Search Results (326)

Background/Objectives: Women who exclusively pump (EP) expend more time and effort on pumping and feeding infants their expressed breast milk (EBM) than those who directly breastfeed. However, little is known about their 24 h milk production (MP) or pumping and feeding dynamics. Methods: An online survey of 195 EP women explored the experiences, characteristics, MP and pumping and feeding dynamics of EP women. Quantitative data were compared by prematurity/neonatal unit admission, milk supply and parity, and self-reported MP was compared with MP in a predominantly breastfeeding reference group. Results: The 24 h pumping frequency was 6.0 [5.0, 8.0] with a pumping session duration of 25 [20, 30] min and a total pumping time of 2.3 [1.8, 3.0] h/24 h. The self-reported MP was 900 [600, 1100] mL/24 h, which was higher than that of the reference group (p = 0.012). Infant cues (58%), as well as advice from the child health nurse (39%), lactation consultant (34%) and pediatrician (34%) informed EBM feed volumes. Fifty-six percent of the infants were fed on demand, with 37% using a feeding schedule. Infants were fed 7.0 [6.0, 8.0] times/24 h; the average individual EBM feed and total 24 h volume were 120 [100, 150] and 750 [600, 900] mL, respectively. Conclusions: This study confirms it is possible to sustain lactation with EP and provide sufficient EBM volumes for the infant. However, the time commitment of EP and infant feeding represents a serious burden. These families would benefit from enhanced professional support and accessible evidence-based pumping and feeding guidelines. Full article

Objective: To investigate the effect of milk minerals on bone mineral density (BMD) and bone quality in growing rats and explore the underlying mechanisms related to calcium absorption, bone metabolism, and the gut–bone axis. Methods: Sixty healthy 4-week-old male Sprague-Dawley (SD) rats were randomly divided to five groups based on their body weight: Low-Calcium Control Group (Control), Low-Dose milk mineral Group (Low), Medium-Dose milk mineral Group (Medium), High-Dose milk mineral Group (High) and Calcium Carbonate Control Group (CaCO₃), which received the same dose level (elemental calcium) as the High group. The milk mineral dosage was set at 5, 10, and 15 times the human recommended intake of elemental calcium. After 12 weeks of intervention, femurs were collected for analysis of BMD, bone microstructure, and bone mechanical strength. Additionally, analyses included calcium levels in the femur, feces, and diet; serum bone metabolism biomarkers; tissue protein expression; as well as gut microbiota composition and short-chain fatty acid content. Result: Milk mineral exhibited non-inferior efficacy to CaCO₃ in increasing femoral calcium content, enhancing BMD, and improving bone microarchitecture. Notably, the Medium group achieved comparable bone-protective effects to the CaCO₃ group despite a 20.8% lower calcium content, which was accompanied by a relatively high calcium absorption rate (90.9% vs. 86.5%). With respect to serum markers, milk mineral maintained bone formation while suppressing bone resorption, resulting in a net anabolic state comparable to that of CaCO₃. Milk mineral significantly upregulated the protein expression of renal CYP27B1 and intestinal calcium ion transporters, and increased serum IGF-I levels. Furthermore, milk mineral promoted the enrichment of certain specific gut microbial genera, which showed a significant positive correlation with IGF-I, bone calcium content and BMD. Conclusions: Milk mineral supplementation appears to promote bone formation and mineralization in growing rats, accompanied by enhanced intestinal calcium absorption, enrichment of characteristic gut microbes and elevated microbial metabolite concentrations. Full article

Milk-derived extracellular vesicles (EVs) transport microRNAs (miRNAs) that are unusually stable and have been proposed to survive digestion and modulate gene expression in the consumer, although their dietary bioavailability and physiological relevance remain debated. How the predicted regulatory potential of these miRNAs differs among the milks of different animals most relevant to human nutrition has not been systematically compared. Here, we performed an integrative in silico analysis of publicly available small-RNA sequencing data from 29 milk and milk-cell samples of human, cow, goat, and donkey origin. miRNAs were quantified against human (hsa) miRBase references—thereby restricting the analysis to evolutionarily conserved miRNAs with human orthologs—and their predicted effect on the human transcriptome was modeled by integrating predicted (mirDIP database) and experimentally supported (TarBase v9 database) miRNA–target interactions into a per-gene, per-species weighted targeting score. Because miRNAs act predominantly as repressors, this score is read as a prediction of which genes would be post-transcriptionally down-regulated in a recipient. miR-148a-3p dominated the exosomal spectrum of all four species (human, cow, goat, and donkey; ≈21.5% of pooled abundance), and the twenty most abundant miRNAs accounted for roughly three quarters of the signal. Of 4577 robustly targeted genes, a 1809-gene conserved “pan-milk” core showed the highest cross-species targeting and was enriched for transcriptional regulation, PI3K–Akt, MAPK, and TGF-β/SMAD signaling, autophagy and—strikingly—the components of the RNA-interference machinery itself. Species-restricted gene sets recapitulated biologically plausible programs, including a human-biased neuronal/axon-guidance and chromatin module, a donkey-biased transcriptional, epithelial, and immune (CD47) module, and a ruminant lipid/cholesterol and insulin–mTOR module. Across categories, we observed a reproducible confidence–exclusivity trade-off. We emphasize that these results are computational predictions that assume dietary miRNA uptake and do not constitute experimental validation. We provide the complete targetome as a hypothesis-generating resource to prioritize candidate genes, pathways, and milk types for future functional, nutritional, and epigenetic investigation. Across the 29 samples from the four species, miRNA composition segregated by species (silhouette width = 0.82, a cluster-separation measure ranging from −1 to 1, with values near 1 indicating well-separated groups) and the category structure exceeded a permutation null, indicating that the between-species signal is robust to differences in dataset origin and milk state. Full article

The chemical composition of donkey and sheep milk showed significant differences, with donkey milk containing less fat and protein and a higher level of lactose, while sheep milk had more dry matter and a higher proportion of casein and saturated, unsaturated, monounsaturated, and polyunsaturated fatty acids. This study investigated the biological effects of in vitro-digested donkey and sheep milk, and their blends at different ratios, on differentiated human intestinal Caco-2 cells. Gene expression analysis showed that treatment with digested donkey milk (100%) significantly altered the expression of the autophagy-related gene (SQSTM1), tight junction (CLDN4), and innate defense genes (DEFB1, MUC2, and MUC5), showing a non-cytotoxic response and a profile consistent with a barrier-related transcriptional response. In contrast, sheep milk (100%) and certain milk blends (70% and 50% sheep’s milk) down-regulated genes involved in the autophagy process (ULK1, AMBRA, BECN1, ATG5, GABARAP, and SQSTM1), tight junction genes (OCLN and CDH1), and innate defense genes (DEFB1 and MUC2), suggesting a distinct epithelial response that warrants further confirmation. Metabolomic profiling revealed clear compositional and functional distinctions among the digested milks, identifying 166 metabolites that were differentially regulated (136 compounds were down-regulated and 30 up-regulated). Donkey milk digestion yielded a metabolite profile enriched in polar peptides and compounds with reported antioxidant associations, whereas sheep milk digestion showed a different pattern dominated by hydrophobic peptides and lipid-related metabolites. These findings indicate that, under the tested in vitro conditions, donkey milk digestion is associated with gene expression and metabolomic patterns consistent with epithelial barrier support. Full article

Fusobacterium nucleatum (F. nucleatum) is a key periodontal pathobiont associated with oral inflammation. This bacterium forms biofilms and expresses adhesins that facilitate its adhesion to and invasion of gingival epithelial cells. These processes disrupt the epithelial barrier and trigger oral inflammation, and in some cases, systemic inflammation. Conventional antimicrobial strategies predominantly depend on the utilization of antibiotics. Nevertheless, this can result in the proliferation of drug-resistant strains and the disruption of the oral microbiome equilibrium. As the predominant human milk oligosaccharide, 2′-Fucosyllactose (2′-FL) demonstrates considerable promise in inhibiting pathogenic bacterial adhesion and fortifying epithelial barrier function, mediated by its characteristic structural and bioactive attributes. In this study, we showed that 2′-FL attenuates the expression of virulence genes in F. nucleatum, reduces biofilm formation, and suppresses the bacterium’s ability to adhere to human gingival epithelial cells (HGECs). Furthermore, at the transcriptional level, 2′-FL suppressed F. nucleatum-induced inflammatory cytokine overexpression in both HGECs and RAW 264.7 macrophages, and upregulated barrier-related proteins (ZO-1, Occludin) and MUC-1 gene expression in HGECs. In vivo studies demonstrated the inhibitory effect of 2′-FL on F. nucleatum-induced periodontal injury in Balb/c mice. Furthermore, 16S rRNA sequencing analysis demonstrated that 2′-FL modulated oral microbiota composition of healthy volunteers and significantly reduced the abundance of Fusobacterium. Full article

Background: Human breast milk (HBM), as the initial food for humans, is quite essential for infant development and also for health throughout the lifespan. Exosomes are bioactive components in HBM, yet their nutritional role remains poorly recognized. Objectives: This study investigates how HBM exosomes change with lactation and their potential role in infant growth and development. Methods: HBM samples were obtained at 2 and 6 months postpartum from a well-established birth cohort. Purified exosomes were detected using transcriptomic, lipidomic, and proteomic approaches. Then, multi-omics data were analyzed to compare differentially expressed miRNAs, lipids, and proteins along with different lactation periods and their association with the infant growth process. Results: Compared with the 2-month postpartum group, the expression levels of miR-214-3p, miR-199a-5p, miR-126-3p, miR-127-5p, miR-144-3p, and miR-4787-5p were down-regulated in the 6-month postpartum group. In addition, 190 lipids and 269 proteins were up-regulated in the 6-month postpartum group, whereas 15 lipids and 244 proteins were down-regulated. Enrichment analysis revealed that the predicted target genes of differentially expressed miRNAs were primarily involved in cell communication and axon guidance. In parallel, the differentially expressed proteins were enriched in biosynthesis of unsaturated fatty acids and fatty acid metabolism pathway, implying a potential role in adipogenesis and neurodevelopment. Conclusions: This study reveals that the cargo contents of HBM exosomes change with the lactation period and may adapt to the needs of infant growth and development, particularly adipogenesis and neurodevelopment. HBM exosomes may play an important role in transferring genetic information from mothers to infants and be related to infants’ development. The underlying mechanisms warrant further investigation and validation. Full article

Human milk is highly rich in miRNAs, with differential expression amongst its fractions, including cells, fat, and skim milk. Various factors, such as the stage of lactation or milk removal during breastfeeding, have been shown to influence the miRNA content of. Here, we sought to determine the effect of maternal and/or infant infection on the miRNA profile of cell and fat fractions analyzed using next-generation sequencing. Breastfeeding mother/infant dyads (n = 18) were followed during one or more infection episodes as well as upon recovery. Cells and fat together contain 1780 known miRNA species, which is the highest number of known miRNAs assayed in human body fluids to date. In addition, 592 novel miRNAs were predicted, of which 95 were of high confidence. Comparisons between samples collected when the participants were healthy and when infected yielded 453 differentially expressed (p < 0.05) known miRNAs. Of these, 70 were highly expressed and differentially regulated during infection, with 62 upregulated and 8 downregulated known miRNAs during infection. Most of the highly and differentially expressed miRNAs are known to play critical roles in immunity and immune system development. These findings support the use of miRNAs as biomarkers of the health status of the lactating breast and the breastfeeding mother/infant dyad. Full article

Chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) is a complex and debilitating disorder with limited treatment options. Camel milk (CM), known for its rich nutrients and anti-fatigue properties, may offer multi-target benefits for managing this condition. This study utilized an integrated approach combining metabolomics, network pharmacology, and animal experiments. CM metabolites were profiled and screened via ADME. Potential targets were predicted and intersected with CFS/ME-associated genes. Male BALB/c mice were subjected to chronic restraint and forced swimming to evaluate the effects of CM (1000 mg/kg) on behavioral, inflammatory, neuroendocrine, and metabolic parameters. CM administration significantly improved exhaustive swimming time and reduced immobility. It attenuated systemic inflammation (restored IL-10), normalized brain CREB and DRD2/OPRM1 mRNA, and enhanced skeletal muscle AKT/GLUT4 expression and glycogen levels. Camel milk alleviates CFS/ME symptoms through the multi-component, multi-target regulation of neuroendocrine, inflammatory, and energy metabolism pathways. These preclinical findings suggest that CM may have potential as a supportive nutritional intervention for alleviating chronic fatigue, pending validation in human studies. Full article

Background/Objectives: The prevalence of women who exclusively pump (EP) their breast milk to feed their infants is increasing; however, this group is underrepresented in research. This study aimed to examine maternal and infant characteristics and the experiences of EP women. Methods: An online survey explored the experiences and characteristics of EP women with <24-month-old infants. Quantitative data included demographics and maternal and infant characteristics; qualitative data included perspectives on how support for EP women can be improved. Results: The survey of 195 EP women revealed that while most had intended to exclusively breastfeed (50%) or breastfeed and pump (26%) their milk for an average of 12 months, the average time of EP cessation was 6 months postpartum. Compared with the general population, EP women had higher rates of pregnancy complications (p < 0.001) and lactation/breastfeeding challenges (p < 0.001). Themes developed from the qualitative data relating to how health professionals/support people could better assist EP women were: ‘Respect for the EP Journey’, ‘EP Information and Logistical Needs’ and ‘Mental and Physical Load’. Conclusions: Most EP women had originally intended to breastfeed but utilised EP because of latching issues, breast refusal and/or neonatal unit admission. They face unique challenges associated with EP, yet current professional acceptance and support are lacking. Targeted education for health professionals on EP is needed so that they can better support women with tailored, evidence-based guidelines aimed at extending lactation and improving maternal well-being. Full article

This study focuses on Lacto-N-neotetraose (LNnT), a core component of human milk oligosaccharides. Although LNnT has been demonstrated to promote early intestinal development and maintain gut homeostasis, its protective mechanism against D-galactose-induced intestinal injury and associated cognitive impairment remains unclear. This investigation systematically examined the protective effects and underlying mechanisms of LNnT against D-gal-induced colonic damage and cognitive impairment in mice. The results demonstrated that LNnT not only significantly improved systemic physiological phenotypes and upregulated the expression of colonic tight junction proteins to repair the intestinal barrier, but also effectively enhanced learning and memory abilities in mice. Concurrently, LNnT reduced serum proinflammatory factor levels, elevated the anti-inflammatory factor IL-10, and alleviated oxidative stress. Furthermore, LNnT remodeled the gut microbiome structure by increasing microbial diversity, enhancing beneficial bacteria abundance, and promoting short-chain fatty acid production. Untargeted metabolomics analysis further revealed that LNnT corrected metabolic disturbances by regulating key sphingolipid molecules (ceramide, sphingosine, S1P) and the expression of related metabolic enzymes (ACER2, SphK2). In summary, this study suggests that LNnT mitigates intestinal injury and improves cognitive function, potentially through modulation of the gut microbiota–sphingolipid metabolism axis, although further causal validation is warranted. These findings provide a mechanistic foundation for future studies exploring its potential as a functional dietary ingredient. Full article

To date, the problem of mastitis in cattle remains relevant for both the industrial sector and scientific research. Despite numerous active investigations, the causes of this disease have not been fully established. It is postulated that several factors may be involved, such as bacterial pathogens, animal husbandry practices, and weather and climatic conditions. In this study, we selected cows from farms in Yakutia to investigate microbial isolates present in the milk of cows affected by mastitis and treated with antibiotics. Five identified Staphylococcus aureus isolates were investigated using whole-genome sequencing (Illumina sequencing and nanopore sequencing), followed by analysis of virulence factors in the genomes and cultural properties of the isolates. The profile of S. aureus virulence genes (exotoxins, cytotoxins, superantigen-like proteins, adhesins) was identified via WGS. Hemolysin gene (hla) was detected in all isolates. An investigation of the cultural properties of the isolates, specifically through hemolysis of rabbit erythrocytes and Western blot analysis of the culture liquid of S. aureus, revealed different expression levels of alpha-hemolysin among the strains. One isolate (17-21) exhibited the highest secretion level of about 320 ± 37 ng, both in the hemolysis test and immunoblotting assay. An investigation of the isolates’ antibiotic resistance showed that all isolates exhibited multidrug resistance, as confirmed by the presence of antibiotic resistance genes in these isolates. One isolate (7-7) exhibited the broadest range of phenotypic resistance and was resistant to all tested antibiotics (except clindamycin). Phylogenetic analysis suggested that the evolution of these isolates occurred independently in their respective ecological niches, although their transfer from cattle to humans, and vice versa, is possible. Isolates 7-7, 18-22, 33-40, and 35-42 are most typical to Yakutian cattle, while isolate 17-21 might have been introduced from a different region. To the best of our knowledge, this is the first in-depth study into a range of S. aureus isolates associated with mastitis infection in Yakutian cattle. Full article

Background/Objectives: Hand, foot, and mouth disease (HFMD) is a major public health concern primarily caused by human enterovirus A71 (EV-A71), coxsackievirus A16 (CVA16), coxsackievirus A6 (CVA6), and certain coxsackievirus B serotypes. Currently available EV-A71 vaccines lack cross-protective efficacy against other serotypes, highlighting the urgent need for multivalent and broadly effective enterovirus vaccines. Methods: Immunoinformatics approaches were used to predict highly immunogenic B-cell and T-cell epitopes, which were assembled to construct a novel multivalent epitope vaccine, rCV-A3V, followed by in silico validation. Recombinant protein expression was confirmed by Western blotting and immunofluorescence assays. The immunogenicity was evaluated in Balb/c mice following intranasal immunization. Results: A preliminary safety evaluation demonstrated that the rCV-A3V vaccine was well tolerated in the mouse model, with no abnormal changes in body weight observed after immunization. In addition, the target protein was successfully expressed. Intranasal immunization induced a strong Th1-biased immune response, robust serum neutralizing and IgG antibody responses, and pronounced mucosal immunity, including elevated sIgA and IgG levels in nasal lavage fluid, sIgA in feces, and substantial sIgA responses in milk. Dominant epitope peptides were also identified. Conclusions: The intranasal live attenuated rCV-A3V vaccine successfully induced humoral, mucosal, and cellular immune responses against EV-A71, CVA16, CVA6, and CVB3, demonstrating broad immunogenicity. These findings provide experimental evidence supporting its potential as a candidate vaccine for HFMD. Full article

Human breast milk evolves beyond simple nutrition to function as a complex signaling system that promotes neonatal development. This review analyzes the bioactive components, delineating how its specific constituents compensate for the physiological vulnerabilities of the neonate. Additionally, the distinct roles of colostral and mature milk are in fortifying the immature immune system and promoting gastrointestinal maturation. Focus is placed on the prevention of necrotizing enterocolitis, where milk oligosaccharides and microbiome function to maintain mucosal integrity and symbiosis, while preventing pathogens’ adhesion. Furthermore, how breastfeeding duration is linked to long-term metabolic and immunological programming is evaluated. MicroRNAs and bioactive lipids actively modulate gene expression and immune responses, thereby reducing the incidence of metabolic diseases and childhood malignancies. By integrating findings, this article underscores the irreplaceable role of breast milk in clinical dietetics and pediatric care. Full article

Background/Objectives: Alzheimer’s disease (AD) is characterized by progressive neurodegeneration driven by interconnected mechanisms, including oxidative stress, mitochondrial dysfunction, neuroinflammation, synaptic impairment, and abnormal protein aggregation. MicroRNAs (miRNAs) have emerged as post-transcriptional regulators of these complex pathways; however, efficient delivery remains a major limitation. Small extracellular vesicles (sEVs) have been proposed as biologically compatible carriers for miRNA delivery. Methods: In this study, milk-derived sEVs were isolated, characterized, and loaded with microRNA-137-5p (miR-137-5p). Their effects were evaluated in an amyloid-β (Aβ)-induced in vitro AD model using SH-SY5Y human neuroblastoma cells. Oxidative stress markers, including reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), lactate dehydrogenase (LDH), and glutathione peroxidase 1 (GPX1), were assessed. Inflammation- and neuroprotection-related gene expression analyses included intercellular adhesion molecule 1 (ICAM1), tumor necrosis factor alpha (TNF-α), and brain-derived neurotrophic factor (BDNF). Cytoskeletal injury was evaluated using neurofilament light chain (NfL). Mitochondrial stress markers included cytochrome c (Cyt-c), 8-hydroxy-2′-deoxyguanosine (8-OHdG), PTEN-induced kinase 1 (PINK1), dynamin-1-like protein (DNM1L), and mitochondrial transcription factor A (TFAM). Synaptic and extracellular matrix-associated proteins, including complexin-2 (CPLX2), SPARC-related modular calcium-binding protein 1 (SMOC1), and receptor tyrosine kinase-like orphan receptor 1 (ROR1), as well as AD-related biomarkers, including total tau, phosphorylated tau at threonine 181 (pTau-181), phosphorylated tau at threonine 217 (pTau-217), and amyloid-β 1–40 (Aβ1–40), were evaluated using molecular and biochemical approaches. Results: Aβ exposure was associated with increased oxidative stress, inflammatory activation, mitochondrial and cytoskeletal alterations, synaptic-related disturbances, and elevations in tau- and amyloid-associated proteins. Treatment with unloaded sEVs was associated with partial modulation of several parameters, whereas miR-137-5p-loaded sEVs were consistently associated with normalization of multiple pathological markers toward control levels. Conclusions: These findings indicate that miR-137-5p-enriched sEVs may represent a useful experimental platform for multi-target modulation of AD-related cellular alterations. Further mechanistic and in vivo studies are required to clarify translational relevance. Full article

Human breast milk harbors commensal lactic acid bacteria with probiotic potential, and microbial fermentation may enhance the bioactivity of plant-derived exosome-like nanovesicles (EVs); this study evaluated whether L. plantarum BMSE-HMP251 isolated from breast milk could safely ferment Hordeum vulgare L. and improve the anti-inflammatory activity of derived EVs. BMSE-HMP251 was identified by 16S rRNA sequencing and characterized by biochemical, safety, and genomic analyses. EVs derived from Hordeum vulgare L. extract and BMSE-HMP251-fermented broth were evaluated for physicochemical properties, antioxidant activity, cytotoxicity, and anti-inflammatory activity in LPS-stimulated HT-29 and RAW 264.7 cells. EVs derived from Hordeum vulgare L. fermentation exhibited a distinct size distribution and significantly enhanced bioactivity, including higher DPPH radical scavenging activity and greater suppression of nitric oxide production and proinflammatory cytokine (TNF-α and IL-1β) mRNA expression, compared with EVs from unfermented extracts. These effects were observed following fermentation with the human breast milk-derived strain L. plantarum BMSE-HMP251, which showed species-consistent phenotypic and genomic characteristics and no safety concerns. Overall, fermentation markedly enhances the anti-inflammatory potential of plant-derived EVs, supporting fermentation as a safe and effective strategy to improve their functional value. Full article