Human Milk Samples Research Articles

Comparing the human milk antibody response after vaccination with four COVID-19 vaccines: A prospective, longitudinal cohort study in the Netherlands

SummaryBackgroundVaccination of lactating women against COVID-19 may protect not only themselves but also their breastfed infant through human milk. Therefore, it is important to gain insight into the human milk antibody response after immunization with the various vaccines that are currently widely used. The aim of this study is to determine and compare the antibody response in human milk following vaccination with mRNA- and vector-based vaccines up to over two months post-vaccination.MethodsThis prospective cohort study was conducted in the Netherlands between January 06, 2021 and July 31, 2021. Participants were recruited through social media. Human milk samples were collected longitudinally during a period of 70 days from women receiving one of the four different severe acute respiratory coronavirus 2 (SARS-CoV-2) vaccines: Pfizer-BioNTech (BNT162b2), Moderna (mRNA-1273), Oxford/AstraZeneca (AZD1222) and Johnson&Johnson (Ad26.COV2.S). SARS-CoV-2-specific antibodies were measured using an enzyme-linked immunosorbent assay. The area under the curve (AUC) of the Immunoglobulins A (IgA) and G (IgG) antibody response was determined over 15 and 70 days following the first vaccination and compared between the different vaccines.FindingsThis study enrolled 134 vaccinated lactating women of whom 97 participated the entire study period. In total, 1887 human milk samples were provided. The human milk antibody response differed between SARS-CoV-2 vaccines over the study period. The mean AUC of SARS-CoV-2-specific IgA, but not IgG, in human milk over 15 days was higher after vaccination with an mRNA-based vaccine than a vector-based vaccine (AUC with respect to ground [AUCg] ± the standard error of the mean [SEM] for IgA was 6·09 ± 0·89 in the BNT162b2 group, 7·48 ± 1·03 in the mRNA-1273 group, 4·17 ± 0·73 in the AZD1222 group, and 5·71 ± 0·70 in the Ad26.COV2.S group). Over a period of 70 days, the mean AUCg of both IgA and IgG was higher after vaccination with an mRNA-based vaccine than a vector-based vaccine (AUCg ± SEM for IgA was 38·77 ± 6·51 in the BNT162b2 group, 50·13 ± 7·41 in the mRNA-1273 group, 24·12 ± 5·47 in the AZD1222 group, and 28·15 ± 6·69 in the Ad26.COV2.S group; AUCg ± SEM for IgG was 40·43 ± 2·67 in the BNT162b2 group, 37·01 ± 2·38 in the mRNA-1273 group, 16·04 ± 5·09 in the AZD1222 group, and 10·44 ± 2·50 in the Ad26.COV2.S group).InterpretationOverall, maternal vaccination during lactation with an mRNA-based vaccine resulted in higher SARS-CoV-2 antibody responses in human milk compared to vector-based vaccines. Therefore, vaccination with mRNA-based vaccines, preferably with the mRNA-1273 vaccine, might not only provide better immunological protection for the mother but also for her breastfed infant.FundingStichting Steun Emma Kinderziekenhuis and the Amsterdam Infection and Immunity Institute (grant 24175).

Validation and application of a quantitative liquid chromatography tandem mass spectrometry assay for the analysis of rifapentine and 25-O-desacetyl rifapentine in human milk

A robust analytical method based on liquid chromatography coupled to tandem mass spectrometry was developed and validated to quantify rifapentine and 25-O-desacetyl rifapentine in human breast milk to aid in determining the breastfed infant risk to the excreted drug in human milk. Samples were extracted by a combination of protein precipitation and solid phase extraction using rifampicin-d3 as an internal standard. An Agilent® Poroshell 120 EC-C18 (4.6 mm × 50 mm, 2.7 µm) column was used for chromatographic separation employing an isocratic mobile phase consisting of acetonitrile: methanol: 0.1% formic acid (55/5/40, v/v/v) at a flow rate of 450 µL/min, and with a total run time of four minutes. Mass detection was on an AB Sciex API 4000 mass spectrometer using electrospray ionization in the positive mode and based on multiple reaction monitoring data acquisition. Rifapentine was accurately quantified across a concentration range of 2.00–2000 ng/mL and 25-O-desacetyl rifapentine from 4.00 to 2000 ng/mL. During validation, the inter- and intra-day accuracy and precision at the tested QC concentrations (N = 18) for rifapentine were between 97.4% and 100.6%, and 3.1% and 8.3%, respectively. The inter- and intra-day accuracy and precision for 25-O-desacetyl rifapentine were between 96.4% and 106.3%, and 6.7% and 11.8%, respectively. No significant matrix effects were observed, and the method was shown to be specific for rifapentine and 25-O-desacetyl rifapentine. Human milk samples (N = 22) generated during a phase I/II clinical trial were successfully analysed for rifapentine and 25-O-desacetyl rifapentine using this validated method. Concentrations for rifapentine and 25-O-desacetyl rifapentine in human milk samples (N = 22) ranged from 11.2-1180 ng/mL and 7.11–573 ng/mL, respectively.

Lactational Changes of Phospholipids Content and Composition in Chinese Breast Milk.

Phospholipids are pivotal polar lipids in human milk and essential for infants’ growth and development, especially in the brain and cognitive development. Its content and composition are affected by multiple factors and there exist discrepancies in different studies. In this study, we determined five major phospholipids classes (phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylcholine, and sphingomyelin) in 2270 human milk samples collected from 0 to 400 days postpartum in six regions of China. The high-performance liquid chromatography coupled with an evaporative light scattering detector (HPLC-ELSD) was performed to quantify the phospholipids. Total phospholipid median (IQR) content was in a range between 170.38 ± 96.52 mg/L to 195.69 ± 81.80 mg/L during lactation and was higher concentrated in colostrum milk and later stage of lactation (after 200 days postpartum) compared with that in the samples collected between 10 to 45 days postpartum. Variations in five major sub-class phospholipids content were also observed across lactation stages (phosphatidylethanolamine: 52.61 ± 29.05 to 59.95 ± 41.74 mg/L; phosphatidylinositol: 17.65 ± 10.68 to 20.38 ± 8.55 mg/L; phosphatidylserine: 15.98 ± 9.02 to 22.77 ± 11.17 mg/L; phosphatidylcholine: 34.13 ± 25.33 to 48.64 ± 19.73 mg/L; sphingomyelin: 41.35 ± 20.31 to 54.79 ± 35.26 mg/L). Phosphatidylethanolamine (29.18–32.52%), phosphatidylcholine (19.90–25.04%) and sphingomyelin (22.39–29.17%) were the dominant sub-class phospholipids in Chinese breast milk during the whole lactation period. These results updated phospholipids data in Chinese human milk and could provide evidence for better development of secure and effective human milk surrogates for infants without access to breast milk.

First application of microwave-assisted extraction in the analysis of polybrominated diphenyl ethers in human milk

As a method, microwave-assisted extraction (MAE) has a number of advantages over other extraction methods, such as lower solvent consumption and analyte loss, shorter processing time, and higher sample throughput. So far, however, MAE has mostly been applied with solid samples like sediments and soil. This paper describes the first MAE of organic pollutants from human milk samples, the influence of sample preparation and pretreatment, and how extraction conditions (type and volume of solvent, extraction temperature) were optimized. The most efficient way to extract polybrominated diphenyl ethers (PBDEs) from human milk using MAE was dissolving a freeze-dried sample in ultrapure water and applying a solvent mixture of n-hexane and acetone (1:1, v/v) with formic acid and 2-propanol as denaturing agents at 105 °C for 20 min. Extracts were cleaned up with concentrated sulfuric acid and on SPE column containing neutral and acidified silica with n-hexane and dichloromethane (4:1, v/v) as elution solvent. The method was validated at two concentration levels through parallel analysis by gas chromatography with two detection techniques and application in real samples and a baby formula (similar matrix). It has shown great potential for the analysis of organic pollutants in important biological matrices such as human milk. Further optimization could extend its application to other classes of persistent organic pollutants (POPs).

Perfluorinated alkyl substances in Canadian human milk as part of the Maternal-Infant Research on Environmental Chemicals (MIREC) study

Perfluorinated alkyl substances (PFAS) were determined in human milk samples (n = 664) from participants in the Maternal-Infant Research on Environmental Chemicals (MIREC) study. ΣPFAS concentrations (sum of seven PFAS) ranged from 3.1 ng L−1 to 603 ng L−1, with a median concentration of 106 ng L−1 in the Canadian mothers' milk analyzed. These data comprise the first pan-Canadian dataset of PFAS in human milk. Perfluorooctanoic acid (PFOA) and linear perfluorooctanesulfonate (L-PFOS) were the dominant contributors to ΣPFAS in human milk samples. An inverse relationship between ΣPFAS concentrations and age was observed (Spearman correlation - 0.184). Primiparous women had elevated PFAS concentrations in milk relative to women who had children previously (p < 0.001). In contrast, the region of maternal birth did not influence ΣPFAS concentrations (p = 0.156). Although China and Norway have observed consistently detectable levels of perfluoroundecanoic acid (PFUdA) in human milk, PFAS with long carbon chains (n ≥ 11) were not present above method detection limits in Canadian human milk samples analyzed as part of the MIREC study. In conclusion, despite the presence of low levels of environmental contaminants in human milk, Health Canada supports breastfeeding due to the benefits to both infants and mothers.

Restricting Cow's Milk in the Maternal Diet Reduces the Titers of β-Lactoglobulin-Specific IgG Antibodies in Human Milk.

Background: Human milk antibodies specific to allergen enhance immunological tolerance in neonates by educating their immature mucosal immunity. The impact of restricting food allergens in diet and maternal factors on the levels of allergen-specific antibodies in human milk remains unclear. We aimed to evaluate the influence of the maternal avoidance diet of cow's milk on the titers of IgG, SIgA/IgA, and IgM specific to β-lactoglobulin (BLG) in human milk. Materials and Methods: Human milk samples were collected from 26 women nonrestricting cow's milk and 11 women restricting cow's milk. The titers of IgG, SIgA/IgA, and SIgM/IgM specific to BLG were measured using ELISA. Results: BLG-specific IgG titers were 2.9-fold higher in women nonrestricting cow's milk than those restricting cow's milk in their diet (p = 0.026), but BLG-specific SIgA/IgA and SIgM/IgM titers were comparable between these two groups. BLG-specific IgG was positively correlated with BLG-specific SIgA/IgA titers in milk from mothers nonrestricting cow's milk (p = 0.0007) but did not correlate for mothers restricting cow's milk. BLG-specific SIgA/IgA titer decreased with increasing postpartum time in milk from women restricting cow's milk (p = 0.019). Type of delivery, infant gender, maternal age, and probiotic intake did not influence the BLG-specific antibody titers. Conclusions: This study reveals that the secretion of BLG-specific IgG in human milk increases in women nonrestricting cow's milk compared with women restricting cow's milk. The role of breast milk allergen-specific antibodies on the neonatal gut (crosstalk with immune and epithelial cells) remains to be investigated.

Lactobacillus paracasei BD5115-Derived 2-Hydroxy-3-Methylbutyric Acid Promotes Intestinal Epithelial Cells Proliferation by Upregulating the MYC Signaling Pathway.

Metabolites of probiotics that are beneficial to human health have been isolated from the intestinal tract and natural dairy products. However, many studies on probiotics and prebiotics are limited to the observation of human cohorts and animal phenotypes. The molecular mechanisms by which metabolites of probiotics regulate health are still need further exploration. In this work, we isolated a strain of Lactobacillus Paracasei from human milk samples. We numbered it as Lactobacillus Paracasei BD5115. The mouse model of high-fat diet confirmed that the metabolites of this strain also promotes intestinal epithelial cells (IECs) proliferation. Single-cell sequencing showed that a bZIP transcription factor MAFF was specifically expressed in some IECs. We found that MAFF interacted with MBP1 to regulate the expression of MYC. Analysis of the active components in BD5115 metabolites confirmed that 2-hydroxy-3-methylbutyric acid promotes the expression of the MYC gene. This promotes the proliferation of IECs. Our findings indicate that 2-hydroxy-3-methylbutyric acid regulate MYC gene expression mediated by MAFF/MBP1 interaction. This study not only screened a strain with promoted IECs proliferation, but also discovered a new signal pathway that regulates MYC gene expression.

Maternal Factors Affecting the Macronutrient Composition of Transitional Human Milk.

This study investigated the influence of selected maternal factors on the macronutrient composition and energy of human milk (HM). The study enrolled 159 breastfeeding mothers from five postpartum care centers in Seoul, Korea. Their gestational weeks were 37–42 weeks, they had no complications before and after childbirth, and were at 7–14 days postpartum. They provided data using structured questionnaires on general characteristics, stress, sleep quality, spousal support, and dietary intake. Breastfeeding assessment (LATCH) was investigated by qualified nurses, and each mother provided one sample of HM. The HM composition was analyzed using the Miris® HM analyzer. The relationships between variables were analyzed using Pearson’s correlation analysis, and a linear regression analysis was performed to verify the main variables. It was found that maternal dietary intake was related to HM composition as the %energy from carbohydrates (β = 0.86, p < 0.01) and %energy from fat (β = 0.77, p < 0.05) showed positive relationships with HM energy. The LATCH score was positively related to HM energy (β = 0.17, p < 0.05). In contrast, postpartum stress, sleep quality, and spousal support were not associated with HM macronutrient composition. In conclusion, HM macronutrients and energy content were associated with maternal dietary intake and LATCH scores, but not with postpartum stress, sleep quality, and spousal support.

Preconcentration and determination of five antidepressants from human milk and urine samples by stir bar filled magnetic ionic liquids using liquid-liquid-liquid microextraction-high-performance liquid chromatography.

A sensitive and straightforward liquid-liquid-liquid microextraction method was developed to preconcentrate and cleanup antidepressants, including mirtazapine, venlafaxine, escitalopram, fluoxetine, and fluvoxamine, from biological samples before analyzing with high-performance liquid chromatography. The essential novelty of this study is using magnetic ionic liquids as the extraction phase in the lumen of hollow fiber and preparing a liquid magnetic stir bar. In this method, polypropylene hollow fiber was utilized as the permeable membrane for the analyte extraction. Six magnetic ionic liquids consisting ofthe transition metal and rare earth compounds were synthesized and then hollow fiber lumen was injected as acceptor phase to extract the antidepressants. Besides, 3-pentanol as a water-immiscible solvent was impregnated in the hollow fiber wall pores. The effective factors in the method were optimized with the central composition design. The resultant calibration curves were linear over the concentration range of 0.8-400.0ng mL-1 (R2 ≥ 0.996). The method displayed the proper detection limit (0.11-0.24ng mL-1 ), the reasonable limit of quantification (≤0.79ng mL-1 ), wide linear ranges, high preconcentration factors (≥294.3), and suitable relative standard deviation (2.31-5.47%) for measuring antidepressant medications. Analysis of human milk and urine samples showed acceptable recoveries of 96.5-103.8% with excellent relative standard deviations lower than 5.95%.

POPs monitoring in Mongolia – Core matrices

Persistent organic pollutants (POPs) are of global concern due to their negative effects on humans and the environment, and globally are regulated in the Stockholm Convention on POPs. The present study had Mongolia as a partner in a multinational project funded and coordinated by the United Nations Environment Programme with the aim to monitor POPs in core media, including air (with passive samplers), water, and a pooled human milk sample. Project implementation and all sampling were undertaken by the Institute of Chemistry and Chemical Technology whereas POPs analysis was performed in laboratories abroad. Brominated and chlorinated POPs were analyzed by gas chromatography/mass spectrometry, perfluorinated compounds (PFAS) and isomers of hexabromocyclododecane (HBCD) by liquid chromatography/mass spectrometry. The monitoring results found very low concentrations of PFAS in river water. Toxic equivalents (TEQs) for PCDD/PCDF were low in air and in human samples but had a relatively higher presence of TEQ from dioxin-like PCB than in other countries. With respect to chlorinated POPs, drins, chlordanes, DDTs, heptachlors, and mirex were very low in air and human milk. Elevated levels were found for polychlorinated biphenyls (PCB), hexachlorobenzene (HCB), hexachlorocyclohexanes (HCHs), pentachlorobenzene (PeCBz), and HBCD in air with extreme values for hexachlorobutadiene. The abundance of HCHs and HCB was confirmed in the human milk sample but not for PCB, PeCBz or HBCD. Short-chain chlorinated paraffins (170 ng/g lipid) were the highest among all human milk pools. It is recommended to continue POPs monitoring in the future, especially for those where high concentrations were found.

Human Milk Oligosaccharides and Bacterial Profile Modulate Infant Body Composition during Exclusive Breastfeeding.

Human milk is a complex and variable ecosystem fundamental to the development of newborns. This study aimed to investigate relationships between human milk oligosaccharides (HMO) and human milk bacterial profiles and infant body composition. Human milk samples (n = 60) were collected at two months postpartum. Infant and maternal body composition was measured with bioimpedance spectroscopy. Human milk bacterial profiles were assessed using full-length 16S rRNA gene sequencing and 19 HMOs were quantitated using high-performance liquid chromatography. Relative abundance of human milk bacterial taxa were significantly associated with concentrations of several fucosylated and sialylated HMOs. Individual human milk bacteria and HMO intakes and concentrations were also significantly associated with infant anthropometry, fat-free mass, and adiposity. Furthermore, when data were stratified based on maternal secretor status, some of these relationships differed significantly among infants born to secretor vs non-secretor mothers. In conclusion, in this pilot study the human milk bacterial profile and HMO intakes and concentrations were significantly associated with infant body composition, with associations modified by secretor status. Future research designed to increase the understanding of the mechanisms by which HMO and human milk bacteria modulate infant body composition should include intakes in addition to concentrations.

Exclusively Breastfed Infant Microbiota Develops over Time and Is Associated with Human Milk Oligosaccharide Intakes.

Temporal development of maternal and infant microbiomes during early life impacts short- and long-term infant health. This study aimed to characterize bacterial dynamics within maternal faecal, human milk (HM), infant oral, and infant faecal samples during the exclusive breastfeeding period and to document associations between human milk oligosaccharide (HMO) intakes and infant oral and faecal bacterial profiles. Maternal and infant samples (n = 10) were collected at 2–5, 30, 60, 90 and 120 days postpartum and the full-length 16S ribosomal RNA (rRNA) gene was sequenced. Nineteen HMOs were quantitated using high-performance liquid chromatography. Bacterial profiles were unique to each sample type and changed significantly over time, with a large degree of intra- and inter-individual variation in all sample types. Beta diversity was stable over time within infant faecal, maternal faecal and HM samples, however, the infant oral microbiota at day 2–5 significantly differed from all other time points (all p < 0.02). HMO concentrations and intakes significantly differed over time, and HMO intakes showed differential associations with taxa observed in infant oral and faecal samples. The direct clinical relevance of this, however, is unknown. Regardless, future studies should account for intakes of HMOs when modelling the impact of HM on infant growth, as it may have implications for infant microbiota development.

Microbiological Quality of Milk Donated to the Regional Human Milk Bank in Warsaw in the First Four Years of Activity.

As the survival rate for preterm infants increases, more emphasis is placed on improving health-related quality of life through optimal nutritional management. Human Milk Banks (HMBs) provide bioactive nutrients and probiotic microorganisms to premature newborns, especially in the first year of life. Donated milk screening and selection of potential donors ensures the quality and microbiological safety of the donated milk. Therefore we reviewed the basic characteristics of donors and the amounts and contamination of breast milk donated to the Regional Human Milk Bank (RHMB) in Warsaw. In four years, the RHMB collected 1445.59 L of milk, of which 96.60% was distributed among hospitalised infants. Additionally, breastmilk from donor candidates (139 samples from 96 women) was tested at least once in the first year of lactation. First analyses showed that 18 women’s milk samples were microbiologically pure, and 78 samples had one or more species of commensal and/or potentially pathogenic bacteria. In human milk samples from 10 women, the bacteria level was above the standard required by the RHMB; therefore, donors were re-educated, and further samples were tested. Most women followed the recommendations on hygienic expression and storage of milk before transfer to the RHMB. Our analysis will help to increase the accessibility and quality of raw donor milk and to meet the needs of more newborns.

The effect of Holder pasteurization on the lipid and metabolite composition of human milk

Human milk (HM) is the gold standard for newborn nutrition. When own mother’s milk is not sufficiently available, pasteurized donor human milk becomes a valuable alternative. In this study we analyzed the impact of Holder pasteurization (HoP) on the metabolic and lipidomic composition of HM. Metabolomic and lipidomic profiles of twelve paired HM samples were analysed before and after HoP by liquid chromatography–mass spectrometry (MS) and gas chromatography-MS. Lipidomic analysis enabled the annotation of 786 features in HM out of which 289 were significantly altered upon pasteurization. Fatty acid analysis showed a significant decrease of 22 out of 29 detectable fatty acids. The observed changes were associated to five metabolic pathways. Lipid ontology enrichment analysis provided insight into the effect of pasteurization on physical and chemical properties, cellular components, and functions. Future research should focus on nutritional and/or developmental consequences of these changes.

Study on the characteristics of glycerides and phospholipids in human milk from Tibet

The unique geographical characteristics and food culture of Tibet can affect the nutrition of human milk lipids. But little has been done in the comparison of the lipids between Tibet and other areas. This study gives in-depth analysis of the species, concentration and composition of lipid subclasses at the molecular level of the Tibetan human milk. There were averagely 132 ± 30 species of lipids, among which triglycerides (TAGs), phosphatidylethanolamine (PE) and sphingomyelin (SM) accounted for 79.78% of the total species number in the Tibetan human milk samples. The contents of TAG, SM, phosphatidylcholine (PC), and PE in the Tibetan human milk were 85.84%, 17.79%, 25.94% and 55.81% of those in the comparative human milk of China, respectively. The contents of TAGs and diglycerides (DAGs) with PUFAs in Tibetan human milk were significantly lower than those in the comparative group. However, the content and percentage of TAGs and DAGs with odd-chain saturated fatty acids were both higher in the Tibetan human milk than those in the comparative human milk. In total, 18 molecular species of lipids were downregulated and 5 ones were upregulated in the Tibetan human milk compared with those in the comparative human milk of China. The profile of lipids in the Tibetan human milk at the molecular level provided the scientific basis for maternal diet and supplemented the Chinese human milk lipids database.

Analysing the protection from respiratory tract infections and allergic diseases early in life by human milk components: the PRIMA birth cohort

BackgroundMany studies support the protective effect of breastfeeding on respiratory tract infections. Although infant formulas have been developed to provide adequate nutritional solutions, many components in human milk contributing to the protection of newborns and aiding immune development still need to be identified. In this paper we present the methodology of the “Protecting against Respiratory tract lnfections through human Milk Analysis” (PRIMA) cohort, which is an observational, prospective and multi-centre birth cohort aiming to identify novel functions of components in human milk that are protective against respiratory tract infections and allergic diseases early in life.MethodsFor the PRIMA human milk cohort we aim to recruit 1000 mother–child pairs in the first month postpartum. At one week, one, three, and six months after birth, fresh human milk samples will be collected and processed. In order to identify protective components, the level of pathogen specific antibodies, T cell composition, Human milk oligosaccharides, as well as extracellular vesicles (EVs) will be analysed, in the milk samples in relation to clinical data which are collected using two-weekly parental questionnaires. The primary outcome of this study is the number of parent-reported medically attended respiratory infections. Secondary outcomes that will be measured are physician diagnosed (respiratory) infections and allergies during the first year of life.DiscussionThe PRIMA human milk cohort will be a large prospective healthy birth cohort in which we will use an integrated, multidisciplinary approach to identify the longitudinal effect human milk components that play a role in preventing (respiratory) infections and allergies during the first year of life. Ultimately, we believe that this study will provide novel insights into immunomodulatory components in human milk. This may allow for optimizing formula feeding for all non-breastfed infants.

Discovering Efficient Method Extracting Arsenic from Human Milk Samples

In this article, the arsenic levels in drinking water were investigated. Hair and nail arsenic is strongly related, however, according to principal component analysis (PCA). According to our findings, lactating women with hypertension higher levels. Arsenic levels in infants' blood, nails, and urine were examined, and the findings indicated a substantially high body burden of arsenic in regions. The hair and nail arsenic levels were shown to be age-dependent utilizing PCA. The methods for extracting arsenic from human milk samples are effective in both literatures. These results would help in the development of a faster and more efficient technique for removing arsenic from milk samples. Although the procedure will take longer than anticipated, it does not exceed 5-6 hours. Furthermore, such studies reveal the degree of arsenic exposure in individuals living in complicated settings. People are exposed to less arsenic as a result of this, which may lead to severe cases of chronic arsenic poisoning. Prenatal exposure to babies may possibly cause genetic abnormalities as well as physical and mental problems.

Chrononutrition and Human Milk

BackgroundBreastfed infants have a reduced risk of infections and allergies. The study of chrononutrition in human milk seeks to understand the circadian variation of various human milk immune factors.MethodsEmpirical studies on human milk, chrononutrition, and immune factors were searched through PUBMED, Google Scholar, and SCOPUS. Keywords included “chrononutrition,” “breastmilk composition,” “human milk,” “day-night cycles,” “sleep-wake cycles” and the names of various immune factors. After excluding duplicate articles, animal studies, studies looking at other human milk components, studies that did not collect human milk samples over a 24 hour period, and studies that were not in English, eleven studies on the topic remained and ten studies were included in the review. The excluded study had a sample size of two.ResultsThis review identified the circadian variation of certain immune factors found in human milk such as antibodies, complement proteins, cytokines, by-products of phagocyte activity, nucleotides, microRNAs, and antioxidants.ConclusionThe circadian variation observed in some human milk components highlights the unique ability of human milk to vary in composition based on the circadian rhythms of mothers and infants. The limited number of studies makes it difficult to make conclusive recommendations and creates an opportunity for further research in this growing field.

Human milk oligosaccharide profiles and child atopic dermatitis up to 2years of age: The Ulm SPATZ Health Study.

Human milk oligosaccharides (HMOs) have several biological functions. Yet, very few studies have investigated the effect of HMOs on the development of allergies and even fewer on their specific associations with atopic dermatitis (AD) during early childhood. This study investigated whether individual HMO concentrations, measured at two time points of lactation, were associated with reported diagnosis of AD in children up to twoyears of age. Outcome data were available for HMOs measured in human milk samples collected at 6weeks (n=534) and 6months (n=356) of lactation. Associations of HMOs with AD, ascertained from parents and pediatricians at ages one and two years, were assessed in crude and adjusted logistic regression models. Few associations were statistically significant at the conventional level (p<.05), for example, 6-week Lacto-N-neotetraose with 2-year AD [OR 95%CI: 0.82 (0.66, 1.00)] and 6-month 3'-sialyllactose among non-secretor mothers with 1-year AD [2.59 (1.53, 6.81)]. Importantly, accounting for multiple testing, these and all further associations were not statistically significant (all p>.0031, which is the threshold for statistical significance after correction for multiple testing). Our findings suggest that the intake of different levels (or even absence) of the individual HMOs measured at 6weeks and 6months of lactation, in the current study, is not significantly associated with the development of AD in early childhood. Given the exploratory nature of our study and the limited sample size, these results should be interpreted with caution. The specific HMOs for which we show plausible associations at conventional level may warrant further research and investigation.

High-Temperature Short-Time Preserves Human Milk's Bioactive Proteins and Their Function Better Than Pasteurization Techniques With Long Processing Times.

Donor human milk is generally processed by holder pasteurization (HoP) at 62. 5°C for 30 min. This temperature-time combination is sufficient for eliminating pathogens in donor milk, but also negatively affects several bioactive milk components. Long heating up times may further affect the bioactive properties of pasteurized milk. High-Temperature-Short-Time (HTST), a treatment with shorter processing times (72°C for 15 sec), was investigated as a suitable alternative to HoP. In addition, pasteurization methods that follow the same temperature regime but with varying heating up times were compared. Human milk samples from four different donors were combined into one pool, which was then used to perform all analyses. The effects of these methods on the levels and functionality of immunoglobulin A, lactoferrin, lysozyme and bile salt-stimulated lipase, were evaluated with LC-MS/MS-based proteomics and activity assays, while the pasteurization efficacy was evaluated with an alkaline phosphatase test. HoP, a treatment with long processing times, times, caused the highest reduction in all proteins studied (reduced by 50–98%). Compounds such as lactoferrin and bile salt-stimulated lipase that are more sensitive to heat treatments were better retained with HTST, but their levels and functionality were still significantly lower than those of untreated donor milk (52 and 81% reduction of lactoferrin and bile salt-stimulated lipase activity, respectively). Our findings showed that a treatment with considerably shorter processing times, such as HTST, may reduce the thermal damage caused to the bioactive proteins compared to HoP, without affecting pasteurization efficacy. Since the vast majority of the donor human milk banks that are currently operating on a global level apply HoP to donor milk, our findings may provide relevant information for the optimization of donor milk processing.