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.

Maternal folic acid supplementation reduces the severity of cleft palate in Tgf-ß3 null mutant mice.

BACKGROUND: Cleft palate (CP) constitutes the most frequently seen orofacial cleft and is often associated with low folate status. Folate plays an essential role in the human body as a major coenzyme in one-carbon metabolism, including DNA synthesis, repair, and methylation. Whether the administration of isolated folic acid (FA) supplements prevents the CP caused by genetic mutations is unknown, as is its effect on the mechanisms leading to palate fusion. METHODS: FA was administered to females from two different strains of transforming growth factor ß3 heterozygous mice. Null mutant progeny of these mice exhibit CP in 100% of cases of varying severity. We measured cleft length, height of palatal shelf adhesion, and the number of proliferating mesenchymal cells. Immunohistochemistry was also carried for collagen IV, laminin, fibronectin, cytokeratin-17, and EGF. RESULTS: FA supplementation significantly reduced CP severity and improved palatal shelf adhesion in both strains both in vivo and in vitro. Medial edge epithelium proliferation increased, and its differentiation was normalized as indicated by the presence and disposition of collagen IV, laminin, fibronectin, and cytokeratin-17. CONCLUSIONS: A maternal FA supplementation reduces the CP appearance by improving the mechanisms leading to palatal shelf adhesion.

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.

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.

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.

Dietary Intake, Nutritional Adequacy, and Food Sources of Selected Antioxidant Minerals and Vitamins; and Their Relationship with Personal and Family Factors in Spanish Children Aged 1 to <10 Years: Results from the EsNuPI Study

Minerals and vitamins involved in the antioxidant defense system are essential for healthy growth and proper development during infancy. Milk and dairy products are of particular importance for improving the supply of these nutrients to children. Indeed, the present study aimed to evaluate the nutrient intake and food sources of zinc (Zn), selenium (Se), retinol and carotenoids (sources of vitamin A), and vitamins C and E, and to analyze their relationships with personal and familiar factors in Spanish children from the EsNuPI study. One subpopulation representative of the Spanish population from 1 to <10 years old (n = 707) (reference group, REF) who reported consuming all types of milk over the last year, and another subpopulation of the same age who reported consuming fortified milk formulas (FMFs) (including follow-on formula, young child formula, growing up milk, toddler's milk, and enriched and fortified milk) (n = 741) (fortified milk consumers, FMCs) completed two 24 h dietary recalls used to estimate their nutrient intakes and to compare them to the European Food Safety Authority (EFSA) Dietary Reference Values (DRVs). The REF reported higher median intakes than FMCs for Se (61 µg/kg vs. 51 µg/kg) and carotenoids (1079 µg/day vs. 998 µg/day). Oppositely, FMCs reported higher intakes than REF for Zn (7.9 mg/day vs. 6.9 mg/day), vitamin A (636 µg/day vs. 481 µg/day), vitamin E (8.9 mg/day vs. 4.5 mg/day), vitamin C (113 mg/day vs. 71 mg/day), and retinol (376 µg/day vs. 233 µg/day). In the REF group, more than 50% of the children met the EFSA recommendations for Zn (79.6%), Se (87.1%), vitamin A (71.3%), and vitamin C (96.7%), respectively. On the other hand, 92.2% were below the EFSA recommendations for vitamin E. In the FMC group, more than 50% of the children met the EFSA recommendations for Zn (55.2%), Se (90.8%), vitamin A (75.7%), vitamin E (66.7%), and vitamin C (100%). We found statistically significant differences between subpopulations for all cases except for Se. In both subpopulations, the main sources of all antioxidant nutrients were milk and dairy products. For carotenoids, the main sources were vegetables and fruits followed by milk and dairy products. A high percentage of children had vitamins A and E intakes below the recommendations, information of great importance to stakeholders. More studies using intakes and biomarkers are needed, however, to determine an association with diverse factors of oxidative damage.

Hearing Function and Nutritional Status in Aviation Pilots from Spain Exposed to High Acoustic Damage.

Noise-induced hearing loss is the most frequent and preventable occupational disease. Aviation pilots are a vulnerable population, as they spend many hours exposed to noise pollution in their working environment. Different studies suggest that certain dietary compounds may play a key role in the etiology and prevention of this pathology. We aimed to study the relationship linking auditory function, dietary intake, and some serum biomarkers in pilots, exposed to noise in the work environment. A total of 235 pilots, who undergo a periodic medical examination at the Centro de Instrucción de Medicina Aeroespacial (Madrid, SPAIN), were evaluated. Auditory function was assessed by tonal audiometry. Energy and nutrient intakes were estimated by 24 h recall (DIALTM program). Serum homocysteine (Hcy) was determined in a Cobas 6000TM multi-analyzer while folate, vitamin B12, and D were analyzed in a Cobas e411TM. The prevalence of hearing loss (HL) was 64.3%. HL was significantly related to age (r = 0.588, p ≤ 0.001) and flight hours (r = 0.283, p ≤ 0.001). A multiple linear regression model was used to assess the relationship among HL, flight hours, serum folate, and Hcy serum levels. HL was significantly (p < 0.050) associated with flight hours (ß = 0.246), serum folate (ß = −0.143), and serum Hcy (ß = 0.227). Nutritional interventions would be of great interest to monitor and slow down the HL progression in populations exposed to noise pollution in their workplace.

Dietary Intake of Individual (Intrinsic and Added) Sugars and Food Sources from Spanish Children Aged One to <10 Years­Results from the EsNuPI Study

Currently, in Spain there are no studies assessing the intakes and sources of intrinsic and added sugars by both children consuming standard milks and children regularly consuming adapted milk formulas. Our goal was to evaluate current sugar intake levels (intrinsic and added) and their major dietary sources within the EsNuPI study participants by applying two 24-h dietary recalls that were completed by 1448 children (1 to <10 years) divided into two subsamples: One "Spanish Reference Sample" (SRS) of the general population (n = 707) and another sample which included children consuming adapted milks including follow-on milk, toddler's or growing up milk and fortified and enriched milks, here called "Adapted Milk Consumers Sample" (AMS) (n = 741). Estimates of intrinsic and added sugar intakes from the Spanish EsNuPI population as well as the adherence to recommendations varied notably according to age segment, but no major differences between subsamples were found. Younger children (1 to <3 years) showed the highest added sugar contribution to total energy intake (TEI) (SRS: 12.5% for boys and 11.7% for girls; AMS: 12.2% for boys and 11.3% for girls) and the lowest adherence to recommendations set at <10% TEI (SRS: 27.4% for boys and 37.2% for girls; AMS: 31.3% for boys and 34.7% for girls). Adherence increased with age but remains inadequate, with approximately one in two children from the older age segment (6 to <10 years) exceeding the recommendations. Main food sources of intrinsic sugars for both subsamples were milk and dairy products, fruits, vegetables and cereals, while for added sugars, these were milk and dairy products (mainly yogurts), sugars and sweets (mainly sugary cocoa and nougat), bakery products (mainly cookies) and cereals (mainly bread and wheat flour). However, for the AMS, the groups milk and dairy products and cereals showed a significantly lower contribution to intrinsic sugar intake but a significantly higher contribution to that of added sugars. These results demonstrate that sugar intake and the adherence to recommendations in the studied population varied notably according to age but not to the type of milk consumed. In addition, our results highlight the need to monitor the consumption of added sugars by the infant population, as well as the need to make efforts to facilitate this task, such as harmonizing the recommendations regarding free/added sugars and the inclusion of information on their content on the nutritional labeling of products in order to incorporate them into food composition databases.

Occurrence of cleft-palate and alteration of Tgf-ß(3) expression and the mechanisms leading to palatal fusion in mice following dietary folic-acid deficiency.

Folic acid (FA) is essential for numerous bodily functions. Its decrease during pregnancy has been associated with an increased risk of congenital malformations in the progeny. The relationship between FA deficiency and the appearance of cleft palate (CP) is controversial, and little information exists on a possible effect of FA on palate development. We investigated the effect of a 2-8 weeks' induced FA deficiency in female mice on the development of CP in their progeny as well as the mechanisms leading to palatal fusion, i.e. cell proliferation, cell death, and palatal-shelf adhesion and fusion. We showed that an 8 weeks' maternal FA deficiency caused complete CP in the fetuses although a 2 weeks' maternal FA deficiency was enough to alter all the mechanisms analyzed. Since transforming growth factor-ß(3) (TGF-ß(3)) is crucial for palatal fusion and since most of the mechanisms impaired by FA deficiency were also observed in the palates of Tgf-ß(3)null mutant mice, we investigated the presence of TGF-ß(3) mRNA, its protein and phospho-SMAD2 in FA-deficient (FAD) mouse palates. Our results evidenced a large reduction in Tgf-ß(3) expression in palates of embryos of dams fed an FAD diet for 8 weeks; Tgf-ß(3) expression was less reduced in palates of embryos of dams fed an FAD diet for 2 weeks. Addition of TGF-ß(3) to palatal-shelf cultures of embryos of dams fed an FAD diet for 2 weeks normalized all the altered mechanisms. Thus, an insufficient folate status may be a risk factor for the development of CP in mice, and exogenous TGF-ß(3) compensates this deficit in vitro.

Low and no calorie sweeteners, diet and health: an updated overview.

INTRODUCTION: Introduction: currently, there is a great deal of controversy surrounding the potential health benefits and risks associated with the use of low and/or no calorie sweeteners (LNCS). Objective: in the present work, the objective was to briefly address the current role of LNCS consumption in the diet and its effects on health. Methods: a brief narrative review of the most recent tudies and policies available was carried out. Results: a limited number of representative studies on the consumption of LNCS and their effect on health are presently available. However, these mostly indicate that the consumption of LNCS can be a useful tool along with other nutritional strategies in the treatment of overweight, obesity, diabetes and the prevention of caries when used appropriately in the context of a balanced diet and physical activity. Still, it is necessary to be cautious with the consumption of certain sweeteners since the effects of LNCS on the intestinal microbiota or its effect on premature deliveries, among others, have not been fully elucidated. Conclusions: it is essential to carry out further studies in order to clarify/establish the safety and value of sweeteners as food ingredients/additives in the medium/long term, in a model of increasing consumption as a consequence of the reformulation of many foods.

INTRODUCCIÓN: Introducción: en la actualidad, existe una gran controversia en torno a los beneficios y riesgos potenciales asociados al uso de los edulcorantes bajos en/o sin calorías (LNCS) en el modelo alimentario y su repercusión en la salud. Objetivo: en el presente trabajo, el objetivo fue abordar brevemente el papel actual del consumo de LNCS y sus efectos en la dieta y salud. Métodos: se llevó a cabo una revisión narrativa de los estudios más recientes disponibles. Resultados: se observa un número limitado de estudios representativos sobre el consumo de LNCS y su efecto en la dieta y la salud. No obstante, los estudios disponibles indican que su consumo puede constituir una herramienta útil junto con otras estrategias nutricionales en el tratamiento del sobrepeso, la obesidad, la diabetes y la prevención de las caries cuando se utilizan adecuadamente en el contexto de una dieta equilibrada y ejercicio. Sin embargo, hay que aplicar el principio de precaución con el consumo de ciertos edulcorantes, ya que los efectos de los LNCS en la microbiota intestinal o su efecto en los partos prematuros, entre otros, no han sido completamente dilucidados. Conclusiones: resulta imprescindible realizar más estudios para poder aclarar/establecer la seguridad de los edulcorantes como ingredientes/aditivos alimentarios a medio/largo plazo, en un modelo de potencial consumo creciente como consecuencia de la emergente reformulación de muchos alimentos.

Updated Database and Trends of Declared Low- and No-Calorie Sweeteners From Foods and Beverages Marketed in Spain.

Background: The past few years have witnessed an increase in the availability of food products containing one or more low- and no-calorie sweeteners (LNCS) in the Spanish market, mostly due to the new massive reformulation plan. However, these are not included in food composition tables or databases, and, therefore, assessment of their intake among the population is complex. This study aims to update a database including commercialized foods and beverages. Method: A systematic search of ingredients information from the different food and beverage categories was undertaken during 2019 by recording the availability and type of LNCS declared in the information of the product from labels and online shopping platforms of retailers from Spain to update a previous food composition database compiled in 2017. Results: A total of 1,238 products were identified. The major groups were sugar and sweets (24%), non-alcoholic beverages (21%), cereals and grains (19%), and milk and dairy products (14%) accounting for >70% of total products. The mainly declared LNCS were sorbitol (19.5%), sucralose (19.5%), and acesulfame K (19.2%). Conclusion: There is a wide variety of products that include LNCS as a main ingredient with higher availability than when compared with the results of database of 2017, consequently, it might be expected that LNCS are commonly consumed at present in the Spanish diet.

Maternal Folic Acid Deficiency Is Associated to Developing Nasal and Palate Malformations in Mice.

Craniofacial development requires extremely fine-tuned developmental coordination of multiple specialized tissues. It has been evidenced that a folate deficiency (vitamin B9), or its synthetic form, folic acid (FA), in maternal diet could trigger multiple craniofacial malformations as oral clefts, tongue, or mandible abnormalities. In this study, a folic acid-deficient (FAD) diet was administered to eight-week-old C57/BL/6J female mouse for 2-16 weeks. The head symmetry, palate and nasal region were studied in 24 control and 260 experimental fetuses. Our results showed a significant reduction in the mean number of fetuses per litter according to maternal weeks on FAD diet (p < 0.01). Fetuses were affected by cleft palate (3.8%) as well as other severe congenital abnormalities, for the first time related to maternal FAD diet, as head asymmetries (4.6%), high arched palate (3.5%), nasal septum malformed (7.3%), nasopharynx duct shape (15%), and cilia and epithelium abnormalities (11.2% and 5.8%). Dysmorphologies of the nasal region were the most frequent, appearing at just four weeks following a maternal FAD diet. This is the first time that nasal region development is experimentally related to this vitamin deficiency. In conclusion, our report offers novel discoveries about the importance of maternal folate intake on midface craniofacial development of the embryos. Moreover, the longer the deficit lasts, the more serious the consequent effects appear to be.