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Despite advances in lactation skills and knowledge, insufficient milk production still continues to mystify mothers and lactation consultants alike. Based on 3 cases with similar threads, a connection is proposed between polycystic ovary syndrome (PCOS) and insufficient milk supply. Described are the etiology and possible symptoms of PCOS such as amenorrhea/oligomenorrhea, hirsutism, obesity, infertility, persistent acne, ovarian cysts, elevated triglycerides, and adult-onset diabetes, along with possible pathological interference with mammogenesis, lactogenesis, and galactopoiesis. Clinical suggestions include guidelines for screening mothers and careful monitoring of babies at risk. Further research is necessary to confirm the proposed association and to develop therapies with the potential to improve lactation success. [ABSTRACT FROM AUTHOR]

Cocoa butter equivalents (CBE) are mixtures of triglycerides from multiple sources (e.g., sunflower oil, mango kernel and sal), which resemble cocoa butter (CB) in both physical and chemical properties. Despite being widely used to replace CB in chocolate products, the crystallization behavior of many CBEs is still poorly understood. The aim of this work was to develop a fundamental understanding, at the molecular level, of the crystallization behavior of selected CBEs, and compare it with that of CB. Chromatography was used to determine the composition of CBEs, in terms of fatty acids and triacylglycerides (TAGs), while their thermodynamic behavior and crystallization kinetics were studied using polarized microscopy, differential calorimetry and three different synchrotron X-ray scattering setups. CBEs of different origin and chemical composition (e.g., different ratios of the main CB TAGs, namely POP, SOS and POS) crystallized in different polymorphs and with different kinetics of nucleation, growth and polymorphic transformation. SOS rich CBEs presented showed more polymorphs than CB and POP rich samples; whereas, CBEs with high concentration of POP showed slow kinetic of polymorphic transformation towards the stable β(3L) form. Additionally, it was observed that the presence of small amounts (<1% w/w) of specific TAGs, such as OOO, PPP or SSS, could significantly affect the crystallization behavior of CBEs and CBs in terms of kinetics of polymorphic transformation and number of phases detected (multiple high melting β(2L) polymorphs were identified in all samples studied). Finally, it was found that, regardless of the CBE composition, the presence of shear could promote the formation of stable β polymorphs over metastable β' and γ forms, and reduced the size of the crystal agglomerates formed due to increased secondary nucleation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

This study describes the identification and quantification of fatty acids in the sn -2 position of triacylglycerols (TAG) and of the most abundant TAG regioisomers in human milk by liquid chromatography coupled with high-resolution mass spectrometry (HPLC-HRMS). Over 300 individual TAG species were observed and 1,3-olein-2-palmitin (OPO) was identified as the most abundant TAG regioisomer. Validation of the HPLC-HRMS method showed repeatability and intermediate reproducibility values ranging from 3.1 to 16.6% and 4.0 to 20.7%, respectively, and accuracy ranging from 75 to 97%. Results obtained by the HPLC-HRMS method were comparable to results from the ISO 6800 method for the quantification of palmitic acid in the sn -2 position of TAG (81.4 and 81.8 g 100 g -1 total palmitic acid, respectively). Processing the data obtained with the HPLC-HRMS method is extremely time consuming and, therefore, a targeted method suitable for the quantification of OPO in human milk samples by ultra-performance (UP) LC coupled with triple quadrupole (QQQ) MS was developed and validated. OPO identification and quantification by UPLC-QQQ were based on nominal mass and a fragmentation pattern obtained by multiple reaction monitoring experiments. The method was validated in terms of accuracy and precision by analyzing different aliquots of the same human milk sample over time and comparing the results with values obtained by HPLC-HRMS. Intermediate reproducibility was <15% and trueness comparable to HPLC-HRMS. Quantification of OPO in human milk samples collected at 30, 60 and 120 days postpartum showed that OPO content varies between 333 ± 11.8 and 383 ± 18.0 mg 100mL -1 .

Background & Aims: Polyphenol intake has been linked to improvements in human vascular function, although data on hydroxycinnamates, such as chlorogenic acid (CGA) have not yet been studied. We aimed to investigate the impact of coffee intake rich in chlorogenic acid on human vascular function and whether CGAs are involved in potential effects., Methods: Two acute randomized, controlled, cross-over human intervention trials were conducted. The impact of coffee intake, matched for caffeine but differing in CGA content (89, and 310 mg) on flow-mediated dilatation (FMD) was assessed in 15 healthy male subjects. In a second intervention trial conducted with 24 healthy male subjects, the impact of pure 5-caffeoylquinic acid (5-CQA), the main CGA in coffee (5-CQA; 450 mg and 900 mg) on FMD was also investigated., Results: We observed a bi-phasic FMD response after low and high polyphenol, (89 mg and 310 mg CGA) intake, with increases at 1 (1.10 ± 0.43% and 1.34 ± 0.62%, respectively) and 5 (0.79% ± 0.32 and 1.52% ± 0.40, respectively) hours post coffee consumption. FMD responses to coffee intake was closely paralleled by the appearance of CGA metabolites in plasma, notably 3-, 4- and 5-feruloylquinic acid and ferulic-4'-O-sulfate at 1 h and isoferulic-3'-O-glucuronide and ferulic-4'-O-sulfate at 5 h. Intervention with purified 5-CQA (450 mg) also led to an improvement in FMD response relative to control (0.75 ± 1.31% at 1 h post intervention, p = 0.06) and concomitant appearance of plasma metabolites., Conclusions: Coffee intake acutely improves human vascular function, an effect, in part, mediated by 5-CQA and its physiological metabolites., Study Registration: The National Institutes of Health (NIH) on ClinicalTrials.govNCT01813981 and NCT01772784., (Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Fatty acids (FA), phospholipids (PL), and gangliosides (GD) play a central role in infant growth, immune and inflammatory responses. The aim of this study was to determine FA, PL, and GD compositional changes in human milk (HM) during lactation in a large group of Chinese lactating mothers (540 volunteers) residing in Beijing, Guangzhou, and Suzhou. HM samples were collected after full expression from one breast and while the baby was fed on the other breast. FA were assessed by direct methylation followed by gas chromatography (GC) analysis. PL and GD were extracted using chloroform and methanol. A methodology employing liquid chromatography coupled with an evaporative light scattering detector (ELSD) and with time of flight (TOF) mass spectrometry was used to quantify PL and GD classes in HM, respectively. Saturated FA (SFA), mono-unsaturated FA (MUFA), and PL content decreased during lactation, while polyunsaturated FA (PUFA) and GD content increased. Among different cities, over the lactation time, HM from Beijing showed the highest SFA content, HM from Guangzhou the highest MUFA content and HM from Suzhou the highest n -3PUFA content. The highest total PL and GD contents were observed in HM from Suzhou. In order to investigate the influence of the diet on maternal milk composition, a careful analyses of dietary habits of these population needs to be performed in the future., Competing Interests: Conflicts of InterestThis study was funded by Nestlé Nutrition Institute, China, and Nestlé Research Centre, Beijing, China. F.G., C.C.-H., E.B., P.F., I.M.E., I.T., C.M., B.S.-B., S.K.T., C.A.D.-C and G.V.-P. were all employees of Nestec Ltd. when this work was done. The opinions expressed in the article are those of the authors alone and do not necessarily reflect the views or recommendations of their respective affiliations.

It has been reported that dietary gangliosides may have an important role in preventing infections and in brain development during early infancy. However, data related to the evolution of their concentration over the different stages of lactation are scarce. Liquid chromatography coupled with electrospray ionization high resolution mass spectrometer (LC/ESI-HR-MS) has been optimized to quantify the two major ganglioside classes, i.e., aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer (GD3) and aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer (GM3) in human milk. Gangliosides were extracted using chloroform and methanol, further purified by solid-phase extraction and separated by reversed-phase liquid chromatography. Repeatability, intermediate reproducibility, and recovery values were assessed to validate the method. In human milk, GD3 and GM3 could be quantified at the level of 0.1 and 0.2 μg/mL, respectively, with relative standard deviation of repeatability [CV(r)] and intermediate reproducibility [CV(iR)] values ranging from 1.9 to 15.0 % and 1.9 to 22.5 %, respectively. The described method was used to quantify GD3 and GM3 in human milk samples collected from 450 volunteers between 0 and 11 days and at 30, 60 and 120 days postpartum, providing for the first time the concentration of these minor lipids in a large cohort. The content of total gangliosides ranged from 8.1 and 10.7 μg/mL and the mean intake of gangliosides in infants 30, 60 and 120 days postpartum could be estimated at about 5.5, 7.0 and 8.6 mg of total gangliosides per day, respectively, when infants were exclusively breastfed.

Scope: Coffee contains phenolic compounds, mainly chlorogenic acids (CGAs). Even though coffee intake has been associated with some health benefits in epidemiological studies, the bioavailability of coffee phenolics is not fully understood., Objective and Study Design: We performed a dose-response study measuring plasma bioavailability of phenolics after drinking three increasing, but still nutritionally relevant doses of instant pure soluble coffee. The study design was a one treatment (coffee) three-dose randomized cross-over design, with a washout period of 2 wks between visits., Results: CGAs, phenolic acids, and late-appearing metabolites all increased with increasing ingested dose. Hence, the sum of area under the curve was significantly higher for the medium to low dose, and high to medium dose, by 2.23- and 2.38-fold, respectively. CGAs were not well absorbed in their intact form, regardless of the dose. CGA and phenolic acids appeared rapidly in plasma, indicating an early absorption in the gastrointestinal tract. Late-appearing metabolites were the most abundant, regardless of the dose., Conclusion: This study confirmed previous findings about coffee bioavailability but also showed that coffee phenolics appear in a positive dose-response manner in plasma when drank at nutritionally relevant doses., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Chlorogenic acids and derivatives like phenolic acids are potentially bioactive phenolics, which are commonly found in many foods. Once absorbed, chlorogenic and phenolic acids are highly metabolized by the intestine and the liver, producing glucuronidated and/or sulphated compounds. These metabolites were analyzed in human plasma using a validated liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method. After protein precipitation, phenolic acids and their metabolites were extracted by using ethanol and chromatographic separation was achieved by reversed-phase using an Acquity UPLC BEH C18 column combined with a gradient elution system using 1% acetic acid aqueous solution and 1% acetic acid with 100% acetonitrile. The method was able to quantify 56 different compounds including 24 phenolic acids, 4 lactones, 15 sulfates and 13 glucuronides metabolites between 5 and 1000nM in plasma for most of them, except for m-dihydrocoumaric acid, 5-ferulloylquinic-glucuronide, 4-methoxycinnamic acid, 3-phenylpropionic acid, 3-(4-methoxyphenyl)propionic acid (25 to 1000nM) and p-dihydrocoumaric acid (50-1000nM). Values of repeatability and intermediate reproducibility were below 15% of deviation in general, and maximum 20% for the lowest concentrations. The validated method was successfully applied to quantify phenolic acids and their metabolites in plasma obtained after oral ingestion of soluble coffee. In conclusion, the developed and validated method is proved to be very sensitive, accurate and precise for the quantification of these possible dietary phenols., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Scope: Tea is an infusion of the Camellia sinensis leaves. The most prevalent bioactive compounds in green tea are catechins (C), which are of great interest for their potential health-promoting effects. However, metabolism and bioavailability of C are not fully understood., Methods and Results: This study investigates the human bioavailability (plasma appearance) of C after drinking three doses of infused green tea in a randomized cross-over design. The sum of area under the curve increased between the small (0.75% w/v, 180 mg total C) and medium (1.25%) dose of ingested green tea but not between the medium and the high (1.75%) dose. The overall pattern for the sum of C did not reflect the fate of individual C. While (-)-epigallocatechin and 4'-O-Me-epigallocatechin showed saturation in plasma between the medium and high green tea doses, (-)-epigallocatechin gallate and (-)-epicatechin did not "saturate" and increased proportionally with the ingested dose. Regardless of the dose, C appeared rapidly in plasma as monophasic curves, suggesting absorption in the small intestine and minimal entero-hepatic circulation., Conclusion: As a conclusion, when studying dose response of polyphenols and metabolites, one must look not only at the overall pattern of plasma appearance, but also at data specific for each metabolite., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Scope: Until now, the question of how the ingested doses of chlorogenic acids (CGA) from coffee influence their absorption and metabolism remains unresolved. To assess absorption in the small intestine, we performed a dose-response study with a randomized, double-blinded, crossover design with ileostomist subjects., Methods and Results: After a polyphenol-free diet, the volunteers consumed, on three separate occasions, coffee with different total CGA contents (high 4525 μmol; medium 2219 μmol; low 1053 μmol). CGA concentrations in plasma, ileal effluent, and urine were subsequently determined by HPLC-DAD-ESI-MS and -ESI-MS/MS. The results show that the consumption of higher CGA concentrations leads to a faster ileal excretion. This corresponds to a renal excretion of 8.0 ± 4.9% (high), 12.1 ± 6.7% (medium), and 14.6 ± 6.8% (low) of total CGA and metabolites. Glucuronidation of CGA became slightly greater with increasing dose. After enzyme treatment, the area under the curve (AUC)(0-8h) for CGA metabolites in plasma was 4412 ± 751 nM × h(0-8) (-1) (high), 2394 ± 637 nM × h(0-8) (-1) (medium), 1782 ± 731 nM × h(0-8) (-1) (low), respectively. Additionally, we were able to identify new metabolites of CGA in urine and ileal fluid., Conclusion: We conclude that the consumption of high CGA concentrations via coffee might influence the gastrointestinal transit time and consequently affect CGA absorption and metabolism., (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Tea is an infusion of the leaves of the Camellia sinensis plant and is the most widely consumed beverage in the world after water. Green tea contains significant amounts of polyphenol catechins and represents a promising dietary component to maintain health and well-being. Epidemiological studies indicate that polyphenol intake may have potential health benefits, such as, reducing the incidence of coronary heart disease, diabetes and cancer. While bioavailability of green tea bioactives is fairly well understood, some gaps still remain to be filled, especially the identification and quantification of conjugated metabolites in plasma, such as, sulphated, glucuronidated or methylated compounds., Aim of the Study: In the present study, we aimed to quantify the appearance of green tea catechins in plasma with particular emphasis on their methylated forms., Results: After feeding 400 mL of green tea, 1.25% infusion to 9 healthy subjects, we found significant amounts of EC, EGC and EGCg in plasma as expected. EGC was the most bioavailable catechin, and its methylated form (4'-O-Me-EGC) was also present in quantifiable amounts. Its kinetics followed that of its parent compound. However, the relative amount of the methylated form of EGC was lower than that of the parent compound, an important aspect which, in the literature, has been controversial so far. The quantitative results presented in our study were confirmed by co-chromatography and accurate mass analysis of the respective standards. We show that the relative abundance of 4'-O-Me-EGC is ~40% compared to the parent EGC., Conclusion: 4'-O-Me-EGC is an important metabolite derived from catechin metabolism. Its presence in significant amounts should not be overlooked when assessing human bioavailability of green tea.

Coffee and green tea are two of the most widely consumed hot beverages in the world. Their respective bioavailability has been studied separately, but absorption of their respective bioactive phenolics has not been compared. In a randomised cross-over design, nine healthy subjects drank instant coffee and green tea. Blood samples were collected over 12 h and at 24 h to assess return to baseline. After green tea consumption, (-)-epigallocatechin (EGC) was the major catechin, appearing rapidly in the plasma; (-)-EGC gallate (EGCg) and (-)-epicatechin (EC) were also present, but (-)-EC gallate and C were not detected. Dihydroferulic acid and dihydrocaffeic acid were the major metabolites that appeared after coffee consumption with a long time needed to reach maximum plasma concentration, suggesting metabolism and absorption in the colon. Other phenolic acid equivalents (caffeic acid (CA), ferulic acid (FA) and isoferulic acid (iFA)) were detected earlier, and they peaked at lower concentrations. Summations of the plasma area under the curves (AUC) for the measured metabolites showed 1.7-fold more coffee-derived phenolic acids than green tea-derived catechins (P = 0.0014). Furthermore, we found a significant correlation between coffee metabolites based on AUC. Inter-individual differences were observed, but individuals with a high level of CA also showed a correspondingly high level of FA. However, no such correlation was observed between the tea catechins and coffee phenolic acids. Correlation between AUC and maximum plasma concentration was also significant for CA, FA and iFA and for EGCg. This implies that the mechanisms of absorption for these two classes of compounds are different, and that a high absorber of phenolic acids is not necessarily a high absorber of catechins.

Previous studies on coffee examined absorption of phenolic acids (PA) in the small intestine, but not the contribution of the colon to absorption. Nine healthy volunteers ingested instant soluble coffee ( approximately 335 mg total chlorogenic acids (CGAs)) in water. Blood samples were taken over 12 h, and at 24 h to assess return to baseline. Many previous studies, which used glucuronidase and sulfatase, measured only PA and did not rigorously assess CGAs. To improve this, plasma samples were analyzed after full hydrolysis by chlorogenate esterase, glucuronidase and sulfatase to release aglycone equivalents of PA followed by liquid-liquid extraction and ESI-LC-ESI-MS/MS detection. Ferulic, caffeic and isoferulic acid equivalents appeared rapidly in plasma, peaking at 1-2 h. Dihydrocaffeic and dihydroferulic acids appeared in plasma 6-8 h after ingestion (T(max=)8-12 h). Substantial variability in maximum plasma concentration and T(max) was also observed between individuals. This study confirms that the small intestine is a significant site for absorption of PA, but shows for the first time that the colon/microflora play the major role in absorption and metabolism of CGAs and PA from coffee.

Chlorogenic acids (CGA) are antioxidants found in coffee. They are becoming of interest for their health-promoting effects, but bioavailability in humans is not well understood. We hypothesized that adding whole milk or sugar and nondairy creamer to instant coffee might modulate the bioavailability of coffee phenolics. Nine healthy participants were asked to randomly drink, in a crossover design, instant coffee (Coffee); instant coffee and 10% whole milk (Milk); or instant coffee, sugar, and nondairy creamer already premixed (Sugar/NDC). All 3 treatments provided the same amount of total CGA (332 mg). Blood was collected for 12 h after ingestion and plasma samples treated using a liquid-liquid extraction method that included a full enzymatic cleavage to hydrolyze all CGA and conjugates into phenolic acid equivalents. Hence, we focused our liquid chromatography-Electrospray ionization-tandem MS detection and quantification on caffeic acid (CA), ferulic acid (FA), and isoferulic acid (iFA) equivalents. Compared with a regular black instant coffee, the addition of milk did not significantly alter the area under the curve (AUC), maximum plasma concentration (C(max)), or the time needed to reach C(max) (T(max)). The C(max) of CA and iFA were significantly lower and the T(max) of FA and iFA significantly longer for the Sugar/NDC group than for the Coffee group. However, the AUC did not significantly differ. As a conclusion, adding whole milk did not alter the overall bioavailability of coffee phenolic acids, whereas sugar and nondairy creamer affected the T(max) and C(max) but not the appearance of coffee phenolics in plasma.

Based on this more thorough assessment, the lactation consultant may be able to identify all of the factors contributing to this complex case. In some situations, her skilled interventions will suffice once the underlying problem is addressed. Occasionally, she will identify a factor that falls outside of her area of expertise; when this happens, she must make the appropriate referrals. For example, a referral to a physician for a frenotomy or suspected neurological or other medical problem is appropriate. It is clear that because Baby E's problems were not resolved after 6 weeks of concerned effort, something was missed. It would certainly be appropriate for the lactation consultant to refer the dyad to another lactation consultant who has more expertise in handling clinically challenging breastfeeding problems. If possible, the referring lactation consultant should accompany the dyad so that she can improve her clinical skills. Assuming Baby E does not have underlying medical problems, the most likely causes of Baby E's difficulties are anatomical variation and/or sucking dysfunction. Because the baby is so fussy, it also would be wise to consider the possibility of allergies or food tolerance. Our first rule is " Feed the baby." The second rule is " Correct or work on correcting the problem or problems." Our goal is to achieve exclusive breastfeeding or as close an approximation as possible. We almost never give up on this goal, but we do educate the mother and work professionally with her choices. Until the baby is breastfeeding well, the lactation consultant will probably need to instruct the mother to continue using a pump ( preferably a hospital-grade, electric, bilateral pump). The mother should use the pump physiologically, pumping as many times a day as the baby would breastfeed. As soon as the situation improves, the mother should be instructed to wean gradually from the pump and any other breastfeeding equipment she is using. The goal should always be to help the mother and baby acheive a breastfeeding relationship, preferably without the use of any devices. We usually suggest that the mother avoid all rubber nipples and pacifiers during this learning period. Babies have a strong need to suck. Correct sucking helps the baby organize and be soothed. Whenever possible, we prefer infants to use their mother's breasts for pacification, warmth, love, smell, and food rather than artificial nipples and devices. Mothers almost always want to know how much work and time is involved before committing to following suggested treatment plans. As a general rule, we have found that it will take approximately the same number of weeks as the baby's age to solve the problems completely. In this case, it will probably take about 6 weeks until mother and baby graduate from "breastfeeding school." The first 2 weeks would most likely be very intense for the whole family, with the mother getting very little sleep. VJ is likely to cry when talking to the lactation consultant during this period of intense change. It is helpful during these times to listen to the mother, reinforce that you know how hard she is working and that what she is feeling is normal. Giving the mother a hug and complimenting her mothering efforts go a long way toward encouraging her to continue. It is not a time to give up. The second 2 weeks typically are easier, as everybody is used to the workload and required skills. The focus becomes refining skills. The last 2 weeks is usually a time to reduce and then wean off the equipment and exercises. This timing is just a guideline and must always be individualized. Although it is a tremendous amount of work for the mother, baby, family, and lactation consultant to correct well-established but incorrect breastfeeding behaviors, we have never met a mother who was sorry that she chose to tackle the problem. Even if she tries and then gives up or achieves only a partial milk supply or partial breastfeeding relationship, she can take pleasure in knowing that she left no stone unturned. Unfortunately, mothers and babies with presentations similar to that of VJ and Baby E all too often fail to establish an exclusive breastfeeding relationship. Not only are patience, dedication, time, and skills needed, but there are often multiple underlying problems that need to be solved. With a thorough assessment and appropriate use of skills and equipment by the lactation consultant, success is much more likely. This particular dyad should be able to acheive an exclusive breastfeeding relationship.

Despite advances in lactation skills and knowledge, insufficient milk production still continues to mystify mothers and lactation consultants alike. Based on 3 cases with similar threads, a connection is proposed between polycystic ovary syndrome (PCOS) and insufficient milk supply. Described are the etiology and possible symptoms of PCOS such as amenorrhea/oligomenorrhea, hirsutism, obesity, infertility, persistent acne, ovarian cysts, elevated triglycerides, and adult-onset diabetes, along with possible pathological interference with mammogenesis, lactogenesis, and galactopoiesis. Clinical suggestions include guidelines for screening mothers and careful monitoring of babies at risk. Further research is necessary to confirm the proposed association and to develop therapies with the potential to improve lactation success.

It seems appropriate for the lactation consultant to be the advocate for infants and children. We must act ourselves if we want change to occur. When individuals cause even small changes to be made, the cumulative effect can be profound.

A review of cases of short frenulum (tongue-tie) seen in a recent year at the Lactation Institute and Breastfeeding Clinic provides data about its relationship to sucking and breastfeeding problems such as insufficient infant weight gain and reduced milk supply, sore nipples and repeat bouts of mastitis in the mother. Frenotomy was recommended for ten of 13 babies who appeared to have a short frenulum. Three mothers chose not to hae the frenulum clipped and either gave up breastfeeding or continued to experience problems. Breastfeeding was successfully established by the five healthy babies whose frenulum was clipped. The two babies for whom frenotomy did not completely correct breastfeeding problems had severe birth defects.