Adults aged > 50 years, which make up to 40% of the European population, are vulnerable to low vitamin B12 (B12) status due to age-related factors that impair digestion and absorption of protein-bound B12 from natural food sources. Older adults are recommended to meeting their dietary B12 intake through the consumption of B12-fortified foods or supplements because these products contain free B12. B12 seems most bioavailable from milk products compared to other food sources, showcasing dairy as a potential vehicle for B12 food fortification. Yogurt is a versatile, popular dairy product, making it a promising food vehicle for B12 fortification to enhance the availability of B12-fortified foods for population groups at risk of B12 deficiency. With the overall goal to develop a novel, B12-fortified dairy product, the objective of this project was to compare the shelf-life stability of different chemical forms of B12 added to yogurt either in isolated or in encapsulated form. For both fortification strategies, we compared methylcobalamin (MeCB), a naturally-occurring B12 form, and cyanocobalamin (CnCB), the synthetic form of B12. Encapsulated microparticles were created by spray-drying a maize starch-derived polymeric material (EUDRAGUARD Natural®) with 1% (w/V) MeCB or CnCB. The release of B12 from encapsulated microparticles was confirmed by in vitro simulated gastrointestinal digestion; maximum B12 release (103% recovery) was achieved under conditions simulating the small intestine, where B12 is absorbed in the human body. Yogurts were produced by incubating whole milk (3.25% milk fat) with yogurt starter bacteria at 43°C overnight; after fermentation and cooling down, isolated (I-MeCB or I-CnCB) or encapsulated B12 powders (E-MeCB or E-CnCB) in the concentration of 50μg of B12/175 g of yogurt were added, creating stirred yogurt. Yogurts were stored at 4°C for eight weeks, with intermittent sampling for B12 stability testing using RIDASCREEN immunoassay. For CnCB-fortified yogurts, I-CnCB and E-CnCB had similar B12 concentrations at baseline (48.3μg and 48.2μg, respectively) and week 8 (53.9μg and 51.4μg, respectively). Difference in B12 content between I-CnCB and E-CnCB was < 5% for both time-points. For MeCB, similar concentrations were found for I-MeCB (64.1μg) and E-MeCB (65.0μg) at baseline; however, at week 8, E-MeCB remained stable (69.8μg) while a pronounced decrease in B12 was observed with I-MeCB (33.3μg). At week 8, I-MeCB B12 concentrations were 52% lower than E-MeCB. In conclusion, CnCB was shown to be the more stable fortificant throughout shelf-life. Encapsulation techniques are a viable option to increase MeCB stability in fortified yogurts.

When obesity arises, adipose tissue (AT) expands and shifts to an influx of pro-inflammatory cells, leading to a state of chronic AT-inflammation.Furthermore, a western diet (WD) modulates the gut microbiome, increasing intestinal permeability. This facilitates the translocation of endotoxins and even entire bacteria into the blood stream, further contributing to the pro-inflammatory state. Even though it is evident a WD, high in fat and carbohydrates (CHO), can cause AT-inflammation, it is still unclear if fat or CHO is the main inducer. Therefore, we are currently investigating the effect of different CHO-types on AT-inflammation.

During 15 weeks, male C57BL/6JRj mice were kept on several diets, consisting of high-fructose (HFRD), high-sucrose-high-fructose (HCFD), high-starch (HSTD) or a WD (n = 10 per group). Weekly monitoring of body weight and food intake was followed by analyses of visceral AT-inflammation. Kruskal-wallis tests were used for statistical analysis.

Mice on HSTD and HCD had gained significantly less body weight compared to mice on WD after 15 weeks of diet. Mice on HSTD also gained significantly less body weight compared to mice on HFRD and HCFD. Moreover, mice on HSTD and HCD also had significantly smaller AT-depots as compared to mice on WD. Total plasma cholesterol as well as HDL and LDL levels were significantly lower in mice on HSTD and on HCD as compared to mice on WD. Gene expression analysis revealed a significantly lower expression of several pro-inflammatory markers (F4/80, Arg1, Mannose receptor, TNF, MCP1 and Saa3) in AT of mice on HSTD and on HCD compared to a WD. A HSTD also induced lower AT-expression of MCP1 and Saa3 than a HCFD and Saa3 expression was also significantly lower in the HSTD-group compared to the HFRD-group. Furthermore, Foxp3 expression, a marker for anti-inflammatory Treg cells, was significantly increased in AT of all CHO-diet fed mice as compared to the WD-group.

In conclusion, these data suggest that certain dietary carbohydrates, in contrast to a WD, do not induce obesity or AT-inflammation, including lower gene expression of Saa3. It is stated that a WD induces Saa3 expression not only in AT, but also in the colon. Furthermore, since Saa3 is able to bind bacteria and is associated with inflammation, further research is necessary to investigate Saa3 as a possible link between disturbances in the gut microbiota and AT-inflammation.

The popularity of smartphone apps is rising globally. However, the quality of widely available health apps and their effectiveness to promote behaviour change, by incorporating behavioural change techniques (BCTs) that have been suggested to result in behaviour change, is questionable. The aim of this study was to conduct a content analysis of smartphone apps that utilise the Mediterranean diet (MD), a dietary pattern linked to numerous health benefits, to promote healthy dietary behaviours. The specific objectives were to evaluate the quality of these apps and their use of BCTs. The iTunes Store and Google Play were searched for all available apps (free and requiring payment to download) promoting the MD. Apps were excluded if they were not available in English, did not focus only on the MD or the download process was corrupt. The Mobile App Rating Scale (MARS) was used to assess app engagement, functionality, aesthetics and information quality (the average of which represents a quality mean score (QMS)) and subjective quality. The Abraham & Michie behaviour change taxonomy was adapted to create a 20-item checklist to assess the presence of BCTs in the eligible apps. Thirty nine apps (64.1% of which were free to download) were analysed. The overall QMS was 2.96 ± 0.46. The lowest and highest scored QMS constructs were engagement (2.42 ± 0.62) and functionality (3.95 ± 0.45), respectively, whilst the overall mean subjective quality score was 1.71 ± 0.51 (scores’ range 1–5). Apps were scored higher for aesthetics if they required payment (3.14 vs. 2.78, p = 0.047). Apps incorporated an average of 3.10 ± 1.87 BCTs, with no difference in the number of BCTs present according to platform or cost. From the five BCTs that have been suggested to promote behaviour change, ‘self-monitoring’ was present in six apps, ‘intention formation’ in 15 and ‘goal setting’ in one. None of the apps incorporated the ‘review of behavioural goals’ and ‘feedback on performance’ BCTs. The total QMS score was positively associated with the presence of BCTs (r = 0.409, p = 0.008). Based on the MARS assessment, apps promoting the MD that are available in the public domain were of moderate quality. In contrast, the low presence, particularly of efficacious, BCTs and low subjective quality scores suggest that apps are actually of poor quality and potentially not effective in promoting behaviour change towards an MD diet. These findings provide important insights for the future development of high-quality apps to promote the MD at population level.

Alternative teaching methods focusing on learner-centered and competency-based education lead to an increase in knowledge and skills students will have to apply after graduation(1); it has been shown that expecting to teach improves learning(2). Flipped classroom is an instructional approach combining individual and team work with active learning strategies(3). Its potential in nutrition courses has been formerly demonstrated, but the results focused on students' satisfaction(1). Aim of this study was to evaluate flipped classroom efficacy for Dietetics students’ achievement in dietetics and nutritional therapy. The study was conducted at University of Pavia, Italy.

Sixteen students (8 males) participated in flipped learning during diet therapy course in the third year. They attended 32 hours of in-class activities, case reports discussions, group-based problem solving activities facing 10 different physiological and clinical conditions implementing the Nutrition Care Process approach. Students were assessed during each lesson and with a low stake test. A mid-term anonymous questionnaire using the 5 Likert scale items was submitted in order to evaluate any critical issues besides the degree of satisfaction. Data were analyzed by frequency.

Both teamwork and individual initiative and problem solving skills were assessed: 12 students out of 16 were positively evaluated; 38% of students contributed greatly to the discussion in class (more than 5 times during the course). Students showed a high level of frequency in using scientific evidence, authoritative websites, guidelines and position papers (80%). All improved their slideshow contribution, becoming more effective as course progressed, scoring high (7/10) at the end of the course. Students were satisfied with the flipped learning class overall (68%), referring that flipped learning was helpful for self-directed learning (80%) and motivation for learning (81%), they enjoyed the use of training videos and would have liked to be able to use more (86%).

The Flipped classroom is a valid approach when applied to final year students who already have reached autonomy in scientific literature search and identification of the appropriate sources for problem solving and clinical cases discussion. It was observed that flipped learning is helpful for the interaction and collaboration in the classroom. However conclusions are difficult to be drawn because this study was conducted only once in only one course, so it is hard to generalize the results. Besides further studies are recommended to compare an experimental group with a control group.

Introduction

For nutrition and behaviour experiments, researchers often desire to change visual properties of food items. However, such changes can lead to unwanted side-effects in textural or sensory properties. With VR technology, research in food science and nutrition can draw from a novel range of possibilities that allow specific changes in visual features without altering other properties.

Method

Study 1 (99 participants): While wearing VR glasses, participants received two banana juices. In the virtual environment, one was coloured yellow (its real appearance) and one was brownish (indicating aging). All participants viewed both juices, either one or the other first, and the juice they tasted was the same in both cases.

Study 2 (100 participants): Participants were divided into a disgust and control condition. Wearing VR glasses, those in the control condition saw chocolate appear on the table and were asked to try it. Those in the disgust condition saw a dog excrete the chocolate as feces before being asked to try it.

Study 3 (50 participants): In the virtual environment, participants saw a disgusting buffet containing carrots, pasta, and meat. They were then asked to serve themselves a plate for lunch.

Study 4 (100 participants): Building on previous sensory studies, participants saw in the virtual environment two juices and a cake, either in the correct or wrong colour. They were asked to try the product, identify the dominant flavour, and rate it for liking and sweetness.

Results and Discussion

Study 1: With increasing presence, participants tended to give higher disgust scores to the brown juice (r = .24, p < 0.05).

Study 2: Four percent of participants refused consumption in the neutral condition compared to 26% in the disgust condition. With χ2(1) = 9.49, p > 0.01, the chi-square test revealed a significant association between the experimental condition and participants’ willingness to eat the chocolate.

Study 3: The total quantity of food that participants scooped onto their plates was significantly correlated with their presence (r = -0.37, p < 0.01) and their state disgust (r = -0.35, p < 0.05).

Study 4: We were able to replicate previous findings.

In summary, our studies assessed a new technology for usability in food research. Our results provided evidence that VR is a promising tool in the study of nutrition and behaviour.

A study by Kent et al (Lancet Public Health, 2017), found a clear relationship between body weight and hospital admissions. In a recent observational study by Agarwal et al (Clinical Nutrition, 2018), one-in-three non-critically ill malnourished obese adults had poor food intake in hospital, and malnourished obese were least likely to receive nutrition support. The aim of this review was to describe the characteristics of obese inpatients referred to the dietitian and the range of nutritional care plans required. Data was collected retrospectively from dietetic records of inpatients with Body Mass Index (B.M.I.) > 35 kg / m2, who were nutritionally assessed by the registered dietitian, in a 12 month period. Data included age, sex, weight, height, B.M.I. and length of stay, as well as diagnosis, comorbidities and their nutritional management. Institutional approval was obtained for this data review. 45 patients with B.M.I. > 35 kg / m2 (30 men and 15 women) were referred to the dietitian during the time period, admitted under both medical (78%) and surgical (22%) specialities. Mean age was 58yrs (range 27–83yrs), mean BMI was 43 kg / m2 (range 35–59.4 kg / m2), 55% were morbidly obese (BMI > 40 kg / m2). Patients had multiple comorbidities requiring nutritional intervention to optimise management. 78% had Diabetes Mellitus: 57 % were on insulin. One third had renal impairment. One third had Congestive Cardiac Failure. 62% had respiratory complications, with 22% having spent time in intensive care. Following nutritional assessment, 78% required nutrition support to meet their nutritional requirements, of which 51 % required nasogastric feeding and 86% were on oral nutritional supplements. Of patients requiring nutrition support, 63% (22 patients) were also on a therapeutic diet, and 18% (8 patients) required texture modified diets. Of the patients not requiring nutrition support, 22% (n = 10):15% received nutritional education regarding diabetes and only 7% received weight management advice as inpatients. Median length of stay was 14.5 days (range 4–193 days), compared to a mean length of stay for medical and surgical patients of 10 days and 7.1 days respectively. Obese inpatients are a complicated cohort, who typically present with multiple comorbidities. They can have complex, multifocal nutritional needs and may struggle to meet their nutritional requirements as inpatients, leading to unintentional weight loss in the acute setting. This review highlights the importance of screening obese patients for malnutrition risk, and the need for early referral to the dietitian for nutritional assessment and individualised nutrition care plans.

Traditional malnutrition screening instruments, such as the Malnutrition Universal Screening Tool (MUST), strongly rely on criteria regarding low BMI and critical weight loss for risk assessment. In an increasingly overweight and obese population, this may result in underdetection of malnutrition. The Patient-Generated Subjective Global Assessment (PG-SGA) was developed in the broader context of anabolic competence and includes assessment of other aspects of malnutrition, such as nutrition impact symptoms and metabolic demand. We therefore aimed to compare MUST and PG-SGA in assessing prevalence of malnutrition in overweight and obese hospitalized patients. We measured risk for malnutrition (MUST score: 0 = low risk, 1 = medium risk, ≥ 2 = high risk) and nutritional status (PG-SGA Categories: A = well nourished, B = moderate/suspected malnutrition, C = severe malnutrition) at admission, in adult patients, on four hospital wards of a university hospital. We compared results for patients with BMI < 25 kg/m2 and overweight or obese patients (BMI ≥ 25 kg/m2). Data were obtained in 387 patients (age 59 ± 16 years, 52% male). BMI was 27.1 ± 5.6 kg/m2, 35% was overweight and 26% was obese. According to MUST, 16% of patients had risk for malnutrition (MUST score ≥ 1), while according to PG-SGA 31% of patients were malnourished (PG-SGA Stage B/C). In patients with BMI < 25 kg/m2 (N = 153), MUST identified 31% as at risk for malnutrition, while PG-SGA assessed 39% as malnourished. In patients with BMI ≥ 25 kg/m2 (N = 234), MUST identified 6% as at risk for malnutrition, while PG-SGA assessed 26% as malnourished. Of the 60 malnourished overweight or obese patients according to the PG-SGA, 52 (87%) were categorized as low risk by the MUST. According to assessment by PG-SGA, a quarter of overweight or obese patients is malnourished at hospital admission. Most of these patients are not identified as having risk for malnutrition by the screening tool MUST. Future research is needed to evaluate the current results in relation to predictive value for patient outcomes, and to further optimize hospital nutritional care policies.

Coeliac disease affects 1% of the population globally, with no cure, the only treatment is a lifelong gluten free (GF) diet consisting of naturally GF foods (e.g., potatoes, rice, unprocessed meat, fruit, and vegetables) and manufactured GF versions of breads, pasta, pastry, cakes and biscuits. Adhering to a GF diet can be very challenging, it requires knowledge, skills, and modified behaviours to undertake substantial changes to dietary habits, including within cultural and social set up. The study aimed to determine whether a healthcare professional led intervention has an impact on GF knowledge and dietary adherence.

Adults diagnosed with coeliac disease were recruited from a hospital database. All participants completed a GF knowledge questionnaire and the validated Coeliac Dietary Adherence Test (CDAT) at baseline and three months later; where a score < 13 indicates dietary adherence. The intervention group received a coeliac disease information leaflet and a healthcare professional led telephone clinic which aimed to improve participants’ knowledge of coeliac disease and gluten-free foods, as well as discussing behaviour change. The control group received no intervention. Data is presented as mean and standard deviation, and median and interquartile range (IQR) where appropriate.

The intervention group consisted of 30 adults not adhering to the GF diet (CDAT score;16, 14–17), mean age 50.5 ± 17.7 years, 77% female and 100% Caucasians. The control group consisted of 86 adults adhering to the GF diet (CDAT ; 9, 8–10), mean age 51.3 ± 17.6 years, 77% female, 76% Caucasians. At baseline the knowledge of GF foods score was significantly lower in the intervention group compared with the control group (13.5 (12.0–14.0) and 15.0 (14.0–16.0) respectively; p < 0.01). GF knowledge significantly improved at 3 months in the intervention group (15.0 (14.0–16.0); p < 0.01) and in the control group (16.0 (15.0–16.0; p < 0.01). Dietary adherence score remained similar in the control group (9 (8–10) and 9 (8–11); NS), whereas there was a significant improvement in the intervention group (16 (14–17) and 13 (12–14); P < 0.001). The component scores for ‘How important are accidental gluten exposures’ and ‘how many times have you eaten gluten containing foods on purpose’ significantly improved (both P < 0.001) in the intervention group only.

This is the first study to report how a healthcare professional led telephone clinic can improve gluten-free dietary adherence in adults with coeliac disease. This evidence also suggests follow up via remote access to patients is effective.

Breast cancer is the most diagnosed cancer among women worldwide. Studies regarding diet and blood levels of vitamins and minerals in the breast cancer aetiology are limited and the results are inconclusive. We investigated the association between serum vitamin-mineral profiles (V-MPs) and breast cancer (BC) risk with including dietary patterns (DPs) and use of supplements. This case-control study involved 420 women aged 40–79 years from north-eastern Poland, including 190 newly-diagnosed breast cancer cases. The serum concentrations of vitamins (folate, cobalamin, 25(OH) vitamin D) and minerals (iron, calcium, magnesium) were marked in 129 post-menopausal women (82 controls, 47 cases) using an immune-analyser Cobas e411 and a Cobas Integra 400plus auto-analyser (Roche Diagnostics®), respectively. A posteriori V-MPs were derived with a Principal Component Analysis (PCA). Three PCA-driven DPs: ‘Non-Healthy’, ‘Prudent’, and ‘Margarine and Sweetened Dairy’ were described previously. A logistic regression analysis was performed. The odds ratios (ORs) and 95% confidence interval (95% CI) were calculated. Two serum V-MPs were identified. The ‘Folate-Cobalamin-Vitamin D’ profile was loaded heavily by the folate (factor loading 0.82), cobalamin (0.81) and vitamin D (0.49). The ‘Iron-Calcium’ profile was loaded heavily by the iron (0.81) and calcium (0.77). The ‘Magnesium-vitamin D’ profile was loaded heavily by the magnesium (0.92) and vitamin D (0.39). The risk of BC was lower by 88% (OR: 0.12; 95%Cl: 0.02–0.88; p < 0.05) in the upper tertile of the serum ‘Iron-Calcium’ profile compared to the bottom tertile. The risk of BC was lower by 67% (OR: 0.33; 95%Cl: 0.11–0.97; p < 0.05) at the level of serum 25(OH) vitamin D ≥ 24.6 ng/mL and lower by 68% (OR: 0.32; 95%Cl: 0.11–0.91; p < 0.05) at the level of serum calcium ≥ 9.6 mg/dL. There was the inverse association of the serum ‘Magnesium-Vitamin D’ profile with the risk of BC, which disappeared after adjustment for DPs. No significant association between BC risk and the serum ‘Folate-Cobalamin-Vitamin D’ profile and also folate, cobalamin, iron or magnesium considered separately was revealed. These findings highlight that the higher-adequate serum concentrations of both iron and calcium as well as close-to-adequate concentrations of calcium and vitamin D considered separately may protect against breast cancer among postmenopausal women, independently of dietary patterns and use of supplements. Therefore, women should be screened for blood concentrations of multiple vitamins and minerals in the breast cancer prevention.

Recent studies suggest that environmental pollutants play a role in the continuous increase of the worldwide prevalence of obesity. To our knowledge, there is only one study investigating the impacts of methylmercury, a common and more toxic form of mercury, mainly present in fish and shellfish, on adipose tissue. It shows that methylmercury can cause changes in the structure and function of 3T3-L1 preadipocytes. In this context, we decided to study the effects of methylmercury on primary cultured adipocytes of rainbow trout. This species is a well-known model to study the impact of pollution. In addition, fish adipose tissue metabolism is close to the one of humans. Using an in vitro model allows us to focus on adipocyte differentiation and lipid metabolism.

Preadipocytes were obtained after isolating precursor cells from perivisceral adipose tissue by enzymatic digestion. Cells were cultured at 19°C until confluence before being incubated for 6 days in growth medium supplemented or not with a hormonal cocktail (insulin, dexamethasone, ciglitizone) enriched with 0, 0.3, 1.7 or 3.8 mM of methylmercury and lipid mixture. Cytotoxicity was evaluated every two days via LDH assay. At day 6, quantification of total mercury in cells was performed with Direct Mercury Analyser-80. Phospholipid and neutral lipid fractions were analyzed by gas chromatography and expression of adipocyte-related genes was evaluated by RT-PCR. Three biological replicates were carried out.

No cytotoxicity was observed during the methylmercury treatment. Methylmercury was partially accumulated but in a dose-dependent manner. At day 6, all fatty acids (FA) from neutral lipid fraction significantly increased at the highest methylmercury concentration. However, the rate of increase was different for each FA. In particular, n-3 polyunsaturated FA were preferentially accumulated. The total amount of phospholipids was not impacted by methylmercury but a decrease of n-6 polyunsaturated FA was observed. Surprisingly, methylmercury decreased the expression of three adipocyte markers, FA transport protein 1 and glycerol-3-phosphate dehydrogenase in presence or absence of hormonal cocktail, and CCAAT/enhancer binding protein delta in presence of hormonal cocktail. On the other hand, methylmercury increased the expression of FA synthase and perilipin.

As a conclusion, methylmercury appears to be a candidate obesogen as it increased the cellular lipid content in primary cultured trout adipocytes. Additional genes involved in adipogenesis, FA de novo synthesis and FA intracellular transport are being investigated. Further experiments currently conducted in our lab should allow us to better clarify the mechanisms involved.

Coffee contains several components other than caffeine such as chlorogenic acids (CGAs), which are a family of polyphenols. Non-caffeinated coffee has been shown to reduce the risk of Type 2 diabetes, but it is unclear whether this effect is primarily due to a beneficial action on glucose regulation in peripheral tissues or whether it is partly mediated via a direct, functional modulation of insulin-secreting beta cells from the pancreas. This study aims to explore the specific role of coffee compounds derived from the polyphenolic family of CGAs (caffeic acid (CA) and ferulic acid (FA)) and their metabolites (dihydroferulic acid (diFA) and ferulic acid 4-O-sulphate (FA-4-OS)) in the regulation of beta cell survival and secretory function. To investigate this role, the cells were initially exposed to conditions of glucotoxicity (30mmol/l glucose), lipotoxicity (0.5mmol/l palmitate), glucolipotoxicity (30mmol/l glucose + 0.5mmol/l palmitate) and cytokine-induced cell toxicity (25U/ml IL1b + 500U/ml TNFα) for 20 and 48 h. INS1 beta cells were subsequently treated with or without 100nmol/l of the CGA compounds for 48 h followed by 20 h exposure to glucolipotoxicity to measure cellular ATP content and 3/7 caspase activity, respectively, as an indication of cell viability and apoptosis. Additionally, insulin release was assessed by radioimmunoassay following 1 h static incubations with or without CA, FA and metabolites. Data were analysed by One-Way ANOVA using GraphPad prism software (version 8). Glucotoxicity, lipotoxicity and glucotoxicity or glucolipotoxicity combined with cytokines significantly reduced INS1 cell viability compared to control at 20 and 48 h (p < 0.001 vs 11mmol/l glucose, p < 0.05; Glucotoxicity vs 11mmol/l glucose at 20 h, n = 6), whereas cytokines alone did not significantly affect cellular ATP content. Moreover, pre-treatment for 48 h with CGAs alone or in combination did not affect INS1 beta-cell viability under basal conditions (n = 3, p > 0.2). Additional exposure to glucolipotoxicity for 20 h significantly decreased beta cell viability and survival and was not alleviated by pre- or co-treatment with CGAs (n = 3, p > 0.05). However, insulin release in response to 1 h incubation with 20 mmol/l glucose + 10μmol/l forskolin (FSK) + 100μmol/l 3-isobutyl-1-methylxanthine (IBMX) was significantly higher from cells pre-treated with CA and FA combined compared to controls (7.64 + 2.89pg insulin/5,000 cells/h vs 2.17 + 0.35pg insulin/5,000 cells/h; p < 0.01 vs. 20mmol/l glucose + 10μmol/l FSK + 100μmol/l IBMX only, n = 5). The results suggest that CGA derivatives from coffee do not directly modulate INS1 beta cell viability and apoptosis whereas the compounds may play a role in the regulation of beta cell secretory function.

The consumption of fish is encouraged in dietary guidelines because of its association with health benefits; however, fish processing generates by-products (gills, bones, skin, head), which can rise up to 50–60% of fish weight after filleting. The investigation of bioactivity of fish by-products and subsequent utilization for the development of high added-value food ingredients and/or supplements will reduce waste and create new products of high nutritional value. We studied the antiproliferative activity of fish by-product protein hydrolysates (FPH) on human colon and breast cancer cells. A total of 6 Meagres “Argirosomus Regius” (1256.45 ± 232.32g) and 16 Gilthead Sea Breams “Sparus Aurata” (403.47 ± 72.92g) were obtained by HCMR Institute in Crete, Greece. The fins, skin, bones, head, and gills were isolated, lyophilized, homogenized and stored separately at -80oC. The protein hydrolysates from each byproduct were prepared using a pH extraction method. Human colorectal adenocarcinoma cells (COLO320 cells) and human breast adenocarcinoma cells (MCF7A cells) were treated with different FPH for 48 hours, and cellular proliferation was measured using the MTT assay. Results are given as mean percentage of triplicate independent analyses. Statistical significance of growth inhibition was estimated following Wilcoxon Signed-Ranks test with α = 0.05. In COLO320 cells, FPH induced a the significant antiproliferative activity at 0.5g/L. Specifically, FPH from bones, gills and skin of Meagre showed a growth inhibition of 23.7%, 9.9% and 26.4%, respectively, after 48 h of treatment. The inhibition by the same by-products from the Gilthead Sea bream was 24.6%, 21.9% and 26.4%, respectively after 48 h. In MCF7A cells, FPH exerted a significant growth inhibition at 1g/L. The skin and fins of Meagre induced a growth inhibition of 25.5% and 39.0% respectively; the gills, skin and fins of Gilthead Sea Bream showed a growth inhibition of 28.5%, 47.1% and 39.6%, respectively, after 48 h. These values were in the same range with etoposide (reference anticancer molecule) in the concentration of 3.1μM for COLO320 cells and 50μM for MCF7A. These preliminary data suggest that certain fish by-products could represent a source of anticancer peptides and may cause growth inhibition of human cancer cells. The isolation of responsible bioactive peptides from fish byproducts and the integration of them into food supplements can have beneficial effects on human health.

Health effects of orally ingested bioactive compounds can only occur if bioactive molecules are absorbed and transported to target tissues. Paradoxically, many foods are rich in micronutrients but intestinal absorption is often limited. Immediately after absorption, lipid soluble nutrients are packaged into chylomicrons (CM), therefore quantification of the micronutrient content in CM has been used as tool to evaluate bioavailability(1). For bioavailability studies the isolated CM fraction must not contain liver derived lipoproteins (VLDL, LDL) since only intestine derived CM carry recently absorbed lipids and lipophilic micronutrients. As isolated CM can be contaminated with liver derived lipoproteins, this study evaluated the purity of collected CM and VLDL fractions. Each CM contains one apolipoprotein B-48 (apoB-48) and each VLDL contains one apolipoprotein B-100 (apoB-100) on its surface(2) therefore purity of CM and VLDL fractions was evaluated via the presence/absence of apoB-100 and apoB-48.

CM and VLDL fractions were isolated as previously described(1,3)from whole plasma collected at 0, 2, 4 or 6 h after participants consumed a lipid (15.4g) and carotenoid (36mg) rich smoothie (480mL). Plasma density was adjusted to 1.1g/mL with KBr (0.14g/mL), placed in ultracentrifugation tubes (38 mL, thickwall) and overlaid with 3 solutions containing NaCl, KBr and Na-EDTA with densities of 1.020, 1.065 and 1.006g/mL.CM fraction was collected after ultracentrifugation at 25,000 rpm for 34 min in a Beckman Coulter Optima XE-90 with SW32Ti swinging bucket rotor. VLDL fraction was collected after additional 102 min at 25,000 rpm.

ELISA analysis revealed that apoB-48 and apoB-100 were present in both CM and VLDL fractions. Less than 0.1% of plasma apoB-48 (< 98pmol/L) was present in CM or VLDL fraction. Less than 0.3% (< 2,425pmol/L) and 4% (< 54,124pmol/L) of plasma apoB-100 was present in CM and VLDL fractions, respectively. Low recoveries of apoB-48 and apoB-100 in CM and VLDL fraction suggests that ultracentrifugation neither concentrated nor isolated CM or VLDL. While ourapoB-48 and apoB-100 results agree with previously reported values in plasma, CM and VLDL(3,4), no previous study evaluated apoB-48 and apoB-100 recovery from plasma.