Using either freshly pulped or preserved seaweed biomass for the extraction of protein can have a great effect on the amount of protein that can be extracted. In this study, the effect of four preservation techniques (frozen, freeze-dried, and air-dried at 40 and 70 °C) on the protein extractability, measured as Kjeldahl nitrogen, of four seaweed species, Chondrus crispus (Rhodophyceae), Ascophyllum nodosum, Saccharina latissima (both Phaeophyceae) and Ulva lactuca (Chlorophyceae), was tested and compared with extracting freshly pulped biomass. The effect of preservation is species dependent: in all four seaweed species, a differenttreatment resulted in the highest protein extractability. The pellet (i.e., the non-dissolved biomass after extraction) was also analyzed as in most cases the largest part of the initial protein ended up in the pellet and not in the supernatant. Of the four species tested, freeze-dried A. nodosum yielded the highest overall protein extractability of 59.6% with a significantly increased protein content compared with the sample before extraction. For C. crispus extracting biomass air-dried at 40 °C gave the best results with a protein extractability of 50.4%. Preservation had little effect on the protein extraction for S. latissima; only air-drying at 70 °C decreased the yield significantly. Over 70% of the initial protein ended up in the pellet for all U. lactuca extractions while increasing the protein content significantly. Extracting freshly pulped U. lactuca resulted in a 78% increase in protein content in the pellet while still containing 84.5% of the total initial total protein. These results show the importance of the right choice when selecting a preservation method and seaweed species for protein extraction. Besides the extracted protein fraction, the remainingpellet also has the potential as a source with an increased protein content.
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Trends in eiwittransitie kunnen regionaal verschillen. In groeiende economieën verschuiven diëten wereldwijd van plantaardige naar dierlijke eiwitten. In veel economisch ontwikkelde regio's gebeurt echter het tegenovergestelde vanwege de zorg voor milieu en gezondheid. Wij onderzochten de relatie tussen vijf drijvende krachten en eiwittransitietrends zoals deze worden ervaren door jongvolwassenen in ontwikkelde regio's in China (Shanghai) en Nederland (Amsterdam, lees: de Randstad). De onderzochte drijvende krachten waren: milieubewustzijn; het beleid; cultuur; geld; en gezondheid. De gegevens zijn verkregen door 200 vragenlijsten te laten beantwoorden in beide regio's. De resultaten geven aan dat jongvolwassenen in Shanghai meer dierlijke eiwitten consumeren dan plantaardige eiwitten, maar dat er een verandering naar plantaardige eiwitten is ingezet, terwijl de trend van jongvolwassenen in Amsterdam om plantaardig eiwit te consumeren al verder ontwikkeld is. De rangschikking van de drijvende krachten in Shanghai was Geld＞ Milieubewustzijn＞ Gezondheid＞ Cultuur＞ Beleid, en in Amsterdam Gezondheid＞ Milieubewustzijn＞ Geld＞ Beleid＞ Cultuur. Eiwitkeuzes in de voeding van jongvolwassenen worden dus in Shanghai door andere drijvende krachten bepaald dan in Amsterdam
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Optimizing protein intake is a novel strategy to prevent age associated loss of muscle mass and strength in older adults. Such a strategy is still missing for older adults from ethnic minority populations. Protein intake in these populations is expected to be different in comparison to the majority of the population due to several socio-cultural factors. Therefore, the present study examined the dietary protein intake and underlying behavioral and environmental factors affecting protein intake among older adults from ethnic minorities in the Netherlands. We analyzed frequency questionnaire (FFQ) data from the Healthy Life in an Urban Setting (HELIUS) cohort using ANCOVA to describe dietary protein intake in older adults from ethnic minorities in the Netherlands (N = 1415, aged >55 years, African Surinamese, South Asian Surinamese, Moroccan, and Turkish). Additionally, we performed focus groups among older adults from the same ethnic minority populations (N = 69) to discover behavioral and environmental factors affecting protein intake; 40-60% of the subjects did not reach minimal dietary protein recommendations needed to maintain muscle mass (1.0 g/kg bodyweight per day (BW/day)), except for Turkish men (where it was 91%). The major sources of protein originated from animal products and were ethnic specific. Participants in the focus groups showed little knowledge and awareness about protein and its role in aging. The amount of dietary protein and irregular eating patterns seemed to be the major concern in these populations. Optimizing protein intake in these groups requires a culturally sensitive approach, which accounts for specific protein product types and sociocultural factors.
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In werkpakket A, Zeewierteelt, werd onderzocht wat het effect is van de nitraatconcentratie op de groei en eiwitgehalte van de zeewiersoorten Saccharina latissima en Ulva lactuca. Bij de laatste soort werd ook gekeken naar de aminozuursamenstelling. Hogere nitraatconcentraties zorgden bij beide zeewiersoorten voor een hogere groeisnelheid en een hogere eiwitgehalte. De totale aminozuurhoeveelheid van Ulva lactuca was hoger bij blootstelling aan een hogere nitraatconcentratie. Alle gemeten aminozuurgehaltes waren hoger, behalve die van methionine, die gelijk was ten opzichte van de Ulva lactuca die gekweekt werd onder lage nitraatconcentraties. Het is dus mogelijk om tijdens het groeiproces, de aminozuur- en eiwitgehalte van zeewier te verhogen. De toename in eiwitgehalte was zelfs zo snel, dat het mogelijk is om zeewier te verrijken door het twee weken voor de oogst onder verhoogde nitraatconcentraties te laten groeien.
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tIn this study we aimed to identify genes that are responsive to pertussis toxin (PTx) and might eventu-ally be used as biological markers in a testing strategy to detect residual PTx in vaccines. By microarrayanalysis we screened six human cell types (bronchial epithelial cell line BEAS-2B, fetal lung fibroblastcell line MRC-5, primary cardiac microvascular endothelial cells, primary pulmonary artery smooth mus-cle cells, hybrid cell line EA.Hy926 of umbilical vein endothelial cells and epithelial cell line A549 andimmature monocyte-derived dendritic cells) for differential gene expression induced by PTx. Imma-ture monocyte-derived dendritic cells (iMoDCs) were the only cells in which PTx induced significantdifferential expression of genes. Results were confirmed using different donors and further extendedby showing specificity for PTx in comparison to Escherichia coli lipopolysaccharide (LPS) and Bordetellapertussis lipo-oligosaccharide (LOS). Statistical analysis indicated 6 genes, namely IFNG, IL2, XCL1, CD69,CSF2 and CXCL10, as significantly upregulated by PTx which was also demonstrated at the protein levelfor genes encoding secreted proteins. IL-2 and IFN- gave the strongest response. The minimal PTx con-centrations that induced production of IL-2 and IFN- in iMoDCs were 12.5 and 25 IU/ml, respectively.High concentrations of LPS slightly induced IFN- but not IL-2, while LOS and detoxified pertussis toxindid not induce production of either cytokine. In conclusion, using microarray analysis we evaluated sixhuman cell lines/types for their responsiveness to PTx and found 6 PTx-responsive genes in iMoDCs ofwhich IL2 is the most promising candidate to be used as a biomarker for the detection of residual PTx.
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Background: Lung fibroblasts are implicated in abnormal tissue repair in chronic obstructive pulmonary disease (COPD). The exact mechanisms are unknown and comprehensive analysis comparing COPD- and control fibroblasts is lacking. Aim: To gain insight in the role of lung fibroblasts in COPD pathology using unbiased proteomic and transcriptomic analysis. Methods: Protein and RNA was isolated from cultured parenchymal lung fibroblasts of 17 stage IV COPD patients and 16 non-COPD controls. Proteins were analyzed using LC-MS/MS and RNA through RNA sequencing. Differential protein and gene expression in COPD was assessed via linear regression, followed by pathway enrichment, correlation analysis and immunohistological staining in lung tissue. Proteomic and transcriptomic data was compared to investigate the overlap and correlation between both levels of data. Results: We identified 40 differentially expressed (DE) proteins and zero DE genes between COPD and control fibroblasts. The most significant DE proteins were HNRNPA2B1 and FHL1. Thirteen of the 40 proteins were previously associated with COPD, including FHL1 and GSTP1. Six of the 40 proteins were related to telomere maintenance pathways, and were positively correlated with the senescence marker LMNB1. No significant correlation between gene and protein expression was observed for the 40 proteins. Conclusions: The 40 DE proteins in COPD fibroblasts include previously described COPD proteins (FHL1, GSTP1) and new COPD research targets like HNRNPA2B1. Lack of overlap and correlation between gene and protein data supports the use of unbiased proteomics analysis and indicates that different types of information are generated with both methods.
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Development of novel testing strategies to detect adverse human health effects is of interest to replace in vivo-based drug and chemical safety testing. The aim of the present study was to investigate whether physiologically based kinetic (PBK) modeling-facilitated conversion of in vitro toxicity data is an adequate approach to predict in vivo cardiotoxicity in humans. To enable evaluation of predictions made, methadone was selected as the model compound, being a compound for which data on both kinetics and cardiotoxicity in humans are available. A PBK model for methadone in humans was developed and evaluated against available kinetic data presenting an adequate match. Use of the developed PBK model to convert concentration–response curves for the effect of methadone on human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) in the so-called multi electrode array (MEA) assay resulted in predictions for in vivo dose–response curves for methadone-induced cardiotoxicity that matched the available in vivo data. The results also revealed differences in protein plasma binding of methadone to be a potential factor underlying variation between individuals with respect to sensitivity towards the cardiotoxic effects of methadone. The present study provides a proof-of-principle of using PBK modeling-based reverse dosimetry of in vitro data for the prediction of cardiotoxicity in humans, providing a novel testing strategy in cardiac safety studies.
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Even in a less eventful year, it’s no easy feat: working to make our food supply healthy and sustainable. But 2020 brought a spate of new challenges. It was the year of Brexit, Black Lives Matter, and the COVID-19 pandemic. A year of hope and loss and solidarity, of masks and worries and Zoom calls. Of infection sweeping through the meatpacking industry and sometimes, of empty supermarket shelves. It was also the year that brought us the glimmering realisation that everything could be different. When so much has changed – how we work, who we spend time with, how far we venture from home – what all might be possible for food and for farming? In Flevo Campus’s latest collection of essays, thirteen journalists, scholars, and thought leaders from the US, the Netherlands, and the UK share insight into the question: How can we build resilience into our food supply – and grow more resilient ourselves? Every year, Flevo Campus publishes the best work on feeding the cities of today and tomorrow. This year’s edition includes essays by Stephen Satterfield, Charles C. Mann, Herman Lelieveldt, Hester Dibbits, Kelly Streekstra, Sigrid Wertheim-Heck, Anke Brons, Joris Lohman, Sebastiaan Aalst, Marian Stuiver, Frank Verhoeven, Emily Whyman, and Lenno Munnikes.
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IL22 is an important cytokine involved in the intestinal defense mechanisms against microbiome. By using ileum-derived organoids, we show that the expression of anti-microbial peptides (AMPs) and anti-viral peptides (AVPs) can be induced by IL22. In addition, we identified a bacterial and a viral route, both leading to IL22 production by T cells, but via different pathways. Bacterial products, such as LPS, induce enterocyte-secreted SAA1, which triggers the secretion of IL6 in fibroblasts, and subsequently IL22 in T cells. This IL22 induction can then be enhanced by macrophage-derived TNFα in two ways: by enhancing the responsiveness of T cells to IL6 and by increasing the expression of IL6 by fibroblasts. Viral infections of intestinal cells induce IFNβ1 and subsequently IL7. IFNβ1 can induce the expression of IL6 in fibroblasts and the combined activity of IL6 and IL7 can then induce IL22 expression in T cells. We also show that IL22 reduces the expression of viral entry receptors (e.g. ACE2, TMPRSS2, DPP4, CD46 and TNFRSF14), increases the expression of anti-viral proteins (e.g. RSAD2, AOS, ISG20 and Mx1) and, consequently, reduces the viral infection of neighboring cells. Overall, our data indicates that IL22 contributes to the innate responses against both bacteria and viruses.
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