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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Novel COX-2 products of n-3 polyunsaturated fatty acid-ethanolamine-conjugates identified in RAW264.7 macrophages
Bus, Ian de; Zuilhof, Han ; Witkamp, Renger ; Balvers, Michiel ; Albada, Bauke - \ 2019
Journal of Lipid Research 60 (2019)11. - ISSN 0022-2275 - p. 1829 - 1840.
cyclooxygenase - cyclooxygenase 2 - fatty acid amides - fatty acid oxidation - high-performance liquid chromatography - inflammation - mass spectrometry - prostaglandins

Cyclooxygenase 2 (COX-2) plays a key role in the regulation of inflammation by catalyzing the oxygenation of PUFAs to prostaglandins (PGs) and hydroperoxides. Next to this, COX-2 can metabolize neutral lipids, including endocannabinoid-like esters and amides. We developed an LC-HRMS-based human recombinant (h)COX-2 screening assay to examine its ability to also convert n-3 PUFA-derived N-acylethanolamines. Our assay yields known hCOX-2-derived products from established PUFAs and anandamide. Subsequently, we proved that eicosapentaenoylethanolamide (EPEA), the N-acylethanolamine derivative of EPA, is converted into PGE3-ethanolamide (PGE3-EA), and into 11-, 14-, and 18-hydroxyeicosapentaenoyl-EA (11-, 14-, and 18-HEPE-EA, respectively). Interestingly, we demonstrated that docosahexaenoylethanolamide (DHEA) is converted by hCOX-2 into the previously unknown metabolites, 13- and 16-hydroxy-DHEA (13- and 16-HDHEA, respectively). These products were also produced by lipopolysaccharide-stimulated RAW267.4 macrophages incubated with DHEA. No oxygenated DHEA metabolites were detected when the selective COX-2 inhibitor, celecoxib, was added to the cells, further underlining the role of COX-2 in the formation of the novel hydroxylated products. This work demonstrates for the first time that DHEA and EPEA are converted by COX-2 into previously unknown hydroxylated metabolites and invites future studies toward the biological effects of these metabolites.

A Diet Rich in Fish Oil and Leucine Ameliorates Hypercalcemia in Tumour-Induced Cachectic Mice
Plas, Rogier L.C. ; Poland, Mieke ; Faber, Joyce ; Argilès, Josep ; Dijk, Miriam van; Laviano, Alessandro ; Meijerink, Jocelijn ; Witkamp, Renger F. ; Helvoort, Ardy van; Norren, Klaske van - \ 2019
International Journal of Molecular Sciences 20 (2019)20. - ISSN 1661-6596
cachexia - fish oil - hypercalcemia - leucine - PTHrP

BACKGROUND: Dietary supplementation with leucine and fish oil rich in omega-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) has previously been shown to reduce cachexia-related outcomes in C26 tumour-bearing mice. To further explore associated processes and mechanisms we investigated changes in plasma Ca2+ levels, the involvement of parathyroid hormone related protein (PTHrP), and its possible interactions with cyclooxygenase 2 (COX-2). METHODS: CD2F1 mice were subcutaneously inoculated with C26 adenocarcinoma cells or sham treated and divided in: (1) controls, (2) tumour-bearing controls, and (3) tumour-bearing receiving experimental diets. After 20 days, body and organ masses and total plasma Ca2+ levels were determined. Furthermore, effects of DHA, EPA and leucine on production of PTHrP were studied in cultured C26 cells. RESULTS: The combination of leucine and fish oil reduced tumour-associated hypercalcemia. Plasma Ca2+ levels negatively correlated with carcass mass and multiple organ masses. DHA was able to reduce PTHrP production by C26 cells in vitro. Results indicate that this effect occurred independently of COX-2 inhibition. CONCLUSION: Our results suggest that cancer-related hypercalcemia may be ameliorated by a nutritional intervention rich in leucine and fish oil. The effect of fish oil possibly relates to a DHA-induced reduction of PTHrP excretion by the tumour.

Urinary Taurine Excretion and Risk of Late Graft Failure in Renal Transplant Recipients
Post, Adrian ; Said, Yusof ; Gomes-Neto, Antonio W. ; Krogt, Jennifer van der; Blaauw, Pim de; Berger, Stefan P. ; Geleijnse, Johanna M. ; Borgonjen, Karin ; Berg, Else van den; Goor, Harry van; Rimbach, Gerald ; Kema, Ido P. ; Tsikas, Dimitrios ; Heiner-Fokkema, Rebecca ; Bakker, Stephan J.L. - \ 2019
Nutrients 11 (2019)9. - ISSN 2072-6643
graft survival - renal transplant recipients - taurine - taurine excretion

Taurine is a sulfur containing nutrient that has been shown to protect against oxidative stress, which has been implicated in the pathophysiology leading to late graft failure after renal transplantation. We prospectively investigated whether high urinary taurine excretion, reflecting high taurine intake, is associated with low risk for development of late graft failure in renal transplant recipients (RTR). Urinary taurine excretion was measured in a longitudinal cohort of 678 stable RTR. Prospective associations were assessed using Cox regression analyses. Graft failure was defined as the start of dialysis or re-transplantation. In RTR (58% male, 53 ± 13 years old, estimated glomerular filtration rate (eGFR) 45 ± 19 mL/min/1.73 m2), urinary taurine excretion (533 (210-946) µmol/24 h) was significantly associated with serum free sulfhydryl groups (β = 0.126; P = 0.001). During median follow-up for 5.3 (4.5-6.0) years, 83 (12%) patients developed graft failure. In Cox regression analyses, urinary taurine excretion was inversely associated with graft failure (hazard ratio: 0.74 (0.67-0.82); P < 0.001). This association remained significant independent of potential confounders. High urinary taurine excretion is associated with low risk of late graft failure in RTR. Therefore, increasing taurine intake may potentially support graft survival in RTR. Further studies are warranted to determine the underlying mechanisms and the potential of taurine supplementation.

N-Eicosapentaenoyl Dopamine, A Conjugate of Dopamine and Eicosapentaenoic Acid (EPA), Exerts Anti-inflammatory Properties in Mouse and Human Macrophages
Augimeri, Giuseppina ; Plastina, Pierluigi ; Gionfriddo, Giulia ; Rovito, Daniela ; Giordano, Cinzia ; Fazio, Alessia ; Barone, Ines ; Catalano, Stefania ; Andò, Sebastiano ; Bonofiglio, Daniela ; Meijerink, Jocelijn ; Witkamp, Renger - \ 2019
Nutrients 11 (2019)9. - ISSN 2072-6643
cyclooxygenase-2 - cytokines - endocannabinoid - inflammation - N-acyl dopamine - N-eicosapentaenoyl dopamine - polyunsaturated fatty acids (PUFAs)

A large body of evidence suggests that dietary n-3 polyunsaturated fatty acids (PUFAs), including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), contribute to a reduced inflammatory tone thereby lowering the risk for several chronic and degenerative diseases. Different mechanisms have been proposed to explain these anti-inflammatory effects, including those involving endocannabinoids and endocannabinoid-like molecules. In this context, fatty acid amides (FAAs), conjugates of fatty acids with amines or amino acids, are an emerging class of compounds. Dopamine conjugates of DHA (N-docosahexaenoyl dopamine, DHDA) and EPA (N-eicosapentaenoyl dopamine, EPDA) have previously been shown to induce autophagy, apoptosis, and cell death in different tumor lines. Additionally, DHDA has displayed anti-inflammatory properties in vitro. Here, we tested the immune-modulatory properties of EPDA in mouse RAW 264.7 and human THP-1 macrophages stimulated with lipopolysaccharide (LPS). EPDA suppressed the production of monocyte chemoattractant protein-1 (MCP-1), and interleukin-6 (IL-6) in both cell lines, and nitric oxide (NO), and macrophage-inflammatory protein-3α (MIP3A) in RAW 264.7 macrophages. At a transcriptional level, EPDA attenuated cyclooxygenase-2 (COX-2) expression in both cell lines and that of MCP-1, IL-6, and interleukin-1β (IL-1β) in THP-1 macrophages. Although further research is needed to reveal whether EPDA is an endogenous metabolite, our data suggest that this EPA-derived conjugate possesses interesting immune-modulating properties.

Characterisation of the effect of day length, and associated differences in dietary intake, on the gut microbiota of Soay sheep
Thomas, Nadine A. ; Olvera-Ramírez, Andrea M. ; Abecia, Leticia ; Adam, Clare L. ; Edwards, Joan E. ; Cox, Georgina F. ; Findlay, Patricia A. ; Destables, Elodie ; Wood, Tracy A. ; McEwan, Neil R. - \ 2019
Archives of Microbiology 201 (2019)7. - ISSN 0302-8933 - p. 889 - 896.
Anaerobic fungi - Bacteria - Ciliated protozoa - Day length - Digestive tract - Soay sheep

Differences in the rumen bacterial community have been previously reported for Soay sheep housed under different day length conditions. This study extends this previous investigation to other organs of the digestive tract, as well as the analysis of ciliated protozoa and anaerobic fungi. The detectable concentrations of ciliated protozoa and anaerobic fungi decreased with increased day length in both the rumen and large colon, unlike those of bacteria where no effect was observed. Conversely, bacterial community composition was affected by day length in both the rumen and large colon, but the community composition of the detectable ciliated protozoa and anaerobic fungi was not affected. Day length-associated differences in the bacterial community composition extended to all of the organs examined, with the exception of the duodenum and the jejunum. It is proposed that differences in rumen fill and ruminal ‘by-pass’ nutrients together with endocrinological changes cause the observed effects of day length on the different gut microbial communities.

Corrigendum: Exposure to antibiotics affects Saponin immersion-induced immune stimulation and shift in microbial composition in Zebrafish larvae
Nadal, Adrià López ; Peggs, David ; Wiegertjes, Geert F. ; Brugman, Sylvia - \ 2019
Frontiers in Microbiology 10 (2019). - ISSN 1664-302X

In the original article, there was an error. The name of the transgenic line used was incorrect. The correct name of the line is "mpeg1:mCherry/mpx:eGFPi114" Corrections have been made to the Materials and Methods subsection Animals: "Adult Tg(mpeg1:mCherry/mpx:eGFPi114) (Renshaw et al., 2006; Bernut et al., 2014) zebrafish (kindly provided by Prof. Meijer, Leiden University), expressing mCherry under the macrophage-specific mpeg1 promotor and GFP under the neutrophil-specific mpx promotor were housed in Zebtec family tanks (Tecniplast, Buguggiate, Italy) under continuous flow-through at 28°C (14/10-hour light/dark cycle) at Carus facilities (WUR, Wageningen, Netherlands). Zebrafish were fed with a mixture of Artemia 230.000 npg (Ocean Nutrition Europe, Essen, Belgium) and Tetramin Flakes (Tetra, Melle, Germany) twice per day. Embryos were obtained by natural spawning and raised with E3 water (0.10 mM NaCl in demineralized water, pH 7.6) in petri dishes at 28°C (12/12-hour light/dark cycle) (Westerfield, 2007). Dead or fungus-infected embryos were identified by microscopy and discarded in tricaine/E3 solution [8.4% (v/v) 24 mM Tricaine (Sigma-Aldrich, DL, United States) stock solution in E3]. Larval ages are expressed in days post-fertilization (dpf). From 5 dpf onward larvae were fed with live daily cultured Tetrahymena pyriformis." Materials and Methods, subsection Dose-Response Experiment Saponin Exposure: "Double Tg(mpeg1:mCherry /mpx:eGFPi114) zebrafish larvae were randomly distributed in 6 well plates (n = 20 fish/well) and exposed to different concentrations [0, 0.5, 0.7 and 1.0 mg/ml] of saponin [ultrapure Soy Saponin 95%, kindly provided by Trond Kortner NMBU Oslo Norway, origin: Organic Technologies, Coshocton, OH (Krogdahl et al., 2015)] dissolved in the E3 (10 ml solution/well) from 6-9 dpf. Mortality was registered and all media were refreshed daily. At 24 h (7 dpf) and 72 h (9 dpf) after the start of the immersion, zebrafish (n = 6-11/group) were anaesthetized embedded and imaged using fluorescent microscopy (as described below). Per time point several larvae were euthanized for further analysis with an overdose MS-222 (8.4 ml of 24 mM Tricaine (Sigma-Aldrich, DL, United States) in 100 ml E3). Pools of 5 larvae were used for RNA extraction (3 pools per group at 24 h, 7-9 pools per group at 72 h) and gene expression was measured on cDNA by Real Time PCR (as described below). Two independent experiments were performed and data were combined." Materials and Methods, subsection Fluorescent in vivo imaging: "Tg(mpeg1:mCherry/mpx:eGFPi114) zebrafish larvae were anaesthetized with tricaine/E3 solution (4.2 ml of 24 mM Tricaine (Sigma-Aldrich, DL, United States) in 100 ml E3) and embedded in 1% low melting point agarose (Thermo Fisher Scientific, MA, United States). Larvae were imaged as whole mounts with a Leica M205 FA Fluorescence Stereo Microscope. After image acquisition, pictures were analyzed with ImageJ® software (United States National Institutes of Health, Bethesda, United States). The intestinal regions were manually selected per fish on the basis of the bright light picture and subsequently copied to the green and red channel pictures (Supplementary Figure S1). Within this intestinal region individual cells were counted for each fish. Furthermore, corrected total cell fluorescence (CTCF) was measured in ImageJ® on total fish larvae by using the following formula: Integrated density-(area of total fish x mean fluorescence of the background reading)." Material and Methods, subsection Experimental Design and Sampling Strategy Antibiotics and Saponin Exposure: A graphical representation of the experimental design and analysis performed per time-point is displayed in Figure 1. To assess the effect of antibiotics, 4 dpf Tg(mpeg1:mCherry/mpx:eGFPi114) fish were randomly distributed in five 6 well-plates (n = 20 fish/well) and 3 treatment conditions were established: (1) control (E3), (2) ciprofloxacin 5 μg/L (Sigma-Aldrich, DL, United States) or (3) oxytetracycline hydrochloride 5 μg/L (Sigma-Aldrich, DL, United States) (10 ml solution/well). The dose of antibiotics was based on several reviews and experimental papers summarizing environmental concentrations of antibiotics in water environments (Ding and He, 2010; Carvalho and Santos, 2016; Watts et al., 2017; Patrolecco et al., 2018; Zhou et al., 2018b) to be at a low dose (ng-μg/L range) and not acute dose (mg/L range). At 6 dpf, 4 pools of 5 larvae were sampled to assess changes in gene expression at baseline. Moreover, at 6 dpf DNA was isolated from 3 pools of 5 larvae to investigate microbiome composition at baseline. In vivo imaging was performed on n = 10 larvae/group to visualize innate immune cells. Subsequently, after sampling, at 6 dpf ultrapure soy saponin was applied to half of the remaining larvae at a concentration 0.5 mg/ml (to induce mild immune stimulation) so each treatment group was split into two, resulting in 6 treatment groups: (1) control, (2) ciprofloxacin (5 μg/L), (3) oxytetracycline hydrochloride (5 μg/L), (4) saponin (0.5 mg/ml), (5) ciprofloxacin + saponin (5 μg/L + 0.5 mg/ml), and (6) oxytetracycline hydrochloride + saponin (5 μg/L + 0.5 mg/ml). All treatment media were refreshed daily. At 9 dpf in vivo imaging was performed on n = 10 larvae/group to visualize innate immune cells. Gene expression was performed on 4 pools of 5 larvae to investigate immune gene expression and from 3 pools of 5 larvae DNA was isolated for microbiological analysis. Because of the error reported above, corrections have also been made to the Figure legends of Figure 2 and Figure 4. The correct legends appear below. Figure 2: Effect of saponin immersion on zebrafish larvae. (A) Percent survival of zebrafish exposed to control (E3), 0.5 mg/ml saponin, 0.7 mg/ml saponin and 1 mg/ml saponin from 6-9 dpf (n = 40 fish/treatment) (Log-rank Mantel-Cox Test for Chi-square, ∗∗∗p < 0.0005). (B) Representative pictures of the saponin-treated Tg(mpeg1:mCherry/mpx:eGFPi114) fish displaying green neutrophils and red macrophages. (C) Quantification of neutrophils and macrophages in the intestinal area (n = 6-11 fish/group) (one way ANOVA Kruskal-Wallis test with Dunn's Multiple comparison Post-Test, mean ± SEM, ∗p < 0.05 ∗∗p < 0.01). Top: counted cells in intestinal area. Bottom: Corrected Total Cell Fluorescence (CTCF, measure for total fluorescent pixels in the whole fish). Two independent experiments were performed and data are combined. Figure 4: Effect of antibiotic exposure on saponin-immune-stimulation. (A) Percent survival of zebrafish exposed to control (E3), ciprofloxacin (4-9 dpf) (5 ug/L) or oxytetracycline (4-9 dpf) (5 ug/ml) + /- saponin (0.5 mg/ml) from 6-9 dpf (n = 100 fish / treatment) (Log-rank Mantel-Cox Test for Chi-square). (B) Representative pictures of the antibiotic/saponin-treated Tg(mpeg1:mCherry/mpx:eGFPi114) fish displaying green neutrophils and red macrophages. (C) Quantification of neutrophils and macrophages in the intestinal area (n = 10 fish/ group) (one way ANOVA Kruskal-Wallis test with Dunn's Multiple comparison Post-Test, mean ± SEM, ∗p < 0.05). Two independent experiments were performed and one representative experiment is shown. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.

The role of n-3 PUFA-derived fatty acid derivatives and their oxygenated metabolites in the modulation of inflammation
Bus, Ian de; Witkamp, Renger ; Zuilhof, Han ; Albada, Bauke ; Balvers, Michiel - \ 2019
Prostaglandins and Other Lipid Mediators 144 (2019). - ISSN 1098-8823
Chemical probes - Endocannabinoid - Inflammation - Oxygenation - PUFA

Notwithstanding the ongoing debate on their full potential in health and disease, there is general consensus that n-3 PUFAs play important physiological roles. Increasing dietary n-3 PUFA intake results in increased DHA and EPA content in cell membranes as well as an increase in n-3 derived oxylipin and -endocannabinoid concentrations, like fatty acid amides and glycerol-esters. These shifts are believed to (partly) explain the pharmacological and anti-inflammatory effects of n-3 PUFAs. Recent studies discovered that n-3 PUFA-derived endocannabinoids can be further metabolized by the oxidative enzymes CYP-450, LOX and COX, similar to the n-6 derived endocannabinoids. Interestingly, these oxidized n-3 PUFA derived endocannabinoids of eicosapentaenoyl ethanolamide (EPEA) and docosahexaenoyl ethanolamide (DHEA) have higher anti-inflammatory and anti-proliferative potential than their precursors. In this review, an overview of recently discovered n-3 PUFA derived endocannabinoids and their metabolites is provided. In addition, the use of chemical probes will be presented as a promising technique to study the n-3 PUFA and n-3 PUFA metabolism within the field of lipid biochemistry.

Colorectal cancer survivors’ beliefs on nutrition and cancer; correlates with nutritional information provision
Veen, Merel R. van; Mols, Floortje ; Smeets, Lian ; Kampman, Ellen ; Beijer, Sandra - \ 2019
Supportive Care in Cancer (2019). - ISSN 0941-4355 - 9 p.
Colorectal cancer survivor - Health professionals - Information provision - Nutrition

Purpose: To investigate CRC survivors’ beliefs on nutrition and cancer and the association with nutritional information provision by (kind and number) of health professionals and to inquire about foods that CRC survivors believed either had a positive or negative influence on their cancer. Methods: A total of 326 CRC survivors of an ongoing prospective cohort study filled out questionnaires 1 month after surgery on whether they had received nutritional information from health professionals. Also, their beliefs that nutrition influences (1) feelings of well-being, (2) complaints after treatment, (3) recovery and (4) cancer recurrence were investigated. Prevalence ratios were calculated (using Cox proportional hazard regression analysis) to study associations between information provision and the four beliefs adjusted for age, gender and cancer stage. Results: Sixty-two percent of respondents received information about nutrition from one or more health professionals. Most respondents who received information strongly believe nutrition influences feelings of well-being (59%) and recovery after cancer (62%). Compared with those who did not receive information, respondents who received information from three professionals showed the strongest beliefs on the influence of nutrition on complaints after treatment (PR 3.4; 95% CI 1.6–7.4), recovery after treatment (PR 2.0; 95% CI 1.2–3.3) and recurrence (PR 2.8; 95% CI 1.3–6.2). Conclusion: Nutritional information provision by health professionals positively influences the beliefs of CRC survivors on the influence of nutrition on cancer outcomes: stronger beliefs occur when respondents received information from three health professionals.

Sensor based eating time variables of dairy cows in the transition periodrelated to the time to first service
Hut, P.R. ; Mulder, A. ; Broek, J. van den; Hulsen, Jan ; Hooijer, G.A. ; Stassen, E.N. ; Eerdenburg, F.J.C.M. van; Nielen, M. - \ 2019
Preventive Veterinary Medicine 169 (2019). - ISSN 0167-5877 - 8 p.
In dairy cattle, reproductive diseases and infertility are some of the most important reasons for culling, where postpartum negative energy balance (NEB) reduces reproductive performance. This single cohort observational study reports the association between eating time and the interval between calving and first service in 2036 dairy cows on 17 commercial farms in The Netherlands. Cows were equipped with a commercially available neck sensor (Nedap, Groenlo, The Netherlands), that measured the time cows spent eating, from 28 days (d) before until 28 d after parturition. Primiparous cows spent a mean of +45 minutes (min) eating time per day ante partum and +15 min eating time post partum more than multiparous cows. A Cox proportional hazard model was used to analyze eating time variables in relation to the interval between calving and first service. From 4 weeks before until 4 weeks after calving eating time variables per week were used. Weeks -4, -3 + 3 and +4 were used as weeks with stable eating time patterns and therefore the mean eating time per week and the standard deviation of the mean eating time per week were used. Weeks -2, -1, +1 and +2 were addressed as periods with unstable eating patterns and therefore the slope in eating time per week and the residual variance of the slope per week were modeled. Significant results were the mean eating time in week -4 and +3 where in both weeks higher eating time lead to a higher hazard for first service. Difference between primiparous and multiparous cows were also significant with a higher hazard for first service for primiparous cows. Week 4 post partum presented a significant difference between eating time of primiparous cows and multiparous cows. These results display how eating time variables in the transition period could be related to the interval between calving and first service, and that there is a relation between mean eating time in week -4, +3, +4 and the interval between calving and first insemination.
High Dietary Intake of Vegetable Protein Is Associated With Lower Prevalence of Renal Function Impairment: Results of the Dutch DIALECT-1 Cohort
Oosterwijk, Milou M. ; Soedamah-Muthu, Sabita S. ; Geleijnse, Johanna M. ; Bakker, Stephan J.L. ; Navis, Gerjan ; Binnenmars, S.H. ; Gant, Christina M. ; Laverman, Gozewijn D. - \ 2019
Kidney International Reports 4 (2019)5. - ISSN 2468-0249 - p. 710 - 719.
diabetes mellitus type 2 - diet - kidney function - lifestyle

Introduction: Dietary protein intake may influence development of renal function impairment in diabetes mellitus type 2 (T2DM). We assessed the association between sources of protein and prevalence of renal function impairment. Methods: Cross-sectional analyses were performed in baseline data of 420 patients of the DIAbetes and LifEstyle Cohort Twente-1 (DIALECT-1)study. Protein intake was assessed using a Food Frequency Questionnaire, modified for accurate assessment of protein intake, including types and sources of protein. Renal function impairment was defined as estimated glomerular filtration rate (eGFR)<60 ml/min per 1.73 m 2 (Chronic Kidney Disease Epidemiology Collaboration formula). Results: Among 420 patients with T2DM, 99 renal function impairment cases were identified. Multivariate Cox proportional hazard models were used and adjusted for the main lifestyle and dietary factors. The prevalence ratios in the fully adjusted model were 1 (reference), 0.74 (95% confidence interval [CI]: 0.44–1.27; P = 0.28)and 0.47 (95% CI: 0.23–0.98; P = 0.04)according to increasing tertiles of vegetable protein intake. For animal protein intake the prevalence ratios were 1 (reference), 1.10 (95% CI: 0.64–1.88; P = 0.74)and 1.06 (95% CI: 0.56–1.99; P = 0.87)according to increasing tertiles of intake. Theoretical replacement models showed that replacing 3 energy percent from animal protein by vegetable protein lowered the prevalence ratio for the association with renal function impairment to 0.20 (95% CI: 0.06–0.63; P = 0.01). Conclusion: In conclusion, we found that higher intake of vegetable protein was associated with a lower prevalence of renal function impairment, and theoretical replacement of animal protein with vegetable protein was inversely associated with renal function impairment among patients with T2DM.

Identification of hydroxytyrosyl oleate, a derivative of hydroxytyrosol with anti-inflammatory properties, in olive oil by-products
Plastina, Pierluigi ; Benincasa, Cinzia ; Perri, Enzo ; Fazio, Alessia ; Augimeri, Giuseppina ; Poland, Mieke ; Witkamp, Renger ; Meijerink, Jocelijn - \ 2019
Food Chemistry 279 (2019). - ISSN 0308-8146 - p. 105 - 113.
COX-2 - Hydroxytyrosyl ester - Inflammation - Macrophages - NO - Olive mill waste water - Olive oil - PGE

Hydroxytyrosyl esters with short, medium and long acyl chains were evaluated for their ability to reduce nitric oxide (NO) production by lipopolysaccharide-stimulated RAW264.7 macrophages. Among the compounds tested, C18 esters, namely hydroxytyrosyl stearate (HtySte) and hydroxytyrosyl oleate (HtyOle), were found to decrease NO production in a concentration-dependent manner, while the other compounds, including the parent hydroxytyrosol, were ineffective in the tested concentration range (0.5–5 μM). Further study of the potential immune-modulating properties of HtyOle revealed a significant and concentration-dependent suppression of prostaglandin E2 production. At a transcriptional level, HtyOle inhibited the expression of inducible NO synthase, cyclooxygenase-2 and interleukin-1β. Moreover, HtyOle was identified for the first time in olive oil by-products by means of high performance liquid chromatography coupled with mass spectrometry. By contrast, HtyOle was not found in intact olives. Our results suggest that HtyOle is formed during oil processing and represents a significant form in which hydroxytyrosol occurs.

The Birth, Growth and Death of Intertidal Soft-Sediment Bivalve Beds: No Need for Large-Scale Restoration Programs in the Dutch Wadden Sea
Meer, Jaap van der; Dankers, Norbert ; Ens, Bruno J. ; Stralen, Marnix van; Troost, Karin ; Waser, Andreas M. - \ 2019
Ecosystems 22 (2019)5. - ISSN 1432-9840 - p. 1024 - 1034.
Crassostrea gigas - demography - Mytilus edulis - overfishing - proportional hazard - recovery - restoration - survival analysis

Recruitment and fate of all 1436 mussel and oyster beds of the Dutch Wadden Sea were studied for the period 1999–2013. Cox’s proportional hazard rate model with covariates such as orbital speed, exposure time and bed size and type showed that large, low-lying beds that experience a low orbital speed live longer. Yet the most striking result was that oyster and mixed beds have a much lower hazard rate than pure mussel beds. Simulation studies, using the observed recruitment series, which was very variable, and the estimated survival curves, showed large variability in total bed area, implying that the present area, though lower than before, does not point to a systematic deviation from the pre-1990 situation, that is, the situation before intensive fisheries on these intertidal beds and the disappearance of them around 1990. Claims that bivalve bed recovery is impossible without restoration efforts are premature and not supported by our analysis. On the contrary, the observed high survival rate of mixed and oyster beds and the expectation that such beds will predominate in the near future can easily result in larger future bed coverage than what has been measured before.

Does clinical mastitis in the first 100 days of lactation 1 predict increased mastitis occurrence and shorter herd life in dairy cows?
Hertl, J.A. ; Schukken, Y.H. ; Tauer, L.W. ; Welcome, F.L. ; Gröhn, Y.T. - \ 2018
Journal of Dairy Science 101 (2018)3. - ISSN 0022-0302 - p. 2309 - 2323.
generalized linear mixed models - herd life - mastitis - survival analysis

The objectives of this study were to estimate the direct effects of clinical mastitis (CM) occurring in early productive life (defined as the first 100 d of the first lactation) of Holstein dairy cows on the future rate of CM occurrence and on the length of total productive lifetime. Information on CM cases and other data occurring in 55,144 lactations in 24,831 cows in 5 New York State Holstein herds was collected from January 2004 until February 2014. For the first objective, a generalized linear mixed model with a Poisson distribution was used to study the effects of CM cases occurring in the first 100 d of a cow's first lactation, as well as farm indicator and number of days in the cow's lifetime, on the future lifetime rate of CM. Only cows that had completed their productive life [i.e., all had been culled (or sold) or had died; n = 14,440 cows] were included in this analysis. For the second objective, a Cox proportional hazards model was used to study the effects of CM cases occurring in the first 100 d of a cow's first lactation on the length of total productive lifetime. The model was stratified by farm. All 24,831 cows were included in this analysis with right censoring. Cows experienced between 0 and 4 CM cases in the first 100 d of lactation 1. Over their lifetime, cows experienced between 0 and 25 CM cases. During the study period, 10% of all cows died and nearly half of all cows were culled. The average length of productive life, including censored observations, was 2.0 yr after first calving. Compared with cows having no CM cases in the first 100 d of lactation 1, cows with 1 CM case in that time period had a 1.5 times higher rate of total number of CM cases over their lifetime. Cows with 2 (or 3 or more) CM cases in the first 100 d of lactation 1 had a 1.7 times (or 2.6 times) higher rate of total number of CM cases over their lifetime. For each additional CM case occurring in the first 100 d of lactation 1, the hazard rate of culling increased by 34%. Given economic conditions for preferentially culling mastitic cows, the study findings may help farmers make optimal decisions with regard to culling of such cows.

Drug use is associated with lower plasma magnesium levels in geriatric outpatients; possible clinical relevance
Orten-Luiten, A.C.B. van; Janse, A. ; Verspoor, E. ; Brouwer-Brolsma, E.M. ; Witkamp, R.F. - \ 2018
Clinical Nutrition (2018). - ISSN 0261-5614
Adverse drug reaction - Cardiovascular disease - Diabetes - Drug-food interaction - Magnesium deficiency - Osteoporosis

Background: Hypomagnesemia has been associated with diabetes, cardiovascular disease, and other disorders. Drug use has been suggested as one of the risk factors for low magnesium (Mg) levels. In the elderly population, prone to polypharmacy and inadequate Mg intake, hypomagnesemia might be relevant. Therefore, we aimed to investigate associations between drug use and plasma Mg. Methods: Cross-sectional data of 343 Dutch geriatric outpatients were analysed by Cox and linear regression, while adjusting for covariates. Drug groups were coded according to the Anatomical Therapeutic Chemical classification system; use was compared to non-use. Hypomagnesemia was defined as plasma Mg < 0.75 mmol/l and <0.70 mmol/l. Results: Prevalence of hypomagnesemia was 22.2% (Mg < 0.75 mmol/l) or 12.2% (Mg < 0.70 mmol/l); 67.6% of the patients used ≥5 medications (polypharmacy). The number of different drugs used was inversely linearly associated with Mg level (beta −0.01; p < 0.01). Fully adjusted Cox regression showed significant associations of polypharmacy with hypomagnesemia (Mg < 0.75 mmol/l) (prevalence ratio (PR) 1.81; 95%CI 1.08–3.14), proton pump inhibitors (PR 1.80; 95%CI 1.20–2.72), and metformin (PR 2.34; 95%CI 1.56–3.50). Moreover, stratified analyses pointed towards associations with calcium supplements (PR 2.26; 95%CI 1.20–4.26), insulins (PR 3.88; 95%CI 2.19–6.86), vitamin K antagonists (PR 2.01; 95%CI 1.05–3.85), statins (PR 2.44; 95%CI 1.31–4.56), and bisphosphonates (PR 2.97; 95%CI 1.65–5.36) in patients <80 years; selective beta blockers (PR 2.01; 95%CI 1.19–3.40) if BMI <27.0 kg/m2; and adrenergic inhalants in male users (PR 3.62; 95%CI 1.73–7.56). Linear regression supported these associations. Conclusion: As polypharmacy and several medications are associated with hypomagnesemia, Mg merits more attention, particularly in diabetes, cardiovascular disease, and in side-effects of proton pump inhibitors and calcium supplements.

Assignment of a dubious gene cluster to melanin biosynthesis in the tomato fungal pathogen Cladosporium fulvum
Griffiths, Scott A. ; Cox, Russell J. ; Overdijk, Elysa J.R. ; Mesarich, Carl H. ; Saccomanno, Benedetta ; Lazarus, Colin M. ; Wit, Pierre J.G.M. de; Collemare, Jérôme - \ 2018
PLoS ONE 13 (2018)12. - ISSN 1932-6203

Pigments and phytotoxins are crucial for the survival and spread of plant pathogenic fungi. The genome of the tomato biotrophic fungal pathogen Cladosporium fulvum contains a predicted gene cluster (CfPKS1, CfPRF1, CfRDT1 and CfTSF1) that is syntenic with the characterized elsinochrome toxin gene cluster in the citrus pathogen Elsinoë fawcettii. However, a previous phylogenetic analysis suggested that CfPks1 might instead be involved in pigment production. Here, we report the characterization of the CfPKS1 gene cluster to resolve this ambiguity. Activation of the regulator CfTSF1 specifically induced the expression of CfPKS1 and CfRDT1, but not of CfPRF1. These co-regulated genes that define the CfPKS1 gene cluster are orthologous to genes involved in 1,3-dihydroxynaphthalene (DHN) melanin biosynthesis in other fungi. Heterologous expression of CfPKS1 in Aspergillus oryzae yielded 1,3,6,8-tetrahydroxynaphthalene, a typical precursor of DHN melanin. Δcfpks1 deletion mutants showed similar altered pigmentation to wild type treated with DHN melanin inhibitors. These mutants remained virulent on tomato, showing this gene cluster is not involved in pathogenicity. Altogether, our results showed that the CfPKS1 gene cluster is involved in the production of DHN melanin and suggests that elsinochrome production in E. fawcettii likely involves another gene cluster.

Association Between Oropharyngeal Dysphagia and Malnutrition in Dutch Nursing Home Residents: Results of the National Prevalence Measurement of Quality of Care
Huppertz, Viviënne A.L. ; Halfens, Ruud J.G. ; Helvoort, Ardy van; Groot, C.P.G.M. de; Baijens, L.W.J. ; Schols, J.M.G.A. - \ 2018
Journal of Nutrition, Health and Aging 22 (2018)10. - ISSN 1279-7707 - p. 1246 - 1252.
Objectives: Nursing home residents often suffer from multi-morbidities and geriatric syndromes leading to lower quality of life or mortality. Oropharyngeal dysphagia (OD) and malnutrition are profound conditions in this complex profile of multi-morbidities and are associated with deprived mental –and physical health status, e.g. aspiration pneumonia or dehydration. This study aimed to assess the association between OD and malnutrition in Dutch nursing home residents. Design: Data for this cross-sectional study were obtained from the annual National Prevalence Measurement of Quality of Care (LPZ). Setting: The National Prevalence Measurement of Quality of Care was conducted in Nursing Homes in The Netherlands. Participants: Participants were nursing home residents age 65 or older and admitted to psychogeriatric- or somatic wards. Measurements: The measurements were taken by trained nurses from the participating nursing homes. Anthropometric measurements and unintended weight loss (%) were assessed to determine nutritional status (malnutrition). OD was assessed by means of a standardized questionnaire assessing clinically relevant symptoms of OD such as swallowing problems or sneezing/coughing while swallowing. Cox regression was applied to assess the association between malnutrition and clinically relevant symptoms of OD in older Dutch nursing home residents. Results: Approximately 12% of the residents suffered from swallowing problems and 7% sneezed/coughed while swallowing liquids or solid foods. Approximately 10% of the residents was malnourished. Residents with OD symptoms were more often malnourished compared to residents without OD symptoms. Approximately 17% of the problematic swallowers were concurrently malnourished. Increased risk for malnutrition was found in residents suffering from swallowing problems (PR 1.5, 95%CI 1.2–1.9), as well as in residents that sneezed/ coughed while swallowing (PR 1.3, 95%CI 1.0–1.7). Stratification based on wards revealed that problematic swallowers from somatic wards were at a high risk of malnutrition (PR 1.9, 95%CI 1.3–2.8). Conclusion: Clinically relevant symptoms of oropharyngeal dysphagia, such as swallowing problems and sneezing/coughing while swallowing are associated with increased risk of malnutrition in psychogeriatric and somatic Dutch nursing home residents.
Low radiographic muscle density is associated with lower overall and disease-free survival in early-stage colorectal cancer patients
Baar, Harm van; Beijer, S. ; Bours, M.J.L. ; Weijenberg, M.P. ; Zutphen, M. van; Duijnhoven, F.J.B. van; Slooter, G.D. ; Pruijt, J.F.M. ; Dronkers, J.J. ; Haringhuizen, A. ; Spillenaar Bilgen, E.J. ; Hansson, B.M.E. ; Wilt, J.H.W. de; Kampman, E. ; Winkels, R.M. - \ 2018
Journal of Cancer Research and Clinical Oncology 144 (2018)11. - ISSN 0171-5216 - p. 2139 - 2147.
Colorectal cancer - Mortality - Skeletal muscle density - Survival

Background: In cancer patients with a poor prognosis, low skeletal muscle radiographic density is associated with higher mortality. Whether this association also holds for early-stage cancer is not very clear. We aimed to study the association between skeletal muscle density and overall mortality among early-stage (stage I–III) colorectal cancer (CRC) patients. Furthermore, we investigated the association between skeletal muscle density and both CRC-specific mortality and disease-free survival in a subset of the study population. Methods: Skeletal muscle density was assessed in 1681 early-stage CRC patients, diagnosed between 2006 and 2015, using pre-operative computed tomography images. Adjusted Cox proportional hazard models were used to evaluate the association between muscle density and overall mortality, CRC-specific mortality and disease-free survival. Results: The median follow-up time was 48 months (range 0–119 months). Low muscle density was detected in 39% of CRC patients. Low muscle density was significantly associated with higher mortality (low vs. normal: adjusted HR 1.91, 95% CI 1.53–2.38). After stratification for comorbidities, the association was highest in patients with ≥ 2 comorbidities (HR 2.11, 95% CI 1.55–2.87). Furthermore, low skeletal muscle density was significantly associated with poorer disease-free survival (HR 1.68, 95% CI 1.14–2.47), but not with CRC-specific mortality (HR 1.68, 95% CI 0.89–3.17) in a subset of the study population. Conclusion: In early-stage CRC patients, low muscle density was significantly associated with higher overall mortality, and worse disease-free survival.

Dietary Approach to Stop Hypertension (DASH) diet and risk of renal function decline and all-cause mortality in renal transplant recipients
Osté, Maryse C.J. ; Gomes-Neto, António W. ; Corpeleijn, Eva ; Gans, Rijk O.B. ; Borst, Martin H. de; Berg, Else van den; Soedamah-Muthu, Sabita S. ; Kromhout, Daan ; Navis, Gerjan J. ; Bakker, Stephan J.L. - \ 2018
American Journal of Transplantation 18 (2018)10. - ISSN 1600-6135 - p. 2523 - 2533.
Clinical research/practice - Graft survival - Kidney transplantation/nephrology - Nutrition - Patient survival
Renal transplant recipients (RTR) are at risk of decline of graft function and premature mortality, with high blood pressure as an important risk factor for both. To study the association of the Dietary Approach to Stop Hypertension (DASH) diet with these adverse events, we conducted a prospective cohort study of adult RTR. Dietary data were collected using a validated 177-item food frequency questionnaire and an overall DASH-score was obtained. We included 632 stable RTR (mean ± standard deviation age 53.0 ± 12.7 years, 57% men). Mean DASH score was 23.8 ± 4.7. During median follow-up of 5.3 (interquartile range, 4.1-6.0) years, 119 (18.8%) RTR had renal function decline, defined as a combined endpoint of doubling of serum creatinine and death-censored graft failure, and 128 (20.3%) died. In Cox-regression analyses, RTR in the highest tertile of the DASH score had lower risk of both renal function decline (hazard ratio [HR] = 0.57; 95% confidence interval [CI], 0.33-0.96, P = .03) and all-cause mortality (HR = 0.52; 95%CI, 0.32-0.83, P = .006) compared to the lowest tertile, independent of potential confounders. Adherence to a DASH-style diet is associated with lower risk of both renal function decline and all-cause mortality. These results suggest that a healthful diet might benefit long-term outcome in RTR.
In vitro anti-inflammatory and radical scavenging properties of chinotto (Citrus myrtifolia Raf.) essential oils
Plastina, Pierluigi ; Apriantini, Astari ; Meijerink, Jocelijn ; Witkamp, Renger ; Gabriele, Bartolo ; Fazio, Alessia - \ 2018
Nutrients 10 (2018)6. - ISSN 2072-6643
Antioxidant - Citrus - Inflammation - Macrophages - Nitric oxide

Chinotto (Citrus myrtifolia Raf.) is a widely diffused plant native from China and its fruits have a wide-spread use in confectionary and drinks. Remarkably, only little has been reported thus far on its bioactive properties, in contrast to those of the taxonomically related bergamot (Citrus bergamia Risso). The present study aimed to investigate potential in vitro anti-inflammatory and radical scavenging properties of chinotto essential oils (CEOs) and to establish to what extent their composition and bioactivities are dependent on maturation. Essential oil from half ripe chinotto (CEO2) reduced the production of nitric oxide (NO) and the expression of inflammatory genes, cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), cytokines, including interleukin-1β (IL-1β) and interleukin-6 (IL-6), and chemokine monocyte chemotactic protein-1 (MCP-1) by lipopolysaccharide (LPS)-stimulated RAW264,7 macrophages. Limonene, linalool, linalyl acetate, and γ-terpinene were found to be the main components in CEO2. Moreover, CEO2 showed high radical scavenging activity measured as Trolox equivalents (TE) against both 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS). These findings show that chinotto essential oil represents a valuable part of this fruit and warrants further in vivo studies to validate its anti-inflammatory potential.

The Peroxisome Proliferator-Activated Receptor α is dispensable for cold-induced adipose tissue browning in mice
Defour, M. ; Dijk, W. ; Ruppert, P.M.M. ; Nascimento, Emmani B.M. ; Schrauwen, Patrick ; Kersten, A.H. - \ 2018
Mus musculus - GSE110420 - PRJNA433669
Chronic cold exposure causes white adipose tissue (WAT) to adopt features of brown adipose tissue, a process known as browning. Previous studies have hinted at a possible role for the transcription factor Peroxisome Proliferator-Activated Receptor alpha (PPARα) in cold-induced browning. Here we aimed to investigate the importance of PPARα in driving transcriptional changes during cold-induced browning in mice. Male wildtype and PPARα−/− mice were housed at thermoneutrality (28 °C) or cold (5 °C) for 10 days. Whole genome expression analysis was performed on inguinal WAT. In addition, other analyses were carried out. Whole genome expression data of livers of wildtype and PPARα−/− mice fasted for 24 h served as positive control for PPARα-dependent gene regulation.Cold exposure increased food intake and decreased weight of BAT and WAT to a similar extent in wildtype and PPARα−/− mice. Except for plasma non-esterified fatty acids, none of the cold-induced changes in plasma metabolites were dependent on PPARα genotype. Histological analysis of inguinal WAT showed clear browning upon cold exposure but did not reveal any morphological differences between wildtype and PPARα−/− mice. Transcriptomics analysis of inguinal WAT showed a marked effect of cold on overall gene expression, as revealed by principle component analysis and hierarchical clustering. However, wildtype and PPARα−/− mice clustered together, even after cold exposure, indicating a similar overall gene expression profile in the two genotypes. Pathway analysis revealed that cold upregulated pathways involved in energy usage, oxidative phosphorylation, and fatty acid β-oxidation to a similar extent in wildtype and PPARα−/− mice. Furthermore, cold-mediated induction of genes related to thermogenesis such as Ucp1, Elovl3, Cox7a1, Cox8, and Cidea, as well as many PPAR target genes, was similar in wildtype and PPARα−/− mice. Finally, pharmacological PPARα activation had a minimal effect on expression of cold-induced genes in murine WAT.Cold-induced changes in gene expression in inguinal WAT are unaltered in mice lacking PPARα, indicating that PPARα is dispensable for cold-induced browning.
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