Oral supplementation with probiotics, prebiotics, and synbiotics is a novel potential complementary therapy for addressing overweight and obesity through gut microbiota modulation. This systematic review provides a comprehensive summary of the existing evidence to guide future research. Literature searches were conducted in four databases to identify human trials published until May 2024 that examined the impact of probiotic, prebiotic, or synbiotic interventions on faecal microbiota composition changes in overweight and obese participants from Latin American and Caribbean populations (LACPs). Of the 13,090 identified records, five randomised controlled trials (RCTs) from Brazil, Mexico, and Chile met the inclusion criteria for this review. The included RCTs evaluated different forms of therapies over short-term interventions (6 or 8 weeks), with sample sizes ranging from 21 to 39 participants across the studies. Variations in the reported outcomes were observed due to differences in supplement formulation, dosage, population characteristics, and methodological heterogeneity. The findings indicate that the available data are inadequate to establish definitive conclusions regarding the impact of biotic treatments on gut microbiota profiles in LACP. Further research with larger sample sizes and precise microbiota analysis is required to elucidate the implications of dietary interventions on gut microbiota in obesity and related disorders.

The coronavirus disease 2019 (COVID-19) pandemic has caused health issues worldwide. Studies have suggested that modulation of the gut microbiota could attenuate the severity of COVID-19 symptoms. In light of this, we explored the effects of the prebiotic dietary fibre partially hydrolyzed guar gum (PHGG) on SARS-CoV-2 infection in a Syrian hamster model, hypothesizing that modulation of the gut microbiome and intestinal metabolites through PHGG administration would improve COVID-19 disease outcomes. Eight hamsters each were assigned to the PHGG administration and control groups. The PHGG group was given a diet supplemented with 5% PHGG for two weeks. Consequently, PHGG improved the host survival rate to 100% compared to 25% of the control group (P = 0.003) and attenuated morbid weight loss. Another non-infected set of hamsters was used for the analysis of the gut microbiome composition with 16S rRNA amplicon sequencing, serum, and faecal metabolites with GC–MS and LC–MS. PHGG altered the gut microbiome composition and increased the relative abundances of Ileibacterium, Bifidobacterium, and Prevotella. Furthermore, it elevated the concentrations of faecal valeric acid, propionic acid, ursodeoxycholic acid, and serum deoxycholic acid. Taken together, our data suggest that the prebiotic PHGG modulates gut metabolites and has the potential to reduce COVID-19 morbidity.

With the increase in egg production rate and the coming of peak laying period, some metabolic disorders usually emerge in layers. The current study was conducted to compare the physiological difference between the early laying stage (around 30% laying rate) and peak laying stage (more than 95% laying rate) of laying hens based on hepatic transcriptome, serum metabolomics and caecal microbiota. The results showed that the egg weight and yolk weight were significantly higher in peak laying hens. Further, serum malondialdehyde and total bile acid concentrations were higher, but total anti-oxidant capacity, total bilirubin and low-density lipoprotein cholesterol (LDL-c) concentrations were significantly lower in peak laying hens. Hepatic transcriptome analysis identified 540 up-regulated and 269 down-regulated genes. Consistently, fatty acid biosynthesis, PPAR and insulin signalling pathways were significantly enriched. Subsequently, the result of serum metabolomics identified 74 up- and 77 down-regulated metabolites. Among down-regulated metabolites, hesperetin, apigenin and betaine related to anti-oxidant function were down-regulated. In addition, western blotting result showed BCL2 and p53 proteins expressions were decreased in the peak laying period, whereas hepatic CEBPα protein level was increased. On the other hand, gut microbiota analysis revealed that Chao index was decreased in peak laying hens. And the LEfSe analysis showed the dominant microflora including Ruminococcus, Oxalobacter, Paracoccus and so on was found in peak laying hens. These findings indicated that the hepatic lipid metabolism of peak laying hens is enhanced and the decline in anti-oxidant performance of hens also implies its importance during the early stage of egg production.

The latest Yakult Science Study Day was held virtually on 2 November 2023. Aimed at healthcare professionals, researchers, and students, a variety of experts explored the latest gut microbiome research and what it means in practice. The morning sessions discussed the role of the microbiome in health and disease, the rapid advancements in DNA sequencing and implications for personalised nutrition, the current state of evidence on health benefits associated with fermented foods, prebiotics and probiotics and the challenges involved in interpreting research in this area. The afternoon session considered the emerging research on the microbiota–gut–brain axis in mediating effects of food on mood, the bidirectional impact of menopause on the gut microbiota, and the interplay between the gut and skin with implications for the treatment of rare and common skin disorders. The session ended with an update on the use of faecal microbiota transplant in both research and clinical practice. Undoubtedly, the gut microbiome is emerging as a key conductor of human health, both in relation to gastrointestinal and non-gastrointestinal outcomes. As research continues to elucidate mechanisms of action and confirm their effects in human trials, the gut microbiome should be a key consideration within a holistic approach to health moving forward.

The gut microbiome is impacted by certain types of dietary fibre. However, the type, duration and dose needed to elicit gut microbial changes and whether these changes also influence microbial metabolites remain unclear. This study investigated the effects of supplementing healthy participants with two types of non-digestible carbohydrates (resistant starch (RS) and polydextrose (PD)) on the stool microbiota and microbial metabolite concentrations in plasma, stool and urine, as secondary outcomes in the Dietary Intervention Stem Cells and Colorectal Cancer (DISC) Study. The DISC study was a double-blind, randomised controlled trial that supplemented healthy participants with RS and/or PD or placebo for 50 d in a 2 × 2 factorial design. DNA was extracted from stool samples collected pre- and post-intervention, and V4 16S rRNA gene sequencing was used to profile the gut microbiota. Metabolite concentrations were measured in stool, plasma and urine by high-performance liquid chromatography. A total of fifty-eight participants with paired samples available were included. After 50 d, no effects of RS or PD were detected on composition of the gut microbiota diversity (alpha- and beta-diversity), on genus relative abundance or on metabolite concentrations. However, Drichlet’s multinomial mixture clustering-based approach suggests that some participants changed microbial enterotype post-intervention. The gut microbiota and fecal, plasma and urinary microbial metabolites were stable in response to a 50-d fibre intervention in middle-aged adults. Larger and longer studies, including those which explore the effects of specific fibre sub-types, may be required to determine the relationships between fibre intake, the gut microbiome and host health.

To investigate the associations between dietary patterns and biological ageing, identify the most recommended dietary pattern for ageing and explore the potential mediating role of gut microbiota in less-developed ethnic minority regions (LEMRs). This prospective cohort study included 8288 participants aged 30–79 years from the China Multi-Ethnic Cohort study. Anthropometric measurements and clinical biomarkers were utilised to construct biological age based on Klemera and Doubal’s method (KDM-BA) and KDM-BA acceleration (KDM-AA). Dietary information was obtained through the baseline FFQ. Six dietary patterns were constructed: plant-based diet index, healthful plant-based diet index, unhealthful plant-based diet index, healthy diet score, Dietary Approaches to Stop Hypertension (DASH), and alternative Mediterranean diets. Follow-up adjusted for baseline analysis assessed the associations between dietary patterns and KDM-AA. Additionally, quantile G-computation identified significant beneficial and harmful food groups. In the subsample of 764 participants, we used causal mediation model to explore the mediating role of gut microbiota in these associations. The results showed that all dietary patterns were associated with KDM-AA, with DASH exhibiting the strongest negative association (β = −0·91, 95 % CI (–1·19, −0·63)). The component analyses revealed that beneficial food groups primarily included tea and soy products, whereas harmful groups mainly comprised salt and processed vegetables. In mediation analysis, the Synergistetes and Pyramidobacter possibly mediated the negative associations between plant-based diets and KDM-AA (5·61–9·19 %). Overall, healthy dietary patterns, especially DASH, are negatively associated with biological ageing in LEMRs, indicating that Synergistetes and Pyramidobacter may be potential mediators. Developing appropriate strategies may promote healthy ageing in LEMRs.

Newborn calf diarrhea has led to widespread overuse of antibiotics. Therefore, it is crucial to find effective solutions for calf diarrhea. In this study, we aimed to evaluate the impact of the synthetic organic zinc-chelating-peptide glycine-glutamine-Zn (GQ-Zn) on the microbiota and metabolites in the gut of calves with diarrhea. The results showed that GQ-Zn alleviated diarrhea in calves. Additionally, 16S rDNA sequencing and metabolomic analysis revealed that GQ-Zn improved antioxidant capacity, relieved inflammation, altered the gut microbiota by decreasing the number of harmful bacteria Prevotella denticola, Fusobacterium necrophorum and influenced metabolomic profiles via the linoleic acid metabolic pathway in calves. In conclusion, GQ-Zn supplementation alleviated diarrhea through regulating the gut microbiota and metabolites in pre-weaning Holstein calves.

Calorie restriction plays a role in reducing food intake and weight gain, and improving health and lifespan. We hypothesized that calorie restriction would affect body weight (BW), serum indices, gut microbiota, metabolites and short-chain fatty acids of finishing pigs. Castrated male (Landrace × Yorkshire) pigs (86.13 ± 3.50 kg) were randomly assigned into two groups indicated as control (Con) and calorie restriction (CR) (eight pigs/group), respectively. Pigs in the Con group consumed feed ad libitum, whereas pigs in the CR group were fed 70% of the amount of feed in the Con group. The trial lasted for 38 days. Blood and colonic contents were collected for serum parameters, and microbiota and metabolome analysis, respectively. Main effects were tested by Student’s t-test. We found that for finishing pigs, calorie restriction reduced the cumulative food intake, BW gain, serum total cholesterol, triglyceride, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, alanine aminotransferase and aspartate aminotransferase levels. Calorie restriction did not change the α and β diversity of intestinal microbiota. However, calorie restriction significantly increased the abundance of Romboutsia and unclassified_c_Bacilli, and significantly reduced the abundance of Lachnospiraceae_XPB1014_group, Candidatus_Saccharimonas, Escherichia-–Shigella and Gastranaerophilales. Calorie restriction also simultaneously changed the structure of intestinal metabolites and increased the concentration of isobutyric acid, isovaleric acid and valeric acid. In conclusion, calorie restriction may affect metabolism, reduce obesity and improve intestinal microbiota, which may be a healthy diet treatment that can reduce obesity and improve metabolism.

This study investigated the impact of regular consumption of fermented vegetables (FVs) on inflammation and the composition of the gut microbiota in adults at increased risk for cardiovascular disease. Eighty-seven adults ages 35–64 were randomized into an FV group, who consumed 100 g FVs daily at least five times per week for eight weeks, or a usual diet (UD) group. Blood and stool samples were obtained before and after the intervention. Dependent samples t tests and adjusted linear models were used for within- and between-group comparisons. The mean age and body mass index of participants were 45 years and 30 kg/m2, and 80% were female. Bloating or gas was the most common side effect reported (19.3% FV group vs. 9.4% UD group). There were no changes in C-reactive protein, oxidized low-density lipoprotein-receptor 1, angiopoietin-like protein 4, trimethylamine oxide, and lipopolysaccharide-binding protein or bacterial alpha diversity between groups. Our findings indicate that consuming 100 g of FVs for at least five days per week for eight weeks does not change inflammatory biomarkers or microbial alpha diversity as measured by the Shannon index. It is possible that higher doses of FVs are necessary to elicit a significant response by gut bacteria.

Hospital placement is essential training for medical interns, involving shift work and high-pressure environments. This can increase physiological and psychological stress, which may be mediated by metabolites of microbial digestion(1). Nutrients of interest include those accessible to microbial digestion and associated with altered signalling within the microbiota-gut-brain axis (MGBA)(1). Fibre is fermented by gut microbes to produce short-chain fatty acids(2) and is associated with improved psychological outcomes(3). Tryptophan, a precursor to gut-derived serotonin(2), has been negatively associated with anxiety(4). Processed foods contain food additives, excess sugars, and saturated fats that may disrupt gut homeostasis(1) and impact psychological well-being(4). Lastly, total energy intake may determine the level of substrate available for microbial fermentation(2). Therefore, this research explores how microbiota-accessible food components interact with physical and psychological well-being in a cohort of medical interns undertaking their first-year of hospital placement. Participants were healthy medical interns, during first-year hospital placement (n = 21) from the Hunter New England Local Health District, NSW, Australia. Participants completed diet and wellbeing surveys at baseline and every 2 months over a 10-month period. 24-hour diet diaries were self-recorded from participants using a mobile application (Easy Diet Diary) and analysed using AusNut and the NOVA classification system of ultra-processed foods (ULP). Wellbeing surveys include depression, anxiety, stress scale (DASS), and PROMIS survey for mental (M), physical (P), and sleep well-being. Current data represents an ‘in-progress’ of the longitudinal data collection. This study utilised Spearman correlation and Tukey’s post hoc test for mixed methods analysis. From baseline to timepoint 3 (T3, 4 months) daily energy intake was consistent with cohort estimated energy requirements (EER). However, consumption ranged from 37% to 167% of EER, indicating a large variation of intakes. Energy consumed from ULP ranged from 30% to 34% (p = 0.6875). Baseline tryptophan intake (x¯ = 1139mg) was within the suggested target, whilst fibre intake (x¯ = 23g) was below the recommended intake. Neither saw significant changes from baseline to T3. Fibre intake was positively correlated with mental and physical well-being at baseline (x¯ = 23.1g, M: r = 0.474, p = 0.04, P: r = 0.608, p = 0.007), and timepoint 2 (x¯ = 31.5g, M: r = 0.647,p = 0.026, P: r = 0.780, p = 0.004) but not at T3. In addition, baseline consumption of sugar (x¯ = 18g) and poly-unsaturated fats (x¯ = 15g) were both negatively correlated with mental and physical well-being. Overall, no significant dietary changes were evident from baseline to mid-year collection in a first-year medical intern cohort during hospital placements. Fibre was significantly associated with mental and physical well-being, building on current understanding of fibre’s role in the MGBA. Planned metabolite analysis will explore the mechanisms of proposed microbiome-accessible nutrients alongside diet, well-being, and microbiota data. Findings from this study will identify how diet-microbiome interactions change under stress, with wider positive implications on intense workplace environments with the aim to preserve individual wellbeing.

MicroRNAs (miRNAs) have emerged as important regulators of lipid metabolism. Recent studies have suggested synbiotics may modulate miRNA expression and lipid metabolism. This study aimed to investigate the effects of synbiotic supplementation on circulating miR-27a, miR-33a and lipid parameters in patients with dyslipidaemia. Fifty-six eligible participants were randomly allocated to receive either synbiotic or placebo sachets twice a day for 12 weeks. Each synbiotic sachet contained 3 × 1010 colony forming unit six species of probiotic microorganisms and 5 g of inulin and fructooligosaccharide as prebiotics. Serum miR-27a and miR-33a expression levels, serum lipids and apolipoproteins, the fecal concentration of short-chain fatty acids (SCFA) and Firmicutes and Bacteroidetes phyla were assessed before and after the study. Real-time PCR was used to determine the relative expression levels of miRNAs. The results showed synbiotic supplementation significantly downregulated the expression levels of miR-27a and miR-33a compared with the placebo group (P = 0·008 and P = 0·001, respectively). Furthermore, the intervention group exhibited significant improvements in serum HDL-cholesterol, small dense LDL (sdLDL-cholesterol), apoA-I and apoB-100 (P = 0·008, P = 0·006, P = 0·003, P = 0·001, respectively). The results showed a significant negative correlation between miR-33a expression levels with HDL-cholesterol, butyrate, propionate and a significant positive correlation with total cholesterol, LDL-cholesterol and sdLDL-cholesterol in the intervention group. Fecal bacteria and SCFA were significantly increased in the intervention group. This study provides evidence that synbiotic supplementation can modulate miR-27a and miR-33a expression and improve lipid metabolism in patients with dyslipidaemia.

Restriction of dietary carbohydrates, fat and/or protein is often used to reduce body weight and/or treat (metabolic) diseases. Since diet is a key modulator of the human gut microbiome, which plays an important role in health and disease, this review aims to provide an overview of current knowledge of the effects of macronutrient-restricted diets on gut microbial composition and metabolites. A structured search strategy was performed in several databases. After screening for inclusion and exclusion criteria, thirty-six articles could be included. Data are included in the results only when supported by at least three independent studies to enhance the reliability of our conclusions. Low-carbohydrate (<30 energy%) diets tended to induce a decrease in the relative abundance of several health-promoting bacteria, including Bifidobacterium, as well as a reduction in short-chain fatty acid (SCFA) levels in faeces. In contrast, low-fat diets (<30 energy%) increased alpha diversity, faecal SCFA levels and abundance of some beneficial bacteria, including Faecalibacterium prausnitzii. There were insufficient data to draw conclusions concerning the effects of low-protein (<10 energy%) diets on gut microbiota. Although the data of included studies unveil possible benefits of low-fat and potential drawbacks of low-carbohydrate diets for human gut microbiota, the diversity in study designs made it difficult to draw firm conclusions. Using a more uniform methodology in design, sample processing and sharing raw sequence data could foster our understanding of the effects of macronutrient restriction on gut microbiota composition and metabolic dynamics relevant to health. This systematic review was registered at https://www.crd.york.ac.uk/prospero as CRD42020156929.

Evidence of the relationship between fecal short-chain fatty acids (SCFA) levels, dietary quality and type 2 diabetes mellitus (T2DM) in rural populations is limited. Here, we aimed to investigate the association between fecal SCFA levels and T2DM and the combined effects of dietar quality on T2DM in rural China. In total, 100 adults were included in the case–control study. Dietary quality was assessed by the Alternate Healthy Eating Index 2010 (AHEI-2010), and SCFA levels were analysed using the GC-MS system. Generalised linear regression was conducted to calculate the OR and 95 % CI to evaluate the effect of SCFA level and dietary quality on the risk of T2DM. Finally, an interaction was used to study the combined effect of SCFA levels and AHEI-2010 scores on T2DM. T2DM participants had lower levels of acetic and butyric acid. Generalised linear regression analysis revealed that the OR (95 % CI) of the highest acetic and butyric acid levels were 0·099 (0·022, 0·441) and 0·210 (0·057, 0·774), respectively, compared with the subjects with the lowest tertile of level. We also observed a significantly lower risk of T2DM with acetic acid levels > 1330·106 μg/g or butyric acid levels > 585·031 μg/g. Moreover, the risks of higher acetic and butyric acid levels of T2DM were 0·007 (95 % CI: 0·001, 0·148), 0·005 (95 % CI: 0·001, 0·120) compared with participants with lower AHEI-2010 scores (all P < 0·05). Acetate and butyrate levels may be important modifiable beneficial factors affecting T2DM in rural China. Improving dietary quality for body metabolism balance should be encouraged to promote good health.

(Poly)phenols are plant secondary metabolites widely abundant in plant foods and beverages comprising a very large number of compounds with diverse structure and biological activities. Accumulating evidence indicates that these compounds exert beneficial effects against cardiometabolic diseases, and this review will provide a summary of current knowledge in this area. Epidemiological and clinical data collectively suggest that intake of flavonoids reduces the risk of cardiovascular disease (CVD), with the evidence being particularly strong for the flavan-3-ol subclass. However, to provide adequate dietary recommendations, a better understanding of their estimated content in foods and intake among the general public is needed. Regarding mechanisms of action, we now know that it is unlikely that (poly)phenols act as direct antioxidants in vivo, as it was hypothesised for decades with the popularity of in vitro antioxidant capacity assays. One of the reasons is that upon ingestion, (poly)phenols are extensively metabolised into a wide array of circulating metabolites with different bioactivities than their precursors. Well-conducted in vitro and in vivo studies and human nutrigenomic analysis have revealed new molecular targets that may be underlying the health benefits of (poly)phenols, such as the nitric oxide pathway. Recently, a bi-directional relationship was established between (poly)phenols and the gut microbiota, suggesting that individual gut microbial metabolising capacity may be a key factor explaining the variability in the cardiometabolic response to (poly)phenols. Future research is needed to elucidate which are the key factors affecting such capacity, and whether it can be modulated, along with the mechanisms of action.

Ulcerative Colitis (UC), a type of Inflammatory Bowel Disease (IBD), is a chronic, relapsing gastrointestinal condition with increasing global prevalence. The gut microbiome profile of people living with UC differs from healthy controls and this may play a role in the pathogenesis and clinical management of UC. Probiotics have been shown to induce remission in UC; however, their impact on the gut microbiome and inflammation is less clear. Anthocyanins, a flavonoid subclass, have shown anti-inflammatory and microbiota-modulating properties; however, this evidence is largely preclinical. To explore the combined effect and clinical significance of anthocyanins and a multi-strain probiotic, a 3-month randomised controlled trial will be conducted in 100 adults with UC. Participants will be randomly assigned to one of four groups: anthocyanins (blackcurrant powder) + placebo probiotic, probiotic + placebo fruit powder, anthocyanin + probiotic, or double placebo. The primary outcome is a clinically significant change in the health-related quality-of-life measured with the Inflammatory Bowel Disease Questionnaire-32. Secondary outcomes include shotgun metagenomic sequencing of the faecal microbiota, faecal calprotectin, symptom severity, and mood and cognitive tests. This research will identify the role of adjuvant anti-inflammatory dietary treatments in adults with UC and elucidate the relationship between the gut microbiome and inflammatory biomarkers in this disease, to help identify targeted individualised microbial therapies. ANZCTR registration ACTRN12623000630617.

The aim of this study is to determine to what extent the addition of chitinase to black soldier fly (BSF) larval meal enriched or not with long-chain PUFA (LC-PUFA) could improve growth, protein digestion processes and gut microbial composition in Nile tilapia. Two different types of BSF meal were produced, in which larvae were reared on substrates formulated with vegetable culture substrate (VGS) or marine fish offal substrate (FOS). The BSF raised on VGS was enriched in α-linolenic acid (ALA), while that raised on FOS was enriched in ALA + EPA + DHA. Six BSF-based diets, enriched or not with chitinase, were formulated and compared with a control diet based on fishmeal and fish oil (FMFO). Two doses (D) of chitinase from Aspergillus niger (2 g and 5 g/kg feed) were added to the BSF larval diets (VGD0 and FOD0) to obtain four additional diets: VGD2, VGD5, FOD2 and FOD5. After 53 d of feeding, results showed that the BSF/FOS-based diets induced feed utilisation, protein efficiency and digestibility, as well as growth comparable to the FMFO control diet, but better than the BSF/VGS-based diets. The supplementation of chitinase to BSF/FOS increased in fish intestine the relative abundance of beneficial microbiota such as those of the Bacillaceae family. The results showed that LC-PUFA-enriched BSF meal associated with chitinase could be used as an effective alternative to fishmeal in order to improve protein digestion processes, beneficial microbiota and ultimately fish growth rate.

Periplaneta americana, one of the most widely distributed insects all over the world, can survive and reproduce in harsh environment which may be closely related to the critical roles of intestinal microorganisms in its multiple physiological functions. However, the composition and structure of gut microbiota throughout different life stages and its effects on the strong resilient and environmental adaptability of P. americana remain unclear. In this study, the gut microbiota across life stages including ootheca (embryos), nymph and adult of P. americana were investigated by 16S rRNA high-throughput sequencing. Multivariate statistical analysis showed the richness and diversity of bacterial communities were significantly different among ootheca, nymph and adult stage of P. americana. Taxonomic analysis showed Blattabacterium was the dominant genus in bacterial community of ootheca while the nutrient absorption-related genera including Christensenellaceae and Ruminococcaceae showed high relative abundance in nymph samples. Moreover, functional prediction analysis showed the metabolic categories in ootheca might have more influence on the basic life activities of the host than improved production and viability, while it was more associated to the society activities, reproduction and development of host in nymph and adult. It was suggested that the gut microbiota in each life stage might meet the requirements for environmental adaptability and survival of P. americana via transforming the composition and structure with specific metabolic capabilities. Overall, these results provided a novel sight to better understand the strong vitality and adaptability throughout life stages of P. americana.

Early life is a sensitive period when microbiota-gut-brain interactions may have important impact on development. This study investigated the associations of the gut microbiota in the first three years of life (two, six, and 12 weeks, and one and three years) with problem behavior and executive functions in N = 64 three-year-old children. Higher relative abundance of Streptococcus at the age of two weeks, as well as its trajectory over time (including ages two, six and 12 weeks, and one and three years), was related to worse executive functions. Higher relative abundance of [Ruminococcus] torques group at the age of three years, as well as its trajectory from one to three years, was associated with less internalizing behavior. Besides, several robust age-specific associations were identified: higher Bifidobacterium relative abundance (age three years) was associated with more internalizing and externalizing issues; higher Blautia relative abundance (age three years) was linked to less internalizing behavior; and increased relative abundance of an unidentified Enterobacteriaceae genus (age two weeks) was related to more externalizing behavior. Our findings provide important longitudinal evidence that early-life gut microbiota may be linked to behavioral and cognitive development in low-risk children.