Betaine-homocysteine S-methyltransferase deficiency causes increased susceptibility to noise-induced hearing loss associated with plasma hyperhomocysteinemia.

Betaine-homocysteine S-methyltransferases (BHMTs) are methionine cycle enzymes that remethylate homocysteine; hence, their malfunction leads to hyperhomocysteinemia. Epidemiologic and experimental studies have revealed a correlation between hyperhomocysteinemia and hearing loss. Here, we have studied the expression of methionine cycle genes in the mouse cochlea and the impact of knocking out the Bhmt gene in the auditory receptor. We evaluated age-related changes in mouse hearing by recording auditory brainstem responses before and following exposure to noise. Also, we measured cochlear cytoarchitecture, gene expression by RNA-arrays and quantitative RT-PCR, and metabolite levels in liver and plasma by HPLC. Our results indicate that there is an age-dependent strain-specific expression of methionine cycle genes in the mouse cochlea and a further regulation during the response to noise damage. Loss of Bhmt did not cause an evident impact in the hearing acuity of young mice, but it produced higher threshold shifts and poorer recovery following noise challenge. Hearing loss was associated with increased cochlear injury, outer hair cell loss, altered expression of cochlear methionine cycle genes, and hyperhomocysteinemia. Our results suggest that BHMT plays a central role in the homeostasis of cochlear methionine metabolism and that Bhmt2 up-regulation could carry out a compensatory role in cochlear protection against noise injury in the absence of BHMT.-Partearroyo, T., Murillo-Cuesta, S., Vallecillo, N., Bermúdez-Muñoz, J. M., Rodríguez-de la Rosa, L., Mandruzzato, G., Celaya, A. M., Zeisel, S. H., Pajares, M. A., Varela-Moreiras, G., Varela-Nieto, I. Betaine-homocysteine S-methyltransferase deficiency causes increased susceptibility to noise-induced hearing loss associated with plasma hyperhomocysteinemia.

Adequate or elevated dietary folate does not ameliorate the reduced antioxidant capacity induced by vitamin B12 deficiency in aged rats.

Folate could have an antioxidant role but also may be detrimental under vitamin B12 deficiency. The aim was to investigate the effect of different dietary folic acid (FA) levels, on oxidative stress in B12 induced deficient aged rats. Thirty-five male aged Sprague-Dawley rats, were fed either a vitamin B12 deficient (n = 27) or a control diet (n = 8) during eight weeks. Then, animals were divided into four groups: B12 and FA deficient diet (DBDF), B12 deficient diet and FA control diet (DBCF), B12 deficient diet and FA supplemented diet (DBSF), and control diet (CBCF) for a 30 days period. Methionine metabolism and antioxidant status were evaluated. Both vitamins deficiencies elevated serum homocysteine (Hcy) (7.7 vs. 4.3 µmol/L, p < 0.05) and reduced S-adenosylmethionine hepatic content (283.7 vs. 581.9 µg/g protein, p < 0.05), the total antioxidant capacity (155.7 vs. 189.3 µmol/L, p < 0.05), glutathione (GSH) (120.5 vs. 419.9 µg/mg protein, p < 0.05) and oxidized glutathione (0.9 vs. 2.6 µg/mg protein, p < 0.05) compared to control. Activities of glutathione peroxidase and glutathione reductase enzymes or damage to macromolecules were unaffected. Adequate or elevated dietary FA in B12 deficiency rats decreased Hcy (5.7 and 6.3 µmol/L, respectively) and increased total antioxidant capacity (189.8 and 192.6 µmol/L, respectively) to values similar to control group, whereas GSH concentration was significantly lower than control (209.1 and 208.0 µg/mg protein respectively, p < 0.05). In conclusion, in a vitamin B12 deficiency status, adequate or elevated FA prevented impairment in one-carbon metabolism, but does not fully reverse the decrease in antioxidant capacity.

Low Adherence to Dietary Guidelines in Spain, Especially in the Overweight/Obese Population: The ANIBES Study.

OBJECTIVE: The objective of this study was to analyze the dietary intake of the Spanish population according to ponderal status and body fat distribution. METHODS: Data were obtained from ANIBES (Anthropometry, Intake, and Energy Balance in Spain), a cross-sectional study of a nationally representative sample (1013 men, 996 women) of the Spanish population (18-64 years). The final fieldwork was carried out from mid-September to November (three months) 2013. A 3-day dietary record provided information about food and beverage consumption. Height, weight, and waist circumference were assessed, and body mass index (BMI) and waist-height ratio (WHtR) calculated. RESULTS: The Spanish population had a low consumption of fruits and vegetables, cereals, whole cereals, and dairy and high consumption of meat products. Individuals with overweight/obesity (BMI ≥ 25 kg/m2) and abdominal adiposity (WHtR ≥ 0.5) showed lower compliance with dietary guidelines. In the male group, adjusting by age, inadequate consumption of cereals (<4 servings/day) and vegetables and fruit (<5 servings/day) was associated with higher risk of overweight (odds ratio [OR] = 1.704, 95% confidence interval [CI], 1.187-2.447, p = 0.001, for cereals and OR = 3.816, 95% CI, 1.947-7.480, p = 0.001, for vegetables and fruits) and abdominal adiposity (OR = 2.081, 95% CI, 1.419-3.053, p = 0.000 and OR = 4.289, 95% CI, 2.108-8.726, p = 0.001, respectively). CONCLUSION: Nutritional campaigns should be conducted to improve the dietary habits of the Spanish population in general, especially men, who have poorer ponderal status and abdominal adiposity, due to their lower adherence to dietary guidelines.

Differences in meal patterns and timing with regard to central obesity in the ANIBES ('Anthropometric data, macronutrients and micronutrients intake, practice of physical activity, socioeconomic data and lifestyles in Spain') Study.

OBJECTIVE: To study the association of meal patterns and timing with central obesity to identify the best dietary strategies to deal with the increasing obesity prevalence. DESIGN: A cross-sectional study performed on data from a representative sample of the Spanish population. Height and waist circumference were measured using standardized procedures and waist-to-height ratio (WHtR) was calculated. The sample was divided into those without central obesity (WHtR<0·5) and those with central obesity (WHtR≥0·5). SETTING: ANIBES ('Anthropometric data, macronutrients and micronutrients intake, practice of physical activity, socioeconomic data and lifestyles in Spain') Study. SUBJECTS: Adults aged 18-64 years (n 1655; 798 men and 857 women). RESULTS: A higher percentage of people ate more than four meals daily in the group without central obesity and those with central obesity more frequently skipped the mid-afternoon snack than those without. Breakfasts containing >25 % of total energy intake and lunches containing >35 % of total energy intake were associated with increased likelihood of central obesity (OR=1·874, 95 % CI 1·019, 3·448; P15 % of total energy were associated with decreased likelihood of central obesity (OR=0·477, 95 % CI 0·313, 0·727; P<0·001 and OR=0·650, 95 % CI 0·453, 0·932; P<0·05, respectively). The variety of cereals, wholegrain cereals and dairy was higher in the population without central obesity. CONCLUSIONS: Our results suggest that 'what and when we eat' should be considered dietary strategies to reduce central obesity.

Sedentary behavior among Spanish children and adolescents: findings from the ANIBES study.

BACKGROUND: An increase of sedentary behaviors far from the Mediterranean lifestyle is happening in spite of the impact on health. The aims of this study were to describe sedentary behaviors in children and adolescents. METHODS: A representative sample of 424 Spanish children and adolescents (38% females) involved in the ANIBES study was analyzed regarding their sedentary behaviors, together with the availability of televisions, computers, and consoles by means of the HELENA sedentary behavior questionnaire. RESULTS: For the total sample of children, 49.3% during weekdays and 84% during weekends did not meet the recommendation of less than 2 hours of screen viewing per day. The use of TV was higher during weekdays (p < 0.05) and there were significant differences between adolescents and children (16.9 vs. 25.1%, p < 0.05). The use of computer, console games and of internet for non-study reasons was higher during weekends (p < 0.001). Adolescents played more computer games and used more internet for non-study reasons than children during both weekdays and weekends (p < 0.05 and p < 0.001, respectively). The use of internet for academic reasons was lower in children (p < 0.001) than adolescents during weekends; however, no significant differences were found between sexes. In addition, more than 30% of the children and adolescents had at least one electronic device in their bedrooms. CONCLUSIONS: Spanish children and adolescents are not meeting the recommendations regarding the maximum of screen viewing (<2 h/day), especially during the weekend, for all of sedentary behaviors. Urgent strategies and intervention studies are needed to reduce sedentary behavior in young people.

Folic acid deficiency induces premature hearing loss through mechanisms involving cochlear oxidative stress and impairment of homocysteine metabolism.

Nutritional imbalance is emerging as a causative factor of hearing loss. Epidemiologic studies have linked hearing loss to elevated plasma total homocysteine (tHcy) and folate deficiency, and have shown that folate supplementation lowers tHcy levels potentially ameliorating age-related hearing loss. The purpose of this study was to address the impact of folate deficiency on hearing loss and to examine the underlying mechanisms. For this purpose, 2-mo-old C57BL/6J mice (Animalia Chordata Mus musculus) were randomly divided into 2 groups (n = 65 each) that were fed folate-deficient (FD) or standard diets for 8 wk. HPLC analysis demonstrated a 7-fold decline in serum folate and a 3-fold increase in tHcy levels. FD mice exhibited severe hearing loss measured by auditory brainstem recordings and TUNEL-positive-apoptotic cochlear cells. RT-quantitative PCR and Western blotting showed reduced levels of enzymes catalyzing homocysteine (Hcy) production and recycling, together with a 30% increase in protein homocysteinylation. Redox stress was demonstrated by decreased expression of catalase, glutathione peroxidase 4, and glutathione synthetase genes, increased levels of manganese superoxide dismutase, and NADPH oxidase-complex adaptor cytochrome b-245, α-polypeptide (p22phox) proteins, and elevated concentrations of glutathione species. Altogether, our findings demonstrate, for the first time, that the relationship between hyperhomocysteinemia induced by folate deficiency and premature hearing loss involves impairment of cochlear Hcy metabolism and associated oxidative stress.

Low and high dietary folic acid levels perturb postnatal cerebellar morphology in growing rats.

The brain is particularly sensitive to folate metabolic disturbances, because methyl groups are critical for brain functions. This study aimed to investigate the effects of different dietary levels of folic acid (FA) on postnatal cerebellar morphology, including the architecture and organisation of the various layers. A total of forty male OFA rats (a Sprague-Dawley strain), 5 weeks old, were classified into the following four dietary groups: FA deficient (0 mg/kg FA); FA supplemented (8 mg/kg FA); FA supra-supplemented (40 mg/kg FA); and control (2 mg/kg FA) (all n 10 per group). Rats were fed ad libitum for 30 d. The cerebellum was quickly removed and processed for histological and immunohistochemical analysis. Slides were immunostained for glial fibrillary acidic protein (to label Bergmann glia), calbindin (to label Purkinje cells) and NeuN (to label post-mitotic neurons). Microscopic analysis revealed two types of defect: partial disappearance of fissures and/or neuronal ectopia, primarily in supra-supplemented animals (incidence of 80 %, P≤0·01), but also in deficient and supplemented groups (incidence of 40 %, P≤0·05), compared with control animals. The primary fissure was predominantly affected, sometimes accompanied by defects in the secondary fissure. Our findings show that growing rats fed an FA-modified diet, including both deficient and supplemented diets, have an increased risk of disturbances in cerebellar corticogenesis. Defects caused by these diets may have functional consequences in later life. The present study is the first to demonstrate that cerebellar morphological defects can arise from deficient, as well as high, FA levels in the diet.

Contribution of folic acid-fortified foods to fertile women's folate Recommended Nutrient Intake through breakfast simulation models.

OBJECTIVE: To assess the potential contribution of foods fortified with folic acid (FA) to target population intakes when included as part of a healthy breakfast. DESIGN: Breakfast models aligned with the Spanish Dietary Guidelines were studied using the recommended, average and 95th percentile of serving sizes consumed by women of childbearing age. Food composition data were obtained from a database including FA analytical data from sixty-eight products and the Spanish food composition tables. Different scenarios were assessed with the inclusion of one, two or three FA-fortified products and with two different fortification levels: ≤ 33 µg/serving (L1) and ≥ 70 µg/serving (L4). FA contents provided by the different models for each scenario were compared with the Recommended Nutrient Intake (RNI) for folate and the Upper Level (UL) of intake for FA. SETTING: Madrid Region, Spain. SUBJECTS: Women aged 16-49 years were considered. RESULTS: Overall, simulation of ten breakfast models and three scenarios of product inclusion accounted for 20-25% of total daily energy recommendations for women. Unfortified breakfast models provided on average 4-23% of the folate RNI. Inclusion of one L4 FA-fortified food contributed 20-60% of the RNI. Fortified yoghurt and milk had the highest FA contents per serving. Scenarios with two or three fortified products delivered 40-80% of the RNI. None of the evaluated models exceeded the FA UL. CONCLUSIONS: At existing levels of FA fortification, inclusion of fortified products as part of a regular breakfast meal could positively impact the nutritional quality of women's diet without involving a risk of excessive FA exposure.

Nutritional Status of Breastfeeding Mothers and Impact of Diet and Dietary Supplementation: A Narrative Review.

Adequate nutrition during breastfeeding is crucial for ensuring the good health of mothers and babies. Despite the high energy and nutrient demands of breastfeeding, lactating women are often vulnerable from a nutritional perspective. The nutritional focus during breastfeeding tends to be on the newborn, often neglecting the mother's diet. Therefore, in the present narrative review, nutrient intakes were compared with the dietary reference values (DRVs) proposed by the European Food Safety Authority (EFSA) as well as by the World Health Organization/Food and Agriculture Organization (WHO/FAO). In the diets of lactating mothers, dietary inadequacies were observed in the intake of some vitamins, such as folic acid, vitamin B12, vitamin A, and vitamin D, and in the intake of certain minerals like calcium, iron, and iodine; polyunsaturated omega-3 fatty acid deficiencies, primarily in eicosapentaenoic acid and docosahexaenoic acid, were also observed. On the other hand, the debate on the necessity of supplementation during lactation continues; the need for nutritional supplementation during lactation depends on many factors, such us mothers' eating habits. There seems to be a positive association between nutritional supplementation of the lactating mother and the concentration of certain nutrients in human milk. The present narrative review provides an update on the nutritional status (fatty acids and micronutrients) of breastfeeding mothers and the impact of diet and dietary supplementation on human milk composition.

Maternal folic acid-deficient diet causes congenital malformations in the mouse eye.

BACKGROUND: The eye is a very complex structure derived from the neural tube, surface ectoderm, and migratory mesenchyme from a neural crest origin. Because structures that evolve from the neural tube may be affected by a folate/folic acid (FA) deficiency, the aim of this work was to investigate whether a maternal folic acid-deficient diet may cause developmental alterations in the mouse eye. METHODS: Female C57BL/6J mice (8 weeks old) were assigned into two different folic acid groups for periods ranging between 2 and 16 weeks. Animals were killed at gestation day 17. Hepatic folate was analyzed, and the eyes from 287 fetuses were macroscopically studied, sectioned and immunolabeled with anti-transforming growth factor (TGF)-ß2 and anti-TGF-ßRII. RESULTS: Mice exposed to a FA-deficient diet exhibited numerous eye macroscopic anomalies, such as anophthalmia and microphthalmia. Microscopically, the eye was the most affected organ (43.7% of the fetuses). The highest incidence of malformations occurred from the 8th week onward. A statistically significant linear association between the number of maternal weeks on the FA-deficient diet and embryonic microscopic eye malformations was observed. The optic cup derivatives and structures forming the eye anterior segment showed severe abnormalities. In addition, TGF-ß2 and TGF-ßRII expression in the eye was also altered. CONCLUSION: This study suggests that an adequate folic acid/folate status plays a key role in the formation of ocular tissues and structures, whereas a vitamin deficiency is negatively associated with a normal eye development even after a short-term exposure.

Dietary folic acid intake differentially affects methionine metabolism markers and hippocampus morphology in aged rats.

PURPOSE: Folic acid (FA) is an emerging nutritional factor in the pathogenesis of diverse neurodegenerative disorders by still unknown mechanisms. The hippocampus is altered during the loss of cognitive abilities in humans and selectively affected when homocysteine increases. The aim was to evaluate the potential protective role of folic acid in the maintenance of biochemical markers related to the methionine cycle, as well as the integrity of the hippocampus as part of the brain in aged rats. METHODS: Male Sprague-Dawley rats (18 months old) were assigned to four different folic acid groups (0 mg FA/kg diet, deficient; 2 mg FA/kg diet, control; 8 mg FA/kg diet, moderate supplementation; 40 mg FA/kg diet, extra supplementation) for 30 days. We evaluated several parameters related to the methionine cycle. In addition, hippocampus areas were immunostained for specific neuronal markers and astrocytes. RESULTS: Serum folate levels increased according to FA dietary level (p < 0.01). There was a significant increase in the serum homocysteine concentrations in the folic acid-deficient diet group (p < 0.01). However, brain S-adenosylmethionine and S-adenosylhomocysteine did not differ significantly between the folic acid groups. Consequently, the methylation ratio was also unchanged. The morphometric analysis did not show any differences in the number of neurons and astrocytes between groups, except when comparing the folic acid-deficient diet versus folic acid-supplemented diet in the striatum of the hippocampus. CONCLUSIONS: Clearly, the dietary FA deficiency negatively affects the methionine metabolism biomarkers, while excessive supplementation seems to be unnecessary for optimal maintenance of the methylation cycle and hippocampus integrity.

Omega-3 and Omega-6 Polyunsaturated Fatty Acid Intakes, Determinants and Dietary Sources in the Spanish Population: Findings from the ANIBES Study.

The multiple roles of polyunsaturated fatty acids (PUFA) in growth and general health are well documented. However, available intake data for the Spanish population are limited and lack gender and age considerations. Therefore, our goal was to assess dietary intake adequacy of omega-3 and omega-6 PUFA, their determinants and their major food sources among the Spanish population. Due to their influence on various beneficial functions attributed to omega-3 PUFA, combined intake adequacy with folic acid (FA), vitamin B12 and choline was also assessed. Intake data were obtained from the ANIBES cross-sectional study on a representative sample of the Spanish population (9-75 years; n = 2009), where dietary intake was analysed with a three-day dietary record. Median intake of total omega-3 PUFA stood at 0.81 g/day (0.56-1.19 g/day), with α-linolenic acid (ALA) at 0.61 g/day (0.45-0.85 g/day), eicosapentaenoic acid (EPA) at 0.03 g/day (0.01-0.12 g/day) and docosahexaenoic acid (DHA) at 0.06 g/day (0.0-0.20 g/day). Accordingly, 65% of the Spanish population showed insufficient intakes for total omega-3 PUFA; 87% for ALA, and 83% for combined EPA and DHA. Inadequate intakes were significantly higher in children, adolescents, and younger women of childbearing age (18-30 years). In contrast, inadequacy due to excessive intakes was almost negligible. Regarding omega-6 PUFA, total intake was 10.1 g/day (7.0-14.0 g/day), 10.0 g/day (6.9-13.9 g/day) for linoleic acid (LA) and 0.08 g/day (0.05-0.13 g/day) for arachidonic acid (AA). Non-compliance due to either insufficient or excessive intakes of LA stood at around 5% of the sample, with the elderly showing significantly higher degrees of inadequacy due to insufficient intakes (10%; p ≤ 0.05). Median omega-6 to omega-3 ratio was 12:1, and significantly higher in men compared to women (p ≤ 0.05); in children, adolescents and adults compared to the elderly (p ≤ 0.05); and in younger women of childbearing age compared to the older group (31-45 years) (p ≤ 0.001). Oils and fats and meat and meat products were the main dietary sources for the essential fatty acids LA and ALA, respectively. Meat and meat products were as well the main providers of AA, while fish and shellfish were almost exclusively the only sources of EPA and DHA. However, main food sources identified showed important differences across age groups. Finally, the total combined degree of inadequacy observed for omega-3 PUFA, FA, vitamin B12 and choline reached 21.3% of the ANIBES population. The observed degree of inadequacy of omega-3 PUFA intakes among the Spanish population makes it urgent to increase its consumption and to consider the need for supplementation. This should also be the main strategy for the optimization of the omega-6/omega-3 ratio, as the adequacy observed for omega-6 intakes is relatively acceptable. Additional improvement of the dietary intake of FA, vitamin B12 and choline could contribute to the beneficial effects of omega-3 PUFA.

Dietary intake, nutritional adequacy and food sources of vitamins involved in the methionine-methylation cycle from Spanish children aged one to <10 years: results from the EsNuPI study.

Background: Methionine-methylation cycle and the derived critical functions during infancy are key regulated by folates, vitamins B12, and B6. At present in Spain, there is an absence of studies that assess the intakes and dietary sources of total folates and B12 by children consuming all types of milks and those regularly consuming adapted milk formulas. Thus, our aim was to evaluate folates intakes alongside with vitamins B6 and B12 while describing their major dietary contributors in Spanish children aged one to <10 years. Methods: A total of 1,448 children aged between 1 and 10 years (49.7% girls and 50.3% boys) from the EsNuPI, a prospective cross-sectional study, were allocated into two cohorts: one Spanish Reference Cohort (SRS) of the general population (n = 707), and another including children consuming adapted milks called Adapted Milk Consumers Cohort (AMS) (n = 741) completed two 24 h dietary recalls used to estimate their nutrient intakes and to compare them to the European Food Safety Authority (EFSA) Population Reference Intakes. Results: The median intake of vitamin B6 was 1.35 (1.06-1.70) mg/day in the SRS and 1.45 (1.17-1.79) mg/day in the AMS, being significantly higher in the AMS for all age-groups. Prevalence of adequacy for vitamin B6 in the SRS and AMS was 97.7 and 98.7%, respectively. Total folates intakes in the AMS were significantly higher (p ≤ 0.001) in all age groups than in the SRS, independently of age. In addition, the prevalence of adequacy for folates intakes in all groups was more than 60%. Vitamin B12 intake increased with age independently of the type of milk consumed. The prevalence of adequacy for vitamin B12 was highly compliant by all population groups. The major contributors to vitamin B6 were milk and dairy products being significantly higher in AMS than SRS (p ≤ 0.001). The highest contributors to folates intakes were milk and dairy products, cereals, vegetables, and fruits in both groups whereas for vitamin B12 in the SRS sample were milk and dairy products followed by meat and meats products and for adapted milks, were milk and dairy products, followed by eggs, then meat and meats products. Conclusion: A satisfactory prevalence of adequacy for vitamins B6, and B12 amongst the Spanish children population was observed, which was not the case for folates, regardless of the dietary group evaluated. Nevertheless, a possible strategy to increase folate intake among the youngest children is to increase the consumption of milk and dairy products within a healthier dietary pattern, as these may contribute significantly to the vitamin needs of the infant population.

Omega 3 fatty acids in the elderly.

Population ageing affects the entire world population. Also at world level one can observe a sharp increase in the proportion of older people. The challenge posed by population ageing translates into ensuring that the extra years of life will be as good as possible, free from high-cost dependency. Omega-3 fatty acids are now generally recognized as potential key nutrients to prevent the pathological conditions associated to the aging process. Ageing physiological process, its association with quality of life and the impact of omega-3 fatty acids intake and/or status is the focus of the present review. This report deals with the effects of omega-3 fatty acids on normal aging of older adults ( ≥ 65 years) mainly on the effects such as nutritional status itself, cognition, bone health, muscle tonus, and general health status. The preliminary broad search of the literature on the effects of omega-3 fatty acids on normal aging yielded 685 citations. Forty two full text papers were checked for inclusion and thirty six studies were finally included in this review. It may be concluded that paradoxically even though the elderly population is the largest one, the number of studies and the methodology employed clearly lacks of sufficient evidence to establish definite conclusions on the effects of omega-3 fatty acids on aging metabolism without pathological conditions and on quality of life.

Efficacy of adding folic acid to foods.

In the past, food fortification along with nutritional education and the decrease in food costs relative to income have proven successful in eliminating common nutritional deficiencies. These deficiencies such as goiter, rickets, beriberi, and pellagra have been replaced with an entirely new set of "emergent deficiencies" that were not previously considered a problem [e.g., folate and neural tube defects (NTDs)]. In addition, the different nutrition surveys in so-called affluent countries have identified "shortfalls" of nutrients specific to various age groups and/or physiological status. Complex, multiple-etiology diseases, such as atherosclerosis, diabetes, cancer, and obesity have emerged. Food fortification has proven an effective tool for tackling nutritional deficiencies in populations; but today a more reasonable approach is to use food fortification as a means to support but not replace dietary improvement strategies (i. e. nutritional education campaigns). Folic acid (FA) is a potential relevant factor in the prevention of a number of pathologies. The evidence linking FA to NTD prevention led to the introduction of public health strategies to increase folate intakes: pharmacological supplementation, mandatory or voluntary fortification of staple foods with FA, and the advice to increase the intake of folate-rich foods. It is quite contradictory to observe that, regardless of these findings, there is only limited information on food folate and FA content. Data in Food Composition Tables and Databases are scarce or incomplete. Fortification of staple foods with FA has added difficulty to this task. Globally, the decision to fortify products is left up to individual food manufacturers. Voluntary fortification is a common practice in many countries. Therefore, the "worldwide map of vitamin fortification" may be analyzed. It is important to examine if fortification today really answers to vitamin requirements at different ages and/or physiological states. The real impact of vitamin fortification on some key biomarkers is also discussed. An important question also to be addressed: how much is too much? It is becoming more evident that chronic excessive intakes may be harmful and a wide margin of safety seems to be a mandatory practice in dietary recommendations. Finally, the "risk/benefit" dilemma is also considered in the "new" FA-fortified world.

Fish, Seafood, and Fish Products Purchasing Habits in the Spanish Population during COVID-19 Lockdown.

The Mediterranean diet is a healthy dietary pattern in which fish consumption is an important key element. In Spain, fish intake is the second highest in Europe. Dietary guidelines recommend an intake of 1-3 portions a week of fish. However, Spanish fish sales have been decreasing since 2008. The unexpected pandemic spread of the SARS-CoV-2 in 2020 led the Spanish Government to take restrictive measures that had an impact on people's behavior, including food purchases and consumption. The aim of the study was to analyze purchase data of fish, seafood, and fish products during the lockdown in Spain, using data from loyalty card holders (>5,000,000 participants) from a hypermarket chain in Spain. The results show a 45% increase in the purchase of all types of fish, seafood, and fish products, with the highest increment observed in the retiree (+65 years) as compared to the younger populations. Moreover, the retiree, in spite of the digital divide, were also the ones that most increased online shopping. These data should be considered since events like COVID-19 confinement can have a permanent impact on people's dietary habits, a possibility that should be monitored in the future.

A Descriptive Analysis of Macronutrient, Fatty Acid Profile, and Some Immunomodulatory Nutrients in Standard and Disease-Specific Enteral Formulae in Europe.

Foods for special medical purposes (FSMPs) are commercially available formulations used as a source of nutrition when administered orally or by tube feeding. This study examines, for the first time, the nutritional composition of enteral formulae (EFs) according to European nutritional guidelines. We developed a descriptive study on 118 EFs from 2020 to 2021. Formulae were classified as standard (SFs) and disease-specific (DSF). According to the protein-energy content, SFs were classified into G1, normoprotein-normocaloric; G2, normoprotein-hypercaloric; G3, hyperproteic-normocaloric; and G4, hyperproteic-hypercaloric. Disease-related formulae for metabolic stress, renal, cancer, pulmonary, diabetes, malabsorption, and surgery were studied. Macronutrient distribution, fatty acid profile (monounsaturated [MUFA], polyunsaturated [PUFA], saturated [SFA]), derived fat quality indexes, and immuno-modulatory nutrients (omega-3, eicosapentaenoic acid [EPA], docosahexaenoic acid [DHA], arginine and nucleotides) per 1,500 kcal infused were calculated. In total, 53% were SFs, mainly normoproteic (G1, G2) with higher carbohydrate contents in normocaloric vs. hypercaloric SFs. The most balanced fatty acid profiles (MUFA: 17.7%; PUFA: 6.8%; SFA: 9.5%) belonged to G1. The PUFA/MUFA ratio: ≥0.5 was in 85.7% with a higher proportion of EPA+DHA (46%) vs. omega-3 (15.8%) in SFs. In DSFs (46.9%), higher carbohydrate content (>50%) was in malabsorption and surgery, whereas high-fat content (>50%) was in pulmonary and renal formulae. DSFs had higher SFA vs. MUFA content, except for diabetes. EPA and DHA were added in 45.5% (cancer, malabsorption, and surgery). Only 12.7% of DSFs had arginine and nucleotides. A higher proportion of SFs was found, in line with current European guidelines. Results highlighted a wide intra-group variability of nutrients among the formula selected. These findings are useful to evaluate the nutritional composition of EFs from a preventive and/or therapeutic perspective in clinical settings.

Iron Supplementation at the Crossroads of Nutrition and Gut Microbiota: The State of the Art.

Gut microbiota has received significant attention owing to its decisive role in human health and disease. Diet exerts a significant influence on the variety and number of bacteria residing in the intestinal epithelium. On the other hand, as iron is a key micronutrient for blood formation and oxygen supply, its deficiency is highly prevalent worldwide. In fact, it is the most common cause of anemia and thus, iron supplementation is widespread. However, there is concern due to some potential risks linked to iron supplementation. Therefore, we have reviewed the available evidence of the effects that iron supplementation exerts on the gut microbiota as well as its potential benefits and risks. The compiled information suggests that iron supplementation is potentially harmful for gut microbiota. Therefore, it should be performed with caution, and by principle, recommended only to individuals with proven iron deficiency or iron-deficiency anemia to avoid potential adverse effects. In any case, large and long-term population studies are urgently needed to confirm or refute these results, mainly focused on vulnerable populations.

Perception and knowledge of low- and no-calorie sweeteners in multidisciplinary stakeholders from Spain.

Low- and no-calorie sweeteners (LNCS) as a category are one of the most thoroughly evaluated additives, and thus their safety has been largely recognised. However, their potential risks and benefits generate great controversy and discussion within countries' food policies and public debate. The goal of this study was to evaluate the degree of knowledge and the perception of key Spanish stakeholders about the role of LNCS in diet, their safety, regulatory issues and their impact on health and wellness, as well as to complete a SWOT analysis of the Strengths, Weaknesses, Opportunities and Threats regarding this topic from their perspective. Participants (n = 45 stakeholders) completed an anonymous survey about their knowledge and perception of LNCS, their role in the diet, safety and legislation, as well as health issues and completed a SWOT analysis. Most of them agreed with aspects related to safety guarantees and authorisation procedure of LNCS; however, certain disparity in their opinion in relation to several of the topics was observed, especially regarding the possible role of LNCS in diet quality and health. Effective communication strategies to inform professionals and the general population, as well as new research that deepens our knowledge of the role of LNCS in weight management and other health outcomes seem to be urgently needed.