Polyphenol analysis in human milk by a rapid, cost-effective, and accurate micromethod: translational development.

OBJECTIVE: To develop a rapid method for analysing polyphenols, which are potentially active antioxidants against neonatal oxidative stress, from small human milk (HM) volumes. METHODS: Acid and alkaline extractions were compared using two dyes: Folin-Ciocalteu and Fast Blue BB. Linearity, sensitivity, recovery percentage, polyphenol content, precision, and stability were assessed in 14 HM samples and compared using the Kruskal-Wallis H test (p<0.05). The best technique was applied to 284 HM samples to determine their polyphenolic content and its association with maternal diet by multifactorial linear regression. RESULTS: Acidic extraction successfully recovered the gallic acid reference standard, whereas alkaline extraction overestimated it. Calibration curves for all methods were linear (R2>0.96) up to 500 mg/L. All bicarbonate-based Folin-Ciocalteu methods assayed were stable and repeatable, whereas Fast Blue BB-based variants were not. HM polyphenols (mean=94.68 mg/L) positively correlated to the dietary intake of hydroxycinnamic acids, the most consumed polyphenolic family in this population. CONCLUSIONS: A bicarbonate-based Folin-Ciocalteu micromethod allowed the accurate determination of polyphenols in HM, which might be useful for translational research settings and HM banks.

Polyphenol analysis in human milk by a rapid, cost-effective, and accurate micromethod: translational development / Análise de polifenóis no leite humano por meio de um micrométodo rápido, econômico e preciso: desenvolvimento translacional

ABSTRACT Objective: To develop a rapid method for analysing polyphenols, which are potentially active antioxidants against neonatal oxidative stress, from small human milk (HM) volumes. Methods: Acid and alkaline extractions were compared using two dyes: Folin-Ciocalteu and Fast Blue BB. Linearity, sensitivity, recovery percentage, polyphenol content, precision, and stability were assessed in 14 HM samples and compared using the Kruskal-Wallis H test (p<0.05). The best technique was applied to 284 HM samples to determine their polyphenolic content and its association with maternal diet by multifactorial linear regression. Results: Acidic extraction successfully recovered the gallic acid reference standard, whereas alkaline extraction overestimated it. Calibration curves for all methods were linear (R2>0.96) up to 500 mg/L. All bicarbonate-based Folin-Ciocalteu methods assayed were stable and repeatable, whereas Fast Blue BB-based variants were not. HM polyphenols (mean=94.68 mg/L) positively correlated to the dietary intake of hydroxycinnamic acids, the most consumed polyphenolic family in this population. Conclusions: A bicarbonate-based Folin-Ciocalteu micromethod allowed the accurate determination of polyphenols in HM, which might be useful for translational research settings and HM banks.

RESUMO Objetivo: Desenvolver um método rápido para analisar polifenóis, que são antioxidantes potencialmente ativos contra o estresse oxidativo neonatal, em pequenos volumes de leite humano (LH). Métodos: Foram comparadas extrações ácidas e alcalinas usando dois corantes: Folin-Ciocalteu e Fast Blue BB. Foram avaliadas variáveis como linearidade, sensibilidade, percentagem de recuperação, teor de polifenóis, precisão e estabilidade em 14 amostras de LH, comparadas usando o teste de Kruskal-Wallis H (p<0,05). A melhor técnica foi aplicada a 284 amostras de LH para determinar seu teor polifenólico e sua associação com a dieta materna por regressão linear multifatorial. Resultados: A extração ácida recuperou com sucesso o padrão de referência do ácido gálico, enquanto a extração alcalina o superestimou. As curvas de calibração para todos os métodos foram lineares (R2>0,96) até os 500 mg/L. Todos os métodos testados baseados em Folin-Ciocalteu com bicarbonato foram estáveis e repetíveis, enquanto as variantes baseadas em Fast Blue BB não. Os polifenóis do HM (média=94,68 mg/L) correlacionaram-se positivamente com a ingestão dietética de ácidos hidroxicinâmicos, a família de polifenóis mais consumida nesta população. Conclusões: Um micrométodo baseado em bicarbonato de Folin-Ciocalteu permitiu a determinação precisa de polifenóis no HM, o que pode ser útil para configurações de pesquisa translacional e bancos de HM.

Caffeinated non-alcoholic beverages on the postpartum mental health related to the COVID-19 pandemic by a cross-sectional study in Argentina.

Purpose: This work aimed to study postpartum mental outcomes and determinants of the intake of caffeinated beverages during the pandemic in women from Argentina. Methods: This cross-sectional study recruited 619 women who responded to online self-report questionnaires during the first and second waves of COVID-19, including validated instruments (Insomnia Severity Index, Perceived Stress Scale, Postpartum Depression Screening Scale, Memory Complaint Scale, and Breastfeeding Self-Efficacy Scale), and general data. Intake frequency and amount of caffeinated beverages were estimated. Multivariate regression and structural equation models identified associations and effects (p < 0.05). Results: Women were under social restrictions for 60.39 days, with home and essential activities increasing caffeinated intake. They ingested (mL/d): yerba mate (1457.71), coffee (66.85), tea (67.61), and soft drinks (50.95), which provided 646.20 mg/d of caffeine. Intakes of coffee and yerba mate were higher than pre-pandemic ones. Coffee was positively associated with stress and insomnia, and indirectly linked to higher levels of depression and memory complaints, and lower breastfeeding self-efficacy. Tea showed a similar but weaker association. Yerba mate correlated inversely with depression (through direct pathways), insomnia, and memory complaints (through indirect pathways), promoting breastfeeding self-efficacy. Soft drinks and caffeine did not present significant associations. Conclusion: Although findings do not imply causation, results suggest that beverages would exhibit caffeine-independent affective and cognitive roles, which might be anxiogenic in the case of coffee and tea (to a lesser extent). Yerba mate showed antidepressant potential. Given that breastfeeding might be compromised during the pandemic, yerba mate intake is promissory to protect postpartum mental health.

Three food pattern-based indices diagnose lactating women's nutritional inadequacies in Argentina: A clinimetric approach using diet quality indicators and breast milk biomarkers.

Diet is a health foundation that supports breastfeeding and enables lactating women's recovery. This study aimed to develop 3 valid diet adherence indices by clinimetric contrast with different parameters. We hypothesize that the 3 diet adherence-based indices are positively correlated with diet quality and milk biomarkers and directly related to calcium, fiber, and energy intake. A cross-sectional study recorded food frequency, anthropometric and dietary measures, and milk analysis to estimate correlations, sensitivity, and specificity (n = 284). Three reliable indices agreed by >75% with food frequency: macronutritional (MDI), phytochemical (PDI), and energetic (EDI), which correlated positively to the Fat Quality Index and Minimum Dietary Diversity for Women. MDI and EDI correlated negatively to the Healthy Plant-Based Diet Index (H-PBDI), whereas PDI correlated positively. MDI correlated positively to the protein-to-carbohydrate ratio, whereas EDI correlated negatively. Inadequate intakes of calcium, fiber, and energy were indicated by MDI <19, PDI <19, and EDI ≥19, respectively, in a sensitive and specific manner. Body fat was therefore positively associated with EDI. MDI was associated with increased protein and superoxide anion in milk, depending on its high protein-to-carbohydrate ratio and low H-PBDI. These indices allowed us to assess diet to diagnose maternal malnutrition promptly. Index validity was supported by significant dietary correlations and biological responsiveness. MDI and EDI promoted breast bioactivity and body adiposity, respectively, whereas PDI represented a healthier option.

[Yerba mate as a safe polyphenolic source during human lactation]. / La yerba mate como fuente segura de polifenoles durante la lactancia humana.

Background: Yerba mate intake is conditioned by assumptions about its benefits and risks for breastfeeding, with a lack of solid evidence. Objective: To evaluate the safety of yerba mate teas during human lactation. Material and methods: Cross-sectional study with 279 adult lactating women from Argentina (years 2013-2020), during the first six months postpartum, without current pregnancy or active pathologies. Social, reproductive, and food data were compiled. Body mass index and percentage of body fat were also established. The adherence to dietary patterns, intake of yerba mate and its phytochemicals were estimated. Milk biochemistry included nutritional, oxidative and tissue markers. Intake means were compared using the Wilcoxon's test depending on the data recorded, while the multiple nonparametric Kernel regression was used to evaluate the effects on body and milk. Results: It was found a wide range of intake, and tube-sipped tea was the main way of consumption. Concerning its constraints, in-cup tea was more used by women without a partner, whereas tube-sipped tea was consumed mainly by employed women, during late postpartum, and with 12 or more years of formal education. Additionally, these teas were an important source of 18 hydroxycinnamic acids and flavonoids. Moreover, yerba mate provided more than 35% of dietary polyphenols. No anthropometric effects or changes in milk nutrients and biomarkers were found. Conclusions: Yerba mate was a safe source of polyphenols, as it did not compromise lactation and maternal nutritional status.

Introducción: el consumo de yerba mate está condicionado por supuestos sobre sus beneficios y riesgos para la lactancia materna, los cuales carecen de evidencia sólida. Objetivo: evaluar la seguridad de las infusiones de yerba mate durante la lactancia. Material y métodos: estudio transversal con 279 mujeres lactantes adultas de Argentina (años 2013-2020), en los primeros seis meses postparto, sin embarazo ni patologías activas. Se recopilaron datos sociales, reproductivos y alimentarios, índice de masa corporal y porcentaje de grasa corporal. Se estimó adherencia a patrones alimentarios, consumo de mate y sus fitoquímicos. Se analizaron marcadores nutricionales, oxidativos y tisulares en leche. Por la prueba de Wilcoxon, se compararon las medias de consumo según las variables recopiladas y por regresión no paramétrica de Kernel múltiple se evaluaron los efectos corporales y lácteos. Resultados: se encontró un amplio rango en la ingesta de infusiones y el mate cebado fue la principal forma consumida. Respecto a sus condicionantes, el mate cocido fue más bebido por mujeres sin pareja, mientras que el mate cebado fue más consumido por puérperas tardías, con empleo y nivel educativo ≥ 12 años. Además, estas infusiones constituyeron una fuente relevante de 18 ácidos hidroxicinámicos y flavonoides, que proveyeron diariamente más del 35% de los polifenoles dietarios. No se encontraron efectos corporales ni cambios en los niveles de los nutrientes y en los biomarcadores lácteos. Conclusiones: la yerba mate fue una fuente polifenólica segura que no comprometió la lactancia ni el estado nutricional materno.

Dietary Intake of Polyphenols Enhances Executive/Attentional Functioning and Memory with an Improvement of the Milk Lipid Profile of Postpartum Women from Argentina.

Puerperium may lead to memory and executive/attentional complaints that interfere with women's daily life. This might be prevented by dietary compounds, such as neuroprotective polyphenols. Their bioactivity depends on their effects on lipid metabolism in different tissues, such as the brain, fat, and breast. Thus, a polyphenol-related cognitive improvement may be associated with changes of lipids in human milk, which are key for infant neurodevelopment. A cross-sectional study was conducted on 75 postpartum women from Córdoba (Argentina), involving several neuropsychological tests. Diet was registered to identify polyphenol intake and food pattern adherence, with sociodemographic and other psychological variables (insomnia, stress, subjective cognitive complaints) being also studied. Triacylglycerols, cholesterol, and their oxidative forms were analyzed as milk biomarkers. Multivariate statistical methods were applied. Results confirmed that women who consumed polyphenols presented better executive/attentional performance (i.e., higher correct responses, conceptual level responses, complete categories, verbal fluency; lower attentional interferences, and perseverative errors) and word retention with lower interference. Polyphenols were positively associated with milk lipids, which were higher in women with better cognition. Furthermore, they had lower oxidized triacylglycerols. In conclusion, polyphenolic intake during postpartum may improve executive/attentional functioning, memory, and milk lipid profile.

Association of Dietary Intake of Polyphenols with an Adequate Nutritional Profile in Postpartum Women from Argentina.

HJ-Biplot analysis is a multivariate graphic representation that collects data covariation structure between variables and individuals to represent them in a low-dimensional space with the highest quality in the same reference system. Consequently, it is a promising technique for evaluating dietary exposure to polyphenols and accurately characterizing female nutrition. Herein, we hypothesized that polyphenol intake defines specific clusters with dietary impacts, which can be assessed using HJ-Biplot, based on a cross-sectional study in Argentina. The study included 275 healthy postpartum women who provided information about their food frequency intake and other conditions, which were then used to evaluate polyphenolic intake using the Phenol-Explorer database. Outcomes were established using HJ-Biplot for clustering and ANOVA to compare their impact on diet quality indicators. Two HJ-Biplot models were run (for intakes >20 mg/d and 5∼20 mg/d, respectively) to identify three clusters per model with excellent statistical fitness to explain the data. Thus, specific polyphenolic clusters with potentially bioactive and safe compounds were defined despite significant interindividual variability. In fact, women with the lowest polyphenolic intake exhibited worse dietary quality, body fat, and physical activity. As a result, HJ-Biplot proved to be an effective technique for clustering women based on their dietary intake of these compounds. Furthermore, cluster membership improved the intake of antioxidants, water, fiber, and healthy fats. Additionally, women with formal jobs and a higher educational level showed a better diet. Dietary polyphenols are critical during postpartum because they exert beneficial effects on women and breastfed infants.

Insomnia is associated with worry, cognitive avoidance and low academic engagement in Argentinian university students during the COVID-19 social isolation.

An increment in mental health problems related to social isolation strategies was evinced in several populations, however few investigations address this problem in students. This study aimed to analyze the relationship between insomnia, worry, coping strategies, and academic engagement in subjects pursuing a university degree in Argentina during the first month of compulsory social isolation against the COVID-19 pandemic. Self-report online questionnaires were administered to 584 participants: Insomnia Severity Index, Penn State Worry Questionnaire, Coping Responses Inventory, Utrecht Work Engagement Scale for Students. Demographic data was recorded. Statistics included bivariate and multivariate techniques. Prevalences of 45% for subthreshold insomnia, 23% for moderate insomnia, and 4% for severe insomnia were found. The severe insomnia group presented higher levels of worry and cognitive avoidance strategies. Insomnia was also associated with daytime concerns. The group without insomnia was more vigorous in their academic engagement. Being physically active was associated with lower sleep problems. Insomnia prevalence is high in socially isolated university students. Cognitive avoidance coping strategies are associated with worry, which leads to an alert state that makes sleep difficult. These findings should be taken into account by academic institutions to define their pedagogic strategies.

Triggering of postpartum depression and insomnia with cognitive impairment in Argentinian women during the pandemic COVID-19 social isolation in relation to reproductive and health factors.

OBJECTIVE: The 2019 coronavirus disease pandemic (COVID-19) required strict confinement measures that differentially impacted the individual's daily life. Thus, this work aimed to study postpartum women's mental health in Argentina during mandatory social isolation. DESIGN: A cross-sectional survey was conducted from May to July 2020, which included five validated questionnaires to assess postpartum depression (Postpartum Depression Screening Scale-Short Form), insomnia (Insomnia Severity Index), memory complaints (Memory Complaint Scale), metacognition (Brief Metamemory and Metaconcentration Scale), and breastfeeding self-efficacy (Breastfeeding Self-Efficacy Scale-Short Form). Sociodemographic variables, social isolation characteristics, and breastfeeding practices were also collected. This study was conducted in accordance with the Declaration of Helsinki. Statistical analysis included zero-order correlations, multiple logistic regressions, and a set of structural equation models (SEM) to test direct and indirect effects. Goodness-of-fit indices were calculated for SEM. SETTING: Postpartum women were recruited from public hospitals, private health clinics, and online community recruitment in the Cordoba province (Argentina). PARTICIPANTS: 305 postpartum women from Argentina. MEASUREMENTS AND FINDINGS: 37% of women reported postpartum depression, 46% insomnia, 42% memory impairment, 60% low metaconcentration, 50% low metamemory, and 23% low breastfeeding efficacy. Also, significant associations were found demonstrating that social isolation promoted postpartum depression and insomnia were reciprocally related, which compromised female cognition and efficacy. This situation was aggravated in women during late postpartum, with previous children, and by low social support (e.g., family, health professionals), with non-exclusive breastfeeding being increased. KEY CONCLUSIONS: This is the first study addressing postpartum women's mental status during social isolation in Argentina, which was a promoting factor for postpartum depression and insomnia that were reciprocally related. This situation was also aggravated by reproductive factors, such as late postpartum, multiparity, breastfeeding frequency, and non-exclusive breastfeeding. Additionally, breastfeeding self-efficacy depended on mental health status, and euthymia therefore favoured the practice of exclusive breastfeeding.

[Food patterns in Argentinian women related to socioeconomic and health factors during puerperium]. / Patrones alimentarios de mujeres argentinas durante el puerperio en relación con factores socioeconómicos y sanitarios.

Introduction: Food choices and nutritional requirements of women, which vary throughout their life cycle, are unknown for Argentinian women. Objective: To identify Argentinian women's dietary patterns and their relation to socioeconomic and health characteristics, during puerperium. Method: The food intake of 249 puerperal women was assessed for a cross-sectional study. Dietary patterns were determined by factor analysis, whose adherence was associated with different variables through Spearman's correlation and multivariate analyses. Results: Three patterns were revealed: macronutritional (MP), phytochemical (PP) and energetic (EP). Physical activity and family history of cardiometabolic diseases were positively associated to MP adherence (odds ratio [OR] = 2.69 and 2.68, respectively). Women without formal employment adhered less to PP (OR = 0.49). Low educational level increased adherence to EP (OR = 2.20). Body mass index correlated inversely to PP (rho = −0.16). Correspondence analysis revealed that high adherence to MP and PP were associated with formal employment, high educational level, adequate physical activity, normal body mass index and a diverse diet. Conclusion: Food choices were conditioned by family disease, physical activity, employment, and educational level. We identified three dietary patterns in postpartum women, which constitute an original contribution to future nutritional approaches.

Introducción: Las elecciones alimentarias afectan la salud de las mujeres, cuyos requerimientos nutricionales varían durante el ciclo vital. No obstante, en Argentina se desconocen los alimentos elegidos por las mujeres durante el puerperio. Objetivo: Identificar los patrones alimentarios de mujeres argentinas durante el puerperio y su relación con las características socioeconómicas y sanitarias. Método: Se realizó un estudio transversal con 249 participantes, quienes respondieron sobre su frecuencia alimentaria. Se determinaron patrones por análisis factorial, cuya adherencia fue asociada a diferentes variables mediante correlación de Spearman y análisis multivariados. Resultados: El análisis reveló tres patrones: macronutricional (PM), fitoquímico (PF) y energético (PE). Tener antecedentes familiares de enfermedades cardiometabólicas y realizar actividad física se asociaron con mayor adherencia al PM (razón de momios [RM] = 2.69 y 2.68, respectivamente). Las mujeres sin empleo formal tuvieron menor adherencia al PF (RM = 0.49). Un bajo nivel educacional incrementó la adherencia al PE (RM = 2.20). El índice de masa corporal (IMC) se asoció inversamente al PF (rho = −0.16). El análisis de correspondencia reveló que la alta adherencia al PM y al PF, junto con una baja adherencia al PE, se asociaron con empleo formal, mayor nivel educativo, adecuada actividad física, lMC normal y una dieta diversa. Conclusiones: Las elecciones alimentarias estuvieron condicionadas por los antecedentes familiares patológicos, la actividad física, el empleo y el nivel educacional, identificándose tres patrones alimentarios propios de las mujeres, lo que constituye una contribución original para futuros abordajes nutricionales.

Positive correlation between proteasome activity and polyphenols in the telencephalon of adult female mice.

INTRODUCTION: Proteasome regulates proteostasis, and can be compromised in neurodegenerative diseases. Thus, our aim was to correlate the activity of proteasome to the level of polyphenols in telencephalon during murine adulthood. METHODS: Proteasome activity, polyphenols and other variables (glucose and hydroperoxides) were analysed in Balb/c female telencephala (n = 20, age = 4-12 months), using multivariate methods. RESULTS: The following values were found: proteasome activity = 3.1 ± 0.6 FI/µg of tissue proteins, glucose = 0.1 ± 0.0 µg/µg, hydroperoxides = 363.4 ± 96.6 OD/µg, and polyphenols = 0.1 ± 0.0 ng/µg. Polyphenols reduced during aging showed a direct correlation with proteasome (Pearson's coefficient = 0.43, p = 0.0590, and a multivariate linear regressive coefficient = 17.85, p = 0.0216), with glucose and hydroperoxides being not involved (p>0.1). This correlation was confirmed by partial least square regression (beta = 0.67). CONCLUSION: Proteasome activity can be affected during ageing, and promoted by telencephalic polyphenol levels. Thus, these diet compounds might exert benefits in adult brain.

The Effect of Freeze-Drying on the Nutrient, Polyphenol, and Oxidant Levels of Breast Milk.

OBJECTIVES: Human milk banks need to extend the suitability of milk for breastfeeding, and for this technological advances are required. Our aim was to establish the capacity of freeze-drying to conserve milk properties without further oxidative deterioration. METHODS: One hundred sixteen healthy women participated from the city of Cordoba (Argentina). Proteins, glucose, triglycerides, polyphenols, and markers (nitrites, superoxide anion, hydroperoxides, lipoperoxides, and γ-glutamyl transpeptidase) were measured in their fresh milk. Samples were then separated for three treatments as follows: freezing and conservation for 6 months at -80°C (F: positive control); freeze-drying for 24 hours at ≤-70°C and ≤1.33 Pa and conservation for 6 months at 4°C (FD: treatment of interest); and freeze-drying for 24 hours at ≤-70°C and ≤1.33 Pa and conservation for 6 months at -80°C (FD+F). Next, analyses were repeated and compared by ANOVA and Tukey tests. RESULTS: Fresh milk showed these values per L as follows: proteins: 12.62 ± 2.51 g, glucose: 4.44 ± 0.25 g, triglycerides: 34.26 ± 0.59 g, polyphenols: 53.27 ± 8.67 mg, nitrites: 62.40 ± 19.09 mg, superoxide: 3,721.02 ± 198.80 OD, hydroperoxides: 7,343.76 ± 294.53 OD, lipoperoxides: 7,349.72 ± 398.72 OD, and γ-glutamyl transpeptidase: 4.66 ± 0.55 IU. Glucose was decreased after F treatment (p < 0.05), all variables were conserved by FD and were not improved by the FD + F combination. CONCLUSIONS: Freeze-drying achieved suitable conservation and may improve bank functioning, by protecting nutritional properties, polyphenol-related functionality, and oxidative integrity of human milk through a 1-day treatment with easy maintenance.