Human milk is rich in bioactive components beyond nutrition, including RNAs—especially miRNAs. It is the most RNA-rich biofluid. While storage effects on other milk components are well studied, RNA preservation has been minimally explored, primarily focusing on miRNAs in exosomes under refrigeration. The goal of this clinical study was to determine optimal collection and storage methods to preserve RNA quantity and quality in human milk, predominantly focusing on RNA stability in research laboratory settings when isolation of milk fat globules is not feasible. The study recruited 30 lactating women in central Arkansas. Milk samples were collected in-clinic and processed immediately for baseline RNA data. Remaining aliquots were stored at 4 °C, -20 °C, or -80 °C with or without RNAlater. RNA was extracted at 4 h, 24 h, 1 week, 4 weeks, 12 weeks, and 24 weeks using Qiagen miRNeasy and MinElute kits. RNA quality (RINe) and concentration (ng/µL) were assessed with Agilent TapeStation with additional quality analysis through qPCR. RNA quality declined over time at all storage temperatures in samples without RNAlater. Addition of RNAlater prevented decreases in RINe scores at most time points, maintaining RINe > 7 for up to 6 months, though it reduced total RNA concentration. qPCR also showed improved mRNA stability with RNAlater. miRNAs were analyzed using Agilent Bioanalyzer Small RNA kits with qPCR for quality analysis. RNAlater was not beneficial for miRNA preservation, and miRNAs remained stable when stored frozen without RNAlater. qPCR verified miRNA stability when frozen. In conclusion, RNA in human milk degrades over time. For mRNA analysis, storage with RNAlater at -80 °C is recommended. For miRNA analysis, storage at -20 °C or -80 °C without preservatives is optimal. These findings offer preliminary guidance for HM RNA storage, and researchers are advised to conduct method testing before full study implementation.

Breastfeeding is widely recognized as the gold standard food for infants. However, many families use infant formulas, including soy-based products, which have not been studied for their long-term safety and developmental effects. The Beginnings Study and the Beginnings Follow-Up Study represent one of the most comprehensive prospective cohorts designed to examine how early infant feeding is related to growth, body composition, cardiovascular, microbiome, and skeletal outcomes, neurodevelopment, and reproductive maturation through adolescence. Conducted in Arkansas, United States, the study enrolled 600 healthy, term infants fed soy-based infant formula, cow milk-based infant formula, or human milk during infancy (200 per group). Of these, 385 participants (73.2%) completed the 6-year visit, and 190 (31.7% of enrolled and 49.4% of 6-year visit completers) participated in the 14-year Follow-Up Study. Breastfeeding was associated with slower weight gain velocity during infancy, and consistent lower BMI, fat mass index, and waist circumference extending into adolescence, than formula feeding. Formula-fed infants had comparable results with breastfed infants for skeletal mineralization, most neurocognitive parameters, and reproductive organ development. However, cardiovascular autonomic measures, including heart rate and vagal tone, differed by feeding group, with some sex-specific effects. Novel contributions included analyses of the gut microbiome and metabolomics profiles in early life, which revealed distinct dietary signatures, as well as neurodevelopmental assessments using electroencephalography, which highlighted transient differences in language-related brain responses among feeding groups. Together, these results demonstrate more similarities than differences between soy-based infant formula and cow milk-based infant formula in health outcomes and support the lasting benefits of breastfeeding. This evidence can help guide health care professionals in infant feeding recommendations and highlight critical windows to prevent obesity and promote lifelong health.

BACKGROUND:Although ∼12% of infants in the United States are fed soy protein-based infant formulas (SF), data on the long-term health effects compared with cow's milk-based infant formulas (MF) and human milk (HM) remain limited. OBJECTIVES:This study aims to assess whether SF feeding during infancy influences adolescent growth, adiposity, dietary intake, metabolic health, or pubertal development compared with MF or HM feeding during infancy. METHODS:In the Beginnings follow-up study, 190 participants (SF = 52, HM = 76, MF = 62) were assessed at age 14 y for anthropometry, body composition, dietary intake, cardiometabolic biomarkers, pubertal stage, and reproductive organ volumes and characteristics. Statistical analyses were performed using R (version 4.4.2), and included generalized linear regression models adjusted for age, sex, race/ethnicity, mother's education, and birth characteristics. RESULTS:At 14.10 ± 0.26 (13.47-14.96) y, adjusted models showed that adolescents who were fed SF as infants had similar body weight, BMI-for-age z-score, fat mass index, abdominal adiposity (visceral and subcutaneous adipose tissue areas), and waist circumference compared with participants fed MF and HM as infants. Energy intake, macronutrient intakes, and diet quality (Healthy Eating Index-2020) were comparable across groups, except for a lower fat intake (%kcal) for participants fed SF and HM as infants compared with MF. There were no significant differences between the groups in blood pressure, glucose metabolism, lipid profile, inflammatory biomarkers, pubertal stage, age of menarche and reproductive organ sizes and characteristics at age 14 y. The prevalence of cardiometabolic risk factors was significantly higher in participants who were fed MF (8.1%) compared with SF (3.8%) and HM (0.0%) as infants. CONCLUSIONS:SF feeding during infancy was associated with similar growth, body composition, cardiometabolic health and puberty outcomes at age 14 y as MF and HM feeding. In adjusted models, HM feeding was protective against adolescent adiposity compared with MF feeding. This study was registered at www. CLINICALTRIALS:gov as NCT03108014.

Latina women have higher rates of lactation initiation compared to other minority women in the United States, however, acculturation to the United States is associated with a gradual shift in breastfeeding exclusivity. This study aims to test the effect and acceptability of an intervention to improve breastfeeding outcomes in a Latina population using a Spanish prenatal education course and bilingual postpartum lactation support for the first six months of life. Participants (N = 42) were randomly assigned to an intervention or standard-of-care group. The intervention group attended a prenatal course taught by a bilingual Certified Lactation Counselor (CLC) and received postpartum lactation support in-person and by tele-lactation. The control group received standard-of-care lactation support from their healthcare providers. Data on breastfeeding successes and challenges were collected up to six months postpartum. Participants receiving the intervention had a significantly higher intention to breastfeed scores than the standard-of-care group within one week after delivery (p = 0.0054). The study’s findings showed good participation and acceptance of change from three to two classes. Lastly, the intervention group had significantly fewer days of formula exposure compared to the control group (p = 0.031). Prenatal and postnatal lactation education classes and support were well accepted by participants as reflected in higher Infant Feeding Intention scores among women in the intervention group compared to those receiving the standard of care. These results lay a foundation for larger studies on the effectiveness of both prenatal and postnatal lactation support interventions in the Latina population.

BACKGROUND AND OBJECTIVE:The objective of this study was to examine the associations of breastfeeding beyond infancy (BBI, >1 year) on toddler growth, development, and diet. METHODS:Participants (n = 185) of a prospective longitudinal study were categorized into 3 groups: formula fed (FF, N = 22), breastfed up to 12 months (BF, N = 108), and breastfed up to 24 months (PBF, N = 55). Data collected included demographics, anthropometrics, toddler Healthy Eating Index (HEI), Child Feeding Questionnaire (CFQ), and Bayley scales of infant and toddler development (BSID). RESULTS:At age 24 months, linear modeling showed PBF children had lower weight-for-length z-scores in comparison to both FF and BF children. FF infants had lower HEI scores for total fruit compared to BF and PBF infants, whereas PBF infants had higher seafood HEI scores. Feeding responsibility was lower in BF parents than FF parents. Restriction was lower in BF and FF groups compared to PBF, and pressure feeding was less common in PBF. Developmental testing revealed no significant differences between the three groups. CONCLUSION:These results strengthen evidence that BBI supports adequate childhood growth, nutritional status, and development. Further studies are needed to evaluate the effect of BBI on health status in childhood. CLINICAL TRIAL ID:#NCT01131117 IMPACT: This research supports the current recommendations to continue breastfeeding up to 24 months. This research indicates that breastfeeding beyond infancy supports adequate growth, nutritional status, and overall development. This article adds to the growing body of literature that breastfeeding beyond 12 months should be supported. This article is one of the first prospective longitudinal studies to investigate breastfeeding beyond infancy.

IntroductionMaternal obesity is associated with increased concentrations of human milk (HM) obesogenic hormones, pro-inflammatory cytokines, and oligosaccharides (HMOs) that have been associated with infant growth and adiposity. The objective of this pilot study was to determine if adherence to a Mediterranean meal plan during lactation modulates macronutrients and bioactive molecules in human milk from mothers with obesity.MethodsSixteen healthy, exclusively breastfeeding women with obesity (body mass index ≥30 kg/m2) enrolled between 4 and 5 months postpartum. The women followed a 4-week Mediterranean meal plan which was provided at no cost. Maternal and infant anthropometrics, HM composition, and infant intakes were measured at enrollment and at weeks 2 and 4 of the intervention. Thirteen mother-infant dyads completed the study. Additionally, participants from an adjacent, observational cohort who had obesity and who collected milk at 5 and 6 months postpartum were compared to this cohort.ResultsParticipants’ healthy eating index scores improved (+27 units, p < 0.001), fat mass index decreased (−4.7%, p < 0.001), and daily energy and fat intake were lower (−423.5 kcal/day, p < 0.001 and-32.7 g/day, p < 0.001, respectively) following the intervention. While HM macronutrient concentrations did not change, HM leptin, total human milk oligosaccharides (HMOs), HMO-bound fucose, Lacto-N-fucopentaose (LNFP)-II, LNFP-III, and difucosyllacto-N-tetrose (DFLNT) concentrations were lower following the intervention. Infant intakes of leptin, tumor necrosis factor (TNF)-α, total HMOs, HMO-bound fucose, LNFP-III and DFLNT were lower following the intervention. Specific components of the maternal diet (protein and fat) and specific measures of maternal diet quality (protein, dairy, greens and beans, fruit and vegetables) were associated with infant intakes and growth.DiscussionAdherence to a Mediterranean meal plan increases dietary quality while reducing total fat and caloric intake. In effect, body composition in women with obesity improved, HM composition and infants’ intakes were modulated. These findings provide, for the first time, evidence-based data that enhancing maternal dietary quality during lactation may promote both maternal and child health. Longer intervention studies examining the impact of maternal diet quality on HM composition, infant growth, and infant development are warranted.

SummaryIntroductionClustering of cardiometabolic risk factors in childhood significantly increases the risk of atherosclerotic cardiovascular disease later in life. Identification of modifiable parental factors that contribute to offspring cardiometabolic health is critical for the prevention of disease. The objective was to identify factors associated with child cardiometabolic risk factors at age 5 years.MethodsTriads from a longitudinal cohort were recalled at 5 years (n = 68). Dietary intake, anthropometrics, physical activity and serum‐based risk factors were collected. Best subset selection, linear and logistic regressions were used to identify triad variables associated with increased risk of cardiometabolic risk factor clustering at age 5 years.ResultsIn this cohort, best subset modelling revealed that increased paternal fat mass, serum low‐density lipoproteins and triglycerides, maternal dietary added sugar and being female were associated with increased odds of offspring having two or more cardiometabolic risk factors at age 5 years.ConclusionsDietary and exercise interventions prior to conception targeting paternal adiposity and dyslipidaemia as well as maternal dietary habits could decrease children's cardiometabolic risk in later life.

Background: Breastfeeding is widely recognized as the optimal feeding method for infants. However, breastfeeding goals are often unmet, especially in mothers with excessive weight. Potential factors associated with unmet goals could be disparities in care for women with higher body mass index (BMI) or mental health symptomology. Methods: Women enrolled in a longitudinal study were stratified by BMI into three groups: mothers with normal weight (18.5-24.9 kg/m2, n = 101), with overweight (25-29.9 kg/m2, n = 78), and with obesity (OB; 30-35 kg/m2, n = 48). Breastfeeding intention and standardized mental health questionnaires were administered at gestational weeks 12 and 36. The prevalence of initiation and duration of breastfeeding were determined based on self-reported breastfeeding start and end dates. Wilcoxon tests, pairwise proportion test, Cox proportional hazards regression, and linear regression were used. Results: Higher maternal weight status (OB) was significantly associated with lower breastfeeding intention and duration. As expected, higher breastfeeding intention scores were associated with significantly longer breastfeeding duration. Higher scores on the Beck Depression Inventory (BDI), associated with a greater number of depression symptoms, mediated the negative impact of weight status on breastfeeding intention. Conclusions: breastfeeding outcomes are negatively associated with maternal weight status and prenatal mental health with the relationship between the two being interconnected, despite subclinical scores on the BDI. Further research is needed to explore the role of mental health on breastfeeding outcomes. From these findings, targeted prenatal interventions for women with excessive weight and depressive symptoms would likely promote and improve breastfeeding outcomes. ClinicalTrials.gov: www.clinicaltrials.gov, ID #NCT01131117.

Breastfeeding MedicineVol. 18, No. 5 CorrespondenceResponse to Spatz and Juntereal re: “Education and Experiences of Antenatal Breast Milk Expression: A Systematic Review”Sarah Sobik, Lindsay Blake, and Aline AndresSarah SobikAddress correspondence to: Sarah Sobik, MD, MPH, FAAP, IBCLC, 1 Children's Way, Slot 512-8, Little Rock, AR 72202, USA E-mail Address: [email protected]https://orcid.org/0000-0003-0140-478XDepartment of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, Arkansas, USA.Arkansas Children's Nutrition Center, Little Rock, Arkansas, USA.Search for more papers by this author, Lindsay Blakehttps://orcid.org/0000-0002-8234-8611UAMS Medical Library, University of Arkansas for Medical Sciences, Little Rock, Arkansas, USA. Search for more papers by this author, and Aline AndresDepartment of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, Arkansas, USA.Arkansas Children's Nutrition Center, Little Rock, Arkansas, USA.Search for more papers by this authorPublished Online:17 May 2023https://doi.org/10.1089/bfm.2023.0076AboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View article"Response to Spatz and Juntereal re: “Education and Experiences of Antenatal Breast Milk Expression: A Systematic Review”." Breastfeeding Medicine, 18(5), pp. 405–406FiguresReferencesRelatedDetails Volume 18Issue 5May 2023 InformationCopyright 2023, Mary Ann Liebert, Inc., publishersTo cite this article:Sarah Sobik, Lindsay Blake, and Aline Andres.Response to Spatz and Juntereal re: “Education and Experiences of Antenatal Breast Milk Expression: A Systematic Review”.Breastfeeding Medicine.May 2023.405-406.http://doi.org/10.1089/bfm.2023.0076Published in Volume: 18 Issue 5: May 17, 2023PDF download

Background: Breastfeeding rates have stagnated recently despite recommendations to breastfeed until age 2 years. Antenatal breast milk expression (ABME) is a method used to prepare the breast for breastfeeding. However, there is limited evidence available on the benefits, risks, and impact of ABME on maternal-infant breastfeeding dyads. Methods: This review identified and summarized studies on women who engaged in ABME and their personal experiences. Databases searched included PubMed MEDLINE, Web of Science, Cumulative Index of Nursing and Allied Health Literature (CINAHL), and EMBASE. Initially, abstracts and titles were reviewed, and then, full-text studies were screened for inclusion by two blinded authors. Two authors assessed the quality of the studies using a standardized tool, two authors completed data extraction, and one author completed data harmonization into tables. Results: A total of 1,410 studies were identified (after duplicates removed) and 10 citations qualified for the inclusion criteria. Only two studies received an overall rating of strong quality and low-risk bias. The selected articles varied in primary outcomes; however, main focuses were experiences, knowledge, and perspective after practicing ABME. Data varied on timing of ABME, but most studies started between 34 and 36 weeks. The average amount of expressed milk was reported in four studies but was variable. Conclusions: This systematic review found that the literature is limited regarding ABME, and most studies were focused on women with diabetes. The current limited evidence suggests that ABME may be a helpful tool in improving maternal breastfeeding confidence and breastfeeding outcomes. Negative side effects reported related to ABME included difficulty learning the technique, discomfort, and feeling of awkwardness while expressing. Future research should focus on higher quality studies regarding use of ABME, proper teaching of ABME technique, and the use of ABME to improve breastfeeding outcomes in diverse populations of maternal-infant dyads.

This study determined the relationship between physical activity (PA), circulating lipids throughout pregnancy and infant anthropometric outcomes at birth and 2 weeks of age. Women (N = 234) with normal weight (NW, BMI 18.5–24.9 kg/m2) and with overweight and class I obesity (OW/OB, BMI 25–35 kg/m2) were categorized into high and low PA based on average cohort steps during pregnancy (8099 steps/day). Circulating fasting lipids were measured at each trimester. Standardized methods were used to obtain anthropometrics measures. Infant body composition was estimated by quantitative nuclear magnetic resonance (EchoMRI-AH small; ECHO Medical Systems). Women with NW who had higher activity had lower circulating triglycerides (TG) and total cholesterol (TC) levels at 12 weeks compared to women with NW and low activity (p < 0.05). Women with OW/OB and high activity level throughout pregnancy had lower circulating TG, and low density lipoprotein (LDL), at 12 weeks, lower LDL at 24 weeks, and lower TG at 36 weeks compared to the women with OW/OB who had low activity levels (p < 0.05). For children born to women with OW/OB, maternal circulating TG and LDL were most associated with infant anthropometrics at 2 weeks of age. This study supports that higher PA during pregnancy is associated with lower lipid levels throughout pregnancy with a greater effect size in women with OW/OB. Maternal lipids were associated with anthropometrics and infant body composition at two weeks of life in women with OW/OB.

ObjectiveThis study investigated which antenatal and postnatal factors determine offspring adiposity during the first 2 years of life.MethodsParticipants were mother and child pairs (N = 224). Offspring percent fat mass (%FM) was obtained using quantitative nuclear magnetic resonance at 11 time points between ages 0.5 and 24 months. Independent variables included race, age, gestational weight gain, first‐trimester %FM, delivery mode, gestational measures of resting energy expenditure, respiratory exchange ratio, physical activity, serum cytokines and lipids, and dietary intake for the mothers, as well as sex, birth weight and length, breastfeeding duration, and physical activity at age 2 years for the children. Linear mixed models were used to construct the best‐fitted models for the entire cohort and for each sex.ResultsMaternal %FM (P = 0.006), high‐density lipoprotein (HDL) (P < 0.001), and breastfeeding duration (P = 0.023) were positively associated with female offspring adiposity, whereas maternal dietary fiber intake (P = 0.016) had a negative association. Birth weight (P = 0.004), maternal HDL (P = 0.013), and breastfeeding duration (P = 0.015) were all positively associated with male offspring adiposity.ConclusionsAntenatal and postnatal factors differentially impact male and female offspring adiposity during the first 2 years of life.

Background This study longitudinally characterized the developmental status, growth, and body composition of children who were fed human milk (breastfed, BF), cow’s milk-based (MF), or soy protein-based (SF) infant formula from 3 to 12 months. Methods Standardized anthropometrics and dual-energy X-ray absorptiometry were used to characterize growth and body composition at 3, 6, 9, 12, 24, 36, 48, 60, and 72 months (NCT00616395). Preschool Language Scale-3, Children’s Memory Scale Index (CMS), and Wechsler Preschool and Primary Scale of Intelligence were administered at age 72 months. Mixed-effects models adjusting for gestational age, birth weight, child race and sex, parental education, and maternal IQ were performed. Results Body Mass index (BMI) was significantly lower between 24 and 72 months in BF children compared to SF children. At 3 and 6 months, BF infants had significantly higher fat mass (FM) than SF infants, whereas BF children had significantly lower FM at 36 and 48 months than SF children. Delayed Recognition Index of the CMS was higher for SF than for MF participants ( p = 0.009). There was no other significant difference in developmental outcomes between groups. Conclusions In conclusion, BF, MF, and SF support adequate growth and development up to age 6 years. Impact Although soy protein-based infant formula is reported to support normal infant growth and development compared to cow’s milk-based formula and human milk, there are limited data on the effect of these feeding methods in school-aged children. This study suggests a significant difference in body composition, specifically BMI, after 24 months between infant feeding methods during the first year of life and in early childhood; however, all diets provide adequate nutrients to maintain normal development up to 72 months.

Summary Background Maternal obesity increases offspring's obesity risk. However, studies have not often considered maternal metabolic and exercise patterns as well as paternal adiposity as potential covariates. Objective To assess the relationship between parental and newborn adiposity. Methods Participants were mother‐child pairs (n = 209) and mother‐father‐offspring triads (n = 136). Parental (during gestation) and offspring (2 weeks old) percent fat mass (FM) were obtained using air displacement plethysmography. Maternal race, age, resting energy expenditure (indirect calorimetry), physical activity (accelerometry), gestational weight gain (GWG), gestational age (GA), delivery mode, infant's sex and infant feeding method were incorporated in multiple linear regression analyses. The association between parental FM and offspring insulin‐like growth factor 1 (IGF‐1) was assessed at age 2 years. Results Maternal adiposity was positively‐associated with male (β = 0.11, P = .015) and female (β = 0.13, P = .008) infant FM, whereas paternal adiposity was negatively‐associated with male newborn adiposity (β = ‐0.09, P = .014). Breastfeeding, female sex, GA and GWG positively associated with newborn adiposity. Vaginal and C‐section delivery methods associated with greater adiposity than vaginal induced delivery method. Plasma IGF‐1 of 2‐year‐old boys and girls positively associated with their respective fathers' and mothers' FM. Conclusions Maternal and paternal adiposity differentially associate with newborn adiposity. The mechanisms of this finding remain to be determined.

Clinical depression has been previously associated with lower birth weight and preterm deliveries. Subclinical depression levels may also influence birth outcomes in at risk populations. This study...