Phenotypic, Technological, Safety, and Genomic Profiles of Gamma-Aminobutyric Acid-Producing Lactococcus lactis and Streptococcus thermophilus Strains Isolated from Cow's Milk.

Gamma-aminobutyric acid (GABA)-producing lactic acid bacteria (LAB) can be used as starters in the development of GABA-enriched functional fermented foods. In this work, four GABA-producing strains each of Lactococcus lactis and Streptococcus thermophilus species were isolated from cow's milk, and their phenotypic, technological, and safety profiles determined. Genome analysis provided genetic support for the majority of the analyzed traits, namely, GABA production, growth in milk, and the absence of genes of concern. The operon harboring the glutamate decarboxylase gene (gadB) was chromosomally encoded in all strains and showed the same gene content and gene order as those reported, respectively, for L. lactis and S. thermophilus. In the latter species, the operon was flanked (as in most strains of this species) by complete or truncated copies of insertion sequences (IS), suggesting recent acquisition through horizontal gene transfer. The genomes of three L. lactis and two S. thermophilus strains showed a gene encoding a caseinolytic proteinase (PrtP in L. lactis and PrtS in S. thermophilus). Of these, all but one grew in milk, forming a coagulum of good appearance and an appealing acidic flavor and taste. They also produced GABA in milk supplemented with monosodium glutamate. Two L. lactis strains were identified as belonging to the biovar. diacetylactis, utilized citrate from milk, and produced significant amounts of acetoin. None of the strains showed any noticeable antibiotic resistance, nor did their genomes harbor transferable antibiotic resistance genes or genes involved in toxicity, virulence, or pathogenicity. Altogether these results suggest that all eight strains may be considered candidates for use as starters or components of mixed LAB cultures for the manufacture of GABA-enriched fermented dairy products.

Prenatal iron supplementation adjusted to maternal iron stores reduces behavioural problems in 4-year-old children.

Prenatal iron supplementation improves children's health and cognitive performance, but few studies explore behavioural development. This study assessed the effects of adjusting prenatal iron supplementation to maternal iron stores during early pregnancy on children's behavioural problems. Randomized controlled trial conducted in Tarragona (Spain) involving 230 nonanaemic pregnant women and their children after a 4-year follow-up. Based on haemoglobin (Hb) levels before gestational week (GW) 12, women receive different iron doses: those with Hb = 110-130 g/L were randomized to receive 80 or 40 mg/day and those with Hb > 130 g/L were randomized to receive 20 or 40 mg/day. Maternal iron stores at GW12 were classified using serum ferritin (SF) as low (SF < 15 µg/L), normal (SF = 15-65 µg/L), and normal-high (SF > 65 µg/L). Children's behaviour was assessed by parents using the Child Behaviour Checklist for ages 1.5-5 years and the Behaviour Rating Inventory of Executive Function-Preschool Version, and by teachers using the Teacher's Report Form for ages 1.5-5 years. Multivariable regression models were performed. Taking 80 mg/day of iron improved child behaviour when women had low iron stores but worsened it when mothers had normal-high iron stores, except for depressive and attention/hyperactivity problems. Taking 20 mg/day of iron improved behaviour only in those children whose mothers had SF > 65 µg/L in early pregnancy. Additionally, executive functioning improved at high doses of prenatal iron when maternal baseline SF < 15 µg/L. Adjusting prenatal iron supplementation to both maternal baseline Hb levels and iron stores reduces behavioural problems in 4-year-old children.

Importance of Maternal Iron Status on the Improvement of Cognitive Function in Children After Prenatal Iron Supplementation.

INTRODUCTION: The effectiveness of prenatal iron supplementation improves maternal hematological outcomes, but little research has focused on child outcomes. The objective of this study was to assess whether prenatal iron supplementation adjusted to maternal needs improves children's cognitive functioning. METHODS: The analyses included a subsample of nonanemic pregnant women recruited in early pregnancy and their children aged 4 years (n=295). Data were collected between 2013 and 2017 in Tarragona (Spain). On the basis of hemoglobin levels before the 12th gestational week, women receive different iron doses: 80 vs 40 mg/d if hemoglobin is 110-130 g/L and 20 vs 40 mg/d if hemoglobin >130 g/L. Children's cognitive functioning was assessed using the Wechsler Preschool and Primary Scale of Intelligence-IV and Developmental Neuropsychological Assessment-II tests. The analyses were carried out in 2022 after the completion of the study. Multivariate regression models were performed for assessing the association between different doses of prenatal iron supplementation and children's cognitive functioning. RESULTS: Taking 80 mg/d of iron was positively associated with all the scales of the Wechsler Preschool and Primary Scale of Intelligence-IV and Neuropsychological Assessment-II when mothers had initial serum ferritin <15 µg/L, but it was negatively associated with Verbal Comprehension Index, Working Memory Index, Processing Speed Index, and Vocabulary Acquisition Index from Wechsler Preschool and Primary Scale of Intelligence-IV and verbal fluency index from Neuropsychological Assessment-II when mothers showed initial serum ferritin >65 µg/L. In the other group, taking 20 mg/d of iron was positively associated with Working Memory Index, Intelligence Quotient, verbal fluency, and emotion recognition indices when women had initial serum ferritin >65 µg/L. CONCLUSIONS: Prenatal iron supplementation adjusted to the maternal hemoglobin levels and baseline iron stores improves cognitive functioning in children aged 4 years.

Adapting prenatal iron supplementation to maternal needs results in optimal child neurodevelopment: a follow-up of the ECLIPSES Study.

BACKGROUND: Prenatal prescription of standard iron supplements to prevent iron deficiency appears not to be appropriate for all women and their children, as some women may be at risk of iron deficiency and others at risk of iron excess early in pregnancy. The present study aimed to assess whether prenatal iron supplementation adapted to the needs of each pregnant woman affects their child's neurodevelopment. METHODS: Follow-up of a community-based RCT involving 503 mother-child pairs. Non-anaemic pregnant women recruited in Tarragona (Spain) early in pregnancy were prescribed a daily iron dose based on their initial haemoglobin levels: Stratum 1 (Hb = 110-130 g/L, 80 or 40 mg/d of iron) and Stratum 2 (Hb > 130 g/L, 40 or 20 mg/d of iron). Women receiving 40 mg/d were considered the control group in each Strata. The child's neurodevelopment was assessed at 40 days of age using the Bayley Scales of Infant Development-III (BSID-III). Adjusted multiple regression models were used. RESULTS: Multiple regression analyses showed no association between the intervention and control group within each Strata on the BSID-III scores on any of the developmental scales in children, including cognitive, language, and motor development: Stratum 1 (ß 1.46, 95%CI -2.15, 5.07; ß 1.30, 95%CI -1.99, 4.59; and ß 2.04, 95%CI -3.88, 7.96, respectively) and Stratum 2 (ß -4.04, 95%CI -7.27, 0.80; ß -0.36, 95%CI -3.47, 2.75; and ß -3.76, 95%CI -9.30, 1.78, respectively). CONCLUSIONS: In non-anaemic women in early pregnancy, no differences were found in the cognitive, language and motor development of children at 40 days of age between the dose of iron tested in each case -adjusted to initial Hb levels- compared to the dose of the control group. Further studies are guaranteed to confirm our findings. TRIAL REGISTRATION: The ECLIPSES study was registered at www.clinicaltrialsregister.eu as EudraCT number 2012-005,480-28.

Prenatal folic acid supplementation and folate status in early pregnancy: ECLIPSES study.

This research evaluates the prevalence of inadequate folate status in early pregnancy, the pattern of prenatal folic acid (FA) supplementation and associated factors in Spanish pregnant women from the ECLIPSES study, which included 791 participants prior gestational week 12. A cross-sectional evaluation of erythrocyte folate levels was performed at recruitment and used to calculate the prevalence of folate deficiency (erythrocyte folate < 340 nmol/l) and insufficiency (erythrocyte folate < 906 nmol/l). Sociodemographic and lifestyle data as well as information on prenatal FA supplementation were recorded. Descriptive and multivariate statistical analyses were performed. The prevalence of folate deficiency and insufficiency were 9·6 % and 86·5 %, respectively. Most of women used prenatal FA supplements, but only 6·3 % did so as recommended. Supplementation with FA during the periconceptional period abolished folate deficiency and reduced folate insufficiency. Prenatal FA supplementation with ≥1000 µg/d in periconceptional time and pregnancy planning increased erythrocyte folate levels. The main risk factor for folate insufficiency in early pregnancy was getting prenatal FA supplementation out of the periconceptional time (OR 3·32, 95 % CI 1·02, 15·36), while for folate deficiency they were young age (OR 2·02, 95 % CI 1·05, 3·99), and smoking (OR 2·39, 95 % CI 1·30, 4·37). In addition, social and ethnic differences according to folate status were also identified. As conclusion, periconceptional FA use is crucial for achieving optimal folate levels in early pregnancy. Pregnancy planning should focus on young women, smokers, those with low consumption of folate-rich foods, low socio-economic status or from ethnic minorities.

Pressurized Extraction as an Opportunity to Recover Antioxidants from Orange Peels: Heat treatment and Nanoemulsion Design for Modulating Oxidative Stress.

Orange peel by-products generated in the food industry are an important source of value-added compounds that can be potentially reused. In the current research, the effect of oven-drying (50-70 °C) and freeze-drying on the bioactive compounds and antioxidant potential from Navelina, Salustriana, and Sanguina peel waste was investigated using pressurized extraction (ASE). Sixty volatile components were identified by ASE-GC-MS. The levels of terpene derivatives (sesquitenenes, alcohols, aldehydes, hydrocarbons, and esters) remained practically unaffected among fresh and freeze-dried orange peels, whereas drying at 70 °C caused significative decreases in Navelina, Salustriana, and Sanguina peels. Hesperidin and narirutin were the main flavonoids quantified by HPLC-MS. Freeze-dried Sanguina peels showed the highest levels of total-polyphenols (113.3 mg GAE·g-1), total flavonoids (39.0 mg QE·g-1), outstanding values of hesperedin (187.6 µg·g-1), phenol acids (16.54 mg·g-1 DW), and the greatest antioxidant values (DPPH•, FRAP, and ABTS•+ assays) in comparison with oven-dried samples and the other varieties. Nanotechnology approaches allowed the formulation of antioxidant-loaded nanoemulsions, stabilized with lecithin, starting from orange peel extracts. Those provided 70-80% of protection against oxidative UV-radiation, also decreasing the ROS levels into the Caco-2 cells. Overall, pressurized extracts from freeze-drying orange peel can be considered a good source of natural antioxidants that could be exploited in food applications for the development of new products of commercial interest.

Iron status in mid-pregnancy and associations with interpregnancy interval, hormonal contraceptives, dietary factors and supplement use.

Adequate iron supply in pregnancy is important for both the woman and the fetus, but iron status is often assessed late in first trimester, if assessed at all. Therefore, identification of factors associated with iron status is important to target vulnerable groups with increased risk of deficiency. Our objectives were to (1) describe iron status in mid-pregnancy and (2) identify sociodemographic and lifestyle predictors of pregnancy iron status. This cross-sectional study uses data from The Norwegian Mother, Father and Child Cohort Study (collected 2002-2008) and The Medical Birth Registry of Norway. Iron status was measured as non-fasting plasma ferritin (P-Fe) and transferrin in gestational week (GW) 18 (n 2990), and by lowest reported Hb in GW 0-30 (n 39 322). We explored predictors of iron status with elastic net, linear and log-binomial regression models. Median P-Fe was 33 µg/l, and 14 % had depleted iron stores (P-Fe <15 µg/l). P-Fe below 30 µg/l was associated with reduced Hb. We identified eleven predictors, with interpregnancy interval (IPI) and parity among the most important. Depleted iron stores was more common among women with IPI < 6 months (56 %) and 6-11 months (33 %) than among those with IPI 24-59 months (19 %) and among nulliparous women (5 %). Positively associated factors with iron status included hormonal contraceptives, age, BMI, smoking, meat consumption and multi-supplement use. Our results highlight the importance of ferritin measurements in women of childbearing age, especially among women not using hormonal contraceptives and women with previous and recent childbirths.

The Effectiveness of Different Doses of Iron Supplementation and the Prenatal Determinants of Maternal Iron Status in Pregnant Spanish Women: ECLIPSES Study.

Iron deficiency (ID), anemia, iron deficiency anemia (IDA) and excess iron (hemoconcentration) harm maternal-fetal health. We evaluated the effectiveness of different doses of iron supplementation adjusted for the initial levels of hemoglobin (Hb) on maternal iron status and described some associated prenatal determinants. The ECLIPSES study included 791 women, randomized into two groups: Stratum 1 (Hb = 110-130g/L, received 40 or 80mg iron daily) and Stratum 2 (Hb > 130g/L, received 20 or 40mg iron daily). Clinical, biochemical, and genetic information was collected during pregnancy, as were lifestyle and sociodemographic characteristics. In Stratum 1, using 80 mg/d instead of 40 mg/d protected against ID on week 36. Only women with ID on week 12 benefited from the protection against anemia and IDA by increasing Hb levels. In Stratum 2, using 20 mg/d instead of 40 mg/d reduced the risk of hemoconcentration in women with initial serum ferritin (SF) ≥ 15 µg/L, while 40 mg/d improved SF levels on week 36 in women with ID in early pregnancy. Mutations in the HFE gene increased the risk of hemoconcentration. Iron supplementation should be adjusted to early pregnancy levels of Hb and iron stores. Mutations of the HFE gene should be evaluated in women with high Hb levels in early pregnancy.

Does the fortified milk with high iron dose improve the neurodevelopment of healthy infants? Randomized controlled trial.

BACKGROUND: Since iron plays an important role in several physiological processes, its deficiency but also overload may harm the development of children. The aim was to assess the effect of iron-fortified milk on the iron biochemical status and the neurodevelopment of children at 12 months of age. METHODS: Randomized controlled trial conducted in 133 Spanish children, allocated in two groups to receive formula milk fortified with 1.2 or 0.4 mg/100 mL of iron between 6 and 12 months of age. Psychomotor (PDI) and Mental (MDI) Development Index were assessed by the Bayley Scales before and after the intervention. Maternal obstetrical and psychosocial variables were recorded. The biochemical iron status of children was measured and data about breastfeeding, anthropometry and infections during the first year of life were registered. RESULTS: Children fortified with 1.2 mg/100 mL of iron, compared with 0.4 mg/100 mL, showed higher serum ferritin (21.5 vs 19.1 µg/L) and lower percentage of both iron deficiency (1.1 to 5.9% vs 3.8 to 16.7%, respectively, from 6 to 12 months) and iron deficiency anemia (4.3 to 1.1% vs 0 to 4.2%, respectively, from 6 to 12 months) at the end of the intervention. No significant differences were found on neurodevelopment from 6 to 12 months between children who received high dose of Fe compared with those who received low dose. CONCLUSION: Despite differences on the iron status were observed, there were no effects on neurodevelopment of well-nourished children in a developed country after iron supplementation with doses within dietary recommendations. Follow-up studies are needed to test for long-term neurodevelopmental improvement. TRIAL REGISTRATION: Retrospectively registered in ClinicalTrials.gov with the ID: NCT02690675.

Prevalence of Anemia in Children from Latin America and the Caribbean and Effectiveness of Nutritional Interventions: Systematic Review and Meta⁻Analysis.

Anemia affects 1.62 billion people worldwide. Latin America and the Caribbean (LAC) comprise several developing countries where children are a population at risk. This systematic review and meta-analysis aimed to estimate the prevalence of anemia in this population. Electronic databases, reference lists, and websites of health ministries were searched until December 2018. Stratified analyses were performed using RevMan5.3 to estimate the overall prevalence of anemia in preschool and school-age children. The effectiveness of nutritional interventions was also evaluated. We included 61 studies from the 917 reviewed, which included 128,311 preschool- and 38,028 school-age children from 21 LAC countries. The number of anemic children was 32.93% and 17.49%, respectively, demonstrating a significant difference according to age (p < 0.01). No difference was observed by gender and only school-age children from low/very low socioeconomic status (SES) (25.75%) were more prone to anemia than those from middle SES (7.90%). It was not a concern in the Southern Cone but constituted a serious public health problem in the Latin Caribbean. Nutritional interventions reduced the prevalence from 45% to 25% (p < 0.01). Anemia is still a public health problem for children in LAC countries. National surveys should include school-age children. Further nutritional interventions are required to control anemia.

Review and meta-analysis found that prenatal folic acid was associated with a 58% reduction in autism but had no effect on mental and motor development.

AIM: The impact of prenatal folic acid on children's neurodevelopment and the risk of autism spectrum disorder (ASD) remain unclear and this review and meta-analysis aimed to quantify any associations. METHODS: We systematically searched PubMed, Scopus and The Cochrane Library until June 2018 with no language restrictions. Standardised mean differences and odds ratio with 95% confidence intervals are used to describe any associations between folic acid and mental development, motor development and ASD. RESULTS: The search strategy identified 647 papers and 16 were finally included in the meta-analysis after the application of the exclusion criteria. These provided a total cohort size of 756 365 children aged 11 months to 15 years from 10 countries. The main finding was that prenatal use of folic acid was associated with a 58% reduction in the risk of ASD in children. We were surprised that better scores for mental development were associated with low prenatal exposure to folic acid. CONCLUSION: Although the results should be interpreted with caution, they showed that routine prenatal supplements of folic acid were associated with significantly lower levels of ASD. Further studies are needed to reach a firm conclusion, given the multifactorial aetiology of neurodevelopment.

Ultrafast determination of vitamin E using LC-ESI-MS/MS for preclinical development of new nutraceutical formulations.

AIM: We proposed a rapid and high quality method to determine α-tocopherol (α-T) in different biopharmaceutical samples using liquid chromatography-diode array detector on-line ESI-MS/MS. MATERIALS & METHODS: A working standard solution of α-T and internal standard, phenyl-5,7-dimethyl-d6-α-tocopherol, were used for optimization and validation of the method. Levels of α-T in nanoemulsions, serum and plasma samples were evaluated. RESULTS & CONCLUSION: Precision (1% for retention time, 5% for peak area and 3% for relative peak area), linearity range (among 0.625-20.0 µg ml-1), LOD and LOQ, accuracy and matrix effect were studied. The validated chromatographic method is presented as valuable analytical tool for the determination of α-tocopherol in loaded drug delivery systems and in biodistribution levels in blood samples.

Bioactive Flavonoids, Antioxidant Behaviour, and Cytoprotective Effects of Dried Grapefruit Peels (Citrus paradisi Macf.).

Grapefruit (Citrus paradisi Macf.) is an important cultivar of the Citrus genus which contains a number of nutrients beneficial to human health. The objective of the present study was to evaluate changes in bioactive flavonoids, antioxidant behaviour, and in vitro cytoprotective effect of processed white and pink peels after oven-drying (45°C-60°C) and freeze-drying treatments. Comparison with fresh grapefruit peels was also assessed. Significant increases in DPPH, FRAPS, and ABTS values were observed in dried grapefruit peel samples in comparison with fresh peels, indicating the suitability of the treatments for use as tools to greatly enhance the antioxidant potential of these natural byproducts. A total of thirteen flavonoids were quantified in grapefruit peel extracts by HPLC-MS/MS. It was found that naringin, followed by isonaringin, was the main flavonoid occurring in fresh, oven-dried, and freeze-dried grapefruit peels. In vivo assay revealed that fresh and oven-dried grapefruit peel extracts (45°C) exerted a strong cytoprotective effect on SH-SY5Y neuroblastoma cell lines at concentrations ranging within 0.1-0.25 mg/mL. Our data suggest that grapefruit (Citrus paradisi Macf.) peel has considerable potential as a source of natural bioactive flavonoids with outstanding antioxidant activity which can be used as agents in several therapeutic strategies.

Estado periconcepcional de ácido fólico: revisión sobre su relevancia para la salud de la madre y de la descendencia. Relevancia del estado periconcepcional de ácido fólico sobre el neurodesarrollo / Perioconceptional folic acid status: review about his relevance on the health of the mother and the offspring. Relevance of perioconceptional folic acid status on neurodevelopment

Fundamento: El déficit de ácido fólico durante el embarazo puede derivar en defectos del tubo neural en el feto, bajo peso al nacer y desórdenes del neurodesarrollo como trastornos de la conducta y de la cognición en el niño pero también puede conllevar anemia megaloblástica en la madre así como desprendimiento de la placenta, abortos, partos prematuros, preeclampsia e incluso eclampsia en algunas ocasiones. Objetivo: El objetivo de este trabajo es revisar los principales hallazgos científicos con relación a la importancia fisiológica del ácido fólico durante el embarazo así como los efectos negativos resultantes de un inadecuado nivel de folato periconcepcional, tanto por déficit como por exceso, y la evidencia existente sobre la relación entre suplementación con folato y los diferentes parámetros de salud, tanto de la madre como del recién nacido. Métodos: Se ha realizado una búsqueda amplia en la que se incluyen artículos originales, revisiones y guías de recomendaciones. Resultados/Conclusión: Tanto el déficit como el exceso de ácido fólico durante el embarazo pueden desencadenar efectos adversos para la madre y para el hijo. Por ello, la suplementación de ácido fólico debería ser individualizada para cada mujer teniendo en cuenta sus características fenotípicas, genotípicas y metabólicas (AU)

Background: The folic acid deficit during pregnancy may lead to a neural tube defects in the fetus, low birth weight and neurodevelopmental disorders as behavior disturbance or cognitive disabilities. Also may carry to the megaloblastic anemia in the mother as well as placental abruption, abortion, premature labor, pre-eclampsia and sometimes eclampsia. Objective: The objective of this work is to review the main scientific finds in relation with the physiological importance of folic acid during pregnancy and negative effects resulting of the inadequate level of folate periconceptional, both deficit and excess. Also review the evidence about the relationship between the folate supplementation and the different health parameters in the mother and the newborn. Methods: The search has been wide and this review includes original articles, review articles and recommendation guides. Results/Conclusion: Both deficit and excess folic acid during pregnancy can trigger adverse effects to the mother and the child. Therefore, supplementation with folic acid should be individualized for each woman considering its phenotypic, genotypic and metabolic characteristics (AU)

Identification and determination of 3-deoxyglucosone and glucosone in carbohydrate-rich foods.

BACKGROUND: α-Dicarbonyl compounds (α-DCs) such as 3-deoxyglucosone (3-DG) and glucosone are markers of both Maillard and degradation reactions of sugars and also of certain enzymatic processes. However, quantitation of these compounds is not straightforward when more abundant carbohydrates are present in real samples. Therefore in this work a GC/MS method was developed to separate monosaccharides, 3-DG and glucosone and applied to analyze them in carbohydrate-rich food products. Difructose anhydrides (DFAs), known markers of sugar degradation, were also determined. The effect of time and temperature in the production and storage of these compounds was also evaluated. RESULTS: Under optimized conditions, good separation between monosaccharides and α-DCs was achieved. Must syrups showed the highest concentrations of 3-DG and glucosone (average values 9.2 and 5.8 mg g(-1) respectively). Coffee substitutes based on carob, chicory and blends showed the highest content of DFAs. Heating and storage assays proved that production of 3-DG was influenced by temperature, while glucosone was more affected by storage time. CONCLUSION: The proposed method allows the rapid quantitation of 3-DG and glucosone along with carbohydrates and DFAs in different food products, which is essential to determine their degradation level. Moreover, the α-DC content in several foods is reported for the first time.