Suppression of genes involved in DNA repair, tumor suppression and detoxification through epigenetic modifications has been implicated in the etiology of cancer. As such analysis of promoter methylation patterns in genes frequently down regulated in breast cancer in non-cancerous subjects may serve as an indicator of breast cancer risk. CpG-island hypermethylation of single genes has been detected in cells isolated from nipple aspirate and ductal lavage, yet both isolation methods yield insufficient cells to complete an extensive analysis on any one donor sample. As an alternative we have turned to magnetic separation of human mammary epithelial cells from breast milk. Initial studies with these cells, which are detailed in chapter one, show that a breast milk sample provides sufficient epithelial cells to isolate high quality RNA for gene expression analyses or genomic DNA for methylation analysis of multiple genes. Using quantitative RT-PCR of RNA collected from these samples we detected differences in the mRNA levels for six genes known to be down regulated in breast cancers: BRCA1, p16, CDH1, TMS-1, GSTPi, and SFRP1. Additionally using methyl-specific PCR (MSP) we assayed for a small panel of genes frequently methylated in cancer and found them to be unmethylated in the few breast milk samples examined. However, given the small number of CpG sites which can be assayed by the MSP technique it is not surprising that methylation was not detected in disease-free subjects. With methods for collecting breast milk samples and processing them for genetic material established we turned to a more comprehensive study of DNA methylation in larger population of donors which is detailed in chapter two. Utilizing a highly sensitive and highly quantitative methylation analysis technique known as Pyrosequencing we examined age-related methylation patterns for RASSF1A, TMS-1, CDH1, SFRP1, GSTPi, and CRBP1 in genomic DNA purified from exfoliated epithelial cells magnetically enriched from breast milk (n=111) and whether the protective effects conveyed by early pregnancy could be partly due to decreases in DNA hypermethylation. Although firm answers about early pregnancy were inconclusive based on our sample pool, this body of work lays down a solid foundation for future studies.

Breastfeeding is associated with long-term wellbeing including low risks of infectious diseases and non-communicable diseases such as asthma, cancer, autoimmune diseases and obesity during childhood. In recent years, important advances have been made in understanding the human breast milk (HBM) composition. Breast milk components such as, non-immune and immune cells and bioactive molecules, namely, cytokines/chemokines, lipids, hormones, and enzymes reportedly play many roles in breastfed newborns and in mothers, by diseases protection and shaping the immune system of the newborn. Bioactive components in HBM are also involved in tolerance and appropriate inflammatory response of breastfed infants if necessary. This review summarizes the current literature on the relationship between mother and her infant through breast milk with regard to disease protection. We will shed some light on the mechanisms underlying the roles of breast milk components in the maintenance of health of both child and mother.

Objectives- The purpose of this study was to determine if differences in breast milk (BM) short chain fatty acid (SCFA) and microbiome profiles are correlated to maternal BMI. Our hypothesis is that BM SCFA are a reflection of colonic SCFA distribution and concentration and may reflect microbiome diversity in the maternal gut. Methods- Study design-This was a cohort study in which forty-six Caucasian participants were recruited from BABE Breastfeeding Coalition of Tri-Cities, divided into two groups, one group with normal pre-gravid BMI between 18.5 and 24.9 kg/m2 (n=23) and the other group with overweight or obese pre-gravid BMI greater than 25.0 kg/m2 (n=23). Each participant completed a demographic and health survey and provided 4 ounces of expressed BM. This study was approved by the ETSU IRB (0915.8s-ETSU). 16s rRNA Isolation & Quantification- Microbiome analysis was performed on thirty-four samples (n=13 for overweight/obese, and n=21 for normal weight). Qiagen QIAmp PowerFecal Pro DNA Kit was utilized for isolation of microbiome DNA; Amplicon sequencing of the 16S rRNA region was performed at the University of Tennessee Genomics Core Laboratory utilizing a modified Klindworth et al method. Microbiome Analysis- Operational Taxonomic Unit (OUT) clustering and taxonomic analysis were performed using CLC Genomics Workbench. Alpha diversity indexes were calculated using the Abundance Analysis tool, and the weighted Unifrac metric was used to calculate Beta diversity. Fatty Acid Profile- BM samples were subjected to SCFA extraction and analysis using a modified Schwiertz et al. method. The resulting SCFA profiles were then utilized to determine if there were any significant differences between groups. Results- No significance was observed in BM microbiome between the normal weight and overweight/obese groups for alpha or beta diversity. Significance was detected between the groups for valeric (p=0.02) and isocaproic acids (p=0.05) with the normal weight group higher than the overweight/obese group. No significance was observed for any of the other SCFAs. Conclusions- Although these results are not significant due to low sample size and lack of diversity, they potentially offer insights into the impact of maternal BMI on microbiome and SCFA profiles, which can have implications for infant health and development. Funding Sources- ETSU Small RDC Grant

Mammary gland (MG) is a dynamic organ that is essential for the production and secretion of breast milk. During pregnancy the MG undergoes a critical phase of remodeling, which is accompanied by the differentiation of mammary epithelial cells (MECs). During lactation, the MG requires high level of energy for proper folding of proteins in the secretory pathway, which takes place in the endoplasmic reticulum (ER). Limited evidence has been reported on the impact of cannabis or its components, delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD), on the differentiation of MECs. However, both THC and CBD have been reported to induce ER stress in various cell types, resulting in impacting cellular function. Furthermore, consequences of cannabis use in the perinatal period on breast milk composition have not been reported. Using the HC11 cell line, we investigated whether THC and CBD evoke ER stress in MECs resulting in the impaired cellular function. We also reported on the effects of cannabis use during pregnancy and lactation on the levels of macronutrients and bioactive factors in the breast milk of cannabis users. Relative to control, 10μM THC and 10μM CBD reduced mRNA levels of milk proteins (CSN2 and WAP) and lipid synthesizing enzymes (FASN, FABP4, PLIN2 and LPL), as well as whey acidic protein and lipid levels. In addition, 10μM THC, 10μM CBD, and the combination of 10μM THC + 10μM CBD significantly induced the expression of ER stress genes in HC11 cells. Furthermore, in the milk of women who used cannabis during pregnancy and lactation, the levels of secretory immunoglobulin A (SIgA) were decreased, relative to non-users. Altogether, our findings indicate that cannabis use in the perinatal period may have implications on breast milk composition and infant health. / Thesis / Doctor of Philosophy (Medical Science) / Breast milk is crucial to the nutrition and immunity of the newborn. It is produced by the mammary gland (MG), which is composed of mammary epithelial cells (MEC) that play a key role in producing and secreting proteins and factors into the milk. During pregnancy, the MG undergoes remodeling which is accompanied by differentiation of the MECs (change from unspecialized to a specialized stage that allows the MECs to produce milk proteins and other factors). The disruption in MEC differentiation can result in altered milk production and composition. Cannabis is used by women during pregnancy and breastfeeding. In this study, we investigated the impact of cannabinoids on the MEC differentiation and examined the effect of cannabis use in the perinatal period on the breast milk composition in humans. We reported that cannabinoids reduced the differentiation of MECs and maternal cannabis use during breastfeeding reduced levels of an essential immune factor.

Thesis (MPH.) -- University of Limpopo, 2022 / BACKGROUND: Exclusive breastfeeding in South Africa to date has not seen progress. The data that is available show that most mothers do initiate breastfeeding immediately after delivery of the baby, but its continuation to the first six months is still a challenge. Although its benefits to the mother and baby are mostly known, it is not translated into positive outcomes as recommended by WHO i.e. to feed the baby with breast milk only, no water, no solids except for medicines that have been prescribed for the first six months. OBJECTIVES: To explore and describe perceptions of breastfeeding mothers regarding factors contributing to exclusive breastfeeding for the first six months at Mugodeni Grace Health centre. METHODS: A qualitative, exploratory and descriptive phenomenological method was used in the study, which was conducted at Mugodeni Grace Health Centre. The convenience sampling strategy was used to recruit participants and the sample size was determined by data saturation. Face-to-face in-depth semi-structured interviews were conducted with nine participants in Xitsonga, using an interview guide and were audio taped with the consent of the participants and transcribed verbatim. Data was analysed using Tesch open coding approach. RESULTS: Three themes and nine subthemes emerged from the data analysis, which includes maternal knowledge of breastfeeding, support systems for exclusive breastfeeding and social and cultural influences leading to non-exclusive breastfeeding CONCLUSION: Exclusive breastfeeding for the first six months is a societal issue, which need the involvement of society as a whole to perceive it differently for it to be successful, a family-centred approach in which the father has a role to be part of it to assist and support the mother. Continuous support from the family and health facilities by knowledgeable health practitioners is desired. The study revealed that some nurses were practising wrong breastfeeding practices in front of community members. This made mothers to lose confidence in them as people who can be consulted in case of breastfeeding challenges. Cultural beliefs and myths should be handled in a culturally sensitive manner. Working mothers should be taught to negotiate and make a plan for breastfeeding with the employer during pregnancy and to know of their rights at work viii regarding breastfeeding. Mothers should be taught to breastfeed for eight to twelve times per day. They should also be taught about the causes of crying in babies and shown practical skills on how to latch and how to calm the crying baby.

Studies of the interaction of the relevant metal, metalloid or nanoparticulate species with biological systems are underpinned by the provision of reliable information about chemical composition of the relevant materials. Often, no methods of chemical analysis are available. The work described in this dissertation centers on developing methods to help with studies for a variety of analytes and samples. A method was developed for the determination of 11 trace elements (As, Cd, Co, Cr, Cu, Fe, Mg, Mn, Pb, Ti, and Zn) in human breast milk and infant formulas by inductively coupled plasma optical emission spectrometry (ICP-OES) following microwave-assisted digestion. A method was established for the determination of trace elements, with an emphasis on titanium as titanium dioxide, in snack foods and consumer products. The interactions of some dissolved metals, including rare earth elements, and metallo-nanoparticles (silver, gold, titanium dioxide, aluminum oxide, and iron) with aquatic plants were studied. After exposure in a variety of mesocosms, the partitioning of the elemental species between various compartments was quantified by ICP-MS and ICP-OES following microwave-assisted digestion. An ICP mass spectrometry (MS) method has also been developed to quantify the uptake of gold and silver nanoparticles by C. elegans. Uptake of gold nanoparticles was size dependent, suggesting increased ingestion efficiency with increased particle diameter. The feasibility of discriminating between suspended TiO2 nanoparticles and dissolved titanium by the analysis of the rapid transient signal events obtained from the ICP-MS instrument operated in a rapid response mode was also developed. Data handling parameters were established that allowed a distinction in the signals for nanoparticulate and standard solutions. Spikes in the signal were defined by distinct parameters using the mean and standard deviation, where a spike in the signal was defined as a signal > + ks (k =3). This approach however did lead to a statistical difference in the spike signal events for solutions and nanosuspensions.

Preterm infants are not able to tolerate full enteral feeding of breast milk due to gut immaturity. To fill this nutritional gap, parenteral nutrition is introduced while enteral feeding is gradually increased. Of parenteral nutrition, lipid emulsions supply energy, essential and polyunsaturated fatty acids, which significantly affect short- and long-term health outcomes of growth, visual-and neuro-development for preterm infants. However, elevated plasma triglyceride (TG) levels in preterm infants receiving lipid emulsions have been observed despite less lipid intake compared to breast milk fed infants. We hypothesized that unbalanced fatty acid profiles in lipid emulsion was one factor to cause high plasma TGs for preterm infants. In the multi-center, observational, prospective study, the following samples were analyzed using GC-MS: (1) lipid emulsions (n=5) and breast milk (n=112), (2) plasma (n=294) including normal TG (n=116) and high TG (n=88). Lipoproteins in normal TG (n=18) and high TG (n=24) plasma were measured using gel electrophoresis. Fatty acid profiles in lipid emulsions differed from ones in breast milk. Plasma fatty acid profiles were related to dietary fatty acid intake. Accumulation of all fatty acids except C20:5n3 and relatively high (LDL+VLDL) levels (p<0.001) resulted in high TG plasma compared to normal TG plasma. Overall, this study supports the hypothesis that nutrition of lipid emulsions was associated high TG with differences in fatty acid uptake but it is still unclear if the imbalance of fatty acids directly causes high TG. More research is necessary to investigate other factors such as enzyme activity, lipid clearance rate, or different rate of fatty acid metabolism. / Thesis / Master of Science (MSc)

No description available.

<p>Of the three components of parenteral macronutrient classes (protein, carbohydrate and lipid), the lipid class is the least understood and the fatty acid distribution in lipid emulsion products has the potential to play a critical role in the development of infant morbidities. Breast milk has long been considered the gold standard for infant nutrition, but when infants cannot tolerate enteral feeds, the use of lipid emulsions cannot be avoided despite their adverse side effects.</p> <p>In this study, fatty acid profiles from five commercially available lipid emulsion products were compared to the profile of breast milk. As well, resulting serum on each of these nutrition sources were compared to the profile of either lipid emulsion (Intralipid) or breast milk. Fatty acid profiles for matched pairs of breast milk from mother and resulting serum from infants were compared as well as the profile of normotriglyceridemic serum samples to hypertriglyceridemic ones. Results indicate that not one lipid emulsion product is like another, nor do their profiles closely resemble breast milk even though they are intended to replace the fat portion of the infant’s natural source of nutrition. Serum was not found to directly reflect the fatty acid profile of the nutrition source administered, as was expected based on literature, highlighting that there is a complex web of pathways between nutrition administration and appearance of fatty acids in the serum. Further research is necessary to define the effect of fatty acid chain length and degree of saturation on these metabolic pathways, as the very essence of interrelations such as these can complicate interpretations of results.</p> / Master of Science in Medical Sciences (MSMS)

The WHO Code of Marketing of Breast Milk Substitute is a public health recommendation to reduce preventable causes of infant morbidity and mortality associated with malnutrition. Irresponsible marketing of infant formula in hospitals is a threat to exclusive breastfeeding. Nurses are mandated to support, encourage and protect breastfeeding and to familiarize themselves with their responsibilities under this Code. The researcher explored Geneva nurses' knowledge of the Code and its impact in practice. Eighty seven point seven percent of the participating nurses had poor level of knowledge of the Code. Poor knowledge of the Code impacts on the quality and consistency of information given to mothers in hospitals. Inadvertent violations of the Code were observed among a minority of the respondents: 7.3% indicated that they received gifts; 2.4% received sponsorships to conferences. Training about the Code and its application in counseling is recommended to complement the babyfriendly initiative at hospitals to improve nutrition outcomes. / Health Studies / M.A. (Public Health)