Objectives Norethindrone progestin-only pills (n-POPs) require a narrow time window for intake to maintain efficacy. Drospirenone (d-POPs) may offer improved acceptability, however little is known about d-POPs' effects on human milk supply, composition, and infant growth. We sought to compare type of progestin (drospirenone vs. norethindrone) and timing (early vs. interval) of initiation on a triad of parental, infant, and milk outcomes. Methods We conducted a three-armed, randomized controlled double-blind trial among lactating postpartum people assigned to: (1) early start (1–7 days) d-POPs and (2) n-POPs and (3) interval start (28–32 days) d-POPs. We assessed feeding via eight weekly text message surveys. Milk and infant growth outcomes were assessed at four weeks postpartum. Results Ten participants were assigned to each group (n=30); four stopped exclusive human milk feeding (none in group 1; two in groups 2 and 3) by eight weeks (log rank comparison across groups (X^2=1.7, p=0.4). There were no significant differences in proportions of any human milk macronutrients analyzed (ie, protein, lactose, calorie) by group; mean percent weight increases were 20.9%, 30.8%, and 28.4%, respectively for groups 1, 2, and 3 with no significant difference (p=0.32). The percentages of respondents agreeing or strongly agreeing that they were satisfied with their assigned POP was higher among the early start groups (86% d-POP, 88% n-POP) compared to interval d-POP group (67%) (p=0.98). Conclusions There was no significant difference in the time to lactation cessation, milk composition or infant growth, between three groups. Assessing outcomes across the triad of parent, infant and milk is promising; a larger trial is warranted to assess between-group differences. Norethindrone progestin-only pills (n-POPs) require a narrow time window for intake to maintain efficacy. Drospirenone (d-POPs) may offer improved acceptability, however little is known about d-POPs' effects on human milk supply, composition, and infant growth. We sought to compare type of progestin (drospirenone vs. norethindrone) and timing (early vs. interval) of initiation on a triad of parental, infant, and milk outcomes. We conducted a three-armed, randomized controlled double-blind trial among lactating postpartum people assigned to: (1) early start (1–7 days) d-POPs and (2) n-POPs and (3) interval start (28–32 days) d-POPs. We assessed feeding via eight weekly text message surveys. Milk and infant growth outcomes were assessed at four weeks postpartum. Ten participants were assigned to each group (n=30); four stopped exclusive human milk feeding (none in group 1; two in groups 2 and 3) by eight weeks (log rank comparison across groups (X^2=1.7, p=0.4). There were no significant differences in proportions of any human milk macronutrients analyzed (ie, protein, lactose, calorie) by group; mean percent weight increases were 20.9%, 30.8%, and 28.4%, respectively for groups 1, 2, and 3 with no significant difference (p=0.32). The percentages of respondents agreeing or strongly agreeing that they were satisfied with their assigned POP was higher among the early start groups (86% d-POP, 88% n-POP) compared to interval d-POP group (67%) (p=0.98). There was no significant difference in the time to lactation cessation, milk composition or infant growth, between three groups. Assessing outcomes across the triad of parent, infant and milk is promising; a larger trial is warranted to assess between-group differences.

Microbiota-accessible carbohydrates (MACs) are powerful modulators of microbiota composition and function. These substrates are often derived from diet, such as complex polysaccharides from plants or human milk oligosaccharides (HMOs) during breastfeeding. Host-derived mucus glycans on gut-secreted mucin proteins serve as a continuous endogenous source of MACs for resident microbes; here we investigate the potential role of purified, orally administered mucus glycans in maintaining a healthy microbial community. In this study, we liberated and purified O -linked glycans from porcine gastric mucin and assessed their efficacy in shaping the recovery of a perturbed microbiota in a mouse model. We found that porcine mucin glycans (PMGs) and HMOs enrich for taxonomically similar resident microbes. We demonstrate that PMGs aid recovery of the microbiota after antibiotic treatment, suppress Clostridium difficile abundance, delay the onset of diet-induced obesity, and increase the relative abundance of resident Akkermansia muciniphila . In silico analysis revealed that genes associated with mucus utilization are abundant and diverse in prevalent gut commensals and rare in enteric pathogens, consistent with these glycan-degrading capabilities being selected for during host development and throughout the evolution of the host–microbe relationship. Importantly, we identify mucus glycans as a novel class of prebiotic compounds that can be used to mitigate perturbations to the microbiota and provide benefits to host physiology.

AllergyVolume 73, Issue 10 p. 2070-2073 LETTER TO THE EDITOR Human milk oligosaccharide profiles and food sensitization among infants in the CHILD Study K. Miliku, K. Miliku orcid.org/0000-0002-9614-7191 Manitoba Developmental Origins of Chronic Diseases in Children Network (DEVOTION), Children's Hospital Research Institute of Manitoba, Winnipeg, MB, Canada Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada The Generation R Study Group, Departments of Pediatrics and Epidemiology, Erasmus University Medical Center Rotterdam, Rotterdam, The NetherlandsSearch for more papers by this authorB. Robertson, B. Robertson Department of Pediatrics and Larsson-Rosenquist Foundation Mother-Milk-Infant Center of Research Excellence, University of California San Diego, La Jolla, CA, USASearch for more papers by this authorA. K. Sharma, A. K. Sharma George & Fay Yee Centre for Healthcare Innovation, University of Manitoba, Winnipeg, MB, CanadaSearch for more papers by this authorP. Subbarao, P. Subbarao Departments of Pediatrics & Physiology, University of Toronto, Toronto, ON, CanadaSearch for more papers by this authorA. B. Becker, A. B. Becker Manitoba Developmental Origins of Chronic Diseases in Children Network (DEVOTION), Children's Hospital Research Institute of Manitoba, Winnipeg, MB, Canada Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, CanadaSearch for more papers by this authorP. J. Mandhane, P. J. Mandhane Department of Pediatrics, University of Alberta, Edmonton, AB, CanadaSearch for more papers by this authorS. E. Turvey, S. E. Turvey Department of Pediatrics, University of British Columbia, Vancouver, BC, CanadaSearch for more papers by this authorD. L. Lefebvre, D. L. Lefebvre Department of Medicine, McMaster University, Hamilton, ON, CanadaSearch for more papers by this authorM. R. Sears, M. R. Sears Department of Medicine, McMaster University, Hamilton, ON, CanadaSearch for more papers by this authorthe CHILD Study Investigators, the CHILD Study InvestigatorsCHILD Study Investigators are listed in Appendix 1Search for more papers by this authorL. Bode, L. Bode Department of Pediatrics and Larsson-Rosenquist Foundation Mother-Milk-Infant Center of Research Excellence, University of California San Diego, La Jolla, CA, USASearch for more papers by this authorM. B. Azad, Corresponding Author M. B. Azad meghan.azad@umanitoba.ca orcid.org/0000-0002-5942-4444 Manitoba Developmental Origins of Chronic Diseases in Children Network (DEVOTION), Children's Hospital Research Institute of Manitoba, Winnipeg, MB, Canada Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada Correspondence Meghan Azad, Children's Hospital Research Institute of Manitoba, Winnipeg, MB, Canada. Email: meghan.azad@umanitoba.caSearch for more papers by this author K. Miliku, K. Miliku orcid.org/0000-0002-9614-7191 Manitoba Developmental Origins of Chronic Diseases in Children Network (DEVOTION), Children's Hospital Research Institute of Manitoba, Winnipeg, MB, Canada Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada The Generation R Study Group, Departments of Pediatrics and Epidemiology, Erasmus University Medical Center Rotterdam, Rotterdam, The NetherlandsSearch for more papers by this authorB. Robertson, B. Robertson Department of Pediatrics and Larsson-Rosenquist Foundation Mother-Milk-Infant Center of Research Excellence, University of California San Diego, La Jolla, CA, USASearch for more papers by this authorA. K. Sharma, A. K. Sharma George & Fay Yee Centre for Healthcare Innovation, University of Manitoba, Winnipeg, MB, CanadaSearch for more papers by this authorP. Subbarao, P. Subbarao Departments of Pediatrics & Physiology, University of Toronto, Toronto, ON, CanadaSearch for more papers by this authorA. B. Becker, A. B. Becker Manitoba Developmental Origins of Chronic Diseases in Children Network (DEVOTION), Children's Hospital Research Institute of Manitoba, Winnipeg, MB, Canada Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, CanadaSearch for more papers by this authorP. J. Mandhane, P. J. Mandhane Department of Pediatrics, University of Alberta, Edmonton, AB, CanadaSearch for more papers by this authorS. E. Turvey, S. E. Turvey Department of Pediatrics, University of British Columbia, Vancouver, BC, CanadaSearch for more papers by this authorD. L. Lefebvre, D. L. Lefebvre Department of Medicine, McMaster University, Hamilton, ON, CanadaSearch for more papers by this authorM. R. Sears, M. R. Sears Department of Medicine, McMaster University, Hamilton, ON, CanadaSearch for more papers by this authorthe CHILD Study Investigators, the CHILD Study InvestigatorsCHILD Study Investigators are listed in Appendix 1Search for more papers by this authorL. Bode, L. Bode Department of Pediatrics and Larsson-Rosenquist Foundation Mother-Milk-Infant Center of Research Excellence, University of California San Diego, La Jolla, CA, USASearch for more papers by this authorM. B. Azad, Corresponding Author M. B. Azad meghan.azad@umanitoba.ca orcid.org/0000-0002-5942-4444 Manitoba Developmental Origins of Chronic Diseases in Children Network (DEVOTION), Children's Hospital Research Institute of Manitoba, Winnipeg, MB, Canada Department of Pediatrics and Child Health, University of Manitoba, Winnipeg, MB, Canada Correspondence Meghan Azad, Children's Hospital Research Institute of Manitoba, Winnipeg, MB, Canada. Email: meghan.azad@umanitoba.caSearch for more papers by this author First published: 18 May 2018 https://doi.org/10.1111/all.13476Citations: 39 Edited by: Liam O'Mahony Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Supporting Information Filename Description all13476-sup-0001-Supinfo.docxWord document, 77.9 KB Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article. Volume73, Issue10October 2018Pages 2070-2073 RelatedInformation

Human milk oligosaccharides (HMOs) are structurally diverse unconjugated glycans with a composition unique to each lactating mother. While HMOs have been shown to have an impact on the development of infant gut microbiota, it is not well known if HMOs also already affect milk microbial composition. To address this question, we analysed eleven colostrum samples for HMO content by high-pressure liquid chromatography and microbiota composition by quantitative PCR. Higher total HMO concentration was associated with higher counts of Bifidobacterium spp. (ρ=0.63, P=0.036). A distinctive effect was seen when comparing different HMO groups: positive correlations were observed between sialylated HMOs and Bifidobacterium breve (ρ=0.84, P=0.001), and non-fucosylated/non-sialylated HMOs and Bifidobacterium longum group (ρ=0.65, P=0.030). In addition to associations between HMOs and bifidobacteria, positive correlations were observed between fucosylated HMOs and Akkermansia muciniphila (ρ=0.70, P=0.017), and between fucosylated/sialylated HMOs and Staphylococcus aureus (ρ=0.75, P=0.007). Our results suggest that the characterised HMOs have an effect on specific microbial groups in human milk. Both oligosaccharides and microbes provide a concise inoculum for the compositional development of the infant gut microbiota.

ABSTRACT Human milk oligosaccharides (HMO), which constitute a major component of human milk, promote the growth of particular bacterial species in the infant's gastrointestinal tract. We hypothesized that HMO also interact with the bacterial communities present in human milk. To test this hypothesis, two experiments were conducted. First, milk samples were collected from healthy women (n = 16); culture-independent analysis of the bacterial communities was performed, HMO content was analyzed, and the relation between these factors was investigated. A positive correlation was observed between the relative abundance of Staphylococcus and total HMO content (r = 0.66). In a follow-up study, we conducted a series of in vitro growth curve experiments utilizing Staphylococcus aureus or Staphylococcus epidermidis and HMO isolated from human milk. HMO exhibited stimulatory effects on bacterial growth under various nutritional conditions. Analysis of culture supernatants from these experiments revealed that HMO did not measurably disappear from the culture medium, indicating that the growth-enhancing effects were not a result of bacterial metabolism of the HMO. Instead, stimulation of growth caused greater utilization of amino acids in minimal medium. Collectively, the data provide evidence that HMO may promote the growth of Staphylococcus species in the lactating mammary gland.

We tested the hypothesis that Human Milk Oligosaccharides (HMO) partially account for the inefficiency of HIV transmission through breastfeeding. Using a case‐control design, we measured HMO by HPLC in breast milk samples of 81 HIV‐infected mothers who transmitted HIV via breastfeeding, 86 HIV‐infected mothers who did not transmit and 36 uninfected mothers recruited in Lusaka, Zambia. Only single‐dose nevirapine was used as prophylaxis. Concentration of the individual HMO 3′‐sialyllactose (3′SL) and lacto‐N‐neo‐tetraose was higher in HIV‐infected vs. uninfected mothers. HIV‐infected mothers with concentrations of combined HMO above the median (1.87 g/L) were significantly less likely to transmit via breastfeeding (odds ratio [OR] 0.45[0.21, 0.97], p=0.04) after adjusting for CD4 count and breast milk viral load. Percent HMO that was 3′SL was higher among transmitting vs. non‐transmitting mothers (p=0.005) and correlated with plasma and breast milk viral load and lower CD4 counts. Higher concentrations of HMO and lesser %3′SL were associated with protection against postnatal HIV transmission. Further study of these novel potentially anti‐HIV components of breast milk is warranted.

Human milk oligosaccharides (HMO), un-conjugated complex carbohydrates that are highly abundant in human milk but not in infant formula, have recently received much attention due to their potential benefits for the breast-fed neonate. While it is becoming evident that HMO structure determines their specific function, understanding the metabolic fate of ingested HMO is key in assessing their biological roles. Remarkably little is known about how, when and where they are metabolized. HMO have long been regarded as metabolically "inert" to the host, as significant amounts are excreted with the feces. HMO reach the colon intact where their prebiotic effects promote healthy gut colonization. HMO can also function as soluble decoy receptors and block adhesion of microbial pathogens to epithelial surfaces. Local effects at the mucosal lining include differential cell responses or modulation of the innate immune system. A small percentage of HMO is believed to be absorbed intact in the small intestine and later excreted with the urine, which opens speculations on possible systemic effects, e.g. in the immune system or in the context of neuronal development. Oligosaccharides currently added to infant formula are structurally different from HMO and therefore most likely not functionally equivalent. Selected "authentic" HMO might soon become available for the supplementation of infant formula, but additional preclinical and clinical studies are required to demonstrate efficacy. This review provides an overview about the structural and functional properties of HMO with emphasis on recent findings in metabolism studies.

Protein-losing enteropathy (PLE), the loss of plasma proteins through the intestine, is a symptom in ostensibly unrelated diseases. Emerging commonalities indicate that genetic insufficiencies predispose for PLE and environmental insults, e. g. viral infections and inflammation, trigger PLE onset. The specific loss of heparan sulfate (HS) from the basolateral surface of intestinal epithelial cells only during episodes of PLE suggests a possible mechanistic link. In the first tissue culture model of PLE using a monolayer of intestinal epithelial HT29 cells, we proved that HS loss directly causes protein leakage and amplifies the effects of the proinflammatory cytokine tumor necrosis factor alpha(TNF alpha). Here, we extend our in vitro model to assess the individual and combined effects of HS loss, interferon gamma (IFN gamma), TNF alpha, and increased pressure, and find that HS plays a central role in the patho-mechanisms underlying PLE. Increased pressure, mimicking venous hypertension seen in post-Fontan PLE patients, substantially increased protein leakage, but HS loss, IFN gamma, or TNF alpha alone had only minor effects. However, IFN gamma up-regulated TNFR1 expression and amplified TNF alpha-induced protein leakage. IFN gamma and TNF alpha compromised the integrity of the HT29 monolayer and made it more susceptible to increased pressure. HS loss itself compromises the integrity of the monolayer, amplifying the effects of pressure, but also amplifies the effects of both cytokines. In the absence of HS a combination of increased pressure, IFN gamma, and TNF alpha caused maximum protein leakage. Soluble heparin fully compensated for HS loss, providing a reasonable explanation for patient favorable response to heparin therapy.

MPI encodes phosphomannose isomerase, which interconverts fructose 6-phosphate and mannose 6-phosphate (Man-6-P), used for glycoconjugate biosynthesis. MPI mutations in humans impair protein glycosylation causing congenital disorder of glycosylation Ib (CDG-Ib), but oral mannose supplements normalize glycosylation. To establish a mannose-responsive mouse model for CDG-Ib, we ablated Mpi and provided dams with mannose to rescue the anticipated defective glycosylation. Surprisingly, although glycosylation was normal, Mpi(-/-) embryos died around E11.5. Mannose supplementation even hastened their death, suggesting that man-nose was toxic. Mpi(-/-) embryos showed growth retardation and placental hyperplasia. More than 90% of Mpi(-/-) embryos failed to form yolk sac vasculature, and 35% failed chorioallantoic fusion. We generated primary embryonic fibroblasts to investigate the mechanisms leading to embryonic lethality and found that mannose caused a concentration- and time-dependent accumulation of Man 6-P in Mpi(-/-) fibroblasts. In parallel, ATP decreased by more than 70% after 24 h compared with Mpi(+/+) controls. In cell lysates, Man-6-P inhibited hexokinase (70%), phosphoglucose isomerase (65%), and glucose-6-phosphate dehydrogenase (85%), but not phosphofructokinase. Incubating intact Mpi(-/-) fibroblasts with 2-[(3)H]deoxyglucose confirmed mannose-dependent hexokinase inhibition. Our results in vitro suggest that mannose toxicity in Mpi(-/-) embryos is caused by Man-6-P accumulation, which inhibits glucose metabolism and depletes intracellular ATP. This was confirmed in E10.5 Mpi(-/-) embryos where Man-6-P increased more than 10 times, and ATP decreased by 50% compared with Mpi(+/+) littermates. Because Mpi ablation is embryonic lethal, a murine CDG-Ib model will require hypomorphic Mpi alleles.

Journal of Pediatric Gastroenterology and NutritionVolume 39, Issue S1 p. S59-S59 ABSTRACTS: Poster Session Abstracts P0009 PP AN IN VITRO CELLULAR MODEL OF PROTEIN-LOSING ENTEROPATHY IMPLICATES HEPARAN SULFATE, INFLAMMATORY CYTOKINES, AND HYDROSTATIC PRESSURE AS CRITICAL PLAYERS L. Bode, L. Bode Glycobiology Program, The Burnham Institute, La Jolla, United StatesSearch for more papers by this authorE. Eklund, E. Eklund Glycobiology Program, The Burnham Institute, La Jolla, United StatesSearch for more papers by this authorC. Salvestrini, C. Salvestrini Centre of Paediatric Gastroenterology, Royal Free and University College School of Medicine, London, United KingdomSearch for more papers by this authorS. Murch, S. Murch Centre of Paediatric Gastroenterology, Royal Free and University College School of Medicine, London, United KingdomSearch for more papers by this authorH. Freeze, H. Freeze Glycobiology Program, The Burnham Institute, La Jolla, United StatesSearch for more papers by this author L. Bode, L. Bode Glycobiology Program, The Burnham Institute, La Jolla, United StatesSearch for more papers by this authorE. Eklund, E. Eklund Glycobiology Program, The Burnham Institute, La Jolla, United StatesSearch for more papers by this authorC. Salvestrini, C. Salvestrini Centre of Paediatric Gastroenterology, Royal Free and University College School of Medicine, London, United KingdomSearch for more papers by this authorS. Murch, S. Murch Centre of Paediatric Gastroenterology, Royal Free and University College School of Medicine, London, United KingdomSearch for more papers by this authorH. Freeze, H. Freeze Glycobiology Program, The Burnham Institute, La Jolla, United StatesSearch for more papers by this author First published: 01 June 2004 https://doi.org/10.1002/j.1536-4801.2004.tb12439.x Submitted by: [email protected] Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume39, IssueS1June 2004Pages S59-S59 RelatedInformation