Breastfeeding is associated with enhanced intestinal gluconeogenesis in infants.

BACKGROUND: Breastfeeding (BF) confers metabolic benefits to infants, including reducing risks of metabolic syndrome such as obesity and diabetes later in life. However, the underlying mechanism is not yet fully understood. Hence, we aim to investigate the impacts of BF on the metabolic organs of infants. METHODS: Previous literatures directly studying the influences of BF on offspring's metabolic organs in both animal models and humans were comprehensively reviewed. A microarray dataset of intestinal gene expression comparing infants fed on breastmilk versus formula milk was analyzed. RESULTS: Reanalysis of microarray data showed that BF is associated with enhanced intestinal gluconeogenesis in infants. This resembles observations in other mammalian species showing that BF was also linked to increased gluconeogenesis. CONCLUSIONS: BF is associated with enhanced intestinal gluconeogenesis in infants, which may underpin its metabolic advantages through finetuning metabolic homeostasis. This observation seems to be conserved across species, hinting its biological significance.

A Deep-Learning Enabled Automatic Fetal Thalamus Diameter Measurement Algorithm.

The analysis of maternal factors that impact the normal development of the fetal thalamus is an emerging field of research and requires the retrospective measurement of fetal thalamus diameter (FTD). Unfortunately, FTD is not measured in routine 2D ultrasound (2D-US) screenings of fetuses. Manual measurement of FTD is a laborious, difficult, and error-prone process because the thalamus lacks well-defined boundaries in 2D-US images of the fetal brain as it has a similar echogenicity to the surrounding brain tissue. Traditional methods based on statistical shape models (SSMs) perform poorly in measuring FTD due to the noisy textures and fuzzy edges of the fetal thalamus in 2D-US images of the fetal brain. To overcome these difficulties, we propose a deep learning-based automatic FTD measurement algorithm, FTDNet. FTDNet measures FTD by learning to directly detect the measurement landmarks through supervised learning. The algorithm first detects the region of the brain that contains the thalamus structure, and then focuses on processing that region for FTD landmark detection. Our FTD dataset, developed through a consensus between two ultrasonographers, contains 1,111 pairs of landmark coordinates for measuring FTD and verified bounding boxes surrounding the fetal thalamus. To assess FTDNet's measurement consistency compared to the ground truth, we used the intraclass correlation coefficient (ICC). FTDNet achieved an ICC score of 0.734, significantly outperforming the prior SSM method and other baseline comparison methods. Our findings are an important step forward in understanding the maternal factors which influence fetal brain development.Clinical relevance- This work proposes an end-to-end thalamus detection and measurement algorithm for measuring fetal thalamus diameter. Our work represents a significant step in the research of how maternal factors can impact fetal thalamus development. The development of an automatic and accurate method for measuring FTD through deep learning has the potential to greatly advance this field of study.

Functional profiling of gut microbial and immune responses toward different types of dietary fiber: a step toward personalized dietary interventions.

Dietary fiber plays a crucial role in maintaining gut and overall health. The objective of this study was to investigate whether different types of dietary fiber elicited specific changes in gut microbiota composition and the production of short-chain fatty acids. To test this, a longitudinal crossover study design was employed, in which healthy adult women consumed three distinct dietary fiber supplements: Inulin (fructo-oligosaccharide), Vitafiber (isomalto-oligosaccharide), and Fibremax (mixture of different fiber) during a one-week intervention period, followed by a 2-week washout period. A total of 15 g of soluble fiber was consumed daily for each supplement. Samples were collected before and after each intervention to analyze the composition of the gut microbiota by 16S rRNA sequencing and fecal levels of short-chain fatty acids measured using nuclear magnetic resonance. Phenotypic changes in peripheral blood mononuclear cells were studied in subsets of participants with higher SCFA levels post-intervention using spectral flow cytometry. The results revealed substantial stability and resilience of the overall gut bacterial community toward fiber-induced changes. However, each supplement had specific effects on gut bacterial alpha and beta diversity, SCFA production, and immune changes. Inulin consistently exerted the most pronounced effect across individuals and certain taxa were identified as potential indicators of SCFA production in response to inulin supplementation. This distinguishing feature was not observed for the other fiber supplements. Further large-scale studies are required to confirm these findings. Overall, our study implies that personalized dietary fiber intervention could be tailored to promote the growth of beneficial bacteria to maximize SCFA production and associated health benefits.

Dysbiotic Gut Microbiota-Derived Metabolites and Their Role in Non-Communicable Diseases.

Dysbiosis, generally defined as the disruption to gut microbiota composition or function, is observed in most diseases, including allergies, cancer, metabolic diseases, neurological disorders and diseases associated with autoimmunity. Dysbiosis is commonly associated with reduced levels of beneficial gut microbiota-derived metabolites such as short-chain fatty acids (SCFA) and indoles. Supplementation with these beneficial metabolites, or interventions to increase their microbial production, has been shown to ameliorate a variety of inflammatory diseases. Conversely, the production of gut 'dysbiotic' metabolites or by-products by the gut microbiota may contribute to disease development. This review summarizes the various 'dysbiotic' gut-derived products observed in cardiovascular diseases, cancer, inflammatory bowel disease, metabolic diseases including non-alcoholic steatohepatitis and autoimmune disorders such as multiple sclerosis. The increased production of dysbiotic gut microbial products, including trimethylamine, hydrogen sulphide, products of amino acid metabolism such as p-Cresyl sulphate and phenylacetic acid, and secondary bile acids such as deoxycholic acid, is commonly observed across multiple diseases. The simultaneous increased production of dysbiotic metabolites with the impaired production of beneficial metabolites, commonly associated with a modern lifestyle, may partially explain the high prevalence of inflammatory diseases in western countries.

Heatwaves and wildfires suffocate our healthy start to life: time to assess impact and take action.

Adverse environmental exposures in utero and early childhood are known to programme long-term health. Climate change, by contributing to severe heatwaves, wildfires, and other natural disasters, is plausibly associated with adverse pregnancy outcomes and an increase in the future burden of chronic diseases in both mothers and their babies. In this Personal View, we highlight the limitations of existing evidence, specifically on the effects of severe heatwave and wildfire events, and compounding syndemic events such as the COVID-19 pandemic, on the short-term and long-term physical and mental health of pregnant women and their babies, taking into account the interactions with individual and community vulnerabilities. We highlight a need for an international, interdisciplinary collaborative effort to systematically study the effects of severe climate-related environmental crises on maternal and child health. This will enable informed changes to public health policy and clinical practice necessary to safeguard the health and wellbeing of current and future generations.

Infection and Vaccine Induced Spike Antibody Responses Against SARS-CoV-2 Variants of Concern in COVID-19-Naïve Children and Adults.

Although a more efficient adaptive humoral immune response has been proposed to underlie the usually favorable outcome of pediatric COVID-19, the breadth of viral and vaccine cross-reactivity toward the ever-mutating Spike protein among variants of concern (VOCs) has not yet been compared between children and adults. We assessed antibodies to conformational Spike in COVID-19-naïve children and adults vaccinated by BNT162b2 and ChAdOx1, and naturally infected with SARS-CoV-2 Early Clade, Delta, and Omicron. Sera were analyzed against Spike including naturally occurring VOCs Alpha, Beta, Gamma, Delta, and Omicron BA.1, BA.2, BA.5, BQ.1.1, BA2.75.2, and XBB.1, and variants of interest Epsilon, Kappa, Eta, D.2, and artificial mutant Spikes. There was no notable difference between breadth and longevity of antibody against VOCs in children and adults. Vaccinated individuals displayed similar immunoreactivity profiles across variants compared with naturally infected individuals. Delta-infected patients had an enhanced cross-reactivity toward Delta and earlier VOCs compared to patients infected by Early Clade SARS-CoV-2. Although Omicron BA.1, BA.2, BA.5, BQ.1.1, BA2.75.2, and XBB.1 antibody titers were generated after Omicron infection, cross-reactive binding against Omicron subvariants was reduced across all infection, immunization, and age groups. Some mutations, such as 498R and 501Y, epistatically combined to enhance cross-reactive binding, but could not fully compensate for antibody-evasive mutations within the Omicron subvariants tested. Our results reveal important molecular features central to the generation of high antibody titers and broad immunoreactivity that should be considered in future vaccine design and global serosurveillance in the context of limited vaccine boosters available to the pediatric population.

Diet, commensal microbiota-derived extracellular vesicles, and host immunity.

The gut microbiota has co-evolved with its host, and commensal bacteria can influence both the host's immune development and function. Recently, a role has emerged for bacterial extracellular vesicles (BEVs) as potent immune modulators. BEVs are nanosized membrane vesicles produced by all bacteria, possessing the membrane characteristics of the originating bacterium and carrying an internal cargo that may include nucleic acid, proteins, lipids, and metabolites. Thus, BEVs possess multiple avenues for regulating immune processes, and have been implicated in allergic, autoimmune, and metabolic diseases. BEVs are biodistributed locally in the gut, and also systemically, and thus have the potential to affect both the local and systemic immune responses. The production of gut microbiota-derived BEVs is regulated by host factors such as diet and antibiotic usage. Specifically, all aspects of nutrition, including macronutrients (protein, carbohydrates, and fat), micronutrients (vitamins and minerals), and food additives (the antimicrobial sodium benzoate), can regulate BEV production. This review summarizes current knowledge of the powerful links between nutrition, antibiotics, gut microbiota-derived BEV, and their effects on immunity and disease development. It highlights the potential of targeting or utilizing gut microbiota-derived BEV as a therapeutic intervention.

Patterns of parental online health information-seeking behaviour.

AIM: This study aimed to understand parents' online health information-seeking behaviour and the potential influence of this on their relationship with their child's physician. METHODS: A survey regarding parental online health information-seeking behaviour was administered to parents of children aged under 18 years admitted to an Australian tertiary paediatric hospital, paediatric hospital ward and paediatric clinic, and in their social media networks. Responses were presented as frequencies and percentages. Associations between parents' trust in their child's doctor and survey responses were analysed using χ2 tests. RESULTS: In all, 300 surveys were completed. Most parents (89%) reported searching for online health information when their child was sick. Some (31%) followed online health information instead of going to the doctor. Parents who trusted their child's doctor were more likely to follow the doctor instead of online health information when it contained conflicting advice. Most parents (91%) wanted health-care professionals' help in searching for online health information. CONCLUSION: Almost all parents search for online health information, but most do not act on it. Parents' trust in their child's doctor influences how parents use online health information. Thus, clinicians could recommend trustworthy websites with information that complements their advice to ensure parents access reliable online health information.

Optimal low-cost cooling strategies for infant strollers during hot weather.

The current study aimed to identity the optimal low-cost stroller cooling strategies for use in hot and moderately humid summer weather. A commercially available stroller was instrumented to assess the key parameters of the thermal environment. The cooling efficacy of eight different stroller configurations was examined in a counterbalanced order across 16 hot summer days (air temperature (Ta) = 33.3 ± 4.1 °C; relative humidity = 36.7 ± 15%; black globe temperature = 43.9 ± 4.6 °C). Compared with a standard-practice stroller configuration, combining a moist muslin draping with a battery-operated clip-on fan provided optimal in-stroller cooling, reducing the end-trial air temperature by 4.7 °C and the wet bulb globe temperature (WBGT) by 1.4 °C. In contrast, in-stroller temperatures were substantially increased by draping a dry muslin (Ta = +2.6 °C; WBGT = +0.9 °C) or flannelette (Ta = +3.7 °C; WBGT = +1.4 °C) cloth over the stroller carriage. These findings provide empirical evidence which may inform guidance aimed at protecting infants during hot weather.Practitioner summary: This study examined the efficacy of traditional and novel stroller cooling strategies for use in hot and moderately humid weather. Covering the carriage with a dry muslin cloth substantially increased stroller temperatures and should be avoided. Evaporative cooling methods reduced in-stroller temperatures. A moist muslin cloth draping combined with a fan provided optimal stroller cooling.

Cerebral and splanchnic near-infrared spectroscopic dataset in premature newborns receiving packed red blood cell transfusion.

This article presents the near-infrared spectroscopy (NIRS) dataset of cerebral (StO2c) and splanchnic (StO2s) oxygenation in 29 stable premature infants admitted to a tertiary neonatal intensive care unit who received elective packed red blood cell transfusion (PRBCT) to treat anemia of prematurity. StO2c and StO2s data were prospectively recorded continuously from at least 4 hours before the beginning of PRBCT until 24 hours after its completion, using a 4-wavelength near-infrared spectroscopy (NIRS) monitor (FORE-SIGHT® absolute cerebral oximeter, CASMED, Branford, Connecticut, 06405 USA). StO2 data were downloaded as an analog output at a sampling rate of 1000Hz and aligned along the time axis in LabChart reader format (.adicht files) using a PowerLab data acquisition system [1] (PowerLab®, ADInstruments, Sydney, Australia). The .adicht files were then converted into .mat file format using a Python script (PythonTM version 3.7.3 [2]) and resampled at 1Hz for faster processing. Data that could not be physiologically explained (e.g., the absence of variability, [3] a 30% step change in StO2 between two subsequent data points for StO2[4]), as well as the data during the period of 'cares' were presumed to be artefactual and were replaced with 'NaN' or 'Not a Number' which is recognised by Matlab [5] (MATLAB 9.3, The MathWorks, Inc., Massachusetts, United States) and ignored for all subsequent processing while maintaining the correct time point of the StO2 signals. The data were then exported into Microsoft Excel format. The splanchnic cerebral oxygenation ratio (SCOR) was calculated as the ratio of StO2s/StO2c. A 4-hour mean pre-transfusion values (StO2s 0, StO2c 0, SCOR 0) and post-transfusion hourly mean values (1-28) were determined. Secondary data were derived from a Mixed Models for Repeated Measures (MMRM) analysis with the time point fitted as a fixed effect and the infant fitted as a random effect. The MMRM was used to perform paired comparisons between pre-transfusion and each of the post-baseline values. This article only provides the NIRS data. The secondary data and demography can be found in the article "Splanchnic-Cerebral Oxygenation Ratio associated with Packed Red Blood Cell Transfusion in preterm infants", published in Transfusion Medicine. [6] The data will be of use to researchers in neonatology, transfusion medicine, and physiology to understand changes in cerebral and splanchnic oxygenation associated with PRBCT. Data collection, processing, and analysis can be remodelled in larger multicentric randomised controlled studies to evaluate the effect of transfusion and feeding on transfusion-associated necrotising enterocolitis. The data are also helpful to explore the autoregulatory behaviour of the brain and gut when the oxygen content of blood is increased by administering PRBCT.

Tracking the clonal dynamics of SARS-CoV-2-specific T cells in children and adults with mild/asymptomatic COVID-19.

Children infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) develop less severe coronavirus disease 2019 (COVID-19) than adults. The mechanisms for the age-specific differences and the implications for infection-induced immunity are beginning to be uncovered. We show by longitudinal multimodal analysis that SARS-CoV-2 leaves a small footprint in the circulating T cell compartment in children with mild/asymptomatic COVID-19 compared to adult household contacts with the same disease severity who had more evidence of systemic T cell interferon activation, cytotoxicity and exhaustion. Children harbored diverse polyclonal SARS-CoV-2-specific naïve T cells whereas adults harbored clonally expanded SARS-CoV-2-specific memory T cells. A novel population of naïve interferon-activated T cells is expanded in acute COVID-19 and is recruited into the memory compartment during convalescence in adults but not children. This was associated with the development of robust CD4+ memory T cell responses in adults but not children. These data suggest that rapid clearance of SARS-CoV-2 in children may compromise their cellular immunity and ability to resist reinfection.

PsyNBIOsis: Investigating the Association between Maternal Gestational Diabetes, Mental Health, Diet and Childhood Obesity Risk: Protocol for a Prospective, Longitudinal, Observational Study.

BACKGROUND: Gestational diabetes mellitus (GDM) is associated with poorer maternal mental health (depression and anxiety). Maternal mental health and GDM are likely to influence diet, which in turn impacts the course of GDM. Maternal diet may also be directly or indirectly associated with changes in infant anthropometry. The aims of this study are to (1) examine the associations between maternal GDM, mental health and diet, and (2) evaluate the associations between these maternal factors, breastmilk composition and infant anthropometry. METHODS: This prospective, observational, longitudinal cohort study compares a cohort of women with and without GDM. Maternal mental health and diet are assessed using validated questionnaires. Breastmilk composition is measured with the Human Milk Analyzer, and infant body composition is measured with air displacement plethysmography. SIGNIFICANCE AND IMPACT: Once data have been collected, PsyNBIOsis will provide evidence for the associations between maternal mental health, GDM status and diet, and their impact on breastmilk composition and early infant growth. The results may inform the Developmental Origins of Health and Disease framework and provide data on which to build cost-effective interventions to prevent both the development of mental health issues in mothers and adverse growth patterns in infants.

Bushfires and Mothers' Mental Health in Pregnancy and Recent Post-Partum.

BACKGROUND: The compounding effects of climate change catastrophes such as bushfires and pandemics impose significant burden on individuals, societies, and their economies. The enduring effects of such syndemics on mental health remain poorly understood, particularly for at-risk populations (e.g., pregnant women and newborns). The aim of this study was to investigate the impact of direct and indirect exposure to the 2019/20 Australian Capital Territory and South-Eastern New South Wales bushfires followed by COVID-19 on the mental health and wellbeing of pregnant women and mothers with newborn babies. METHODS: All women who were pregnant, had given birth, or were within three months of conceiving during the 2019/2020 bushfires, lived within the catchment area, and provided consent were invited to participate. Those who consented were asked to complete three online surveys. Mental health was assessed with the DASS-21 and the WHO-5. Bushfire, smoke, and COVID-19 exposures were assessed by self-report. Cross-sectional associations between exposures and mental health measures were tested with hierarchical regression models. RESULTS: Of the women who participated, and had minimum data (n = 919), most (>75%) reported at least one acute bushfire exposure and 63% reported severe smoke exposure. Compared to Australian norms, participants had higher depression (+12%), anxiety (+35%), and stress (+43%) scores. Women with greater exposure to bushfires/smoke but not COVID-19 had poorer scores on all mental health measures. CONCLUSIONS: These findings provide novel evidence that the mental health of pregnant women and mothers of newborn babies is vulnerable to major climate catastrophes such as bushfires.

Splanchnic-cerebral oxygenation ratio associated with packed red blood cell transfusion in preterm infants.

BACKGROUND: Splanchnic-cerebral oxygenation ratio (SCOR), the ratio of splanchnic tissue oxygen (StO2 s) to simultaneously measured cerebral tissue oxygen (StO2 c), has been described as a surrogate to detect impaired splanchnic oxygenation associated with hypoperfusion status such as necrotizing enterocolitis. This concept is based on the presumption that any change in SCOR indicates a corresponding change in splanchnic tissue oxygenation as the numerator, whereas cerebral tissue oxygenation as the denominator remains stable. However, it is questionable to utilise this concept to detect splanchnic oxygenation changes in the context of packed red blood cell transfusion (PRBCT). AIM: The current study examines the contribution of both cerebral and splanchnic oxygenation components to PRBCT-associated SCOR changes in preterm infants. DESIGN: Prospective cohort study. SETTING: Neonatal intensive care. PATIENTS: Hemodynamically stable infants: Gestation <32 weeks; birth weight <1500 g; postmenstrual age <37 weeks: tolerating ≥120 ml/kg/day feed volume. INTERVENTIONS: PRBCT at 15 ml/kg, over 4 h. MAIN OUTCOME MEASURES: Transfusion-associated changes were determined by performing mixed models for repeated measures analysis between the 4-h mean pre-transfusion values (SCOR 0, StO2 s 0, and StO2 c 0) and the post-transfusion hourly mean values for the next 28 h (SCOR 1-28, StO2 s 1-28, and StO2 c 1-28). Dunnett's method was used to adjust for the multiplicity of the p value. RESULTS: Of 30 enrolled infants 14 [46.7%] male; median [IQR] birth weight, 923 [655-1064] g; gestation, 26.4 [25.5-28.1] weeks; enrolment weight, 1549 [1113-1882] g; and postmenstrual age, 33.6 [32.4-35.0] weeks, one infant was excluded because of corrupted NIRS data. With the commencement of PRBCT, SCOR demonstrated a downward trend throughout the study period. This drift was associated with an increasing StO2 c trend, while StO2 s remained unchanged throughout the study period. CONCLUSIONS AND RELEVANCE: PRBCT-associated SCOR decrease suggests improvement in cerebral oxygenation rather than worsening splanchnic oxygenation. Our study underlines that it is necessary to determine individual components of SCOR, namely cerebral and splanchnic StO2 to understand SCOR changes in the context of PRBCT.

Periodontitis and Preeclampsia in Pregnancy: A Systematic Review and Meta-Analysis.

OBJECTIVES: A conflicting body of evidence suggests localized periodontal inflammation spreads systemically during pregnancy inducing adverse pregnancy outcomes. This systematic review and meta-analysis aim to specifically evaluate the relationship between periodontitis and preeclampsia. METHODS: Electronic searches were carried out in Medline, Pubmed, Embase, Lilacs, Cochrane Controlled Clinical Trial Register, CINAHL, ClinicalTrials.gov, and Google Scholar with no restrictions on the year of publication. We identified and selected observational case-control and cohort studies that analyzed the association between periodontal disease and preeclampsia. This meta-analysis was conducted following the PRISMA checklist and MOOSE checklist. Pooled odds ratios, mean difference, and 95% confidence intervals were calculated using the random effect model. Heterogeneity was tested with Cochran's Q statistic. RESULTS: Thirty studies including six cohort- and twenty-four case-control studies were selected. Periodontitis was significantly associated with increased risk for preeclampsia (OR 3.18, 95% CI 2.26 - 4.48, p < 0.00001), especially in a subgroup analysis including cohort studies (OR 4.19, 95% CI 2.23 - 7.87, p < 0.00001). The association was even stronger in a subgroup analysis with lower-middle-income countries (OR 6.70, 95% CI 2.61 - 17.19, p < 0.0001). CONCLUSIONS: Periodontitis appears as a significant risk factor for preeclampsia, which might be even more pronounced in lower-middle-income countries. Future studies to investigate if maternal amelioration of periodontitis prevents preeclampsia might be warranted.

Hierarchical improvement of regional tissue oxygenation after packed red blood cell transfusion.

BACKGROUND: It is well established that counter-regulation to hypoxia follows a hierarchical pattern, with brain-sparing in preference to peripheral tissues. In contrast, it is unknown if the same hierarchical sequence applies to recovery from hypoxia after correction of anemia with packed red blood cell transfusion (PRBCT). OBJECTIVE: To understand the chronology of cerebral and splanchnic tissue oxygenation resulting after correction of anemia by PRBCT in preterm infants using near-infrared spectroscopy (NIRS). DESIGN: Prospective cohort study. SETTING: Neonatal intensive care. PATIENTS INCLUDED: Haemodynamically stable infants: <32 weeks gestation, <37weeks postmenstrual age, <1500 grams birth weight; and ≥120 mL/kg/day feeds tolerated. INTERVENTION: PRBCT at 15 mL/Kg over 4 hours. MAIN OUTCOME MEASURES: Transfusion-associated changes were determined by comparing the 4-hour mean pre-transfusion cerebral and splanchnic fractional tissue oxygen extraction (FTOEc0; FTOEs0) with hourly means during (FTOEc1-4; FTOEs1-4) and for 24 hours after PRBCT completion (FTOEc5-28; FTOEs5-28). RESULTS: Of 30 enrolled infants, 14[46.7%] male; median[IQR] birth weight, 923[655-1064]g; gestation, 26.4[25.5-28.1]weeks; enrolment weight, 1549[1113-1882]g; and postmenstrual age, 33.6[32.4-35]weeks, 1 infant was excluded because of corrupted NIRS data. FTOEc significantly decreased during and for 24 hours after PRBCT (p < 0.001), indicating prompt improvement in cerebral oxygenation. In contrast, FTOEs showed no significant changes during and after PRBCT (p>0.05), indicating failure of improvement in splanchnic oxygenation. CONCLUSION: Improvement in regional oxygenation after PRBCT follows the same hierarchical pattern with a prompt improvement of cerebral but not splanchnic tissue oxygenation. We hypothesise that this hierarchical recovery may indicate continued splanchnic hypoxia in the immediate post-transfusion period and vulnerability to transfusion-associated necrotizing enterocolitis (TANEC). Our study provides a possible mechanistic underpinning for TANEC and warrants future randomised controlled studies to stratify its prevention.

Dietary protein increases T-cell-independent sIgA production through changes in gut microbiota-derived extracellular vesicles.

Secretory IgA is a key mucosal component ensuring host-microbiota mutualism. Here we use nutritional geometry modelling in mice fed 10 different macronutrient-defined, isocaloric diets, and identify dietary protein as the major driver of secretory IgA production. Protein-driven secretory IgA induction is not mediated by T-cell-dependent pathways or changes in gut microbiota composition. Instead, the microbiota of high protein fed mice produces significantly higher quantities of extracellular vesicles, compared to those of mice fed high-carbohydrate or high-fat diets. These extracellular vesicles activate Toll-like receptor 4 to increase the epithelial expression of IgA-inducing cytokine, APRIL, B cell chemokine, CCL28, and the IgA transporter, PIGR. We show that succinate, produced in high concentrations by microbiota of high protein fed animals, increases generation of reactive oxygen species by bacteria, which in turn promotes extracellular vesicles production. Here we establish a link between dietary macronutrient composition, gut microbial extracellular vesicles release and host secretory IgA response.

PrEggNut Study: protocol for a randomised controlled trial investigating the effect of a maternal diet rich in eggs and peanuts from <23 weeks' gestation during pregnancy to 4 months' lactation on infant IgE-mediated egg and peanut allergy outcomes.

INTRODUCTION: Clinical studies supported by immunological data indicate early life intervention strategies to be promising in reducing the growing global burden of food allergies. The events that predispose to food allergy, including the induction of allergen-specific immune responses, appear to be initiated early in development. Early exposure to food allergens in utero and via breast milk is likely to be important in initiating oral tolerance. We aim to determine the effectiveness of higher maternal food allergen consumption during pregnancy and lactation on infant food allergy outcomes. METHODS AND ANALYSIS: This is a multisite, parallel, two-arm (1:1 allocation), single-blinded (outcome assessors, statistical analyst and investigators), randomised controlled trial. Pregnant women (<23 weeks' gestation) whose (unborn) infants have at least two biological family members (mother, father or siblings) with medically diagnosed allergic disease are eligible to participate. After obtaining written informed consent, pregnant women are randomised to either a high egg and peanut diet (at least 6 eggs and 60 peanuts per week) or standard (low) egg and peanut diet (no more than 3 eggs and 30 peanuts per week). The women are asked to follow their allocated diet from <23 weeks' gestation to 4 months' lactation. The primary outcome is food challenge proven IgE-mediated egg and/or peanut allergy in the infants at 12 months of age. Key secondary outcomes include infant sensitisation to egg and/or peanut and infant eczema. Our target sample size is 2136 women. Analyses will be performed on an intention-to-treat basis according to a pre-specified statistical analysis plan. ETHICS AND DISSEMINATION: Ethical approval has been granted from the Women's and Children's Health Network Human Research Ethics Committee (approval number HREC/18/WCHN/42). Trial results will be presented at scientific conferences and published in peer-reviewed journals. TRIAL REGISTRATION NUMBER: Australian New Zealand Clinical Trials Registry ACTRN12618000937213.