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Background: Devices measuring the macronutrient content of human milk are commonly used to assist with clinical decision-making. It is unknown if these devices accurately measure protein content in donor human milk (DHM). Our objective is to quantify the nitrogen sources and protein content in commercial DHM., Methods: The total nitrogen content (Dumas method) and nonprotein nitrogen content (Kjeldahl method) was measured in triplicate from six commercial DHM samples with protein content noted on the labels. In addition, the amino acid content was measured in 15 commercial DHM samples and protein content in each sample was calculated. The calculated protein content for each DHM sample was compared for consistency., Results: The nonprotein nitrogen content in DHM was consistently higher (0.33 ± 0.05 g/g) than previous reports, leading to overreporting of protein content on DHM labels by a median value of 0.15 g/dl (range 0.02-0.23 g/dl). Similarly, calculation of the protein content from the total nitrogen content with an assumption of 20% (grams per gram) nonprotein nitrogen consistently overrepresented the protein content as determined from the amino acid profile for DHM., Conclusion: Common methods for assessing the macronutrient content of human milk may overestimate the protein content of DHM., (© 2024 American Society for Parenteral and Enteral Nutrition.)

Preterm birth disrupts the natural progression of events in the parent-infant relationship and bestows many of the typical parent responsibilities to the clinical care team. In turn, the neonatal intensive care environment (NICU) introduces obstacles to parents that would not otherwise be encountered and forces parents to adapt to this artificial environment as they seek to bond with and care for their newborn. Facilitating parent presence at the bedside and incorporating them into the care of their preterm infant is critical for lessening the immediate burden to both the parent and offspring while also ensuring the best possible outcome for preterm infants. In this review, we explore the impact that parents exert on the neurodevelopmental outcome of preterm infants and identify several barriers and facilitators to parent presence., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to the International Pediatric Research Foundation, Inc.)

Objective: Double-blind randomized control trial of early addition of a bovine milk-derived human milk fortifier (HMF) in very low birth weight (VLBW) infants (NCT05228535)., Methods: VLBW infants were randomized to receive bovine milk-derived HMF with first feedings or delayed fortification at 80 ml/kg/day. Anthropometrics were assessed weekly through 36 weeks postmenstrual age (PMA). Unadjusted and adjusted (race, gender, gestational age, and birth weight) differences between study arms were examined using two-sample t-test and ANCOVA, respectively., Results: Fifty-two VLBW infants (57% female, 60% Black) were enrolled. Baseline demographics did not differ between groups. Weight velocity at DOL 28 did not differ between study arms. Secondary outcomes including NPO occurrence, incidence of metabolic acidosis, NEC, retinopathy, or late-onset sepsis did not differ between groups., Conclusion: Immediate fortification of enteral feedings with a bovine milk-derived HMF appears safe and well-tolerated although no clear growth benefit could be established., (© 2024. The Author(s).)

Infection of lung endothelial cells with pneumococci activates the superoxide-generating enzyme NADPH oxidase 2 (NOX2), involving the pneumococcal virulence factor pneumolysin (PLY). Excessive NOX2 activity disturbs capillary barriers, but its global inhibition can impair bactericidal phagocyte activity during pneumococcal pneumonia. Depletion of the α subunit of the epithelial sodium channel (ENaC) in pulmonary endothelial cells increases expression and PMA-induced activity of NOX2. Direct ENaC activation by TIP peptide improves capillary barrier function -measured by electrical cell substrate impedance sensing in endothelial monolayers and by Evans Blue Dye incorporation in mouse lungs- following infection with pneumococci. PLY-induced hyperpermeability in HL-MVEC monolayers is abrogated by both NOX2 inhibitor gp91dstat and TIP peptide. Endothelial NOX2 expression is assessed by increased surface membrane presence of phosphorylated p47 phox subunit (Western blotting) in vitro and by co-localization of CD31 and gp91 phox in mouse lung slices using DuoLink, whereas NOX2-generated superoxide is measured by chemiluminescence. TIP peptide blunts PMA-induced NOX2 activity in cells expressing ENaC-α, but not in neutrophils, which lack ENaC. Conditional endothelial ENaC-α KO (enENaC-α KO) mice develop increased capillary leak upon i.t. instillation with PLY or pneumococci, compared to wild type (wt) animals. TIP peptide diminishes capillary leak in Sp -infected wt mice, without significantly increasing lung bacterial load. Lung slices from Sp -infected enENaC-α KO mice have a significantly increased endothelial NOX2 expression, as compared to infected CRE mice. In conclusion, endothelial ENaC may represent a novel therapeutic target to reduce NOX2-mediated oxidative stress and capillary leak in ARDS, without impairing host defense.

Background: Urine sodium concentration has been suggested as a marker to guide enteral sodium supplementation in preterm infants; however, no previous data have demonstrated relationships between urine sodium concentration and postnatal growth., Methods: We performed a single-center retrospective cohort study on 224 preterm infants admitted to the neonatal intensive care unit at the Children's Hospital of Georgia between January 2010 and July 2022. Spot urine sodium was measured in preterm infants (<34 weeks postmenstrual age [PMA]) between days of life (DOLs) 7 and 28. Our exposure of interest was spot urine sodium concentration (milliequivalents per liter) obtained between postnatal days 7 and 28, and our primary outcome was weight velocity (grams per kilograms per day) determined at DOL 28. Statistical relationships were assessed by multivariate analysis with subgroup comparisons by Student t test and analysis of variance., Results: In 224 preterm infants (199 ± 17 days, 56% male, 71% Black), urine sodium concentration did not associate with weight velocity at DOL 28 and 36 weeks PMA. Urine sodium concentration was weakly associated with gestational age at birth, and Black preterm infants had higher urine sodium values when compared with "other," but not White preterm infants., Conclusion: Spot urine sodium during the first month of life does not associate with weight velocity at DOL 28 or 36 weeks PMA., (© 2024 American Society for Parenteral and Enteral Nutrition.)

Objective: To determine if an intervention designed to enhance early responsive parenting (RP) practices (e.g., reading infant cues, establishing bedtime routines) and promote infant sleep and soothing among Black families has secondary benefits for mothers' postpartum sleep., Method: This preregistered secondary analysis of the Sleep Strong African American Families randomized controlled trial investigated effects of an RP intervention versus a safety control condition on self-reported maternal sleep difficulties at 8 and 16 weeks postpartum and on actigraph-measured maternal sleep at 8 weeks postpartum., Results: The 212 randomized mothers were Black/African American (100%) and non-Hispanic (98.6%) and averaged 22.7 years ( SD = 4.5) of age. Among 138 mothers with useable actigraph data, RP mothers had a mean 20 [95% CI: 2, 37] minutes longer actigraph-measured total sleep time than controls at 8 weeks postpartum, after adjusting for age and other covariates likely to influence mothers' sleep ( p = .04). Participation in the RP intervention did not significantly impact self-reported sleep difficulties or other actigraph-measured sleep parameters (e.g., efficiency) in either unadjusted or adjusted models, although RP effects on sleep difficulties and sleep efficiency were in the hypothesized directions., Conclusions: Interventions supporting responsive sleep parenting practices to increase infant sleep may also help first-time Black mothers get more sleep themselves during the postpartum period, even without an explicit focus on maternal sleep strategies. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Objectives: Hypothyroxinemia of prematurity (HOP) is characterized by low free thyroxine (FT4) associated with low or normal thyroid stimulating hormone (TSH). The objective of this study is to define FT4 and TSH values in very preterm infants (<32 weeks postmenstrual age, PMA) and correlate hypothyroxinemia and levothyroxine treatment with growth velocity at 28 days and 36 weeks PMA., Methods: Preterm neonates <32 weeks PMA admitted to the regional neonatal intensive care unit (NICU) at the Children's Hospital of Georgia (USA) between January 2010 and July 2022 were routinely screened for hypothyroxinemia. FT4 and TSH values were obtained on 589 eligible neonates between day of life (DOL) 4 and 14. Growth velocity (g/kg/day) from DOL 14 to DOL 28 and 36-weeks PMA were calculated for each neonate and potential explanatory variables (PMA, sex, and race) were incorporated into multivariate regression models to identify associations between HOP and growth velocity., Results: In 589 preterm infants, PMA at birth was strongly associated inversely with FT4 (R=0.5845) and modestly with TSH (R=0.2740). Both FT4 and gestational age, but not TSH or levothyroxine treatment, were associated with growth velocity at 28 days of life and at 36 weeks PMA., Conclusions: We provide a large data set for identifying FT4 and TSH measurements and identify hypothyroxinemia of prematurity as a potential mediator of slow postnatal growth in very preterm infants., (© 2024 Walter de Gruyter GmbH, Berlin/Boston.)

Objective: Many parents use food to soothe their infant, regardless of infant hunger, which can increase risk for rapid weight gain. Interventions promoting alternative soothing strategies may help parents respond more appropriately to crying. This secondary analysis aimed to examine effects of the Sleep SAAF (Strong African American Families) responsive parenting (RP) intervention on maternal responses to infant crying and to explore moderating effects of infant negativity., Methods: Primiparous Black mothers (n = 212) were randomized to an RP or safety control intervention, delivered during home visits at 3 and 8 weeks postpartum. Parents were empowered to first use non-food soothing strategies (eg, white noise, swaddling) when responding to crying. Mothers completed the Babies Need Soothing questionnaire at 8 and 16 weeks, and the Infant Behavior Questionnaire at 16 weeks. Data were analyzed using linear or logistic regression., Results: RP mothers were significantly more likely than controls to use shushing/white noise to soothe their infant at 8 (OR = 4.9, 95% CI: 2.2-10.6) and 16 weeks (OR = 4.8, 95% CI: 2.2-10.5), to go for a walk in stroller/ride in car at 8 weeks (OR = 2.3, 95% CI: 1.2-4.6), and to swing/rock/bounce their infant at 16 weeks (OR = 5.5, 95% CI: 1.2-25.7). RP mothers also reported significantly more frequent use of deep breathing, exercising, and bathing/showering than controls when frustrated with crying. Infant negativity moderated some intervention effects such that the RP intervention was more effective at increasing use of some soothing practices among mothers with less negative infants., Conclusions: An RP intervention positively impacted first-time Black mothers' responses to infant crying., (Copyright © 2023 Academic Pediatric Association. Published by Elsevier Inc. All rights reserved.)

Retinopathy of prematurity (ROP) is a complex neonatal disorder with multiple contributing factors. In this paper we have mounted the evidence in support of the proposal that neonatal sepsis meets all requirements for being a cause of ROP (not a condition, mechanism, or even innocent bystander) by means of initiating the early stages of the pathomechanism of ROP occurrence, systemic inflammation. We use the model of etiological explanation, which distinguishes between two overlapping processes in ROP causation. It can be shown that sepsis can initiate the early stages of the pathomechanism via systemic inflammation (causation process) and that systemic inflammation can contribute to growth factor aberrations and the retinal characteristics of ROP (disease process). The combined contribution of these factors with immaturity at birth (as intrinsic risk modifier) and prenatal inflammation (as extrinsic facilitator) seems to provide a cogent functional framework of ROP occurrence. Finally, we apply the Bradford Hill heuristics to the available evidence. Taken together, the above suggests that neonatal sepsis is a causal inducer of ROP., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Background: We quantify the osmolality of human milk fortified with human milk fortifiers (HMFs), powder infant formulas and protein additives., Methods: Commercial liquid HMFs and powder infant formulas were added to pasteurized pooled donor human milk in triplicate and stirred. The osmolality of unfortified and fortified human milk at 22, 24, 26, 27, 28, and 30 kcal/oz (0.73, 0.8, 0.87, 0.9, 0.93, and 1 kcal/ml, respectively) was determined using freezing-point depression., Results: The osmolality of fortified human milk associated with energy density in a linear relationship regardless of the fortification strategies. Multiple liquid HMFs and every powder infant formula exceeded the osmolality threshold of 450 mOsm/kg H 2 O within the energy densities tested., Conclusion: The osmolality of fortified human milk is highly variable and should be considered when selecting a fortifying agent for human milk., (© 2023 The Authors. Journal of Parenteral and Enteral Nutrition published by Wiley Periodicals LLC on behalf of American Society for Parenteral and Enteral Nutrition.)

Background: This study quantified the displacement of human milk by commercial human milk fortifiers (HMFs) and infant formulas., Methods: Commercial liquid HMFs and powder infant formulas were added to pasteurized pooled donor human milk in triplicate, stirred, and weighed. The difference in weight between unfortified and fortified human milk at 22, 24, 26, 27, 28, and 30 kcal/ounce was calculated., Results: The displacement of human milk by liquid HMFs and powder infant formulas and powder HMF was highly associated with energy density. A human milk-derived HMF displaced significantly more human milk when compared with bovine milk-derived HMFs at equivalent energy densities. Similarly, powder infant formulas displaced less human milk when compared with a powder HMF, and the addition of hydrolyzed powder infant formulas resulted in less human milk displacement when compared with nonhydrolyzed powder infant formulas., Conclusions: The displacement of human milk by commercial liquid HMFs and infant formulas must be considered when selecting a fortifying strategy., (© 2023 American Society for Parenteral and Enteral Nutrition.)

Background: Reduction in oxygen delivery to developing kidneys of premature infants may be an important source for acute kidney injury in premature infants., Purpose: To describe changes in continuous kidney oxygenation (RrSO 2 ) measures before, during, and after routine diaper changes., Methods: Non-a priori analysis of a prospective cohort that received continuous measurement of RrSO 2 with near-infrared spectroscopy (NIRS) over the first 14 days of life demonstrating acute RrSO 2 drops surrounding diaper changes., Results: In total, 26 of 38 (68%) infants (≤1800 g) from our cohort exhibited acute drops in RrSO 2 that temporally correlated with diaper changes. Mean (SD) RrSO 2 baseline prior to each diaper change event was 71.1 (13.2), dropped to 59.3 (11.6) during diaper change, and recovered to 73.3 (13.2). There was a significant difference between means when comparing baseline to diaper change ( P < .001; 95% CI, 9.9 to 13.8) and diaper change to recovery ( P < .001; 95% CI, -16.9 to -11.2). The mean decrease in RrSO 2 during diaper change averaged 12 points (17%) below 15-minute RrSO 2 mean prior to diaper change, with quick recovery to prediaper change levels. No decreases in SpO 2 , blood pressure, or heart rate were documented during the intermittent kidney hypoxic events., Implications for Practice and Research: Routine diaper changes in preterm infants may increase the risk for acute reductions in RrSO 2 as measured by NIRS; however, the impact on kidney health remains unknown. Larger prospective cohort studies assessing kidney function and outcomes related to this phenomenon are needed., (Copyright © 2023 by The National Association of Neonatal Nurses.)

Background: Our goal was to quantify the pH and total acidity of human milk fortified with human milk fortifiers (HMFs), powder infant formulas, and protein additives., Methods: Commercial liquid HMFs and powder infant formulas were added to pasteurized pooled donor human milk in triplicate and stirred. The pH of unfortified and fortified human milk at 22, 24, 26, 27, 28, and 30 kcal/ounce (624, 680, 737, 765, 794, and 850 kcal/g, respectively) was determined using a pH meter. Phenolphthalein acidity at 24 and 30 kcal/ounce (680 and 850 kcal/g, respectively) was determined using diluted sodium hydroxide., Results: The pH of unfortified human milk increased within the first hour (6.52 ± 0.06 vs 6.62 ± 0.05, P < 0.0001). Changes in pH largely correlated with caloric density; however, directional changes varied considerably between HMFs and powder infant formulas. Two liquid HMFs demonstrated modest reductions in pH with increasing caloric density whereas one liquid HMF alkalinized human milk with increasing caloric density (analysis of variance P < 0.0001). Phenolphthalein acidity was significantly higher for five HMFs and lower for one HMF at 30 kcal/ounce (850 kcal/g) but not 24 kcal/ounce (680 kcal/g). Powder infant formulas generally increased pH with increasing caloric density (analysis of variance P < 0.0001), but no differences in phenolphthalein acidity were noted., Conclusion: Changes in acid/base balancefor fortified human milk are variable and may be a consideration when selecting a fortifying agent for human milk., (© 2023 American Society for Parenteral and Enteral Nutrition.)

Background: Human milk is the preferred diet for very low birth weight (VLBW, <1500 g) infants. When mother's own milk is unable to meet the needs of VLBW infants, donor human milk (DHM) is the preferred alternative. Unfortunately, the composition of DHM remains elusive and no comparative studies between preterm human milk and DHM have been performed previously., Objectives: We aimed to analyze the nutrient content of commercial pooled DHM and compare nutrient content in DHM with that of early and mature preterm human milk., Methods: We analyzed nutrient content in 15 DHM samples provided from 7 commercial milk banks including calories, carbohydrate, fat, protein, sodium, chloride, potassium, zinc, calcium, phosphorus, magnesium, and vitamin D and compared each nutrient to early (7 d of life) and mature (28 d of life) preterm human milk samples (n = 28-36 per nutrient, gestational age = 28 ± 3 wk). Protein-to-energy ratio and carbohydrate-to-nonprotein energy ratio were calculated for each sample and compared., Results: Mean values for all macro- and micronutrients in DHM are reported. In comparison to early or mature preterm human milk, DHM had significantly lower protein, sodium, chloride, potassium, and zinc content. Calorie, carbohydrate, calcium, phosphorus, magnesium, and vitamin D content did not differ statistically between DHM and early or mature preterm human milk. Fat content was modestly lower in early but not mature human milk when compared with DHM., Conclusions: We provide mean values for several macro- and micronutrients for DHM and identify key differences between DHM and preterm human milk, which may be considered when designing human milk-based feeding plans. This study was registered at clinicaltrials.gov as NCT05742815., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: Intrauterine growth restriction (IUGR) exerts a negative impact on developing cardiomyocytes and emerging evidence suggests activation of oxidative stress pathways plays a key role in this altered development. Here, we provided pregnant guinea pig sows with PQQ, an aromatic tricyclic o-quinone that functions as a redox cofactor antioxidant, during the last half of gestation as a potential antioxidant intervention for IUGR-associated cardiomyopathy., Methods: Pregnant guinea pig sows were randomly assigned to receive PQQ or placebo at mid gestation and fetuses were identified as spontaneous IUGR (spIUGR) or normal growth (NG) near term yielding four cohorts: NG ± PQQ and spIUGR ± PQQ. Cross sections of fetal left and right ventricles were prepared and cardiomyocyte number, collagen deposition, proliferation (Ki67) and apoptosis (TUNEL) were analyzed., Results: Cardiomyocyte endowment was reduced in spIUGR fetal hearts when compared to NG; however, PQQ exerted a positive effect on cardiomyocyte number in spIUGR hearts. Cardiomyocytes undergoing proliferation and apoptosis were more common in spIUGR ventricles when compared with NG animals, which was significantly reduced with PQQ supplementation. Similarly, collagen deposition was increased in spIUGR ventricles and was partially rescued in PQQ-treated spIUGR animals., Conclusion: The negative influence of spIUGR on cardiomyocyte number, apoptosis, and collagen deposition during parturition can be suppressed by antenatal administration of PQQ to pregnant sows. These data identify a novel therapeutic intervention for irreversible spIUGR-associated cardiomyopathy.

Importance: Black individuals in the US experience sleep disparities beginning in infancy and continuing throughout the lifespan, suggesting early interventions are needed to improve sleep., Objective: To investigate whether a responsive parenting (RP) intervention for Black mothers improves infant sleep and increases responsive sleep parenting practices., Design, Setting, and Participants: This is a post hoc secondary analysis of the Sleep SAAF (Strong African American Families) study, a randomized clinical trial comparing an RP intervention with a safety control condition over the first 16 weeks post partum. Data were collected between spring 2018 and summer 2021. Families were recruited from the mother-infant nursery at Augusta University Medical Center, Augusta, Georgia, and completed home visits at 1, 3, 8, and 16 weeks post partum. Primiparous Black mother-infant dyads were screened for eligibility using medical records. Results were analyzed on an intention-to-treat model. Data were analyzed from March 2022 to January 2023., Interventions: The RP intervention curriculum focused on infant sleep, soothing and crying, and feeding. The control group received a safety intervention. Community research associates delivered the interventions during home visits at 3 and 8 weeks post partum., Main Outcomes and Measures: The primary end point was infant sleep duration at 16 weeks. Mothers reported on bedtime routine and sleep behaviors, infant sleep duration, and nighttime waking and feeding at 8 and 16 weeks post partum using questionnaires., Results: A total of 212 Black mothers (mean [SD] age, 22.7 [4.5] years) were randomized, including 208 mothers (98.6%) who identified as non-Hispanic and 3 mothers (1.4%) who identified as Hispanic; 108 mothers were randomized to the RP group and 104 mothers were randomized to the control group. At 16 weeks post partum, infants in the RP group had longer reported nighttime sleep duration (mean difference, 40 [95% CI, 3 to 77] minutes), longer total sleep duration (mean difference, 73 [95% CI, 14 to 131] minutes), fewer nighttime wakings (mean difference, -0.4 [95% CI, -0.6 to -0.1] wakings), and greater likelihood of meeting guidelines of at least 12 hours of total sleep per day (risk ratio [RR], 1.4 [95% CI, 95% CI, 1.1 to 1.8]) than controls. Relative to controls, mothers in the RP group more frequently reported engaging in some RP practices, including giving the baby a few minutes to fall back asleep on their own (RR, 1.6 [95% CI, 1.0 to 2.6]) and being less likely to feed their baby as the last activity before bed (RR, 0.5 [95% CI, 0.3 to 0.8])., Conclusions and Relevance: This secondary analysis of a randomized clinical trial found that an RP intervention for Black families improved infant sleep and increased some responsive sleep parenting practices., Trial Registration: ClinicalTrials.gov Identifier: NCT03505203.

Objective: Identify associations between cannulation approach and mortality in neonates who received ECMO support for respiratory failure., Study Design: A retrospective analysis of neonates receiving ECMO for respiratory indications at a single quaternary-referral NICU. Associations between cannulation approach and mortality were assessed after adjustment for Neo-RESCUERS score. Cox Proportional Hazards (CPH) model was used to estimate hazard ratios (HR) and 95% confidence intervals (CI) for each variable and outcome., Results: Among 244 neonates, overall survival was 88%, with 71% undergoing VV cannulation. After adjusting for Neo-RESCUERS score, VA cannulation was associated with higher mortality during ECMO when compared with VV cannulation (HR 4.189, 95% CI 1.480-11.851, P = 0.0069). Disease-specific comparisons revealed no statistical difference in Neo-RESCUERS score between VA and VV cohorts; however, VA cannulation was associated with higher ECMO mortality for neonates with congenital diaphragmatic hernia (50% vs. 5.5%, Χ 2  = 8.5965, P = 0.0034) and PPHN (20% vs. 1.8%, Χ 2  = 9.1047, P = 0.0025) when compared with VV cannulation., Conclusion: VA cannulation was associated with increased mortality in neonates while on ECMO for respiratory failure, which was independent of illness severity., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Background: Blood product transfusions are necessary for critically ill neonates on extracorporeal membrane oxygenation (ECMO). Transfusions are administered in response to unstudied arbitrary thresholds and may be associated with adverse outcomes. The objective of this study was to identify relationships between blood product components and mortality in neonates receiving ECMO support for respiratory indications., Study Design and Methods: A retrospective review of neonates receiving ECMO for respiratory indications from 2002 to 2019 from a single quaternary-referral neonatal intensive care unit (NICU). Demographic and outcome data and transfusion volume (ml/kg/day) were harvested from the medical record, and baseline mortality risk was assessed using NEO-RESCUERS scores. The association between volume of red blood cells (RBC), platelet, plasma transfusion rates (ml/kg/day), and mortality on ECMO were assessed after adjustment for NEO-RESCUERS score. Cox proportional hazards (CPH) competing risk model was used to estimate hazard ratios (HR) and 95% confidence intervals (CI) for each variable and mortality outcome., Measurements and Main Results: Among 248 neonates undergoing ECMO for respiratory failure, overall survival was 93%. RBC, platelet, and plasma volume were highly associated with mortality during ECMO in an unadjusted model. After adjusting for NEO-RESCUERS score, RBC volume was associated with increased mortality risk (HR 1.013, 95% CI 1.004-1.022, p = .0043), but platelet and plasma volume were not associated with mortality., Conclusions: RBC, but not platelet or plasma volume, is associated with mortality in neonates on ECMO. Our findings refute previous studies demonstrating an association between platelet volume and mortality for neonates on ECMO., (© 2022 AABB.)

Background/objective: Parents shape children's early experiences with food, influencing what is served, children's food choices, and how much children eat. Responsive parenting (RP) interventions such as INSIGHT have improved maternal infant feeding practices, but have only been tested among predominantly White families. This secondary analysis of data from the Sleep SAAF (Strong African American Families) RCT tests the effects of an RP intervention designed to prevent rapid infant weight gain on African American mothers' infant feeding practices., Methods: Primiparous African American mother-infant dyads (n = 194) were randomized to an RP or safety control intervention delivered by community research associates at infant age 3 and 8 weeks. At 16 weeks, mothers completed the Babies Need Feeding questionnaire, the Infant Feeding Styles Questionnaire, and the Babies Need Soothing questionnaire. Logistic regression and general linear models examined the effect of study group on infant feeding practices. Moderation analyses explored whether effects varied by feeding mode (any breast milk versus exclusive formula), maternal age (≥ 20 years versus < 20 years), and maternal pre-pregnancy BMI (with obesity versus not)., Results: RP mothers reported more responsive feeding (p = 0.005, partial η 2  = 0.02), lower likelihood of using beverages other than breast milk/formula to soothe their infant (p = 0.01, OR = 0.42, 95% CI [0.2-0.8]), and less pressure with cereal than control mothers (p = 0.09, partial η 2  = 0.02). RP mothers also reported less pressure to finish/soothe than controls (p = 0.007, partial η 2  = 0.04); feeding mode (B = 0.74, p = 0.003) and maternal age (B = 0.53, p = 0.04) moderated this effect. There were no significant group differences in bottle-feeding practices (e.g., adding cereal to bottle, using an appropriate nipple/bottle size), or in context-based or emotion-based food to soothe., Conclusions: Responsive parenting education influenced some feeding practices of African American mothers. Mothers reported using less pressure, a control-based feeding practice, and more responsive feeding than controls., Trial Registration: Sleep SAAF: A Strong African American Families Study. www., Clinicaltrials: gov NCT03505203. Registered 3 April 2018., (© 2022. The Author(s).)

Accumulation of lipid-laden foam cells in the arterial wall plays a central role in atherosclerotic lesion development, plaque progression, and late-stage complications of atherosclerosis. However, there are still fundamental gaps in our knowledge of the underlying mechanisms leading to foam cell formation in atherosclerotic arteries. Here, we investigated the role of receptor-independent macropinocytosis in arterial lipid accumulation and pathogenesis of atherosclerosis. Genetic inhibition of fluid-phase macropinocytosis in myeloid cells ( LysMCre + Nhe1 fl/fl ) and repurposing of a Food and Drug Administration (FDA)-approved drug that inhibits macrophage macropinocytosis substantially decreased atherosclerotic lesion development in low-density lipoprotein (LDL) receptor-deficient and Apoe -/- mice. Stimulation of macropinocytosis using genetic ( H-RAS G12V ) and physiologically relevant approaches promoted internalization of unmodified native (nLDL) and modified [e.g., acetylated (ac) and oxidized (ox) LDL] lipoproteins in both wild-type and scavenger receptor (SR) knockout ( Cd36 -/- / Sra -/- ) macrophages. Pharmacological inhibition of macropinocytosis in hypercholesterolemic wild-type and Cd36 -/- / Sra -/- mice identified an important role of macropinocytosis in LDL uptake by lesional macrophages and development of atherosclerosis. Furthermore, serial section high-resolution imaging, LDL immunolabeling, and three-dimensional (3D) reconstruction of subendothelial foam cells provide visual evidence of lipid macropinocytosis in both human and murine atherosclerotic arteries. Our findings complement the SR paradigm of atherosclerosis and identify a therapeutic strategy to counter the development of atherosclerosis and cardiovascular disease.

Current therapies for treatment of proliferative retinopathy focus on retinal neovascularization (RNV) during advanced disease and can trigger adverse side-effects. Here, we have tested a new strategy for limiting neurovascular injury and promoting repair during early-stage disease. We have recently shown that treatment with a stable, pegylated drug form of the ureohydrolase enzyme arginase 1 (A1) provides neuroprotection in acute models of ischemia/reperfusion injury, optic nerve crush, and ischemic stroke. Now, we have determined the effects of this treatment on RNV, vascular repair, and retinal function in the mouse oxygen-induced retinopathy (OIR) model of retinopathy of prematurity (ROP). Our studies in the OIR model show that treatment with pegylated A1 (PEG-A1), inhibits pathological RNV, promotes angiogenic repair, and improves retinal function by a mechanism involving decreased expression of TNF, iNOS, and VEGF and increased expression of FGF2 and A1. We further show that A1 is expressed in myeloid cells and areas of RNV in retinal sections from mice with OIR and human diabetic retinopathy (DR) patients and in blood samples from ROP patients. Moreover, studies using knockout mice with hemizygous deletion of A1 show worsened RNV and retinal injury, supporting the protective role of A1 in limiting the OIR-induced pathology. Collectively, A1 is critically involved in reparative angiogenesis and neuroprotection in OIR. Pegylated A1 may offer a novel therapy for limiting retinal injury and promoting repair during proliferative retinopathy., (© 2022. The Author(s).)

Responsive parenting (RP) interventions reduce rapid infant weight gain but their effect for underserved populations is largely unknown. The Sleep SAAF (Strong African American Families) study is a two-arm randomized clinical trial for primiparous African American mother-infant dyads that compares an RP intervention to a child safety control over the first 16 weeks postpartum. Here we report on intervention effects on rapid infant weight gain and study implementation. Families were recruited from a mother/baby nursery shortly after delivery. Community Research Associates (CRAs) conducted intervention home visits at 3 and 8 weeks postpartum, and data collection home visits at 1, 8, and 16 weeks postpartum. To examine rapid infant weight gain, conditional weight gain (CWG) from 3 to 16 weeks, the primary outcome, and upward crossing of 2 major weight-for-age percentile lines were calculated. Among the 212 mother-infant dyads randomized, 194 completed the trial (92% retention). Randomized mothers averaged 22.7 years, 10% were married, and 49% participated in the Supplemental Nutrition Assistance Program (SNAP). Adjusting for covariates, mean CWG was lower among RP infants (0.04, 95% CI [-0.33, 0.40]) than among control infants (0.28, 95% CI [-0.08, 0.64]), reflecting non-significantly slower weight gain (p = 0.15, effect size d = 0.24). RP infants were nearly half as likely to experience upward crossing of 2 major weight-for-age percentile lines (14.1%) compared to control infants (24.2%), p = 0.09, odds ratio = 0.52 (95% CI [0.24, 1.12]). Implementation data revealed that participating families were engaged in the intervention visits and intervention facilitators demonstrated high fidelity to intervention materials. Findings show that RP interventions can be successfully implemented among African American families while suggesting the need for modifications to yield stronger effects on infant weight outcomes., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: Currently, reference ranges for renal oxygenation measured by near-infrared spectroscopy (NIRS) in preterm infants beyond the first days of life are lacking, especially those born prior to 29 weeks' gestation. Population estimates of renal oxygenation (rSO 2 ) levels among preterm infants over time have yet to be established, leading to reluctance in clinical application., Purpose: To characterize the distribution and estimate population parameters for renal oxygenation measured by NIRS during the first 14 days of life among preterm infants., Methods: We prospectively observed rSO 2 trends of 37 infants before 34 weeks' gestation and 1800-g or less birth weight for the first 14 days of life. Analyses included distribution fit tests, ordinary least squares (OLS) regression, and t tests., Results: Average daily rSO 2 variation steadily increased with 42% difference through the first 14 days of life. For all infants, renal rSO 2 means peaked during the first 3 days of life and plateaued around 7 days. Daily rSO 2 slopes were significantly lower among males and infants 29 weeks' or less gestation., Implications for Practice: Renal rSO 2 during the first 14 days of life reflects normal extrauterine transition reaching stabilization around 7 days of life. Gestational age, birth weight, and gender may predict the early trajectory of rSO 2 patterns. Population estimates provide parameters for renal rSO 2 that may indicate early-onset tissue hypoxia when acute or significant drops from baseline occur., Implications for Research: We present a framework to guide future research using renal NIRS technology in preterm infants to determine deviations from expected trends that may precede renal injury., Competing Interests: The authors of this manuscript declare no potential, real, or perceived conflict of interest related to the submission of this manuscript to Advances in Neonatal Care journal. Dr Marin composed the initial draft and received subsequent input from all coauthors. Dr Marin was the principal investigator of this study and oversaw all aspects of data collection, analysis, and interpretation. Dr Williams performed data analysis; Drs Mann, Mansuri, Stansfield, Mundy, and Cockfield participated in data collection, analysis, and interpretation. Each additional author has viewed and approved the submission of this manuscript version., (Copyright © 2021 by The National Association of Neonatal Nurses.)

Introduction: The objectives of this retrospective cohort study were to examine the effect of vitamin K administration on hemorrhagic and thrombotic complications, blood product utilization, and outcomes in neonatal extracorporeal membrane oxygenation (ECMO)., Methods: In the pilot study, complications, blood product use, and outcome data for neonates who received ( n  = 21) or did not receive ( n  = 18) a single dose of vitamin K (5 mg) immediately after initiation of ECMO for respiratory failure between 2006 and 2010 were compared. In the validation cohort, complications and outcomes were compared for 74 consecutive neonates supported with ECMO for respiratory failure who received ( n  = 45) or did not receive ( n  = 29) additional vitamin K once daily for prothrombin time (PT) ⩾14 seconds during ECMO from 2014 to 2019., Results: In the pilot study, vitamin K at ECMO initiation was associated with fewer thrombotic complications and similar hemorrhagic complications. The volume of fresh frozen plasma was higher in neonates who received vitamin K, but total blood product and other component volume did not differ between groups. ECMO run time, survival off ECMO, survival to discharge, and length of stay did not differ between cohorts. In the validation cohort, neonates who received additional vitamin K during ECMO had longer ECMO run time and length of stay, but no difference in mortality was observed. Further, thrombotic and hemorrhagic complications as well as blood product exposure were similar between cohorts., Conclusions: These data suggest that routine vitamin K administration may have limited or no benefit during neonatal ECMO.

The health benefits of breast milk feeding have been well-established, yet disparities exist, with African American mothers having the lowest breast milk feeding rates in the United States. This prospective, longitudinal study examined infant feeding (breast milk and/or infant formula) from birth to age 16 weeks, predictors of any breast milk feeding by age 1 week, and predictors of cessation of any breast milk feeding by ages 3, 8, and 16 weeks among primiparous African American mothers. This secondary analysis included 185 mother-infant dyads from the Sleep SAAF (Strong African American Families) study, a randomized clinical trial testing a responsive parenting vs. child safety control intervention. Mothers reported sociodemographic and psychosocial characteristics at age 1 week and infant feeding practices at ages 1, 3, 8, and 16 weeks. Rates of any breast milk feeding decreased from 66.5% at 1 week to 23.3% at 16 weeks. Bivariate logistic regression models showed that prepregnancy BMI (OR = 1.09), working prepregnancy (OR = 2.25), and food insecurity (OR = 2.49) significantly increased the odds of mothers feeding any breast milk by 1 week, whereas Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) participation (OR = 0.21) significantly decreased the odds. Bivariate logistic regression models showed that Supplemental Nutrition Assistance Program (SNAP) participation (OR = 2.86) and racial discrimination (OR = 2.14) significantly increased the odds of cessation of any breast milk feeding by 3 weeks. SNAP (OR = 2.33) and WIC (OR = 2.38) participation significantly increased the odds of cessation of any breast milk feeding by 8 weeks, whereas higher prepregnancy BMI (OR = 0.95) decreased the odds. Higher mother's age (OR = 0.92) significantly decreased the odds of cessation of any breast milk feeding by 16 weeks. The findings can be used to inform targeted interventions to promote mothers feeding any breast milk and help reduce breast milk feeding disparities among African American mothers.

Background: The objective of this study was to determine whether human milk supplemented with a novel human milk-based human milk fortifier (Novel HMF), compared with a bovine milk-based HMF (Bovine HMF), supports preterm infant growth through 36 weeks' postmenstrual age (PMA)., Methods: This single-center, prospective trial compared growth and nutrition outcomes of preterm infants provided a human milk-based diet (mother's own milk or donor milk) supplemented with a Novel HMF with historic controls provided Bovine HMF. Preterm infants with an estimated gestational age (EGA) between 23 and 33 weeks' PMA and birth weight between 750 and 1800 g were eligible for study inclusion. Weight, length, and head circumference (HC) were monitored weekly. The occurrence of late-onset sepsis, nil per os (NPO) days, necrotizing enterocolitis, metabolic acidosis, and serious adverse events were monitored., Results: Birth weight, length, HC, and EGA were similar between the Novel HMF (n = 37) and Bovine HMF (n = 49) groups. The days to regain birth weight was shorter in the Novel HMF group (9.4 ± 4.0 vs 11.4 ± 4.8, P = .0343), with similar weight gain (g/day) from birth to 36 weeks' PMA. Adjusted weight growth velocity (g/kg/day) was significantly higher in the Novel HMF group at 14 and 21 days but similar at 36 weeks' PMA. The Novel HMF group experienced fewer NPO days with a similar total number of feeding days., Conclusions: A novel, multinutrient, human milk-based HMF is well tolerated and meets the nutrition needs of preterm infants., (© 2021 American Society for Parenteral and Enteral Nutrition.)

Vascular endothelial growth factor receptor type 2 (VEGFR2, also known as KDR and FLK1) signalling in endothelial cells (ECs) is essential for developmental and reparative angiogenesis. Reactive oxygen species and copper (Cu) are also involved in these processes. However, their inter-relationship is poorly understood. Evidence of the role of the endothelial Cu importer CTR1 (also known as SLC31A1) in VEGFR2 signalling and angiogenesis in vivo is lacking. Here, we show that CTR1 functions as a redox sensor to promote angiogenesis in ECs. CTR1-depleted ECs showed reduced VEGF-induced VEGFR2 signalling and angiogenic responses. Mechanistically, CTR1 was rapidly sulfenylated at Cys189 at its cytosolic C terminus after stimulation with VEGF, which induced CTR1-VEGFR2 disulfide bond formation and their co-internalization to early endosomes, driving sustained VEGFR2 signalling. In vivo, EC-specific Ctr1-deficient mice or CRISPR-Cas9-generated redox-dead Ctr1(C187A)-knockin mutant mice had impaired developmental and reparative angiogenesis. Thus, oxidation of CTR1 at Cys189 promotes VEGFR2 internalization and signalling to enhance angiogenesis. Our study uncovers an important mechanism for sensing reactive oxygen species through CTR1 to drive neovascularization., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Mutations in the NF1 tumor suppressor gene are linked to arteriopathy. Nf1 heterozygosity (Nf1+/-) results in robust neointima formation, similar to humans, and myeloid-restricted Nf1+/- recapitulates this phenotype via MEK-ERK activation. Here we define the contribution of myeloid subpopulations to NF1 arteriopathy. Neutrophils from WT and Nf1+/- mice were functionally assessed in the presence of MEK and farnesylation inhibitors in vitro and neutrophil recruitment to lipopolysaccharide was assessed in WT and Nf1+/- mice. Littermate 12-15 week-old male wildtype and Nf1+/- mice were subjected to carotid artery ligation and provided either a neutrophil depleting antibody (1A8), liposomal clodronate to deplete monocytes/macrophages, or PD0325901 and neointima size was assessed 28 days after injury. Bone marrow transplant experiments assessed monocyte/macrophage mobilization during neointima formation. Nf1+/- neutrophils exhibit enhanced proliferation, migration, and adhesion via p21 Ras activation of MEK in vitro and in vivo. Neutrophil depletion suppresses circulating Ly6C low monocytes and enhances neointima size, while monocyte/macrophage depletion and deletion of CCR2 in bone marrow cells abolish neointima formation in Nf1+/- mice. Taken together, these findings suggest that neurofibromin-MEK-ERK activation in circulating neutrophils and monocytes during arterial remodeling is nuanced and points to important cross-talk between these populations in the pathogenesis of NF1 arteriopathy., (© 2021. The Author(s).)

Background: The human milk-fed preterm infant is at risk for growth failure, micronutrient deficiencies, and neurocognitive delay. Although protective and better tolerated than formula, human milk alone cannot meet the high nutrient requirements of this population, and fortification is necessary. Clinicians use assumptions of preterm human-milk composition to determine the type and quantity of fortification., Objectives: The objectives of this review were to identify evidence of macronutrient and micronutrient concentration in preterm human milk and to identify knowledge gaps regarding composition., Methods: PubMed and the Cumulative Index to Nursing and Allied Health Literature were used to identify original articles published between January 1950 and December 2019., Results: Twenty-seven articles were found containing original data on macronutrients and micronutrients. Most (67%) of the studies published after 2011 measured the macronutrients and included gestational ages from 28 to 36 weeks. Milk collection methods, experimental design, and analytical methods varied between studies. There are 15 countries represented in this review; all of the American studies (n = 7) were published from 1980 to 1984., Conclusions: African American women, or women delivering before 28 weeks' gestation are not represented in the literature. Accurate and targeted human-milk fortification depends on comprehensive, complete, and representative human-milk nutrient data. We have aggregated all available preterm human-milk macronutrient and micronutrient data and reported trends associated with lactation stage and gestational age. This report can aid in the design of feeding plans that are appropriate for the gestational age of the preterm infant and the lactation stage of the breastmilk., (© 2020 American Society for Parenteral and Enteral Nutrition.)

Background: Breast milk feedings are the optimal feeding choice for premature infants. Clinicians depend on accurate nutrient profiles of the breast milk in order to make informed decisions regarding the need for nutrient supplementation. Existing data for nutrient composition of preterm breast milk are dated and not representative of the current population of women delivering prematurely in the United States., Objectives: The purpose of this prospective, longitudinal, single-center observational study was to measure the macronutrient and micronutrient composition of breast milk expressed by mothers, including women who self-identify as black, delivering preterm infants at ≤33 completed weeks of gestation., Methods: We collected breast milk samples from mothers of preterm infants admitted to the neonatal intensive care unit at Augusta University Medical Center from January 2019 through November 2019. Mother's milk samples were collected on postpartum days 7, 14, 21, and 28 and analyzed for macronutrients (energy, fat, protein, and carbohydrates) and micronutrients (sodium, potassium, chloride, calcium, phosphorus, magnesium, vitamin D, and zinc)., Results: Thirty-eight mothers, mean age 27 ± 5.1 y and majority black (66%), provided milk for the study. The mean estimated gestational age and birth weight were 28.2 ± 2.8 weeks of gestation and 1098 ± 347 g, respectively, with 42% of mothers in the cohort delivering before week 28 of pregnancy. Differences in protein, sodium, potassium, calcium, phosphorus, and zinc concentrations based on race, day, and milk volume were identified. Dilution effects for protein, sodium, chloride, and vitamin D concentrations over time were identified., Conclusions: Our study is among the first to characterize breast milk composition from women who delivered extremely preterm infants and adds to the evidence that race, gestational age, and volume influence the composition of preterm mother's milk. These factors should be considered when designing mother's milk-based feeds for premature infants., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society for Nutrition.)

Background: Conflicting statements by stakeholders in obstetric care have suggested different criteria for defining peripartum fever and suspected intraamniotic infection, which have not been evaluated., Methods: A case-control study of pregnancies between 35 and 41 weeks at a single tertiary care center between January 2016 and December 2017. Cases with pathology-confirmed chorioamnionitis were identified, and demographic data, risk factors, and neonatal outcomes were extracted from the medical record. The American College of Gynecology (ACOG) and National Institutes of Health (NIH) Workshop guidelines for identifying isolated maternal fever and suspected intraamniotic infection were applied, retrospectively. Odds ratios, sensitivity/specificity, and predictive value of each guideline for pathology-confirmed chorioamnionitis and for secondary outcomes of interest were determined., Results: 943 mother-infant dyads were evaluated including 41 (4.3%) with pathology-confirmed chorioamnionitis. Among cases, 18 (43.9%) experienced any maternal temperature ≥38°C (100.4°F) with 12 (29.2%) and 8 (19.5%) cases meeting criteria for isolated maternal fever according to the ACOG and Workshop guidelines, respectively. Furthermore, the ACOG and Workshop guidelines correctly identified 6 (14.6%) and 3 (7.3%) of cases of pathology-confirmed chorioamnionitis with high agreement between definitions (κ = 0.63). Laboratory evaluation, antimicrobial exposure, and prolonged length of stay in offspring are substantially higher in cases as compared to controls., Conclusions: Guidelines that rely on maternal fever definitions for the diagnosis of suspected intraamniotic infection exhibit high agreement with low sensitivity, but high specificity and negative predictive value for pathology-confirmed chorioamnionitis. Maternal temperature ≥38°C continues to drive clinical decision-making for both mother and offspring., (© 2021 Wiley Periodicals LLC.)

Aims: Impaired lymphatic drainage of the arterial wall results in intimal lipid accumulation and atherosclerosis. However, the mechanisms regulating lymphangiogenesis in atherosclerotic arteries are not well understood. Our studies identified elevated levels of matrix protein R-spondin 2 (RSPO2) in atherosclerotic arteries. In this study, we investigated the role of RSPO2 in lymphangiogenesis, arterial cholesterol efflux into lesion-draining lymph nodes (LNs) and development of atherosclerosis., Methods and Results: The effect of RSPO2 on lymphangiogenesis was investigated using human lymphatic endothelial cells (LEC) in vitro and implanted Matrigel plugs in vivo. Cellular and molecular approaches, pharmacological agents, and siRNA silencing of RSPO2 receptor LGR4 were used to investigate RSPO2-mediated signalling in LEC. In vivo low-density lipoprotein (LDL) tracking and perivascular blockade of RSPO2-LGR4 signalling using LGR4-extracellular domain (ECD) pluronic gel in hypercholesterolemic mice were utilized to investigate the role of RSPO2 in arterial reverse cholesterol transport and atherosclerosis. Immunoblotting and imaging experiments demonstrated increased RSPO2 expression in human and mouse atherosclerotic arteries compared to non-atherosclerotic controls. RSPO2 treatment inhibited lymphangiogenesis both in vitro and in vivo. LGR4 silencing and inhibition of RSPO2-LGR4 signalling abrogated RSPO2-induced inhibition of lymphangiogenesis. Mechanistically, we found that RSPO2 suppresses PI3K-AKT-endothelial nitric oxide synthase (eNOS) signalling via LGR4 and inhibits activation of the canonical Wnt-β-catenin pathway. ApoE-/- mice treated with LGR4-ECD developed significantly less atherosclerosis compared with control treatment. Finally, increased arterial lymphatic vessel density and improved lymphatic drainage of fluorescently labelled LDL to deep cervical LNs were observed in LGR4-ECD-treated mice., Conclusion: These findings demonstrate that RSPO2 inhibits lymphangiogenesis via LGR4 and downstream impairment of AKT-eNOS-nitric oxide signalling. These results may also inform new therapeutic strategies to promote lymphangiogenesis and improve cholesterol efflux from atherosclerotic arteries., (Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2020. For permissions, please email: journals.permissions@oup.com.)

Background: Length of stay (LOS) is an important measure of quality; however, estimating LOS for rare populations such as gastroschisis is problematic. Our objective was to identify explanatory variables for LOS and build a model to estimate LOS in neonates with simple gastroschisis., Methods: In 73 neonates with simple gastroschisis (47% female, 67% White), statistical correlations for 31 potential explanatory variables for LOS were evaluated using multivariate linear regression. Poisson regression was used to estimate LOS in predetermined subpopulations, and a life table model was developed to estimate LOS for simple gastroschisis., Results: Female sex (-2.4 d), "time to silo placement" (0.9 d), total parenteral nutrition days (0.6 d), need for any nasogastric feedings (11.4 d) and at discharge (-7 d), "feeding tolerance" (0.4 d), days to first postoperative stool (-0.3 d), and human milk exposure (-3.4 d) associated with LOS in simple gastroschisis. Estimated LOS for preterm neonates was longer than term infants (5.4 versus 4.6 wk) but similar for estimates based on sex and race. Based on these associations, we estimate that >50% of neonates with simple gastroschisis will be discharged by hospital day 35., Conclusions: We identified several associations that explained variations in LOS and developed a novel model to estimate LOS in simple gastroschisis, which may be applied to other rare populations., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Neurofibromatosis type 1 (NF1) is a heritable cancer predisposition syndrome resulting from mutations in the NF1 tumor suppressor gene. Genotype-phenotype correlations for NF1 are rare due to the large number of NF1 mutations and role of modifier genes in manifestations of NF1; however, emerging reports suggest that persons with NF1 display a distinct anthropometric and metabolic phenotype featuring short stature, low body mass index, increased insulin sensitivity, and protection from diabetes. Nf1 heterozygous (Nf1+/-) mice accurately reflect the dominant inheritance of NF1 and are regularly employed as a model of NF1. Here, we sought to identify whether Nf1+/- mice recapitulate the anthropometric and metabolic features identified in persons with NF1. Littermate 16-20 week-old male wildtype (WT) and Nf1+/- C57B/6J mice underwent nuclear magnetic resonance (NMR), indirect calorimetry, and glucose/insulin/pyruvate tolerance testing. In some experiments, tissues were harvested for NMR and histologic characterization. Nf1+/- mice are leaner with significantly reduced visceral and subcutaneous fat mass, which corresponds with an increased density of small adipocytes and reduced leptin levels. Additionally, Nf1+/- mice are highly reliant on carbohydrates as an energy substrate and display increased glucose clearance and insulin sensitivity, but normal response to pyruvate suggesting enhanced glucose utilization and preserved gluconeogenesis. Finally, WT and Nf1+/- mice subjected to high glucose diet were protected from diet-induced obesity and hyperglycemia. Our data suggest that Nf1+/- mice closely recapitulate the anthropometric and metabolic phenotype identified in persons with NF1, which will impact the interpretation of previous and future translational studies of NF1., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Aims: Chronic adventitial and medial infiltration of immune cells play an important role in the pathogenesis of abdominal aortic aneurysms (AAAs). Nicotinic acid (niacin) was shown to inhibit atherosclerosis by activating the anti-inflammatory G protein-coupled receptor GPR109A [also known as hydroxycarboxylic acid receptor 2 (HCA2)] expressed on immune cells, blunting immune activation and adventitial inflammatory cell infiltration. Here, we investigated the role of niacin and GPR109A in regulating AAA formation., Methods and Results: Mice were supplemented with niacin or nicotinamide, and AAA was induced by angiotensin II (AngII) infusion or calcium chloride (CaCl2) application. Niacin markedly reduced AAA formation in both AngII and CaCl2 models, diminishing adventitial immune cell infiltration, concomitant inflammatory responses, and matrix degradation. Unexpectedly, GPR109A gene deletion did not abrogate the protective effects of niacin against AAA formation, suggesting GPR109A-independent mechanisms. Interestingly, nicotinamide, which does not activate GPR109A, also inhibited AAA formation and phenocopied the effects of niacin. Mechanistically, both niacin and nicotinamide supplementation increased nicotinamide adenine dinucleotide (NAD+) levels and NAD+-dependent Sirt1 activity, which were reduced in AAA tissues. Furthermore, pharmacological inhibition of Sirt1 abrogated the protective effect of nicotinamide against AAA formation., Conclusion: Niacin protects against AAA formation independent of GPR109A, most likely by serving as an NAD+ precursor. Supplementation of NAD+ using nicotinamide-related biomolecules may represent an effective and well-tolerated approach to preventing or treating AAA., (© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society of Cardiology.)

Central vascular thrombosis (CVT) in critically ill neonates carries significant clinical implications. Neonates with congenital heart disease (CHD) awaiting cardiac intervention might be associated with increased risk of thrombosis. Outcome analysis was undertaken. An analysis of 77 neonates with CHD who were admitted to the NICU prior to cardiac intervention between January 2015 and December 2016 was undertaken. Patients requiring extracorporeal life support prior to any cardiac intervention, or receiving prophylactic anticoagulation not related to central vascular catheter (CVC) were excluded. Diagnosis of CVT was provided based on clinical indication and verified with imaging that warranted anticoagulation therapy. Location of CVC and extent of CVT along with treatments, outcomes, and vascular access types and durations were assessed. Logistic regression multivariate analysis was used to assess predictors of outcome. Neonates with CHD were complicated with CVT in 10.4%. Longer duration of CVC was also associated with thrombosis in neonates with CHD (72.7 days vs. 29.3 days, p < 0.001). Independent predictors of outcome included lower gestational age and CHD with single-ventricle (SV) anatomy (p < 0.05). In neonates with CHD awaiting cardiac intervention risk of CVT is substantial. Duration CVC, lower gestational age and SV anatomy are risk determinants of outcome. Standardized development of customized surveillance protocols tailored to this unique subsets of neonates and adherence to quality guidelines can influence outcome.

Retinopathy of prematurity (ROP) is the leading cause of blindness in infants. We have investigated the efficacy of the secondary bile acid ursodeoxycholic acid (UDCA) and its taurine and glycine conjugated derivatives tauroursodeoxycholic acid (TUDCA) and glycoursodeoxycholic acid (GUDCA) in preventing retinal neovascularization (RNV) in an experimental model of ROP. Seven-day-old mice pups (P7) were subjected to oxygen-induced retinopathy (OIR) and were treated with bile acids for various durations. Analysis of retinal vascular growth and distribution revealed that UDCA treatment (50 mg/kg, P7-P17) of OIR mice decreased the extension of neovascular and avascular areas, whereas treatments with TUDCA and GUDCA showed no changes. UDCA also prevented reactive gliosis, preserved ganglion cell survival, and ameliorated OIR-induced blood retinal barrier dysfunction. These effects were associated with decreased levels of oxidative stress markers, inflammatory cytokines, and normalization of the VEGF-STAT3 signaling axis. Furthermore, in vitro tube formation and permeability assays confirmed UDCA inhibitory activity toward VEGF-induced pro-angiogenic and pro-permeability effects on human retinal microvascular endothelial cells. Collectively, our results suggest that UDCA could represent a new effective therapy for ROP.

Background Neurologic complications including hemorrhage, ischemia, and infarction are often identified in neonates undergoing extracorporeal membrane oxygenation (ECMO) and may contribute to the high morbidity observed in ECMO survivors. Screening for intracranial complications is reliant on bedside transcranial ultrasound (CUS) prior to and during ECMO therapy, and advanced imaging [i.e. computed tomography (CT)/magnetic resonance imaging (MRI)] is recommended after completion of ECMO support. The goal of this study is to describe the correlation of intracranial complications identified on CUS during ECMO and MRI after completion of ECMO. Methods Fifty-five neonates underwent ECMO support at the Children's Hospital of Georgia at Augusta University from January 1, 2012 to December 31, 2017. Forty-four (80%) had a brain MRI performed prior to transfer or discharge. Ultrasound studies were reviewed by a single blinded pediatric radiologist and MRIs were reviewed by a single blinded neuro-radiologist. Results Of the 44 neonates with post-ECMO MRI, CUS during ECMO identified intracranial lesions in nine neonates, which were all confirmed on post-ECMO MRI. Sixteen subjects (46%) with unremarkable CUS during ECMO had identifiable lesions on post-ECMO MRI, yielding a sensitivity of 36% and a specificity of 100% for CUS in the detection of intracranial lesions. Despite the lack of correlation between CUS and MR, 84.6% of survivors exhibited normal development at 24 months of age. Conclusion While necessary for the identification of intracranial lesions during neonatal ECMO, CUS demonstrated low correlation with post-ECMO MRI in the identification of intracranial lesions, which supports Extracorporeal Life Support Organization (ELSO) recommendations.

Background: Neonates receiving extracorporeal membrane oxygenation (ECMO) support are transfused large volumes of red blood cells (RBCs) and platelets (PLTs). Transfusions are often administered in response to specific, but largely unstudied thresholds. The aim of this study is to examine the relationship between RBC and PLT transfusion rates and mortality in neonates receiving ECMO support., Study Design and Methods: We retrospectively examined outcomes of neonates receiving ECMO support in the neonatal intensive care unit (NICU) for respiratory failure between 2010 and 2016 at a single quaternary-referral NICU. We examined the association between RBC and PLT transfusion rate (mL per kg per day) and in-hospital mortality, adjusting for confounding by using a validated composite baseline risk score (Neo-RESCUERS)., Results: Among the 110 neonates receiving ECMO support, in-hospital mortality was 28%. The median RBC transfusion rate (mL/kg/d) after cannulation was greater among non-survivors, compared to survivors: 12.4 (IQR 9.3-16.2) versus 7.3 (IQR 5.1-10.3), p < 0.001. Similarly, PLT transfusion rate was greater among non-survivors: 22.9 (9.3-16.2) versus 12.1 (8.4-20.1), p = 0.02. After adjusting for baseline mortality risk, both RBC transfusion (adjusted relative risk per 5 mL/kg/d increase: 1.33; 95% CI 1.05-1.69, p = 0.02) and PLT transfusion (adjusted relative risk per 5 mL/kg/d increase: 1.12; 95% CI 1.02-1.23, p = 0.02) were both associated with in-hospital mortality., Conclusions: RBC and PLT transfusion rates are associated with in-hospital mortality among neonates receiving ECMO. These data provide a basis for future studies evaluating more restrictive transfusion practices for neonates receiving ECMO support., (© 2019 AABB.)

Neurofibromin, the protein product of the neurofibromatosis type 1 (NF1) tumor suppressor gene, is a negative regulator of Ras signaling. Patients with mutations in NF1 have a strong predisposition for cardiovascular disease, which contributes to their early mortality. Nf1 heterozygous (Nf1 +/- ) bone marrow to wild type chimeras and mice with heterozygous recombination of Nf1 in myeloid cells recapitulate many of the vascular phenotypes observed in Nf1 +/- mutants. Although these results suggest that macrophages play a central role in NF1 vasculopathy, the underlying mechanisms are currently unknown. In the present study, we employed macrophages isolated from either Nf1 +/- or Lysm Cre + /Nf1 f/f mice to test the hypothesis that loss of Nf1 stimulates macropinocytosis in macrophages. Scanning electron microscopy and flow cytometry analysis of FITC-dextran internalization demonstrated that loss of Nf1 in macrophages stimulates macropinocytosis. We next utilized various cellular and molecular approaches, pharmacological inhibitors and genetically modified mice to identify the signaling mechanisms mediating macropinocytosis in Nf1-deficient macrophages. Our results indicate that loss of Nf1 stimulates PKCδ-mediated p47 phox phosphorylation via RAS activation, leading to increased NADPH oxidase 2 activity, reactive oxygen species generation, membrane ruffling and macropinocytosis. Interestingly, we also found that Nf1-deficient macrophages internalize exosomes derived from angiotensin II-treated endothelial cells via macropinocytosis in vitro and in the peritoneal cavity in vivo. As a result of exosome internalization, Nf1-deficient macrophages polarized toward an inflammatory M1 phenotype and secreted increased levels of proinflammatory cytokines compared to controls. In conclusion, the findings of the present study demonstrate that loss of Nf1 stimulates paracrine endothelial to myeloid cell communication via macropinocytosis, leading to proinflammatory changes in recipient macrophages., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Responsive parenting interventions that shape parenting behaviors in the areas of sleep and soothing, appropriate and responsive feeding, and routines represent a promising approach to early obesity prevention and have demonstrated effectiveness in our previous trials. However, this approach has yet to be applied to the populations most at-risk for the development of early obesity, including African Americans. The Sleep SAAF (Strong African American Families) study is a two-arm randomized controlled clinical trial evaluating whether a responsive parenting intervention focused on promoting infant sleeping and self-soothing can prevent rapid weight gain during the first 16 weeks postpartum among first-born African American infants. The responsive parenting intervention is compared to a child safety control intervention., Methods: Three hundred first-time African American mothers and their full-term infants will be enrolled from one mother/baby nursery. Following initial screening and consent in the hospital, mothers and infants are visited at home by Community Research Associates for data collection visits at 1 week, 8 weeks, and 16 weeks postpartum and for intervention visits at 3 weeks and 8 weeks postpartum. The primary study outcome is a between-group comparison of infant conditional weight gain (CWG) scores from 3 weeks to 16 weeks; additional weight-related outcomes include differences in change in infants' weight for age over time and differences in infants' weight outcomes at age 16 weeks. Several other outcomes reflecting infant and maternal responses to intervention (e.g., sleeping, soothing, feeding, maternal self-efficacy, maternal depressive symptoms) are also assessed., Discussion: The Sleep SAAF trial can inform efforts to prevent rapid weight gain and reduce risk for obesity early in the lifespan among African Americans., Trial Registration: NCT03505203 . Registered April 3, 2018 in clinicaltrials.gov .

Alterations in perinatal conditions (such as preterm birth) is linked to adult health and disease, in particular, the cardiovascular system. Neddylation, a novel posttranslational modification through which the ubiquitin-like protein NEDD8 is conjugated to protein substrates, has emerged as an important mechanism regulating embryonic cardiac chamber maturation. However, the importance of neddylation in postpartum cardiac development has not been investigated. Here, we aimed to determine whether transient, postnatal inhibition of neddylation has immediate and prolonged impact on the structure and function of the neonatal and adult hearts. Sprague-Dawley pups were given three intraperitoneal injections of MLN4924 (MLN), a specific neddylation inhibitor, at postnatal days (P)1, 3, and 5. Cardiac structure and function were temporally assessed during aging and after 2 wk of isoproterenol (ISO) infusion in adulthood. MLN treatment resulted in modest reduction of neddylated proteins in neonatal hearts. The MLN-treated rats developed cardiac hypertrophy and dysfunction by P7, which was accompanied by significantly reduced cardiomyocyte proliferation. At 3 mo of age, cardiac contractile function was restored in MLN-treated rats, but MLN-treated hearts displayed hypertrophic phenotype. Whereas ISO infusion triggered compensatory cardiac hypertrophy without impairing cardiac contractility in the control rats, the MLN-treated rats displayed a similar degree of hypertrophy, which quickly progressed to decompensation with ventricular wall thinning, chamber dilatation, and reduced ejection fraction as well as exacerbated pathological cardiac remodeling. Our findings suggest that neddylation is required for postnatal cardiac development and that perturbation of neddylation during development predisposes adult hearts to cardiac failure under stress conditions. NEW & NOTEWORTHY Our study demonstrates that perinatal perturbation of neddylation induces cardiomyopathy, impairs postnatal cardiac development, and increases susceptibility to catecholamine-induced cardiac dysfunction. The results reveal a previously unappreciated role of neddylation in postnatal cardiac maturation and call for close monitoring for the potential cardiotoxicity of MLN4924 (pevonedistat) and other agents that modify neddylation, especially in pregnant women and preadolescents.

Background: Perinatal growth has important implications for cardiac development. Low birth weight is associated with cardiovascular (CV) events and mortality, and animal studies have shown that fetal growth restriction is associated with cardiac remodeling in the perinatal period leading to a permanent loss of cardiomyocyte endowment and compensatory hypertrophy., Aims: To determine associations of birthweight (BW) and multiple proportionality indexes (body mass index (BMI); weight/length 2 and Ponderal index (PI); weight/length 3 ) at birth on one hand, with left ventricular (LV) structure and function during adolescence., Subjects: 379 healthy adolescents aged 14-18 years in Augusta, Georgia., Outcome Measures: LV structure and function parameters, including intraventricular septal thickness in diastole (IVSd), LV internal dimension in diastole (LVIDd), LV internal diameter in systole (LVIDs), LV posterior wall thickness in diastole (LVPWd), relative wall thickness (RWT), midwall fractional shortening (MFS), and ejection fraction, were assessed by echocardiography., Results: When associations of birthweight, birth BMI, and birth PI with LV structure and function parameters were separately evaluated with linear regression adjusting for age, sex, race, Tanner stage, socioeconomic status, and physical activity, significant positive associations of BW with LVIDd (P = 0.004), birth BMI with LV mass index (P = 0.01), and birth PI with IVSd (P = 0.02), LVPWd (P = 0.03), and LV mass index (P = 0.002) were identified. When LV structure and function parameters were compared across PI tertiles, a significant U-shaped trend for LV mass index (P quadratic  = 0.04) was identified., Conclusions: Our adolescent data suggest that proportionality at birth may identify associations between perinatal growth and cardiac remodeling independent of birthweight alone., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Purpose: Perivascular adipose tissue (PVAT) surrounds the arterial adventitia and plays an important role in vascular homeostasis. PVAT expands in obesity, and inflamed PVAT can locally promote endothelial dysfunction and atherosclerosis. Here, using adipose tissue transplantation, we tested the hypothesis that expansion of PVAT can also remotely exacerbate vascular disease., Methods: Fifty milligrams of abdominal aortic PVAT was isolated from high-fat diet (HFD)-fed wild-type mice and transplanted onto the abdominal aorta of lean LDL receptor knockout mice. Subcutaneous and visceral adipose tissues were used as controls. After HFD feeding for 10 weeks, body weight, glucose/insulin sensitivity, and lipid levels were measured. Adipocytokine gene expression was assessed in the transplanted adipose tissues, and the thoracic aorta was harvested to quantify atherosclerotic lesions by Oil-Red O staining and to assess vasorelaxation by wire myography., Results: PVAT transplantation did not influence body weight, fat composition, lipid levels, or glucose/insulin sensitivity. However, as compared with controls, transplantation of PVAT onto the abdominal aorta increased thoracic aortic atherosclerosis. Furthermore, PVAT transplantation onto the abdominal aorta inhibited endothelium-dependent relaxation in the thoracic aorta. MCP-1 and TNF-α expression was elevated, while adiponectin expression was reduced, in the transplanted PVAT tissue, suggesting augmented inflammation as a potential mechanism for the remote vascular effects of transplanted PVAT., Conclusions: These data suggest that PVAT expansion and inflammation in obesity can remotely induce endothelial dysfunction and augment atherosclerosis. Identifying the underlying mechanisms may lead to novel approaches for risk assessment and treatment of obesity-related vascular disease.

Background: Maternal nutrient restriction (MNR) is a widespread cause of fetal growth restriction (FGR), an independent predictor of heart disease and cardiovascular mortality. Our objective was to examine the developmental and long-term impact of MNR-induced FGR on cardiac structure in a model that closely mimics human development., Methods: A reduction in total caloric intake spanning pregestation through to lactation in guinea pig sows was used to induce FGR. Proliferation, differentiation, and apoptosis of cardiomyocytes were assessed in late-gestation fetal, neonatal, and adult guinea pig hearts. Proteomic analysis and pathway enrichment were performed on fetal hearts., Results: Cardiomyocyte proliferation and the number of mononucleated cells were enhanced in the MNR-FGR fetal and neonatal heart, suggesting a delay in cardiomyocyte differentiation. In fetal hearts of MNR-FGR animals, apoptosis was markedly elevated and the total number of cardiomyocytes reduced, the latter remaining so throughout neonatal and into adult life. A reduction in total cardiomyocyte number in adult MNR-FGR hearts was accompanied by exaggerated hypertrophy and a disorganized architecture. Pathway analysis identified genes related to cell proliferation, differentiation, and survival., Conclusions: FGR influences cardiomyocyte development during critical windows of development, leading to a permanent deficiency in cardiomyocyte number and compensatory hypertrophy in a rodent model that recapitulates human development.