrils in mixtures under various conditions. The WPI fibrils are prepared from WPI and the BC microfibrils are extracted from commercial available ‘Nata de Coco’ by high-energy de-agglomeration. In Chapter 1, a general introduction is given, where we introduce two different fibrillar structures that were studied in this thesis. Also, the aim and the outline of the thesis are presented. In Chapter 2, 3, 4 and 5, the behaviour of mixtures containing WPI and BC microfibrils under different conditions are investigated. By varying the concentration ratios, pH, NaCl concentration and further applying heating treatment, their physico-chemical properties in mixed solutions, mixed solutions after heating and further heat-induced mixed gels are investigated and characterized at both pH 2 and pH 7. In general, both mixing WPI and BC microfibrils without heating and subsequently applying heating treatment lead to stable and homogeneous mixtures at pH 7, as long as BC microfibril concentration is above a critical value. Microscopic images showed that the WPI aggregates and BC microfibrils co-existed in the system. WPI denatured and aggregated in the mixture in the same way as when it is heated alone. Upon gelation, the WPI and BC microfibrils form a duplex gel consisting of two independent and homogeneous networks spanning the whole system. At pH 2, the WPI and BC microfibrils also form stable and homogeneous mixtures in the liquid state, both before and after heating. Microscopic images showed two fibrillar structures that are uniformly and independently present. Upon gelation at higher WPI concentration, a bi-fibrillar gel is formed consisting of a WPI fibrilllar gel and BC microfibrillar gel that co-exist. In Chapter 6 and 7, the behaviour of WPI fibrils at pH 2 in dispersions containing spheres, i.e. emulsions and polystyrene latex dispersions are studied. When WPI and spheres are both positively charged (i.e. WPI-stabilized emulsion), we observed depletion flocculation and depletion stabilization when the WPI fibril concentration increases. When WPI and the spheres are oppositely charged (i.e. polystyrene latex dispersions), bridging flocculation and steric/electrostatic stabilization were observed at low WPI fibril concentration, followed by depletion flocculation and depletion stabilization upon increasing WPI fibril concentrations. In Chapter 8 the stability of emulsions at pH 2 in the presence of only BC microfibrils and in the presence of both BC microfibrils and WPI fibrils was studied. When only BC microfibrils added at a sufficiently high concentration, the emulsions are stabilized by the presence of a yield stress as generated by the BC network. When both WPI fibrils and BC microfibrils are added to the emulsions, the networks they form behave in the same way, as when they are added to the emulsions separately. The WPI fibrils induced depletion flocculation and stabilization of the emulsions, despite the presence of the BC microfibrils. However, at high enough BC microfibril concentrations, the emulsions can be stabilized against depletion flocculation as induced by the WPI fibrils. The competition between stabilization and/or de-stabilization induced by the BC microfibrils and the WPI fibrils can lead to emulsions with different microstructures and rheological properties. A general discussion on the results obtained in this thesis is presented in Chapter 9, which includes recommendations for further research and concluding remarks.

Foam properties of proteins, low molecular weight surfactants and their complexes
Lech, F.J. - \ 2016
Wageningen University. Promotor(en): Harry Gruppen; Peter Wierenga; Marcel Meinders. - Wageningen : Wageningen University - ISBN 9789462576247 - 122
surfactants - proteins - bovine serum albumin - beta-lactoglobulin - lysozyme - foams - chemical properties - stability - mixtures - food chemistry - oppervlaktespanningsverlagende stoffen - eiwitten - runderserumalbumine - bèta-lactoglobuline - lysozym - schuim - chemische eigenschappen - stabiliteit - mengsels - voedselchemie

This thesis shows the effects that the addition of low molecular weight surfactants (LWMS) to proteins has on the foam stability of the mixture. For this, the bulk, interfacial, thin liquid films and foam properties are determined for different protein-LWMS mixtures at different molar ratios (MR). It was shown that the MR as well as the charge of the protein and LMWS determine the foam stability of the mixtures. For all mixtures it was found that the proteins have a select number of high affinity binding sites. So, the concentration of free LMWS in the solution is 0 until a critical MR (MRcr), at which all high affinity binding sites are saturated. Above this MRcr, part of the LMWS binds to low affinity binding sites of the proteins. The low affinity binding sites have a binding ratio < 1, which determines the concentration of bound and free LMWS. For similarly charged protein-LMWS mixtures (i.e. b-lactoglobulin (BLG) and sodium dodecyl sulphate (SDS) and bovine serum albumin (BSA) and SDS at pH 7) the foam stability typically decreases from the foam stability of the pure protein solution (MR 0) until MRcr is reached. At MR > MRcr the foam stability is dominated by the amount of free LMWS. For oppositely charged protein-LMWS mixtures, the binding of the LMWS to the proteins can be described in a similar way, although the number of high affinity sites and low affinity binding ratio are different. There is also a regime of MRs in which the protein-LMWS complexes form large aggregates. These aggregates were in some cases found to increase foam stability (lysozyme (LYS) and SDS and BLG-SDS at pH 3), while in another case (BLG and cetyltrimethylammonium bromide (CTAB)) they lead to decreased foam stability. Still, in all cases it was found that above MRD the aggregates dissociate and the foam stability becomes dominated by free surfactants, equivalent to what was observed for similarly charged protein-LMWS mixtures.

A multi-scale model was developed to describe the stability of thin liquid films in terms of rupture time and thickness. Initially, the model was used to predict the stability of surfactant free films of water and electrolyte solutions. Later, it was used to predict the foam stability in LYS-SDS mixtures. For that purpose, the model was combined with a foam drainage model to provide theoretical estimations of foam stability. This model is the basis to understand coalescence of bubbles in foam. Finally, the concept of the critical MRs and the free LMWS was introduced. Using this, the foam properties of protein-LMWS mixtures can partly be predicted by relative charge of the components and the binding to both high and low affinity binding sites.

Production of structured soy-based meat analogues using simple shear and heat in a Couette Cell
Krintiras, G.A. ; Gobel, T.W. ; Goot, A.J. van der; Stefanidis, G.D. - \ 2015
Journal of Food Engineering 160 (2015). - ISSN 0260-8774 - p. 34 - 41.
calcium caseinate dispersions - functional-properties - process parameters - extrusion-cooking - fibrous materials - protein isolate - wheat gluten - microstructure - mixtures - system
A Couette Cell device was employed to provide proof of concept for the production of structured meat analogues by application of simple shear flow and heat to a 31 wt% Soy Protein Isolate (SPI)–Wheat Gluten (WG) dispersion. Three relevant process parameters (temperature, time and rotation rate) were varied over a range of realistic values (90–110 °C, 5–25 min and 5–50 RPM, respectively). Layer- or fibre-structured products with high stress and strain anisotropy indices have been demonstrated. Fibrousness is favoured at temperatures over 90 °C and under 100 °C, whereas the role of process time and rotation rate is not critical. Simultaneous application of simple shear and heat is the key to obtaining structured plant protein-based products. The Couette Cell concept is scalable and can enable continuous operation. On this ground, it appears as a realistic option for production of meat analogues at commercial scale.
Polymer organogelation with chitin and chitin nanocrystals
Nikiforidis, C.V. ; Scholten, E. - \ 2015
RSC Advances : An international journal to further the chemical sciences 5 (2015)47. - ISSN 2046-2069 - p. 37789 - 37799.
mechanical characterization - supramolecular gels - lecithin organogel - molecular-weight - edible oils - chitosan - derivatives - morphology - chemistry - mixtures
In this paper, we show that biodegradable and biocompatible organogels can be formed with chitin as the filler material and triglycerides as the continuous hydrophobic phase. When crude chitin was used, a large degree of aggregation was observed that prevented the formation of stable organogels. Two approaches were used to diminish this degree of aggregation and increase the stability. Either surfactants were used to increase the dispersability of the crude chitin, or the crude chitin was transformed into smaller rod-like nanocrystals by acid hydrolysis. Both approaches led to the formation of stable organogels with storage moduli up to 106 Pa for high chitin concentrations (20 wt%). Three different types of surfactants were used, namely phosphatidylcholine, enzymatically modified phosphatidylcholine and sorbitan monostearate (Span 60). The choice of surfactant has a large influence on the gel strength and the temperature sensitivity of the gels. With chitin nanocrystals, in the presence of surfactants, larger gel strengths were observed for lower concentrations (1-10 wt%), indicating more efficient packing of the particles. Gels were stable even after addition of considerable amounts of water up to 25 wt%. The increase in gel strength in the presence of water (storage modulus) was most likely an effect of the water absorption ability of chitin that increased the effective volume fraction of the fillers.
Surfactant-polymer interactions: molecular architecture does matter
Banerjee, S. ; Cazeneuve, C. ; Baghdadli, N. ; Ringeissen, S. ; Leermakers, F.A.M. ; Luengo, G.S. - \ 2015
Soft Matter 11 (2015). - ISSN 1744-683X - p. 2504 - 2511.
block copolymer adsorption - nonionic surfactants - statistical thermodynamics - chain molecules - phase-behavior - simulation - interface - association - mixtures - water
Polymer–surfactant mixtures are found in many industrial formulations, and hence there is a significant interest in understanding, at a molecular level, how the self-assembly of surfactant is affected by oppositely-charged polyelectrolytes (PEs). We use self-consistent field modeling and show that the modes of interaction of PEs strongly depend on the architecture of the PE on the segmental level. Hydrophilic cationic PEs with their charge proximal to the linear backbone are expected to bind electrostatically to the outsides of the coronas of the spherical micelles of anionic surfactants, such as sodium laureth sulphate (SLES). As a result, the surfactant aggregation number increases, but at the same time the colloidal stability deteriorates, due to bridging of the PEs between micelles. PEs with their charge somewhat displaced from the backbone by way of short hydrophobic spacers, are expected to be present inside a micelle at the core–corona boundary. In this case the aggregation number decreases, yet the colloidal stability is retained. Hence, SLES tends to remove hydrophilic PEs from an aqueous solution, whereas it solubilizes more hydrophobic ones. The binding isotherm shows that the uptake of PEs remains typically below charge compensation and in this case the spherical micelle topology remains the preferred state.
Effects of salt on the expansion of starchy snacks: a multiscale analysis
Sman, R.G.M. van der; Broeze, J. - \ 2014
Food & Function 5 (2014)12. - ISSN 2042-6496 - p. 3076 - 3082.
glass-transition - sodium-chloride - aqueous-solutions - mass-transfer - sucrose - extrusion - potato - water - trehalose - mixtures
We investigate the effect of salt on the expansion of starchy snacks during frying by means of a multiscale simulation model. This model has been developed earlier for starchy snacks without salt. The simulation results are analysed by means of the supplemented state diagram. We have found that the optimal expansion for salty snacks occurs under the same conditions as for snacks without salt. This occurs at the moisture content where the 4 bar boiling line intersects the critical isoviscosity line of 1 MPa s. Salt is shown to influence both the boiling line and the critical isoviscosity line, via a change of the glass transition. The optimal moisture content for salty snacks is lower than that of unsalted snacks. We view our findings as important for reformulations of starchy snacks with lower salt levels. Furthermore, the presented tools of the multiscale simulations and supplemented state diagram can generally be used for reformulation problems in structured foods.
Competitive adsorption of the protein hydrophobin and an ionic surfactant: Parallel vs sequential adsorption and dilatational rheology
Stanimirova, R. ; Marinova, K.G. ; Danov, K.D. ; Kralchevsky, P.A. ; Basheva, E.S. ; Stoyanov, S.D. ; Pelan, E.G. - \ 2014
Colloids and Surfaces. A: Physicochemical and Engineering Aspects 457 (2014). - ISSN 0927-7757 - p. 307 - 317.
air-water-interface - sodium dodecyl-sulfate - beta-casein - air/water interface - fluid interfaces - layers - hfbii - elasticity - stability - mixtures
The competitive adsorption of the protein HFBII hydrophobin and the anionic surfactant sodium dodecyl sulfate (SDS) is investigated in experiments on parallel and sequential adsorption of the two components. The dynamic surface tension and the surface storage and loss dilatational moduli are determined by the oscillating bubble method. A new procedure for data processing is proposed, which allows one to collect data from many different runs on a single master curve and to determine more accurately the dependence of the dilatational elasticity on the surface pressure. Experiments on sequential adsorption are performed by exchanging the HFBII solution around the bubble with an SDS solution. Experiments with separate thin foam films bring additional information on the effect of added SDS. The results indicate that if HFBII has first adsorbed at the air/water interface, it cannot be displaced by SDS at any concentration, both below and above the critical micellization concentration (CMC). In the case of parallel adsorption, there is a considerable difference between the cases below and above the CMC. In the former case, SDS cannot prevent the adsorption of HFBII at the interface, whereas in the latter case adsorption of HFBII is absent, which can be explained with hydrophilization of the hydrophobin aggregates by the SDS in the bulk. The surface dilatational elasticity of the HFBII adsorption layers markedly decreases in the presence of SDS, but it recovers after washing out the SDS. With respect to their dilatational rheology, the investigated HFBII layers exhibit purely elastic behavior, the effect of dilatational viscosity being negligible. As a function of surface tension, the elasticity of the investigated interfacial layers exhibits a high maximum, which could be explained with the occurrence of a phase transition in the protein adsorption layer.
The effect of fibers on coagulation of casein-based enteral nutrition in an artificial gastric digestion model
Luttikhold, J. ; Norren, K. van; Minor, M. ; Buijs, N. ; Braak, C.C.M. van den; Ludwig, T. ; Abrahamse, E. ; Rijna, H. ; Leeuwen, P.A.M. - \ 2014
Food & Function 5 (2014). - ISSN 2042-6496 - p. 1866 - 1871.
critically-ill patients - thermodynamic incompatibility - intestinal-obstruction - in-vitro - motility - proteins - absorption - guidelines - mixtures - feedings
A serious complication seen in critically ill patients is the solidification of enteral nutrition causing gastrointestinal obstruction. It has been suggested that enteral nutrition enriched with insoluble fibers may increase the risk of this complication. Therefore, we investigate the effect of soluble and insoluble dietary fibers on the coagulation of a casein-based enteral nutrition in an artificial gastric digestion model. A 100% casein-based enteral nutrition was enriched with increasing concentrations of soluble fibers (acacia fiber, oligofructose and inulin) and insoluble fibers (soy polysaccharide, resistant starch and alpha cellulose). After digestion in an artificial gastric model, the chyme was poured over sequentially placed sieves, separating the coagulate into size fractions of larger than 2 mm, between 1 and 2 mm, and between 0.25 and 1 mm. Of these fractions we measured wet weight, dry weight and protein content. A significant effect on the fraction larger than 2 mm was considered to be clinically relevant. Addition of high concentrations soy polysaccharide and resistant starch to a casein-based enteral nutrition, did not alter the wet weight, whereas dry weight and protein content of the coagulate was significantly reduced. When high concentrations of soy polysaccharide and resistant starch are added to a 100% casein-based enteral nutrition, the coagulate consist of more water and less proteins, which may lead to an increased protein digestion and absorption in a clinical setting. The suggestion that insoluble fibers increase the risk of gastrointestinal obstruction in critically ill patients is not supported by these data.
Self-assemblies of lecithin and a-tocopherol as gelators of lipid material
Nikiforidis, C.V. ; Scholten, E. - \ 2014
RSC Advances : An international journal to further the chemical sciences 4 (2014)5. - ISSN 2046-2069 - p. 2466 - 2473.
edible oils - rheological properties - reverse micelles - drug-delivery - fatty-acids - organogels - gels - mixtures - agents
Amongst the different mechanisms that have been proposed and used to structure organogels, self-assembly of the gelators into supramolecular structures linked through non-covalent bonds is the most interesting. The gelator activity of LMGOs is often found most effective when micellar or lamellar phases are obtained, which is dependent on the gelator geometry and the specific packing parameter. Gelation can therefore be induced by altering the packing parameter of different gelators, but due to the law restrictions there are only a few edible gelators that can be used to structure edible lipids. Here, we show that a combination of a-tocopherol and phosphatidylcholine (PC) can be used to alter the packing geometry to provide supramolecular structures needed for the organogelation. We have observed that when the gelators were combined at 1:1 ratio in sunflower oil, edible organogels were obtained. The firmness of the solid-like material was enhanced when 1.0 wt% of water was added. The proposed mechanism for this assembly is that most likely cylindrical micellar structures are formed, due to combined assembly of the a-tocopherol and phosphatidylcholine, stabilized through physical interactions. Since these interactions, and the accompanied packing geometry, depends on temperature and application of external stresses, the formation of the organogels showed reversibility when the organogels were subjected to shear or when the temperature was increased to values above 35 oC. Polarized microscopy along with small angle X-ray scattering were used to provide a hypothesis for the mechanism behind the gelation.
Segregation of Granular Particles by Mass, Radius, and Density in a Horizontal Rotating Drum
Arntz, M.M.H.D. ; Beeftink, H.H. ; Otter, W.K. den; Briels, W.J. ; Boom, R.M. - \ 2014
AIChE Journal 60 (2014)1. - ISSN 0001-1541 - p. 50 - 59.
size segregation - particulate systems - flow - simulation - cylinders - mixtures - dynamics - solids - behavior - motion
The impact of particle properties on segregation and mixing of bidisperse granular beds in a rotating horizontal drum have been studied by discrete element method (DEM) simulations. Bidispersities in radius, density, and mass have pronounced influences on the stationary mixing pattern, although they hardly affect the granules’ flow regime. At 50% fill level, all beds mix well for a Froude number of 0.56, corresponding to a flow regime intermediate to cascading and cataracting, while segregation occurs both at lower (rolling and cascading regime) and higher (cataracting/centrifuging regime) Froude numbers. These observations are explained qualitatively by noticing that the angular drum velocity dictates the flow regime, which in turn determines the effectiveness and direction of four competing (de)mixing mechanisms: random collisions, buoyancy, percolation, and inertia. A further dozen particle properties have been varied, including the friction coefficients and elastic modulus, but these proved inconsequential to the steady-state degree of mixing.
Moisture Sorption Isotherms of Broccoli Interpreted with the Flory-Huggins Free Volume Theory
Jin, X. ; Sman, R.G.M. van der; Maanen, J.F.C. van; Deventer, H.C. van; Straten, G. van; Boom, R.M. ; Boxtel, A.J.B. van - \ 2014
Food Biophysics 9 (2014)1. - ISSN 1557-1858 - p. 1 - 9.
water activity - glassy-polymers - drying methods - carrot slices - tissue-damage - quality - vegetables - mixtures - heat - pretreatment
In this work, the Flory Huggins Free Volume theory is used to interpret the sorption isotherms of broccoli from its composition and using physical properties of the components. This theory considers the mixing properties of water, biopolymers and solutes and has the potential to describe the sorption isotherms for varying product moisture content, composition and temperature. The required physical properties of the pure components in food became available in recent years and allow now the prediction of the sorption isotherms with this theory. Sorption isotherm experiments have been performed for broccoli florets and stalks, at two temperatures. Experimental data shows that the Flory Huggins Free Volume (FHFV) theory represents the sorption isotherm of fresh and blanched broccoli samples accurately. The results also show that blanching affects the sorption isotherm due to the change of composition via leaching solutes and the change of interaction parameter due to protein denaturation
Interfacial tension and wettability in water-carbon dioxide systems: Experiments and self-consistent field modeling
Banerjee, S. ; Hassenklover, E. ; Kleijn, J.M. ; Cohen Stuart, M.A. ; Leermakers, F.A.M. - \ 2013
The Journal of Physical Chemistry Part B: Condensed Matter, Materials, Surfaces, Interfaces & Biophysical 117 (2013)28. - ISSN 1520-6106 - p. 8524 - 8535.
block copolymer adsorption - statistical thermodynamics - elevated pressures - mixtures - co2 - temperature - surfaces - density - mpa
This paper presents experimental and modeling results on water–CO2 interfacial tension (IFT) together with wettability studies of water on both hydrophilic and hydrophobic surfaces immersed in CO2. CO2–water interfacial tension (IFT) measurements showed that the IFT decreased with increasing pressure and the negative slopes of IFT–pressure isotherms decreased with increasing temperature. Water contact angle on a cellulose surface (hydrophilic) immersed in CO2 increased with pressure, whereas the water contact angle on a hydrophobic surface such as hexamethyl disilazane (HMDS) coated silicon surface was almost independent of pressure. These experimental findings were augmented by modeling using the self-consistent field theory. The theory applies the lattice discretization scheme of Scheutjens and Fleer, with a discretization length close to the size of the molecules. In line with this we have implemented a primitive molecular model, with just small variations in the molar volume. The theory makes use of the Bragg-Williams approximation and has binary Flory–Huggins interaction parameters (FH) between CO2, water, and free volume. Using this model, we generated the complete IFT–pressure isotherms at various temperatures, which coincided well with the trends reported in literature, that is, the water–CO2 interfacial tension decreased with increasing pressure for pressures =100 bar and became independent of pressure >100 bar. The transition point occurred at higher pressures with increasing temperature. At three-phase coexistence (water–CO2–free volume) and at the water–vapor interface (water–free volume), we always found the CO2 phase in between the water-rich and free volume-rich phases. This means that for the conditions studied, the water–vapor interface is always wet by CO2 and there are no signs of a nearby wetting transition. Calculation of the water contact angle on a solid surface was based on the computed adsorption isotherms of water from a vapor or from a pressurized CO2-rich phase and analysis of surface pressures at water–vapor or water–CO2 coexistence. The results matched reasonably well with the experimental contact angle data. Besides, we also computed the volume fraction profiles of the CO2, H2O, and the V phase, from which the preferential adsorption of CO2 near the hydrophilic surface was deduced.
Editorial: Dynamics of complex fluid-fluid interfaces
Sagis, L.M.C. ; Fischer, P. ; Anderson, P.D. - \ 2013
The European Physical Journal. Special Topics 222 (2013)1. - ISSN 1951-6355 - p. 1 - 5.
in-water emulsions - liquid interfaces - bending rigidity - rheology - fibrils - viscoelasticity - permeability - particles - mixtures - behavior
Generalized surface momentum balances for the analysis of surface dilatational data
Sagis, L.M.C. - \ 2013
The European Physical Journal. Special Topics 222 (2013)1. - ISSN 1951-6355 - p. 31 - 38.
in-water emulsions - interfacial permeability - superficial viscosity - bending rigidity - lipid-bilayers - fluid - dynamics - liquid - microbubbles - mixtures
Dilatational rheological properties of interfaces are often determined using drop tensiometers, in which the interface of the droplet is subjected to oscillatory area changes. A dynamic surface tension is determined either by image analysis of the droplet profile or by measuring the capillary pressure. Both analysis modes tend to use the Young-Laplace equation for determining the dynamic surface tension. For complex fluid-fluid interfaces there is experimental evidence that this equation does not describe the response of the interface to deformations adequately. Generalizations of this equation are available, and in this comment we will discuss these generalizations, and the conditions for which they reduce to the Young-Laplace equation.
Towards a realistic risk characterization of complex mixtures using in vitro bioassays
Montano Garces, M. - \ 2013
Wageningen University. Promotor(en): Tinka Murk, co-promotor(en): A.C. Gutleb. - [S.l.] : s.n. - ISBN 9789461736598
risicoschatting - persistente organische verontreinigende stoffen - verontreinigde sedimenten - mengsels - biotesten - in vitro - toxiciteit - schildklierhormonen - risk assessment - persistent organic pollutants - contaminated sediments - mixtures - bioassays - in vitro - toxicity - thyroid hormones

This thesis aims to better understand and further improve the relevance and reliabilityof in vitro bioassaysfor a biobased risk characterisation of complex mixtures, with special focus on persistent organic pollutants (POPs) in sediments.

In Chapter 1 the importance of complex mixture characterization in modern society is introduced. The methods available, their current advantages and their disadvantages for complex mixture testing are described. With the shift from policy oriented chemical testing towards the inclusion of in vitro bioanalysis, important challenges have to be overcome to ensure a relevant and reliable quantification of the toxic potency of complex mixtures. These challenges are explained in the introduction, including the status of development and validation of those aspects for reliable testing. One of the main advantages that in vitro bioanalysis has to offer is the possibility to quantify the toxic potency of compounds for which chemical analytical methods have not or hardly been developed, for example because standards do not yet exist. Hydroxylated metabolites of POPs are an example of a toxicologically relevant group of compounds that can exert endocrine disrupting effects, but they cannot yet be routinely analysed. A selection of yet unsolved issues are further studied and discussed in this thesis, as outlined in the “approach and structure of the thesis”.

In Chapter 2 a meta-analysis is performed to study the occurrence and relevance of hydroxylated (OH) compounds in humans and wildlife. Reported body burdens of halogenated phenolic contaminants (HPCs), including OH-POP in different tissues from humans and wildlife species, are reviewed in relation to the concentration of their putative parent compounds to be able to reveal relevant exposure routes and sub-populations at risk. Highest OH-POP levels were found in blood plasma, and highly perfused and fetal tissues. Plasma concentrations of analysed known HPCs ranged from 0.1-100 nM in humans and up to 240, 454, 800 and 7650 nM for birds, fish, cetaceans and other mammals, respectively. Reported metabolite blood plasma levels also are compared with relevant toxicological threshold concentrations from toxicological studies, and appeared to fully fall within the in vitro (0.05–10000 nM) and in vivo (3-940 nM) effect concentrations reported for OH-POPs. Given the sensitivity of early developmental stages, and information lacking about the general population, it is advisable to determine HPC background blood levels in children and fetal tissue .

Given the toxicological relevance of the OH-POPs, Chapter 3 aims at providing solutions to the long standing problem of the in vitro production and analysis of OH-POP metabolite thyroid hormone disrupting (THD) potency via binding to plasma thyroid hormone binding proteins (THBPs). In sediments and for example seafood, the POPs occur as parent compounds that would only become THD after metabolisation (hydroxylation). Several methods have shown the competitive thyroxine (T4) T4 displacement potency of pure metabolites. However, in vitro metabolization of, among others, polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers(PBDEs) followed by in vitro quantification of their potency has encountered drawbacks related to the co-extraction of compounds disturbing the T4-TTR competitive binding assay. The present study identifies and quantifies the major co-extractants, cholesterol and saturated and non-saturated fatty acids (SFA and NSFA), at levels above 20 μM (20 nmol per mg protein in the incubation mixture) following various extraction methods. A new method is presented to in vitro metabolise parent compounds into OH-metabolites followed by selective extraction of metabolites while four-fold reducing co-extraction of the disturbing compounds. In addition a microplate-format non-radioactive fluorescence displacement assay was developed to quantify the TTR binding potency of the metabolites formed. The effectiveness of the in vitro metabolism and extraction of the OH-metabolites of the model compounds CB 77 and BDE 47 was chemically quantified with a newly developed chromatographic method analyzing silylated derivatives of the OH-metabolites and co-extractants. Due to the mentioned improvements, it is now possible to make a dose-response curve up to 50% inhibition with OH-metabolites extracted from bioactivated CB 77 and BDE 47. Without taking the toxic potencies of bio-activated POPs into account with bioanalysis, the hazard and risk posed by POPs will be seriously underestimated.

The chapters 4 and 5 are committed to tackle the issues of supramaximal (SPMX) responses and sample extract concentration which are crucial to reliably quantify of the toxic potencies of complex mixtures with in vitro bioassays.

A SPMX effect is the phenomenon that compounds induce a maximum response in an assay that is significantly higher than that of the positive control. As the positive control is used to quantify the toxic potency of a sample, this could result in over-estimation of its toxic potency. As this has been most elaborately reported for in vitro estrogenicity assays, a meta-analysis was performed of such assays, compounds and conditions in which the effect is observed (Chapter 4a).For the 21 natural and industrial chemicals that could be identified as SPMX inducers, the culture and exposure conditions varied greatly among and between the assays. Relevant information on assay characteristics, however, sometimes lacked. Diethylstilbestrol (DES), genistein (GEN) and bisphenol A (BPA) were selected to build a database. The meta-analysis revealed that the occurrence of SPMX effects, could be related to a number of specific assay characteristics: 1) the type and concentration of the serum used to supplement the exposure medium; 2) the endpoint used to quantify the estrogenic potency (endogenous or transfected reporter gene), 3) the number of EREs (estrogen responsive elements) used before the reporter gene, and 4) the nature of the promoter’s. There were no indications that solvent concentration in culture, exposure period or cell model influenced the occurrence of SPMX. It is important to understand the mechanism behind this phenomenon because in vitro assays for estrogenicity are used extensively to characterize and quantify the estrogenic potency of compounds, mixtures and environmental extracts.

Several SPMX inducers also have been reported to block cellular efflux pumps in vivo and in vitro (Anselmo et al. 2012; Georgantzopoulou et al. 2013). Therefore it was hypothesized that efflux pump blockers present in environmental matrices could increase the internal concentration of bioassay agonists and thus cause the SPMX. In Chapter 4b this hypothesis was tested by adapting a 96-well plate cellular efflux pump inhibition assay (CEPIA) to the H4IIE rat hepatoma cell line used for the DR.Luc reporter gene assay for dioxin-like compounds. The influence of various environmentally relevant efflux pump inhibitors on the 2,3,7,8-tetrachlorodibenzo-p-dioxine (TCDD) response was tested. Under the DR.Luc assay conditions there was no evidence that P-gp efflux pump inhibitors modified or potentiated the activity of TCDD. Neither genistein nor quercetin, two potent SPMX inducers on ER-mediated assays, induced any signal on the DR.Luc assay, nor influenced the luciferase induction by TCDD. Future work should be focused on testing the consequences of efflux pump inhibition with an AhR-agonist which is a P-gp substrate, as this could result in intracellular accumulation of this AhR-agonist.

It is standard practice to use a high single stock concentration of extracts to further dilute test concentrations from and perform the analysis. However, a high contaminant load in an extract may oversaturate the solubility of the extracted compounds in carrier solvents and overload the clean-up columns which may reduce the efficiency of polyaromatic hydrocarbons (PAHs) elimination from the extract. These problems may cause respectively under- or over-estimation of the quantified dioxin-like toxic potency. Therefore Chapter 5 focuses on the effects of initial stock concentrations, including sonication assisted dissolution and exposure time, on the quantified dioxin-like potency of cleaned nonpolar sediment extracts. Indeed, more than 20 g sediment equivalents (SEQ)/mL DMSO) as initial stock concentrations resulted in underestimation of bio-TEQ levels in the sediments as observed for cleaned nonpolar sediment extracts from various locations in Luxembourg. An overload of extract on clean-up columns caused an over-estimation of the dioxin-like potency at 24 hours of exposure, probably due to limited removal of PAHs that can induce false positive responses in the in vitro assays. Sonication assisted dissolution of the stock before serial dilution strongly reduced the standard variation of the outcomes. Taking into account the aspects revealed in this study, in addition to already described important issues for quality control, the in vitro bioassays based bio-TEQs can be applied in a comprehensive monitoring program to determine whether sediments comply with health and safety standards for humans and the environment. For the generally applied sediment quality criteria, advices are given about maximum initial stock concentrations to achieve reliable bioassay outcomes.

The methods and concepts developed for metabolic activation of compounds in non-polar sediment extracts and in in vitro analysis of the TTR-competitive binding are applied in Chapter 6 to extracts from highly or less contaminated sediments collected in Luxembourg. Nonpolar fractions of sediment extracts were incubated with S9 rat microsomes, and the metabolites were extracted with a newly developed method that excludes most of the lipids to avoid interference in the non-radioactive 96-well plate transthyretin (TTR) competitive binding assay. Metabolic activation increased the TTR binding potency of nonpolar fractions of POP-polluted sediments up to 100 times, resulting in potencies up to 240 nmol T4 equivalents/g sediment equivalent (nmol T4-Eq/g SEQ). Without bioactivation, medium polar and polar fractions also contained potent TTR-binding compounds with potencies from 1.6 to 17 nmol T4-Eq/g SEQ. This demonstrates that a more realistic in vitro sediment THD risk characterization should also include testing ofboth polar and medium polar sediment extracts for THD, as well as bioactivated nonpolar sediment fractions. Without bioactivation THD potency is not observed in nonpolar sediment extracts, although in in vivo experiments PCBs and PBDEs, and not with dioxins or PAHs, have shown to be thyroid hormone disrupting (THD), demonstrating this bio-activation is toxicologically relevant and therefore required for sediment hazard characterisation.

Chapter 7 discusses the implications of our results to improve the relevance and reliability of in vitro bioassay applied for risk characterisation of complex mixtures from sediments and other matrices. The evidence obtained to support the relevance of POP bio-activation is considered both from the exposure perspective as well as the toxicity perspective. Various features of the newly developed methods and knowledge acquired within this PhD project are discussed in relation to in vitro bioassay risk characterization of sediments towards a realistic in vitro bioassay-based risk characterization of complex mixtures. Some important aspects for the inclusion of metabolizing systems within in vitro bioassay are discussed. In addition, alternatives to deal with the SPMX effect and the definition of suitable sample amounts to improve in vitro bioassay reliability are offered. The suitability of the developed approach application is considered for the risk characterization of sediments. Furthermore, an analysis is made to decide whether this thesis have made in vitro bioassays more reliable and relevant for risk characterization of complex mixtures. Finally, it provides some concluding remarks and aspects for further applications and research.

Colorimetry and photoacoustic spectroscopy as suitable tools for direct determination of cocoa powder in confectionary products
Doka, O. ; Pragai, E. ; Bicanic, D.D. ; Kulcsar, R. - \ 2013
European Food Research and Technology 236 (2013)6. - ISSN 1438-2377 - p. 963 - 968.
phenolic content - theobroma-cacao - powders - antioxidant - quantification - polyphenols - mixtures - capacity - samples - beans
Laser photoacoustic spectroscopy (PAS) and colorimetry have been used to rapidly and accurately determine the content of fat-free cocoa solids in dark chocolates. Both techniques are inexpensive and require only a one-time calibration step versus a method capable of absolute concentration measurement (for example HPLC). Once the response of PAS and/or colorimetry has been obtained, the content of fat-free cocoa solids in dark chocolates can be determined directly (i.e. without any sample preparation including the process of extraction) from the calibration curves and the experimentally measured microphone signal (in PAS studies) and colorimetric indices L* and ¿E* (in colorimetric investigations). Both colorimetric indices and PA signals correlate positively with the content of fat-free cocoa solids. The correlation is highly linear over a wide concentration range (25–50 %).
The influence of the type of oil phase on the self-assembly process of ¿-oryzanol + ß-sitosterol tubules in organogel systems
Sawalha, H.I.M. ; Margry, G. ; Adel, R. den; Venema, P. ; Bot, A. ; Flöter, E. ; Linden, E. van der - \ 2013
European Journal of Lipid Science and Technology 115 (2013)3. - ISSN 1438-7697 - p. 295 - 300.
edible oils - emulsions - mixtures
Mixtures of ¿-oryzanol and ß-sitosterol were used to structure different oils (decane, limonene, sunflower oil, castor oil and eugenol). The ¿-oryzanol and ß-sitosterol mixtures self-assemble into double-walled hollow tubules (~10 nm in diameter) in the oil phase, which aggregate to form a network resulting in firm organogels. The self-assembly of the sterol molecules into tubules was studied using light scattering and rheology. By using different oils, the influence of the polarity of the oil on the self-assembly was studied. The effects of temperature and structurant concentration on the tubular formation process were determined and the thermodynamic theory of self-assembly was applied to calculate the change in Gibbs free energy ( DG0), enthalpy (DH0) and entropy ( DS0) resulting from the aggregation of the structurants was determined. The self-assembly was found to be enthalpy-driven as characterized by a negative DH0 and DS0. A decreasing polarity of the oil promotes the self-assembly leading to formation of tubules at higher temperatures and lower structurants concentrations.
Leaf litter quality drives litter mixing effects through complementary resource use among detritivores
Vos, V.C.A. ; Ruijven, J. van; Berg, M.P. ; Peeters, E.T.H.M. ; Berendse, F. - \ 2013
Oecologia 173 (2013)1. - ISSN 0029-8549 - p. 269 - 280.
species-diversity - ecosystem function - soil processes - decomposition - biodiversity - mixtures - forest - respiration - millipedes - richness
To comprehend the potential consequences of biodiversity loss on the leaf litter decomposition process, a better understanding of its underlying mechanisms is necessary. Here, we hypothesize that positive litter mixture effects occur via complementary resource use, when litter species complement each other in terms of resource quality for detritivores. To investigate this, monocultures and mixtures of two leaf litter species varying in quality were allowed to decompose with and without a single macro-detritivore species (the terrestrial woodlice Oniscus asellus). Resource quality of the mixture was assessed by the mean concentration, the dissimilarity in absolute and relative concentrations, and the covariance between nitrogen (N), phosphorus (P) and calcium (Ca) supply. Our results clearly show that litter mixing effects were driven by differences in their resource quality for detritivores. In particular, complementary supply of N and P was a major driver of litter mixing effects. Interestingly, litter mixing effects caused by the addition of woodlice were predominantly driven by N dissimilarity, whereas in their absence, increased P concentration was the main driver of litter mixing effects. These results show that ultimately, litter diversity effects on decomposition may be driven by complementary resource use of the whole decomposer community (i.e., microbes and macro-detritivores).
The influence of end walls on the segregation pattern in a horizontal rotating drum
Arntz, M.M.H.D. ; Otter, W.K. den; Beeftink, H.H. ; Boom, R.M. ; Briels, W.J. - \ 2013
Granular Matter 15 (2013)1. - ISSN 1434-5021 - p. 25 - 38.
reversible axial segregation - granular-materials - mixtures - simulation - cylinders - dynamics - motion
The influence of end walls on segregation of bidisperse granular beds in a short rotating horizontal drum is studied by a discrete element method. Whereas non-closed periodically continued drums segregate radially, all simulations of drums with end walls resulted in axial segregation with two bands at low friction between the particles and the end-wall, and three bands at high friction. Various simulations show irregular transitions between two approximately equally stable states, with rapid oscillations preceding the conversions. The formation of two axial bands decreases the energy dissipation by the bed, whereas neither radial segregation nor axial segregation into three bands reduced the power absorption at constant angular velocity. Roughening up the end-walls also increased the rate of axial segregation.
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