Exposure to aflatoxin B1 in utero is associated with DNA methylation in white blood cells of infants in The Gambia

BACKGROUND: Exposure to environmental toxins during embryonic development may lead to epigenetic changes that influence disease risk in later life. Aflatoxin is a contaminant of staple foods in sub-Saharan Africa, is a known human liver carcinogen and has been associated with stunting in infants.

METHODS: We have measured aflatoxin exposure in 115 pregnant women in The Gambia and examined the DNA methylation status of white blood cells from their infants at 2-8 months old (mean 3.6 ± 0.9). Aflatoxin exposure in women was assessed using an ELISA method to measure aflatoxin albumin (AF-alb) adducts in plasma taken at 1-16 weeks of pregnancy. Genome-wide DNA methylation of infant white blood cells was measured using the Illumina Infinium HumanMethylation450beadchip.

RESULTS: AF-alb levels ranged from 3.9 to 458.4 pg/mg albumin. We found that aflatoxin exposure in the mothers was associated to DNA methylation in their infants for 71 CpG sites (false discovery rate < 0.05), with an average effect size of 1.7% change in methylation. Aflatoxin-associated differential methylation was observed in growth factor genes such as FGF12 and IGF1, and immune-related genes such as CCL28, TLR2 and TGFBI. Moreover, one aflatoxin-associated methylation region (corresponding to the miR-4520b locus) was identified.

CONCLUSIONS: This study shows that maternal exposure to aflatoxin during the early stages of pregnancy is associated with differential DNA methylation patterns of infants, including in genes related to growth and immune function. This reinforces the need for interventions to reduce aflatoxin exposure, especially during critical periods of fetal and infant development.

Identification of lactic acid bacteria strains modulating incretin hormone secretion and gene expression in enteroendocrine cells

Glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) are incretin hormones released from intestinal enteroendocrine (EE) cells and have well-established glucose-lowering actions. Lactic acid bacteria (LAB) colonise the human intestine, but it is unknown whether LAB and EE cells interact. Acute co-culture of LAB with EE cells showed that certain LAB strains elicit GLP-1 and GIP secretion (13-194-fold) and upregulate their gene expression. LAB-induced incretin hormone secretion did not appear to involve nutrient mechanisms, nor was there any evidence of cytolysis. Instead PCR array studies implicated signalling agents of the toll-like receptor system, e.g. adaptor protein MyD88 was decreased 23-fold and cell surface antigen CD14 was increased 17-fold. Mechanistic studies found that blockade of MyD88 triggered significant GLP-1 secretion. Furthermore, blocking of CD14 completely attenuated LAB-induced secretion. A recent clinical trial clearly shows that LAB have potential for alleviating type 2 diabetes, and further characterisation of this bioactivity is warranted.

Awareness and attitudes towards the emerging use of nanotechnology in the agri-food sector

Nanotechnology has relevance to applications in all areas of agri-food including agriculture, aquaculture, production, processing, packaging, safety and nutrition. Scientific literature indicates uncertainties in food safety aspects about using nanomaterials due to potential health risks. To date the agri-food industry's awareness and attitude towards nanotechnology have not been addressed. We surveyed the awareness and attitudes of agri-food organisations on the island of Ireland (IoI) with regards to nanotechnology. A total of 14 agri-food stakeholders were interviewed and 88 agri-food stakeholders responded to an on-line questionnaire. The findings indicate that the current awareness of nanotechnology applications in the agri-food sector on the IoI is low and respondents are neither positive nor negative towards agri-food applications of nanotechnology. Safer food, reduced waste and increased product shelf life were considered to be the most important benefits to the agri-food industry. Knowledge of practical examples of agri-food applications is limited however opportunities were identified in precision farming techniques, innovative packaging, functional ingredients and nutrition of foods, processing equipment, and safety testing. Perceived impediments to nanotechnology adoption were potential unknown human health and environmental impacts, consumer acceptance and media framing. The need for a risk assessment framework, research into long term health and environmental effects, and better engagement between scientists, government bodies, the agri-food industry and the public were identified as important.

Suitability, efficiency and microbiological safety of novel physical technologies for the processing of ready-to-eat meals, meats and pumpable products

Consumer studies and market reports show an increase in consumption of ready-to-eat (RTE) foods. Although conventional processing technologies can in most cases produce safe products, they can also lead to the degradation of nutritional compounds and negatively affect quality characteristics. Consumers strongly prefer food that is minimally processed with the maximum amount of health-promoting substances. Novel processing technologies as pre- or post-treatment decontamination methods or as substitutes of conventional technologies have the potential to produce foods that are safe, rich in nutrient content and with superior organoleptic properties. Combining novel with conventional processes can eliminate potential drawbacks of novel technologies. This review examines available scientific information and critically evaluates the suitability and efficiency of various novel thermal and nonthermal technologies in terms of microbial safety, quality as well as nutrient content on the production of RTE meals, meats and pumpable products.

Challenging conventional risk assessment with respect to human exposure to multiple food contaminants in food: A case study using maize

Mycotoxins and heavy metals are ubiquitous in the environment and contaminate many foods. The widespread use of pesticides in crop production to control disease contributes further to the chemical contamination of foods. Thus multiple chemical contaminants threaten the safety of many food commodities; hence the present study used maize as a model crop to identify the severity in terms of human exposure when multiple contaminants are present. High Content Analysis (HCA) measuring multiple endpoints was used to determine cytotoxicity of complex mixtures of mycotoxins, heavy metals and pesticides. Endpoints included nuclear intensity (NI), nuclear area (NA), plasma membrane permeability (PMP), mitochondrial membrane potential (MMP) and mitochondrial mass (MM). At concentrations representing legal limits of each individual contaminant in maize (3. ng/ml ochratoxin A (OTA), 1. μg/ml fumonisin B1 (FB1), 2. ng/ml aflatoxin B1 (AFB1), 100. ng/ml cadmium (Cd), 150. ng/ml arsenic (As), 50. ng/ml chlorpyrifos (CP) and 5. μg/ml pirimiphos methyl (PM), the mixtures (tertiary mycotoxins plus Cd/As) and (tertiary mycotoxins plus Cd/As/CP/PM) were cytotoxic for NA and MM endpoints with a difference of up to 13.6% (. p≤. 0.0001) and 12% (. p≤. 0.0001) respectively from control values. The most cytotoxic mixture was (tertiary mycotoxins plus Cd/As/CP/PM) across all 4 endpoints (NA, NI, MM and MMP) with increases up to 61.3%, 23.0%, 61.4% and 36.3% (. p≤. 0.0001) respectively. Synergy was evident for two endpoints (NI and MM) at concentrations contaminating maize above legal limits, with differences between expected and measured values of (6.2-12.4% (. p≤. 0.05-. p≤. 0.001) and 4.5-12.3% (. p≤. 0.05-. p≤. 0.001) for NI and MM, respectively. The study introduces for the first time, a holistic approach to identify the impact in terms of toxicity to humans when multiple chemical contaminants are present in foodstuffs. Governmental regulatory bodies must begin to contemplate how to safeguard the population when such mixtures of contaminants are found in foods and this study starts to address this critical issue.

Site Distribution at the Edge of the Palaeolithic World: A Nutritional Niche Approach

This paper presents data from the English Channel area of Britain and Northern France on the spatial distribution of Lower to early Middle Palaeolithic pre-MIS5 interglacial sites which are used to test the contention that the pattern of the richest sites is a real archaeological distribution and not of taphonomic origin. These sites show a marked concentration in the middle-lower reaches of river valleys with most being upstream of, but close to, estimated interglacial tidal limits. A plant and animal database derived from Middle-Late Pleistocene sites in the region is used to estimate the potentially edible foods and their distribution in the typically undulating landscape of the region. This is then converted into the potential availability of macronutrients (proteins, carbohydrates, fats) and selected micronutrients. The floodplain is shown to be the optimum location in the nutritional landscape (nutriscape). In addition to both absolute and seasonal macronutrient advantages the floodplains could have provided foods rich in key micronutrients, which are linked to better health, the maintenance of fertility and minimization of infant mortality. Such places may have been seen as ‘good (or healthy) places’ explaining the high number of artefacts accumulated by repeated visitation over long periods of time and possible occupation. The distribution of these sites reflects the richest aquatic and wetland successional habitats along valley floors. Such locations would have provided foods rich in a wide range of nutrients, importantly including those in short supply at these latitudes. When combined with other benefits, the high nutrient diversity made these locations the optimal niche in northwest European mixed temperate woodland environments. It is argued here that the use of these nutritionally advantageous locations as nodal or central points facilitated a healthy variant of the Palaeolithic diet which permitted habitation at the edge of these hominins’ range.

Food Colours: Existing and Emerging Food Safety Concerns

Food colours are added to different types of commodities to increase their visual attractiveness or to compensate for natural colour variations. The use of these additives is strictly regulated in the European Union, the United States and many other countries worldwide. There is a growing concern about the safety of some commonly used legal food colourants and there is a trend to replace the synthetic forms with natural products. Additionally, a number of dyes with known or suspected genotoxic or carcinogenic properties have been shown to be added illegally to foods. Robust monitoring programs based on reliable detection methods are required to assure the food is free from harmful colours. The aim of this review is to present an up to date status of the various concerns arising from use of colour additives in food. The most important food safety concerns in the field of food colours are lack of uniform regulation concerning legal food colours worldwide, possible link of artificial colours to hyperactive behaviour, replacement of synthetic colours with natural ones and the presence of harmful illegal dyes - both known but also new, emerging ones in food. The legal status of food colour additives in the EU, US and worldwide is summarized. The reported negative health effects of both legal and illegal colours are presented. The European Rapid Alert System for Food and Feed notifications and US import alerts concerning food colours are analyzed and trends in fraudulent use of colour additives identified. The detection methods for synthetic colours are also reviewed.

An evaluation of portion size estimation aids precision, ease of use and likelihood of future use

OBJECTIVE: The present study aimed to evaluate the precision, ease of use and likelihood of future use of portion size estimation aids (PSEA).

DESIGN: A range of PSEA were used to estimate the serving sizes of a range of commonly eaten foods and rated for ease of use and likelihood of future usage.

SETTING: For each food, participants selected their preferred PSEA from a range of options including: quantities and measures; reference objects; measuring; and indicators on food packets. These PSEA were used to serve out various foods (e.g. liquid, amorphous, and composite dishes). Ease of use and likelihood of future use were noted. The foods were weighed to determine the precision of each PSEA.

SUBJECTS: Males and females aged 18-64 years (n 120).

RESULTS: The quantities and measures were the most precise PSEA (lowest range of weights for estimated portion sizes). However, participants preferred household measures (e.g. 200 ml disposable cup) - deemed easy to use (median rating of 5), likely to use again in future (all scored either 4 or 5 on a scale from 1='not very likely' to 5='very likely to use again') and precise (narrow range of weights for estimated portion sizes). The majority indicated they would most likely use the PSEA preparing a meal (94 %), particularly dinner (86 %) in the home (89 %; all P<0·001) for amorphous grain foods.

CONCLUSIONS: Household measures may be precise, easy to use and acceptable aids for estimating the appropriate portion size of amorphous grain foods.

Food fraud and consumers' choices in the wake of the horsemeat scandal

Purpose – In 2012, the European food industry was hit by a food fraud: horsemeat was found in
pre-prepared foods, without any declaration on the package. This is commonly referred to as the
“horsemeat scandal”. The purpose of this paper is to investigate consumers’ preferences across
Europe for a selected ready meal, ready to heat (RTH) fresh lasagne, to consider whether the effects of
potential food frauds on consumers’ choices can be mitigated by introducing enhanced standards of
RTH products.
Design/methodology/approach – An online survey was administered to 4,598 consumers of RTH
lasagne in six European countries (Republic of Ireland, France, Italy, Spain, Germany and Norway),
applying discrete choice experiments to estimate consumers’ willingness to pay for enhanced food
safety standards and highlight differences between countries.
Findings – Many similarities across countries emerged, as well as some differences. Consumers in
Europe are highly concerned with the authenticity of the meat in ready meals and strongly prefer to
know that ingredients are nationally sourced. Strong regional differences in price premiums exist for
enhanced food safety standards.
Originality/value – This research adds relevant insights in the analysis of consumers’ reaction to
food fraud, providing practical guidelines on the most appropriate practices that producers should
adopt and on the information to reduce food risk perception among consumers. This would prove
beneficial for the food processing industry and the European Union. The survey is based on a
representative sample of European consumers making this the largest cross-country study of this kind.

Beneficial effect of a polyphenol-rich diet on cardiovascular risk: a randomised control trial

OBJECTIVES: There is previous epidemiological evidence that intake of polyphenol-rich foods has been associated with reduced cardiovascular disease risk. We aimed to investigate the effect of increasing dietary polyphenol intake on microvascular function in hypertensive participants.

METHODS: All participants completed a 4-week run-in phase, consuming <2 portions of fruit and vegetables (F&V) daily and avoiding berries and dark chocolate. Subjects were then randomised to continue with the low-polyphenol diet for 8 weeks or to consume a high-polyphenol diet of six portions F&V (including one portion of berries/day and 50 g of dark chocolate). Endothelium-dependent (acetylcholine, ACh) and endothelium-independent (sodium nitroprusside) vasodilator responses were assessed by venous occlusion plethysmography. Compliance with the intervention was measured using food diaries and biochemical markers.

RESULTS: Final analysis of the primary endpoint was conducted on 92 participants. Between-group comparison of change in maximum % response to ACh revealed a significant improvement in the high-polyphenol group (p=0.02). There was a significantly larger increase in vitamin C, carotenoids and epicatechin in the high-polyphenol group (between-group difference p<0.001; p<0.001; p=0.008, respectively).

CONCLUSIONS: This study has shown that increasing the polyphenol content of the diet via consumption of F&V, berries and dark chocolate results in a significant improvement in an established marker of cardiovascular risk in hypertensive participants.

Processo terminográfico: vertentes conceptual, comunicativa e textual

O presente trabalho de investigação visa propor uma metodologia de elaboração de uma base de dados terminológica destinada a um público não- -especialista, e surge como resposta à necessidade de transmissão de informação ao consumidor, fruto de falta de – ou parca – compreensão do mesmo, relativa a géneros alimentícios com alegações de saúde disponíveis no mercado: os denominados alimentos funcionais. A proposta metodológica de segmentação e caracterização do processo terminográfico, baseada no modelo desenvolvido por Gouadec, para organização do processo global de tradução, encontra-se organizada em três fases – pré-terminografia, terminografia e pós-terminografia –, e compreende três vertentes de análise – uma vertente conceptual, uma vertente comunicativa e uma vertente textual. Em termos gerais, na fase de pré-terminografia é desenvolvido um trabalho preparatório – de familiarização com a área de especialidade e de delimitação da subárea de especialidade, de identificação dos contextos comunicativos e de constituição de corpora especializados – essencial à subsequente fase executória – fase de terminografia – de elaboração do recurso terminológico. A última fase – fase de pós-terminografia – compreende o desenvolvimento de esforços com vista à aplicação industrial do recurso, assim como a sua posterior constante actualização. Constituem objecto de análise do presente trabalho as duas primeiras fases supramencionadas e as etapas que as constituem. A consideração de três vertentes de análise é, de igual forma, relevante.Tal facto é demonstrado ao longo do processo terminográfico, designadamente a nível da análise das repercussões, na fase de terminografia, de cada uma destas vertentes, consideradas já na fase de pré-terminografia. Com este trabalho de investigação pretendemos demonstrar o papel social da Terminologia, no contributo que pode prestar na divulgação de ciência, concretamente através da apresentação de uma proposta de uma base de dados terminológica sobre alimentos funcionais para o consumidor – a AlF Beta. Do mesmo modo, temos por objectivo contribuir a nível da reflexão teórica e metodológica em Terminologia, nomeadamente no que concerne a sua vertente aplicada, através da elaboração de recursos terminológicos destinados a públicos não-especialistas.

Volatile terpenoids and C13 norisoprenoids in wines: development of rapid methods of analysis and evaluation of the sesquiterpenoids biological potential

Vitis vinifera L., the most widely cultivated fruit crop in the world, was the starting point for the development of this PhD thesis. This subject was exploited following on two actual trends: i) the development of rapid, simple, and high sensitive methodologies with minimal sample handling; and ii) the valuation of natural products as a source of compounds with potential health benefits. The target group of compounds under study were the volatile terpenoids (mono and sesquiterpenoids) and C13 norisoprenoids, since they may present biological impact, either from the sensorial point of view, as regards to the wine aroma, or by the beneficial properties for the human health. Two novel methodologies for quantification of C13 norisoprenoids in wines were developed. The first methodology, a rapid method, was based on the headspace solid-phase microextraction combined with gas chromatography-quadrupole mass spectrometry operating at selected ion monitoring mode (HS-SPME/GC-qMS-SIM), using GC conditions that allowed obtaining a C13 norisoprenoid volatile signature. It does not require any pre-treatment of the sample, and the C13 norisoprenoid composition of the wine was evaluated based on the chromatographic profile and specific m/z fragments, without complete chromatographic separation of its components. The second methodology, used as reference method, was based on the HS-SPME/GC-qMS-SIM, allowing the GC conditions for an adequate chromatographic resolution of wine components. For quantification purposes, external calibration curves were constructed with β-ionone, with regression coefficient (r2) of 0.9968 (RSD 12.51 %) and 0.9940 (RSD of 1.08 %) for the rapid method and for the reference method, respectively. Low detection limits (1.57 and 1.10 μg L-1) were observed. These methodologies were applied to seventeen white and red table wines. Two vitispirane isomers (158-1529 L-1) and 1,1,6-trimethyl-1,2-dihydronaphthalene (TDN) (6.42-39.45 μg L-1) were quantified. The data obtained for vitispirane isomers and TDN using the two methods were highly correlated (r2 of 0.9756 and 0.9630, respectively). A rapid methodology for the establishment of the varietal volatile profile of Vitis vinifera L. cv. 'Fernão-Pires' (FP) white wines by headspace solid-phase microextraction combined with comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (HS-SPME/GCxGC-TOFMS) was developed. Monovarietal wines from different harvests, Appellations, and producers were analysed. The study was focused on the volatiles that seem to be significant to the varietal character, such as mono and sesquiterpenic compounds, and C13 norisoprenoids. Two-dimensional chromatographic spaces containing the varietal compounds using the m/z fragments 93, 121, 161, 175 and 204 were established as follows: 1tR = 255-575 s, 2tR = 0,424-1,840 s, for monoterpenoids, 1tR = 555-685 s, 2tR = 0,528-0,856 s, for C13 norisoprenoids, and 1tR = 695-950 s, 2tR = 0,520-0,960 s, for sesquiterpenic compounds. For the three chemical groups under study, from a total of 170 compounds, 45 were determined in all wines, allowing defining the "varietal volatile profile" of FP wine. Among these compounds, 15 were detected for the first time in FP wines. This study proposes a HS-SPME/GCxGC-TOFMS based methodology combined with classification-reference sample to be used for rapid assessment of varietal volatile profile of wines. This approach is very useful to eliminate the majority of the non-terpenic and non-C13 norisoprenic compounds, allowing the definition of a two-dimensional chromatographic space containing these compounds, simplifying the data compared to the original data, and reducing the time of analysis. The presence of sesquiterpenic compounds in Vitis vinifera L. related products, to which are assigned several biological properties, prompted us to investigate the antioxidant, antiproliferative and hepatoprotective activities of some sesquiterpenic compounds. Firstly, the antiradical capacity of trans,trans-farnesol, cis-nerolidol, α-humulene and guaiazulene was evaluated using chemical (DPPH• and hydroxyl radicals) and biological (Caco-2 cells) models. Guaiazulene (IC50= 0.73 mM) was the sesquiterpene with higher scavenger capacity against DPPH•, while trans,trans-farnesol (IC50= 1.81 mM) and cis-nerolidol (IC50= 1.48 mM) were more active towards hydroxyl radicals. All compounds, with the exception of α-humulene, at non-cytotoxic levels (≤ 1 mM), were able to protect Caco-2 cells from oxidative stress induced by tert-butyl hydroperoxide. The activity of the compounds under study was also evaluated as antiproliferative agents. Guaiazulene and cis-nerolidol were able to more effectively arrest the cell cycle in the S-phase than trans,trans-farnesol and α-humulene, being the last almost inactive. The relative hepatoprotection effect of fifteen sesquiterpenic compounds, presenting different chemical structures and commonly found in plants and plant-derived foods and beverages, was assessed. Endogenous lipid peroxidation and induced lipid peroxidation with tert-butyl hydroperoxide were evaluated in liver homogenates from Wistar rats. With the exception of α-humulene, all the sesquiterpenic compounds under study (1 mM) were effective in reducing the malonaldehyde levels in both endogenous and induced lipid peroxidation up to 35% and 70%, respectively. The developed 3D-QSAR models, relating the hepatoprotection activity with molecular properties, showed good fit (R2LOO > 0.819) with good prediction power (Q2 > 0.950 and SDEP < 2%) for both models. A network of effects associated with structural and chemical features of sesquiterpenic compounds such as shape, branching, symmetry, and presence of electronegative fragments, can modulate the hepatoprotective activity observed for these compounds. In conclusion, this study allowed the development of rapid and in-depth methods for the assessment of varietal volatile compounds that might have a positive impact on sensorial and health attributes related to Vitis vinifera L. These approaches can be extended to the analysis of other related food matrices, including grapes and musts, among others. In addition, the results of in vitro assays open a perspective for the promising use of the sesquiterpenic compounds, with similar chemical structures such as those studied in the present work, as antioxidants, hepatoprotective and antiproliferative agents, which meets the current challenges related to diseases of modern civilization.

Barrier properties of poly(lactic acid) based films

In recent years, the search for a environmentally friendly products has increased. One of the major challenges has been the demand for biodegradable materials that can replace plastic. If a few decades ago, plastic replaced, for example, the ivory in billiard balls, and in other products, saving the lives of thousands elephants, nowadays a replacement for that plastic is being searched, to prevent the change of the environmental conditions, essential to life in harmonly with the fauna and flora that the human specie has, in recent years, destroyed. Plastic is a petroleum derivate, whose price has been growing exponentially, mainly due to the fact of beind a cheap material and also to enable the production of products that are essential to modern life. Therefore, the petrochemical era is going to come to an end and a new environmentally sustainable era, based on biodegradable materials from renewable sources, will follow. The change to green routes only will be possible with the support of the major companies, and the implementation of drastic governmental law. Poly(lactic acid), PLA, is produced from the lactose present in the corn or sugarcane and has been intensively studied in recent years because if some limitants properties required its extrusion are overcome, it has the potential to replace the traditional polymers. PLA have high brittleness, low toughness and low tensile elongation. In this work, natural antioxidant (alpha-tocopherol) and synthetics antioxidants (BHT ant TBHQ) were added to the PLA with the aim not only to improve their flexibility, but also to create an active packaging to extend the shelf life of the foods and improve the organoleptic properties by preventing food losses. The impact of the addition of antioxidants into the PLA films, in its mechanical, thermal and barrier properties were studied by FTIR, DSC, SEM, AFM, DMA, TGA, QCM and time-lag techniques.

Improved extraction of arabinoxylans from rye bran and production of health beneficial oligosaccharides

Dissertação de mest., Engenharia Biológica, Faculdade de Ciências e Tecnologia, Univ. do Algarve, 2011

Problems in the use of benzoic acid for estimating the internal pH of yeasts

Some yeasts have the peculiar ability to grow in the presence of weak acids at rather low pH. These conditions are predominant in preserved foods and beverages such as fruit concentrates, juices, wine, where these yeasts are responsible for spoilage. The main preservatives currently utilized by food industries are sorbic, propionic, benzoic acids and SO2. It is usually assumed that weak acids diffuse through the plasma membrane in the undissociated form. In the cytoplasm, where the pH is higher, dissociation occurs resulting in accumulation of the lipid-insoluble anion and internal acidification. This is probably a very general mechanism of preventing microbial growth in foods.