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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Review : Converting nutritional knowledge into feeding practices: A case study comparing different protein feeding systems for dairy cows
Lapierre, H. ; Larsen, M. ; Sauvant, D. ; Amburgh, M.E. van; Duinkerken, G. van - \ 2018
Animal (2018). - ISSN 1751-7311 - 10 p.
dairy cows - energy - feeding system - milk protein yield - protein

Improving milk nitrogen efficiency through a reduction of CP supply without detrimental effect on productivity requires usage of feeding systems estimating both the flows of digestible protein, the exported true proteins and from these predict milk protein yield (MPY). Five feeding systems were compared in their ability to predict MPY v. observed MPY in two studies where either protein supply or protein and energy supply were changed. The five feedings systems were: Cornell Net Carbohydrate and Protein System (v6.5.5), Dutch protein evaluation system (1991 and 2007), Institut National de la Recherche Agronomique in France (INRA), National Research Council and NorFor. The key characteristic of the systems with the best predicted MPY was the inclusion of a variable efficiency of utilisation of protein supply taking into account the supply of both protein and energy. The systems still using a fixed efficiency had the highest slope bias in their prediction of MPY. Therefore, the development of new feeding systems or improvement of existing systems should include a variable efficiency of utilisation of the protein related to both the protein and energy supply. The limitation of the current comparison did not allow determining if additional factors, as used in INRA, were beneficial. This concept should also probably be transferred to essential amino acids.

Soya bean meal increases litter moisture and foot pad dermatitis in maize and wheat based diets for turkeys but maize and non-soya diets lower body weight
Hocking, P.M. ; Vinco, L.J. ; Veldkamp, T. - \ 2018
British Poultry Science 59 (2018)2. - ISSN 0007-1668 - p. 227 - 231.
Cereal - dermatitis - diet - electrolyte balance - feed - feed intake - litter moisture - protein
1. A 2 × 2 factorial experiment was conducted to compare the effects of wheat or maize based diets differing in dietary electrolyte balance (DEB) on litter moisture and foot pad dermatitis (FPD) at 4, 8 and 12 weeks of age in heavy-medium turkeys. A second objective was to investigate the effects on foot pad dermatitis of the interaction between dietary composition and artificially increasing litter moisture by adding water to the litter. 2. High DEB diets contained soya as the main protein source whereas low DEB diets did not contain soya bean meal. Diets were formulated to be iso-caloric and iso-nitrogenous in each of 3 successive 4-week phases following recommended dietary compositions. DEB concentrations were 330, 290 and 250 mEq/kg in high DEB diets and 230, 200 and 180 mEq/kg in low DEB diets. 3. Litter moisture and mean FPD score were higher in turkeys fed on high DEB diets compared with low DEB diets whereas there was no difference between maize and wheat. 4. Food intake was similar and body weight was lower after litter moisture was artificially raised in the wet compared with the dry litter treatment and there was no interaction with dietary composition. 5. Mean body weight and feed intake were higher in turkeys fed on wheat compared with maize and in high DEB compared with low DEB diets at 12 weeks of age. 6. Lowering dietary DEB for turkeys may improve litter moisture and lower the prevalence of FPD in commercial turkey flocks.
Sustainable protein technology : An outlook for further research
Voudouris, Panagiotis ; Tamayo Tenorio, Angelica ; Lesschen, Jan Peter ; Kyriakopoulou, Konstantina ; Sanders, Johan P.M. ; Goot, Atze Jan van der; Bruins, Marieke E. - \ 2017
Wageningen : Wageningen Food & Biobased Research - 6 p.
nutrition - biobased economy - protein
Nutritional composition of black soldier fly (Hermetia illucens) prepupae reared on different organic waste substrates
Spranghers, Thomas ; Ottoboni, Matteo ; Klootwijk, Cindy ; Ovyn, Anneke ; Deboosere, Stefaan ; Meulenaer, Bruno De; Michiels, Joris ; Eeckhout, Mia ; Clercq, Patrick De; Smet, Stefaan De - \ 2017
Journal of the Science of Food and Agriculture 97 (2017)8. - ISSN 0022-5142 - p. 2594 - 2600.
black soldier fly - fatty acid, amino acid - feed - protein - vegetable waste processing
BACKGROUND: Black soldier fly larvae are converters of organic waste into edible biomass, of which the composition may depend on the substrate. In this study, larvae were grown on four substrates: chicken feed, vegetable waste, biogas digestate, and restaurant waste. Samples of prepupae and substrates were freeze-dried and proximate, amino acid, fatty acid and mineral analyses were performed. RESULTS: Protein content of prepupae varied between 399 and 431 g kg−1 dry matter (DM) among treatments. Differences in amino acid profile of prepupae were small. On the other hand, the ether extract (EE) and ash contents differed substantially. Prepupae reared on digestate were low in EE and high in ash (218 and 197 g kg−1 DM, respectively) compared to those reared on vegetable waste (371 and 96 g kg−1 DM, respectively), chicken feed (336 and 100 g kg−1 DM, respectively) and restaurant waste (386 and 27 g kg−1 DM, respectively). Prepupal fatty acid profiles were characterised by high levels of C12:0 in all treatments. CONCLUSION: Since protein content and quality were high and comparable for prepupae reared on different substrates, black soldier fly could be an interesting protein source for animal feeds. However, differences in EE and ash content as a function of substrate should be considered.
Undernutrition management and the role of protein-enriched meals for older adults
Ziylan, Canan - \ 2016
University. Promotor(en): Lisette de Groot; Stefanie Kremer; Annemien Haveman-Nies. - Wageningen : Wageningen University - ISBN 9789462579323 - 148
elderly - elderly nutrition - undernutrition - enrichment - protein - eating patterns - feeding behaviour - meals - nursing homes - ouderen - ouderenvoeding - ondervoeding - verrijking - eiwit - eetpatronen - voedingsgedrag - maaltijden - verpleeghuizen

Undernutrition is a major health problem in the growing elderly population. It is estimated that one in ten Dutch community-dwelling older adults is suffering from undernutrition, and one in three Dutch older adults who receive home care. Undernutrition may lead to many negative consequences, ranging from fatigue and falls to impaired immune function and death. This makes undernutrition an obvious target for preventive measures.

Undernutrition can be defined as “a state of nutrition in which a deficiency or excess (or imbalance) of energy, protein, and other nutrients causes measurable adverse effects on tissue/body form (body shape, size and composition) and function, and clinical outcome”. In addition, it is often described as protein energy malnutrition. Adequate protein intake may to some extent prevent and reverse this process. However, throughout ageing, it becomes increasingly difficult to reach adequate protein intake due to higher protein needs and lower protein intakes. Finding solutions to assist older adults in reaching their optimal protein intake is necessary.

In our overall research project, we considered 1.2g protein per kg weight per day (g/kg/d) as adequate protein intake. In Dutch community-dwelling older adults, protein intake is around 1.0 g/kg/d, implying room for improvement. However, it is possible that many of these older adults deal with physiological changes, medical conditions, and physical and mental limitations that impair their appetite and food provision. For these older adults with higher protein needs, merely recommending that they eat more would not be realistic. It would be more realistic to explore strategies that increase protein intake without having to increase food intake. This calls for the exploration of instruments that match the needs and preferences of older adults: protein-enriched regular products.

One particular group that can be identified as a target group for such products, are older adults who receive home care. Undernutrition prevalence is high in this group, which may be explained by their health problems that led to this dependence on home care. Likewise, many of these older adults also depend on meals-on-wheels. These meals-on-wheels recipients, regardless of whether they receive home care or not, often risk undernutrition too. In both these (overlapping) care-dependent groups, difficulties in adhering to energy and protein recommendations can be discerned. For this reason, enriching the readymade meals that these older adults receive may contribute to the prevention of protein undernutrition by increasing protein intake while keeping food intake the same. Here, protein enrichment instruments can be used to prevent undernutrition, but only when implemented in a timely manner. Adequate undernutrition management systems are therefore necessary to facilitate timely intervention, ensuring that the developed protein-enriched meals are actually offered and effective. For this reason, the overall aim of our research project was to gain insight into the current state of undernutrition management in community-dwelling older adults in the Netherlands and explore the role of protein-enriched regular products as a supportive instrument in protein undernutrition management.

In Study 1 (chapter 2) we explored the experiences of 22 Dutch nutrition and care professionals and researchers with undernutrition awareness, monitoring, and treatment among community-dwelling older adults. This qualitative study among, for example, dietitians, general practitioners, nurse practitioners, and home care nurses provided insight into the current bottlenecks within the existing undernutrition management guidelines. In these telephone interviews, these experts also discussed the current dietary behaviour problems of older adults and their impact on undernutrition risk. The experts’ experiences implied that undernutrition awareness is limited, among both older adults and care professionals. In addition, the interviewees were unclear about which professionals are responsible for monitoring and which monitoring procedures are preferred. The dietitians feel that they become involved too late, leading to decreased treatment effectiveness. In general, the interviewees desired more collaboration and a coherent and feasible allocation of responsibilities regarding undernutrition monitoring and treatment. This implied that the available guidelines on undernutrition management require more attention and facilitation.

In the following mixed-methods study (chapter 3), with interviews, we qualitatively explored the dietary behaviour and undernutrition risk of 12 Dutch elderly meals-on-wheels clients, one of the largest at-risk groups. We followed up on this information by quantifying the topics that emerged from the qualitative exploration of experienced bottlenecks in performing adequate dietary behaviour. For this, we used a survey among 333 meals-on-wheels clients. The interviews with elderly meals-on-wheels clients made clear that they have fixed and habitual eating patterns, while at the same time their appetite had decreased throughout the years. This was confirmed by the survey finding that regular portion size meals were perceived as too large by the oldest group aged over 75y. In addition, as the professionals suggested earlier, the interviewed elderly clients indeed showed limited awareness of undernutrition risk. Simultaneously, the survey showed that almost one in four elderly meals-on-wheels clients was undernourished. These findings led to the conclusion that staying close to the identified dietary habits may facilitate small yet effective modifications within these habits to prevent inadequate nutritional intake. Still, the limited awareness of undernutrition risk was expected to play a limiting role in whether clients believe they need dietary modifications. Consequently, informing them about this need could facilitate their motivation to implement modifications.

After learning about the general dietary behaviour of these older adults, we used this information for Study 3 (chapter 4). We developed two kinds of protein-enriched readymade meals that are in line with the needs and preferences of older adults: one of regular size (450g) and one of reduced size (400g). We tested these meals in a lab setting in 120 community-dwelling older adults in a single-blind randomised crossover trial. One day a week at lunchtime, for four weeks, participants had to consume and evaluate a readymade meal. Overall, regardless of portion size, the protein-enriched meals led to higher protein intakes in vital older adults in a lab setting during lunch. In this crossover study, the participants liked the protein-enriched meals and the regular meals equally. However, we did not find the expected lower ratings of satiety after the reduced-size meals, while one reduced-size enriched meal and another regular-size enriched meal led to higher ratings of subsequent satiety. This higher satiety in the enriched meals could lead to compensational behaviour on the remainder of the day.

After establishing that the protein-enriched meals were effective and acceptable in the lab setting, we moved to the homes of older adults to test the meals in a longer-term study in Study 4 (chapter 5). In this double-blind randomised controlled trial of two weeks, we also included protein-enriched bread to assess whether both this bread and the meals could increase daily protein intake to 1.2g/kg/d in 42 community-dwelling older adults to reach optimal protein intake. We found that the enriched products again led to higher protein intakes and a high liking. The mean protein intake per day was 14.6g higher in the intervention group, which amounted to a protein intake of 1.25g/kg/d, compared with 0.99g/kg/d in the control group. In addition, the meals scored 7.7 out of 10, while the bread scored 7.8 out of 10, which both were comparable with their regular counterparts. Lastly, we found no negative effect of compensational behaviour throughout the day. These promising findings indicated that we achieved a good match between older adults’ needs and preferences regarding protein intake.

In the general discussion of this thesis (chapter 6), we combined our learnings from the four studies to reflect on protein undernutrition management in community-dwelling older adults and the possible role of protein-enriched regular products. We have discussed a conceptual framework consisting of three wheels of protein undernutrition management. In the first wheel regarding awareness, we proposed that limited awareness of adequate nutrition and body composition forms the largest bottleneck in undernutrition management. When this awareness is generated among both older adults and professionals, it will benefit the second wheel of monitoring. Here, we argued that a policy and the actual facilitation of that policy are required for this monitoring to succeed. When the monitoring is performed adequately, in the third wheel, the appropriate treatment can be carried out. We discussed that personalisation and evaluation of this treatment are important conditions. All in all, the public health implications that we have discussed on the basis of our findings can be summarised by the three key messages that could help us ace in adequate protein undernutrition management: address awareness in both older adults and professionals, facilitate continuous collaboration between professionals, and offer protein-enriched products expediently.

Older adults, mealtime-related emotions, and functionalities : tailoring protein-enriched meals
Uijl, Louise C. den - \ 2016
University. Promotor(en): Kees de Graaf, co-promotor(en): Stefanie Kremer; Gerry Jager. - Wageningen : Wageningen University - ISBN 9789462578920 - 178
meals - emotions - elderly nutrition - elderly - smell - food preferences - protein - proteins - questionnaires - young adults - chocolate - maaltijden - emoties - ouderenvoeding - ouderen - reuk - voedselvoorkeuren - eiwit - eiwitten - vragenlijsten - jongvolwassenen - chocolade

Background and aim

Dietary proteins are of special interest for the heterogeneous group of older adults, since these people do not always have an adequate protein intake. When protein-rich products are better aligned with the requirements of older persons, an adequate nutrient intake is more likely. In this thesis we therefore explored two approaches for tailoring protein-enriched meals to older consumer subgroups; emotion-based and functionality-based. We expected a better ‘product-cluster fit’ (i.e. a more positive meal experience) when the clusters’ meal associations are congruent to their mealtime expectations.


We conducted an online survey in which vital community-dwelling older adults (n=392) reported their mealtime-related emotions and mealtime functionality. Using a hierarchical clustering analysis we described clusters within our population. Subsequently, we explored the extent to which the expectations of these clusters can be applied for the development of tailored protein-enriched meals. For the emotion-based approach, we conducted two central location tests (CLTs, n=461) to explore older adults’ food-evoked emotions. For the functionality-based approach we conducted in-depth interviews in order to get further insights regarding functional mealtime expectations and attitudes towards proteins and protein-enrichment. Based on the latter insights we tailored PE meal concepts to two functionality-based segments. In a final home-use test, the members of the functionality-based segments (n=91) prepared and evaluated the tailored PE meal concepts.


The emotion-based approach resulted in four clusters; pleasurable averages, adventurous arousals, convivial indulgers, and indifferent restrictives. These emotions that these segments associated with their mealtimes varied along the two dimensions valence and arousal. However, from both CLTs we learned that the variation in valence-arousal as observed for mealtime-related emotions was not observed for emotions related to actual foods. The latter makes it challenging to identify products that evoke emotions congruent to the mealtime expectations of the emotion-based clusters.

With regard to the functionality-based approach, we encountered three clusters; physical nutritioners, cosy socialisers, and thoughtless averages. The cosy socialisers value the social interactions and cosiness during their mealtimes, whereas the physical nutritioners focus more on the health and nutrient aspects of meals. Thoughtless averages have the least distinctive mealtime expectations. We translated these functional mealtime expectations into two PE meal concepts; one tailored to cosy socialisers and one tailored to physical nutritioners. These meal concepts were well-accepted by the participants. However, congruency between mealtime expectations and functional meal associations did not result in a better ‘product-cluster fit’.


Given the challenge to identify congruency between the meal associations and the mealtime expectations of the emotion-based clusters, we consider the emotion-based approach to be not yet actionable enough as a basis for tailoring PE products to older consumers. In contrast, the functionality-based approach appeared to be more promising, since the functional meal expectations could be translated to well-accepted tailored PE meal concepts. However, the effectivity of our functionality-based approach was not yet confirmed in this thesis, since congruency between functional meal associations and functional meal expectations did not necessarily result in a better ‘product-cluster fit’. Future studies, focussing on e.g. other meal types, are recommended to further explore mealtime functionality as a basis for tailoring PE meals to older consumer subgroups.

Cater with Care : impact of protein-enriched foods and drinks for elderly people
Beelen, J. - \ 2016
University. Promotor(en): Lisette de Groot; Frans Kok, co-promotor(en): Nicole de Roos. - Wageningen : Wageningen University - ISBN 9789462578814 - 142 p.
undernutrition - hospital catering - hospitals - protein - elderly - protein intake - food - beverages - diet studies - dietetics - dietitians - randomized controlled trials - ondervoeding - ziekenhuiscatering - ziekenhuizen - eiwit - ouderen - eiwitinname - voedsel - dranken - dieetstudies - diëtetiek - diëtisten - gestuurd experiment met verloting

Protein undernutrition is a major health concern for older adults, especially for those who are ill. There is growing consensus for a protein intake target of 1.2 - 1.5 gram per kg bodyweight per day (g/kg/d) for these older adults. However, this target is not reached by the majority of older adults. Therefore, more effective and novel strategies to increase protein intake are warranted, including the use of protein-enriched foods and drinks. This thesis evaluated the impact of the developed protein-enriched foods and drinks on protein intake and physical performance among older adults. The studies in this thesis were done as part of the Cater with Care® project; a collaboration between the university, care organizations, and partners from the food industry. The industrial partners developed the products, focusing each on different product categories: Carezzo Nutrition developed bread, pastry, and fresh juices and soups; The Kraft Heinz Company focused on long shelf-life and convenience foods; and the Veal Promotion Foundation produced veal meat.

To fit the products to the needs of the target group, interviews with undernourished older adults (at home or hospitalized) and with dietitians were conducted (chapter 2). These interviews showed that undernutrition awareness is low among older adults. To treat undernutrition by changing their eating habits, older adults need to be aware of their health problem, they need to be willing to change, and they need to be able to understand and implement the dietitian’s advices. This process takes time while undernutrition should be treated immediately. For immediate treatment, enriched products could be used, without first creating awareness. According to the interviewees, enriched products should fit within older adults’ eating habits, and have small portion sizes.

To gain insights in food choices of hospitalized older adults (65 years and older) an observational study was conducted. In this study, energy and protein intakes of 80 hospitalized older patients at low and high risk of undernutrition were assessed (chapter 3). Patients who received an energy- and protein-rich menu, because of their risk of undernutrition, were better able to reach the protein and energy targets than patients with a low risk of undernutrition receiving a standard menu. Based on these results we proposed that all hospitalized older adults – both at low and high risk of undernutrition – should receive an energy- and protein-rich menu.

Subsequently, a pilot study was done in a care home and a rehabilitation center with the aim to explore the potential of the developed protein-enriched products to increase protein intake (chapter 4). Participants did not compensate their consumption of regular protein-rich foods (e.g. dairy, cheese) upon the introduction of protein-enriched foods and drinks. The 22 institutionalized elderly (mean age 83 years) consumed 12 gram protein per day more than they did before the intervention. Consequently, more people met the protein target of 1.2 g/kg/d than before the intervention. We concluded that protein-enriched products enabled institutionalized elderly to reach protein intake targets. Furthermore, we gained valuable feedback to improve the assortment of protein-enriched products for the effectiveness study.

In the final study, effects of the protein-enriched products on protein intake and physical performance were studied in a randomized controlled trial during hospitalization and subsequent recovery at home. During the hospital period in which 147 older patients participated, patients that received protein-enriched products increased their protein intake compared to the control group that already received a protein-rich hospital menu (chapter 5). As a result, 79% of the intervention group reached a protein intake of 1.2 g/kg/d, compared to 48% of the control group. Finally, effects of the protein-enriched products were tested at home, for a longer period (chapter 6). Half of the hospital phase participants (n = 75) continued the intervention at home for 12 weeks. The protein-enriched products were successfully implemented in the daily menu of the older adults: the intervention group had a higher average protein intake (1.5 ± 0.6 g/kg/d) than the control group (1.0 ± 0.4 g/kg/d) during the 12-week intervention period. Seventy-two percent of the intervention group reached a protein intake of 1.2 g/kg/d during the 12-week intervention, compared to 31% of the control group. Protein intake of the intervention group was mainly increased by the following protein-enriched products: bread, dairy drinks, dairy desserts, soups, and fruit juices. However, despite the successful improvement of protein intake, we found no added value on physical performance in the first 6 months after hospitalization.

It was concluded that with the protein-enriched familiar foods and drinks, we have a feasible, acceptable, and appetizing long-term strategy to increase protein intake of older adults in various settings. We envisage a beneficial role of these protein-enriched products in combination with physical activity in older adults with lower protein intakes.

How to fulfill EU requirements to feed organic laying hens 100% organic ingredients
Krimpen, M.M. Van; Leenstra, F. ; Maurer, V. ; Bestman, M. - \ 2016
Journal of Applied Poultry Research 25 (2016)1. - ISSN 1056-6171 - p. 129 - 138.
EU requirements - laying hens - methionine - organic - protein

From December 2017 onward, including non-organic protein sources in diets for organic poultry will no longer be allowed in the EU. Moreover, in the EU the use of synthetic amino acids in organic diets is prohibited. The main dietary challenge in European organic egg production is to fulfill the protein requirement, especially the methionine (Met) requirement of the hens. Currently available Met-rich ingredients are discussed. In the group of ingredients of plant origin, expelled sunflower seed has a relatively high digestible Met content and is also commonly available. Met content of plant ingredients can be increased by selection of high Met varieties and by specifically breeding on high Met content, e.g. by crossing different breeds. Plant processing techniques might be helpful to concentrate the protein and digestible Met content of ingredients. Applying the dry fractionation technique on legumes and cereals might result in protein concentrates with CP content of at least 50%. A further development of simple separation techniques, which separate the hulls from the other plant fractions and reduce the fiber content after de-hulling, might be helpful to increase digestible Met content. Energy dilution of the diet, concomitant with a proportional reduction in other nutrients, is an option as well to fulfill the requirement of 100% organic diets. As a consequence, hens have to consume more feed to meet their nutrient requirements. There are options to fulfill the requirement of 100% ingredients of organic origin, but if the practical, economical, and footprint issues are taken into account, the list of options is very small.

Naar 100% regionaal eiwit : kansen en knelpunten voor eiwitrijke veevoergrondstoffen
Zanders, R. ; Cormont, A. ; Krimpen, M.M. van; Prins, Udo ; Ridder, A. de; Kessel, Hans van; Hartog, H. den; Krajenbrink, Wim ; Pluimers, Jacomijn ; Haren, Rob ; Gankema, P. - \ 2016
Raad voor Regionaal Veevoer - 18 p.
veevoeder - eiwit - veevoederindustrie - veevoeding - eiwitwaarde - streekgebonden producten - duurzame veehouderij - fodder - protein - feed industry - livestock feeding - protein value - regional specialty products - sustainable animal husbandry
Ongeveer de helft van het eiwitrijke veevoer in Nederland wordt geïmporteerd van buiten Europa. Het overgrote deel van deze grondstoffen bestaat uit soja- en palmproducten. Op Europees niveau zijn we voor 96% van onze sojabehoefte en voor 70% van onze totale eiwitbehoefte afhankelijk van import van buiten Europa. In de teeltgebieden, met name in Zuid-Amerika, leidt de grootschalige teelt van deze gewassen tot grote ecologische en sociale schade; door ontbossing van natuurgebieden en daarmee gepaard gaande CO2- uitstoot en biodiversiteitsverlies, door uitputting van de bodem, vervuiling van drinkwater, bedreiging van de lokale voedselvoorziening en gedwongen landonteigening. Regionale eiwitteelt biedt een enorme kans voor de Nederlandse landbouw door het sluiten van kringlopen en het verminderen van milieuschade en sociaal onrechtvaardige omstandigheden. Om knelpunten en kennisvragen te identificeren die een (snelle) transitie naar het gebruik van meer regionaal eiwitrijk veevoer in de weg staan, richtte Milieudefensie in 2015 de Raad voor Regionaal Veevoer op. Dit rapport geeft de bevindingen weer van de Raad voor Regionaal Veevoer, inclusief haar aanbevelingen richting overheid, bedrijfsleven en maatschappelijke organisaties.
Measurement errors in dietary assessment using duplicate portions as reference method
Trijsburg, L.E. - \ 2016
University. Promotor(en): Pieter van 't Veer; Anouk Geelen; Jeanne de Vries. - Wageningen : Wageningen University - ISBN 9789462576421 - 128 p.
diet studies - nutritional assessment - questionnaires - reference standards - correction factors - validity - body mass index - regression analysis - food intake - food - protein - potassium - sodium - energy intake - methodology - dieetstudies - voedingstoestandbepaling - vragenlijsten - referentienormen - correctiefactoren - geldigheid - quetelet index - regressieanalyse - voedselopname - voedsel - eiwit - kalium - natrium - energieopname - methodologie

Measurement errors in dietary assessment using duplicate portions as reference method

Laura Trijsburg

Background: As Food Frequency Questionnaires (FFQs) are subject to measurement error, associations between self-reported intake by FFQ and outcome measures should be corrected for measurement error with data from a reference method. Whether the correction is adequate depends on the characteristics of the reference method used in the validation study. The duplicate portion method (DP), compared to the often used 24h recall (24hR), seems a promising reference method as correlated errors between FFQ and DP, such as memory bias, errors in portion size estimations and food composition databases, are not expected.

Aim: This thesis aimed to determine the validity of the DP compared to the 24hR as a reference method for FFQ validation. The second aim was to explore the validity of nutrient densities for DP, 24hR and FFQ. The third aim was to determine the factors associated with misreporting of energy, protein and potassium as estimated by DP, 24hR and FFQ.

Methods: Within the DuPLO-study, a Dutch validation study which is part of the NQplus study, two DPs, two FFQs, two blood and urinary biomarkers and one to fifteen 24hRs (web-based and/or telephone-based) were collected in 198 subjects, within 1.5 years. Also, one or two doubly labelled water measurements were available for 69 participants. Multivariate measurement error models were used to assess proportional scaling bias, error correlations with the FFQ, validity coefficients and attenuation factors. Furthermore linear regression analysis was used to determine the association between misreporting and various factors.

Results: The DP was less influenced by proportional scaling bias, had lower correlated errors with the FFQ and showed higher attenuation factors than the 24hR for potassium, sodium and protein. Also, the DP seemed a better reference method than the 24hR for the assessment of validity coefficients for the FFQ for various fatty acids. The attenuation factors for the FFQ, using either the DP or 24hR as reference method, agreed reasonably well. Furthermore, the DP showed, when using plasma fatty acids as reference, slightly better ranking of participants according to their intake of n-3 fatty acids (0.33) and the n‑3/LA ratio (0.34) than the 24hR (0.22 and 0.24, respectively). Less group level bias was observed for protein and sodium densities compared to their absolute intakes for FFQ, 24hR and DP, but not for potassium. Overall the validity coefficients and attenuation factors for DP, 24hR and FFQ did not improve for nutrient densities compared to absolute intakes, except for the attenuation factor for sodium density. Lastly, BMI proved to be the most consistent determinant associated with misreporting (group level bias) of energy, protein and potassium for DP, 24hR and FFQ. Men tended to underreport protein by the DP, FFQ and 24hR and persons of older age underreported potassium but only by the 24hR and FFQ. Other explorative determinants did not show a consistent association with misreporting of energy or nutrients by the different dietary assessment methods.

Conclusion: With respect to error correlations and attenuation factors the DP performed slightly better than the 24hR as a reference method for validating FFQs in epidemiological research. Furthermore, the use of nutrient densities does not necessarily improve the validity of the dietary intake estimates from DP, 24hR and FFQ. Moreover, it was shown that BMI is an important determinant of misreporting of energy, protein and potassium for these three assessment methods.

Het effect van aminozuuraanbod en -samenstelling van het voer op zoötechnische prestaties van beren gehuisvest onder verschillende sanitaire condities
Meer, Y. van der; Gerrits, W.J.J. ; Jansman, A.J.M. - \ 2016
Wageningen UR Livestock Research (Livestock Research rapport 938) - 32 p.
beren (varkens) - aminozuren - eiwit - voedselsamenstelling - varkensvoeding - mestresultaten - hygiëne - varkenshouderij - zoötechniek - dierlijke productie - boars - amino acids - protein - food composition - pig feeding - fattening performance - hygiene - pig farming - zootechny - animal production
Dit experiment was opgezet om het effect van eiwitniveau (normaal versus verlaagd) en aminozuursamenstelling in het rantsoen te evalueren op de technische prestaties van beren gehuisvest onder een tweetal sanitaire condities.
Grasraffinage en gebruik van grasvezel in de rundveevoeding
Klop, A. ; Durksz, D.L. ; Zonderland, A. ; Koopmans, B. - \ 2015
Wageningen : Wageningen UR, Livestock Research (Livestock Research rapport 790) - 26 p.
veevoeding - melkveevoeding - kalvervoeding - bioraffinage - grasmaaisel - vezels - eiwit - proeven - melkveehouderij - livestock feeding - dairy cattle nutrition - calf feeding - biorefinery - grass clippings - fibres - protein - trials - dairy farming
In 2012 is een proef met melkkoeien uitgevoerd met als doel de waarde van grasvezel te onderzoeken. In het rantsoen van de koeien werd een deel van de graskuil vervangen door grasvezel. De grasvezel kwam beschikbaar na de raffinage van gras. De resultaten van de proef vielen tegen. De voeropname van de koeien die grasvezel kregen was namelijk lager dan van de (controle)koeien die het gangbare rantsoen kregen. De melkgift was eveneens lager op het rantsoen met grasvezel. De oorzaak van de lagere voeropname heeft waarschijnlijk te maken met de versheid en daarmee de smakelijkheid van grasvezel. Daarom is in 2013 besloten om eerst te kijken naar de mogelijkheid om grasvezel te conserveren (in te kuilen), waardoor de kwaliteit en de houdbaarheid mogelijk werd verbeterd. In 2012 is eveneens een oriënterend onderzoek gedaan met graseiwit verstrekt aan kalveren. Graseiwit is het eiwit dat gewonnen wordt uit het grassap en in de proef werd het in gelvorm verstrekt. De resultaten van de proef met kalveren waren uitermate positief. De dieren namen het graseiwit graag op. De groei van de kalveren was vergelijkbaar met de controlegroep.
Drying and hydration of proteins at high concentration
Bouman, J. - \ 2015
University. Promotor(en): Erik van der Linden, co-promotor(en): Renko de Vries. - Wageningen : Wageningen University - ISBN 9789462575509 - 161
eiwit - wei-eiwit - zeïne - drogen - droogmethoden - geneesmiddeltoedieningssystemen - hydratatie - hydrofobiciteit - ph - vacuolen - protein - whey protein - zein - drying - drying methods - drug delivery systems - hydration - hydrophobicity - vacuoles

Proteins are the building blocks of life and serve a wide range of essential functions in organisms. Many metabolic reactions in organisms are catalysed by enzymes, DNA is replicated by proteins and in cells proteins often facilitate active transport of e.g. glucose or ions. Proteins also serve an essential functionality in foods, pharmaceutics, bioplastics and even clothing. Recently, the use of proteins towards higher concentrations is of interest for food, pharmaceutical and medical applications. Nevertheless, the preparation of products with desired product properties can be challenging, when approaching higher protein concentrations. Therefore, in this thesis we investigate proteins at higher concentrations, especially focussing on their drying and hydration behaviour.

In part one of the thesis, the focus is on the dynamics of drying of proteins towards higher concentrations. Dense proteins systems have been scarcely studied compared to proteins at lower concentrations. We address drying behaviour where we focus on the use of whey protein isolate as a model system. In part two of the thesis we focus on the hydration properties of the corn protein zein, where we apply it as a drug excipient. In this part we also investigate the influence of hydration on the release behaviour of drugs into the hydration media.

The drying part (part one) contains two studies. The first study is more fundamental in nature, focussing on the drying of a protein coating. In previous studies mainly the macroscopic properties of protein coatings after drying are investigated, leaving the drying dynamics virtually unexplored. Here we investigate the drying behaviour of the model protein β-lactoglobulin on multiple length scales with an unique combination of in-line techniques. On the microscopic length scale we use dynamic vapour sorption and magnetic resonance imaging while on a smaller length scales, we apply diffusing wave spectroscopy and IR-spectroscopy to monitor the drying process. For all used techniques, the changes in the measured physical properties of the coating as a function of water weight fraction Xw from Xw = 0.8 down to Xw = 0.2 are gradual. However, using dynamic vapour sorption and IR-spectroscopy we measure a sharp change below water weight fractions of Xw = 0.2. We hypothesise that changes in the molecular interactions caused by dehydration of the protein results in a change in the drying kinetics of the film.

In the second study of part one, protein drying is approached on a more applied level, where we study the drying of a spherical droplet. We use single droplet drying as a methods that can model the spray drying process in a simplified and well-controlled way. Sessile droplets are subjected to varying drying conditions such as temperature, initial protein concentration, presence of airflow and droplet rotation. During these experiments the morphological development is monitored by a camera. After drying, scanning electron microscopy and X-ray tomography are used to examine the particles that are formed after complete drying. Irrespective of the conditions used, we observe an initial droplet shrinkage, followed by the nucleation of a hole in the droplet skin, which is followed by the formation of a vacuole. The drying conditions used, strongly influenced the location of the hole and the locking point prior to hole formation. We hypothesise that the location of the hole is caused by local inhomogeneities in protein concentration causing a the nucleation of the hole where the local skin modulus is lowest. Also the locking point of the droplet is found to be due to a inhomogeneity over the whole droplet caused by rapid evaporation. These results can be of importance to understand powder structure and functionality as obtained in spray drying.

In the hydration part (part 2), we investigate the potential of zein as a sole excipient in macroscale caplets obtained by hot melt extrusion (HME) and injection moulding (IM). Zein is good candidate as a sustained release agent, because it is insoluble in two studies. In the first study zein matrices were loaded with the drug paracetamol. Physical mixtures of zein, water and crystalline paracetamol are extruded and injection moulded into caplets. Characterisation of these caplets is performed using differential scanning calorimetry, IR- spectroscopy, scanning electron microscopy and powder X-ray diffraction. The hydration and drug release kinetics from the caplet slices is measured. We find that the drug release kinetics is broadly independent of the dissolution medium and drug loading. The release kinetics is diffusion limited and could be well described by a 2D diffusion model. The results demonstrate that the drug release rate from zein caplet slices can be tuned by its dimensions.

In the second study, a wider range of drugs differing in hydrophobicity is studied. Next to paracetamol, we have used two other model drugs: the hydrophobic indomethacin and the more hydrophilic ranitidine. The zein matrix is capable to stabilize the different dugs in a non-crystalline state, which is promising especially for increasing the bioavailability of poorly water-soluble drugs. Overall crystallinity of the drugs in the caplets increases with its degree of hydrophobicity. For the poorly soluble indomethacin, dissolution rates at low pH were higher from caplet slices, compared to the dissolution rates of indomethacin crystals by themselves. In addition, we found that the electrostatic interactions between zein and drugs can also be used to influence the release kinetics.

Various aspects were found to be of importance both for drying and hydration of concentrated protein systems. The homogeneity during both processes deserves attention as its manipulation can strongly influence final properties if the system. Also the plasticising effect of water on dense proteins is often found essential, when understanding the dynamics of both drying and hydration processes. Finally protein hydrophobicity and its manipulation can provide a window of opportunities in many applications which are involve by drying or hydration.

Microbubble stability and applications in food
Rovers, T.A.M. - \ 2015
University. Promotor(en): Erik van der Linden, co-promotor(en): Marcel Meinders; Guido Sala. - Wageningen : Wageningen University - ISBN 9789462574755 - 138
microbubbles - eiwit - stabiliteit - karakterisering - voedsel - voedseladditieven - oppervlaktespanningsverlagende stoffen - zuurbehandeling - reologische eigenschappen - sensorische evaluatie - tribologie - druk - verwarming - koelen - protein - stability - characterization - food - food additives - surfactants - acid treatment - rheological properties - sensory evaluation - tribology - pressure - heating - cooling

Aeration of food is considered to be a good method to create a texture and mouthfeel of food products that is liked by the consumer. However, traditional foams are not stable for a prolonged time. Microbubbles are air bubbles covered with a shell that slows down disproportionation significantly and arrests coalescence. Protein stabilized microbubbles are seen as a promising new food ingredient for encapsulation, to replace fat, to create new textures, and to improve sensorial properties of foods. In order to explore the possible functionalities of microbubbles in food systems, a good understanding is required regarding the formation of protein stabilized microbubbles as well as their stability in environments and at conditions encountered in food products. The aim of this research was to investigate the key parameters for applications of microbubbles in food systems. In Chapter 1 an introduction to this topic is given.

In Chapter 2, the effect of the microbubble preparation parameters on the microbubble characteristics, like the microbubble yield, size and stability, was investigated. The protein Bovine Serum Albumin (BSA) and the method sonication was used to manufacture the microbubbles. The manufactured number and stability of microbubbles was highest when they were prepared at a pH around 5 to 6, just above the isoelectric point, and at an ionic strength of 1.0 M. This can be related to the protein coverage at the air/water interface of air bubbles formed during sonication. At a pH close to the isoelectric point the BSA molecules is in its native configuration. Also the repulsion between the proteins is minimized at these pH values and ionic strength. Both the native configuration and the limited repulsion between the proteins result in an optimal protein coverage during the first part of sonication. Also a high protein concentration contributes to a higher surface coverage. The surface coverage is proportional to the protein concentration up to a concentration of 7.5% after which an increase in protein concentration did not lead to a substantial increase in the number of microbubble . In the second part of sonication the protein layer around the air bubble becomes thicker and stronger by heat induced protein-protein interactions. We found that and at a preheating temperature of 55-60°C, about 5 °C below the BSA denaturation temperature, and a final solution temperature of 60-65°C most microbubbles were obtained, while at higher temperatures mainly protein aggregates and (almost) no microbubbles are formed. This suggests that at temperature of around 60°C to 65°C protein aggregated mostly at the air-water interface creating a multi-layered shell, while at higher temperature, they also aggregated in bulk. These aggregates cannot form microbubbles. We found that optimal preparation parameters strongly depend on the protein batch. We hypothesize that the differences in microbubble formation between the protein batches is due to (small) differences in the protein molecular and denaturation properties that determine the temperature at which the molecules start to interact at the air-water interface. Microbubbles made with different protein concentration and preheating temperatures shrunk in time to a radius between 300 nm and 350 nm, after which the size remained constant during further storage. We argue that the driving force for the shrinkage was the Laplace pressure, resulting in an air flux from the bubbles to the solution. We argue that the constant final size can be explained by a thickening of the microbubble shell as a result of the microbubble shrinkage, thereby withstanding the Laplace pressure.

In Chapter 3 and Chapter 4, microbubble stability at environments and conditions representative for food products were studies. In Chapter 3 we investigated the stability upon addition of surfactants and acid, When surfactants or acid were added, the microbubbles disappeared in three subsequent steps. The release of air from the microbubble can be well described with the two-parameter Weibull process. This suggests two processes are responsible for the release of air: 1) a shell-weakening process and 2) a random fracture of the weakened shell. After the air has been released from the microbubble the third process is identified in the microbubble disintegration: 3) the shell disintegrated completely into nanometer-sized particles. The probability of fracture was exponentially proportional to the concentration of acid and surfactant, meaning that a lower average breaking time and a higher decay rate were observed at higher surfactant or acid concentrations. For different surfactants, different decay rates were found. The disintegration of the shell into monomeric proteins upon addition of acid or surfactants shows that the interactions in the shell are non-covalent and most probably hydrophobic. After surfactant addition, there was a significant time gap between complete microbubble decay (release of air) and complete shell disintegration, while after acid addition the time at which the complete disintegration of the shell was observed coincided with the time of complete microbubble decay.

In Chapter 4 the stability of the microbubbles upon pressure treatment, upon fast cooling after heating and at different storage temperatures was studied. The microbubble stability significantly decreased when microbubbles were pressurized above 1 bar overpressure for 15 seconds or heated above 50°C for 2 minutes. Above those pressures the microbubbles became unstable by buckling. Buckling occurred above a critical pressure. This critical pressure is determined by the shell elastic modulus, the thickness of the shell, and the size of the microbubble. Addition of crosslinkers like glutaraldehyde and tannic acid increased the shell elastic modulus. It was shown that microbubbles were stable against all tested temperatures (up to 120°C) and overpressures (4.7 bar) after they were reinforced by crosslinkers. From the average breaking time at different storage temperatures, we deduced that the activation energy to rupture molecular bonds in the microbubbles shell is 27 kT.

In Chapter 5, we investigated the effect of microbubbles on the rheological, tribological sensorial properties of model food systems and we compared this effect to the effect on food systems with emulsion droplets and without an added colloid. We investigated the effect in three model food systems, namely fluids with and without added thickener and a mixed gelatine-agar gel. In a sensory test panellists were asked whether they could discriminate between samples containing microbubbles, emulsion droplets or no added colloid. Emulsions could be sensorially well distinguished from the other two samples, while the microbubble dispersion could not be discriminated from the protein solution. Thus, we concluded that at a volume fraction of 5% of these BSA covered microbubbles were not comparable to oil-in-water emulsions. The good discrimination of emulsion might be ascribed to the fact that emulsion had a lower friction force (measured at shear rates form 10 mm/s to 80 mm/s) than that microbubbles dispersions and protein solutions. Upon mixing emulsions and microbubble dispersions the friction value approximated that of emulsions. This effect was already noticed at only 1.25% (v/v) oil, indicating that microbubbles had not a significant contributions to the friction of these samples. Also microbubble dispersions with and without protein aggregates were compared. The microbubble dispersions with and without thickener containing protein aggregates had a higher viscosity than the those samples without protein aggregates. Protein aggregates in the gelled microbubble sample yielded a higher Young’s modulus and fracture stress. The differences between the gelled samples could be well perceived by the panellists. We attribute this mainly to the fracture properties of the gel. In general we concluded that microbubbles, given their size of ~ 1 mm and volume fraction of 5%, did not contribute to a specific mouthfeel.

Finally in Chapter 6, the results presented in the previous chapters are discussed and put in perspective of the general knowledge on microbubbles production, stability, and applications in food. We described the main mechanisms leading to microbubble formation and stability. We showed that the production parameters significantly influence the interactions in the microbubble shell, and the those interactions highly determine the stability of the microbubbles under several conditions. We reported about limitations of sonication as a method to produce microbubbles suitable for food applications and we provided some ways to overcome these limitations. The use of microbubbles in food systems has been explored and we clearly see possible applications for microbubbles in food. We reported about directions for possible further research.

In this work we made significant progress in understanding the interactions in the microbubble shell and their relation to microbubble stability. We also advanced in comprehension towards possible applications of microbubbles in food.

Organizer-Derived WOX5 Signal Maintains Root Columella Stem Cells through Chromatin-Mediated Repression of CDF4 Expression.
Pi, L. ; Graaff, E. van der; Llavata Peris, C.I. ; Weijers, D. ; Henning, L. ; Groot, E. de; Laux, T. - \ 2015
Developmental cell 33 (2015)5. - ISSN 1534-5807 - p. 576 - 588.
histone deacetylase - arabidopsis-thaliana - transcriptional repression - gene-expression - wuschel - meristem - shoot - topless - protein - fate
Stem cells in plants and animals are maintained pluripotent by signals from adjacent niche cells. In plants, WUSCHEL HOMEOBOX (WOX) transcription factors are central regulators of stem cell maintenance in different meristem types, yet their molecular mode of action has remained elusive. Here we show that in the Arabidopsis root meristem, the WOX5 protein moves from the root niche organizer, the quiescent center, into the columella stem cells, where it directly represses the transcription factor gene CDF4. This creates a gradient of CDF4 transcription, which promotes differentiation opposite to the WOX5 gradient, allowing stem cell daughter cells to exit the stem cell state. We further show that WOX5 represses CDF4 transcription by recruiting TPL/TPR co-repressors and the histone deacetylase HDA19, which consequently induces histone deacetylation at the CDF4 regulatory region. Our results show that chromatin-mediated repression of differentiation programs is a common strategy in plant and animal stem cell niches.
A Novel Virus Causes Scale Drop Disease in Lates calcarifer
Groof, A. ; Guelen, L. ; Deijs, M. ; Wal, Y. van der; Miyata, M. ; Ng, K.S. ; Grinsven, L. van; Simmelink, B. ; Biermann, Y. ; Grisez, L. ; Lent, J.W.M. van; Ronde, A. de; Chang, S.F. ; Schrier, C. ; Hoek, L. - \ 2015
PLoS Pathogens 11 (2015)8. - ISSN 1553-7366
red-sea bream - family iridoviridae - pagrus-major - protein - vaccine
From 1992 onwards, outbreaks of a previously unknown illness have been reported in Asian seabass (Lates calcarifer) kept in maricultures in Southeast Asia. The most striking symptom of this emerging disease is the loss of scales. It was referred to as scale drop syndrome, but the etiology remained enigmatic. By using a next-generation virus discovery technique, VIDISCA-454, sequences of an unknown virus were detected in serum of diseased fish. The near complete genome sequence of the virus was determined, which shows a unique genome organization, and low levels of identity to known members of the Iridoviridae. Based on homology of a series of putatively encoded proteins, the virus is a novel member of the Megalocytivirus genus of the Iridoviridae family. The virus was isolated and propagated in cell culture, where it caused a cytopathogenic effect in infected Asian seabass kidney and brain cells. Electron microscopy revealed icosahedral virions of about 140 nm, characteristic for the Iridoviridae. In vitro cultured virus induced scale drop syndrome in Asian seabass in vivo and the virus could be reisolated from these infected fish. These findings show that the virus is the causative agent for the scale drop syndrome, as each of Koch’s postulates is fulfilled. We have named the virus Scale Drop Disease Virus. Vaccines prepared from BEI- and formalin inactivated virus, as well as from E. coli produced major capsid protein provide efficacious protection against scale drop disease.
Zeewier voor de toekomst
Ramaker, R. ; Brandenburg, W.A. ; Wald, J. - \ 2015
Resource: weekblad voor Wageningen UR 10 (2015)1. - ISSN 1874-3625 - p. 12 - 15.
mariene gebieden - zeewierenteelt - zeewieren - voedselproducten - oosterschelde - noordzee - toegepast onderzoek - aquatische biomassa - eiwit - financieren - marine areas - seaweed culture - seaweeds - food products - eastern scheldt - north sea - applied research - aquatic biomass - protein - financing
In 2050 moeten grote zeewierplantages op zee voorzien in onze behoefte aan voedsel en grondstoffen. In de Oosterschelde doen Wageningse onderzoekers nu experimenten met duurzame zeewierteelt.
Protein mixtures: interactions and gelation
Ersch, C. - \ 2015
University. Promotor(en): Erik van der Linden, co-promotor(en): A.H. Martin; Paul Venema. - Wageningen : Wageningen University - ISBN 9789462574212 - 199
eiwit - wei-eiwit - sojaeiwit - gelering - gelatine - gels - reologie - structuur - moleculaire interacties - protein - whey protein - soya protein - gelation - gelatin - rheology - structure - molecular interactions

Gelation is a ubiquitous process in the preparation of foods. As most foods are multi constituent mixtures, understanding gelation in mixtures is an important goal in food science. Here we presented a systematic investigation on the influence of molecular interactions on the gelation in protein mixtures. Gelatin gels with added globular protein and globular protein gels with added gelatin were analyzed for their gel microstructure and rheological properties. Mixed gels with altered microstructure (compared to single gels) also differed in modulus from single gels. Mixed gels with microstructures similar to single gels were rheologically similar to single gels. Alterations in microstructure were attributed to segregative phase separation between proteins which occurred during gelation. Gelation was treated as a growth process from macromolecule to space spanning network. At conditions where electrostatic interactions were screened the occurrence of phase separation was attributed to the molecular size ratio between gelling and non-gelling proteins before gelation and changes of this size ratio during gelation. Here only mixtures that during gelation passed a region of high compatibility (similar molecular sizes) before entering a region of decreasing solubility phase separated. For applications this implies that whenever the gelling molecule is larger than the non-gelling molecule phase separation during gelation is unlikely while reversely, if the gelling molecules is smaller than the non-gelling molecule phase separation during gelation typically does occur

The development of direct extrusion-injected moulded zein matrices as novel oral controlled drug delivery systems
Bouman, J. ; Belton, P. ; Venema, P. ; Linden, E. van der; Vries, R.J. de; Qi, Sheng - \ 2015
Pharmaceutical Research 32 (2015)8. - ISSN 0724-8741 - p. 2775 - 2786.
solid-state - diffusion - protein - acetaminophen - paracetamol - release - history - ftir
Purpose To evaluate the potential of zein as a sole excipient for controlled release formulations prepared by hot melt extrusion. Methods Physical mixtures of zein, water and crystalline paracetamol were hot melt extruded (HME) at 80 degrees C and injection moulded (IM) into caplet forms. HME-IM Caplets were characterised using differential scanning calorimetry, ATR-FTIR spectroscopy, scanning electron microscopy and powder X-ray diffraction. Hydration and drug release kinetics of the caplets were investigated and fitted to a diffusion model. Results For the formulations with lower drug loadings, the drug was found to be in the non-crystalline state, while for the ones with higher drug loadings paracetamol is mostly crystalline. Release was found to be largely independent of drug loading but strongly dependent upon device dimensions, and predominately governed by a Fickian diffusion mechanism, while the hydration kinetics shows the features of Case II diffusion. Conclusions In this study a prototype controlled release caplet formulation using zein as the sole excipient was successfully prepared using direct HME-IM processing. The results demonstrated the unique advantage of the hot melt extruded zein formulations on the tuneability of drug release rate by alternating the device dimensions.
Encapsulation of GFP in complex coacervate core micelles
Nolles, A. ; Westphal, A.H. ; Hoop, J.A. de; Fokkink, R.G. ; Kleijn, J.M. ; Berkel, W.J.H. van; Borst, J.W. - \ 2015
Biomacromolecules 16 (2015)5. - ISSN 1525-7797 - p. 1542 - 1549.
fluorescence correlation spectroscopy - protein - dynamics - behavior - nanocontainers - purification - copolymers - lipase - tag
Protein encapsulation with polymers has a high potential for drug delivery, enzyme protection and stabilization. Formation of such structures can be achieved by the use of polyelectrolytes to generate so-called complex coacervate core micelles (C3Ms). Here, encapsulation of enhanced green fluorescent protein (EGFP) was investigated using a cationic-neutral diblock copolymer of two different sizes: poly(2-methyl-vinyl-pyridinium)41-b-poly(ethylene-oxide)205 and poly(2-methyl-vinyl-pyridinium)128-b-poly(ethylene-oxide)477. Dynamic light scattering and fluorescence correlation spectroscopy (FCS) revealed a preferred micellar composition (PMC) with a positive charge composition of 0.65 for both diblock copolymers and micellar hydrodynamic radii of approximately 34 nm. FCS data show that at the PMC, C3Ms are formed above 100 nM EGFP, independent of polymer length. Mixtures of EGFP and nonfluorescent GFP were used to quantify the amount of GFP molecules per C3M, resulting in approximately 450 GFPs encapsulated per micelle. This study shows that FCS can be successfully applied for the characterization of protein-containing C3Ms.
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