The fatty acid composition of breast tumors may influence disease outcomes, but its use as a biomarker and its relationship with circulating fatty acids remain unclear. This study evaluated the composition of fatty acids in tumors and red blood cells (RBCs) of breast cancer patients to assess the relationship with tumor characteristics, RBC fatty acids, and survival. Paired (n = 122) frozen tumor and RBC samples from breast cancer patients were analyzed by gas-liquid chromatography. In the multivariable-adjusted model, the RBC fatty acid composition of n-3 fatty acids, including eicosapentaenoic acid (EPA, β = 0.13, 95 %CI: 0.05, 0.22), docosahexaenoic acid (DHA, β = 0.17, 95 %CI: 0.05, 0.28), and total n-3 (β = 0.17, 95 %CI: 0.05, 0.29) were positively associated with the fatty acid composition of tumor phospholipids and triglycerides. The relative percentage of linoleic acid and DHA in tumor phospholipids, and DHA and arachidonic acid in tumor triglycerides varied by tumor hormone receptor status. In multivariable-adjusted Cox analyses, the total monounsaturated fatty acid (MUFA) composition of tumor phospholipids was associated with poorer overall survival (hazard ratio = 1.10, 95 %CI: 1.01, 1.20) and disease-free survival (hazard ratio = 1.09, 95 %CI: 1.01, 1.18). This study demonstrates the significance of using circulating n-3 PUFAs to predict tumor fatty acid composition, supporting their use as an accessible indicator of tumor n-3 fatty acid status. Although fatty acids differed by hormone receptor status, their relationship with survival was not confirmed. The MUFA composition of tumors may be clinically relevant to survival outcomes, but requires further investigation.

To investigate whether early-life docosahexaenoic acid (DHA) consumption mitigated the effects of a high-fat diet (HFD) on body composition, endocrine hormones, immune function, inflammatory biomarkers, and identify sex differences. Wistar rats were fed (PND 0-70) diets differing only in fat composition: control low-fat (10% fat, 0% arachidonic acid [ARA], 0% DHA), control high-fat (HF, 20% fat, 0% ARA, 0% DHA), or DHA HF (20% fat, 1% ARA, 2% DHA) (n = 16/group, males/females = 8/8). Plasma hormones (ELISA), inflammatory biomarkers (electrochemiluminescence), splenocytes phenotype (immunofluorescence), and ex vivo cytokine production (ELISA) after mitogen stimulation were measured. HFD did not alter body weight/composition or inflammatory profile but increased plasma glucagon-like peptide 1 (GLP-1, males, p = 0.02). In the spleen, HFD lowered transforming growth factor-beta (TGF-β) and increased interleukin (IL)-10 (females) and IL-2 production (p < 0.03). It also lowered % of regulatory T cells (Treg), B cells (males), conventional type 1dendritic cells (cDC1), and MHC-II and CD86 expression (males) (p < 0.03). DHA mitigated HFD-related changes in plasma GLP-1 (males) andTGF-β, IL-10 (females), and IL-2 production (p ≤ 0.03) and increased % of B cells (males), cDC1, MHC-II, and CD86 expression (males) (p < 0.03). DHA mitigated HFD-induced impaired Treg response and compromised adaptive immunity and antigen-presenting function in a sex-specific manner.

Long-chain polyunsaturated fatty acids (LCPUFA) promote immune development and improve obesity-related inflammation. This study determined the effects of LCPUFA-supplemented diets (age Wk3-10) fed to male and female rats on: (1) anthropometrics and glucose tolerance, (2) splenocyte membrane lipid composition, (3) systemic and intestinal immune function, and (4) systemic, subcutaneous (SAT) and visceral (VAT) adipose inflammation. Wistar rats were fed a high-fat diet (20% w/w fat) with 0% docosahexaenoic acid (DHA)+0% arachidonic acid (ARA) (control), 1% DHA+1% ARA (1% DHA), 2% DHA+1% ARA (2% DHA) or 1% DHA+1% ARA+1% eicosapentaenoic acid (EPA) (fish oil). Diets did not alter anthropometrics or glucose tolerance. Compared to control, all diets increased DHA in splenocyte phosphatidylethanolamine, 2%DHA also increased DHA in phosphatidylcholine; fish oil increased EPA in phosphatidylethanolamine and phosphatidylcholine (P < .01). Experimental diets altered splenocyte phenotypes (P < .05), including higher % of activated cytotoxic T cells (2%DHA), natural killer T cells (2%DHA, fish oil) and dendritic cells/macrophages expressing MHC-II complex (1%DHA) without altering ex vivo responses to mitogens. In mesenteric lymph nodes, experimental diets lowered IL-10 (1%DHA, 2%DHA, female-only) and increased IL-8 (2%DHA) production after mitogen stimulation (P < .01). In plasma, experimental diets lowered IL-8 (1%DHA, male-only), CRP (2%DHA) and TGF-β (2%DHA, female-only) (P < .02). In SAT, 2%DHA increased IL-10 (male-only, P < .05). In VAT, all diets increased IL-10, 2%DHA also increased leptin (P < .01). Collectively, compared to high-fat control, LCPUFA diets reduced systemic and adipose inflammation and altered the splenocyte phenotypes and MLN immune response in a sex-specific manner. These effects may indicate improved immune and adipose function and were more pronounced in rats fed the 2%DHA.

BACKGROUND:Early-life feeding of arachidonic acid (ARA)+docosahexaenoic acid (DHA) has been shown to promote immune changes associated with oral tolerance (OT). Oxylipins have been demonstrated to be modulated by diet and alter immune function. OBJECTIVE:To determine whether early-life feeding of ARA+DHA modulated the ileum and ovalbumin (OVA)-challenged splenocyte oxylipin profile in a way that is beneficial for OT development. METHOD:Allergy-prone BALB/c dams were fed a control (0 %ARA, 0 %DHA) or ARA+DHA (1 %ARA, 1 %DHA) diet during suckling. At 3wks, half of the pups were killed to analyze ileum morphology and oxylipin profile. The remaining pups continued consuming the maternal diets. From day 21-25, pups received daily oral gavage of sucrose or OVA, followed by intraperitoneal OVA injections on day 35 and 41. At 6wks, pups were killed to analyze plasma OVA-specific-IgE and -IgG, ileum morphology, splenocyte phospholipid fatty acid composition and ex vivo splenocyte oxylipin production after OVA stimulation. RESULTS:ARA+DHA supplementation resulted in a 5-fold reduction in plasma OVA-IgE concentration, confirming OT development. At 3wks, ARA+DHA-fed mice had higher ileum levels of 8-HETE, 14,15-DiHETrE, 4-HDHA, 17-HDHA and 19,20-EpDPE and lower levels of 13-HODE and 20-HETE, which suggests better ileum maturation, lower inflammation and enhanced tolerogenic immune regulation to support OT. The longer villi, shorter crypts and higher villus/crypt ratio confirmed the superior ileum maturation. At 6wks, ARA+DHA supplementation increased oxylipin substrates (ARA, DHA, linoleic acid and eicosapentaenoic acid) in splenocyte phospholipids. After OVA stimulation, splenocytes from ARA+DHA-fed mice produced more PGD2, 5-HETE, 15-HETE and 20-HDHA and less TXB2 and 12-HETE, which suggests inhibited Th2 and allergic responses and enhanced tolerogenic immune modulation to support OT. CONCLUSION:Early-life feeding of ARA+DHA beneficially modulated the oxylipin profile in the ileum and OVA-challenged splenocytes to support OT development.

Introduction:Human milk fatty acids and human milk oligosaccharides (HMOs) are milk components inconsistently associated with neurodevelopment. The objective of this research is to examine the link between fatty acids, HMOs and neurodevelopment. Methods:This study includes a subset of 240 parent-infant pairs from the Edmonton site of the CHILD Cohort Study. At 3-4 months post-partum, breastfeeding parents provided a milk sample which was analyzed to identify 20 fatty acids and 19 HMOs. Research assistants administered the Bayley Scales of Infant and Toddler Development at 1 and 2 years of age, comprising cognitive, language and motor development scales (standardized to a mean of 100 and a standard deviation of 15; higher scores indicate better development). Adjusted linear regression was used to estimate the relationships between individual milk components or principal components and neurodevelopment, adjusting for maternal and infant factors. Interactions were tested with infant sex and maternal secretor status. Results:After adjustment, the first fatty acid principal component, characterized by high saturated fat and low n-3 and n-6 fatty acids, was related to higher motor scores (β = 1.59; 95% CI: 0.75, 2.43). Higher concentrations of disialyllacto-N-tetraose were related to lower motor scores (β = -3.91, 95% CI: -5.81, -2.01). Higher concentrations of difucosyllacto-N-hexaose were related to higher language and motor scores for infants of maternal non-secretors, while higher concentrations of 3'-sialyllactose were related to higher scores for infants of maternal secretors. Conclusion:Both fatty acids and HMOs are related to early neurodevelopment. Maternal secretor status moderates the relationship between select HMOs and neurodevelopment.

Clinical trials on docosahexaenoic acid (DHA) supplementation and immune changes during breast cancer neoadjuvant chemotherapy (NAC) are limited. This study evaluated the impact of DHA supplementation during NAC on systemic and tumor immune modulation by assessing plasma inflammatory and cardiac damage markers, tumor-infiltrating lymphocyte (TIL) proportions, and n-6- and n-3-derived oxylipins produced in response to an ex vivo immune challenge. Venous blood was collected at baseline, 9, and 15 weeks during NAC from participants in the DHA for Women with Breast Cancer in the Neoadjuvant Setting (DHA-WIN) trial, which compared DHA-enriched algae (4.4g/day; n=23) with a placebo (n=26) over 18 weeks. Plasma markers were measured using electrochemiluminescence assays. CD4+ and CD8+ TILs were identified in tumor tissue by immunohistochemistry, and oxylipins were quantified in the supernatant of lipopolysaccharide-stimulated peripheral blood mononuclear cells via liquid chromatography-tandem mass spectrometry. DHA supplementation resulted in greater increases in the plasma cytokines IFN-γ and TNF-α compared to placebo (P-interaction < .05). In the DHA group, concentrations of these cytokines increased at 15 weeks compared to baseline (P < .05). No differences were found between groups for other immune markers or the proportion of TILs. Compared to the placebo, DHA led to an overall increase in total oxylipin concentrations (P < .05) and higher production of n-6 fatty acid-derived oxylipins, particularly prostanoids, and n-3 fatty acid-derived oxylipins, including 13-HDoHE. These results suggest that DHA may enhance immune responses by promoting an increase in oxylipin and cytokine concentrations, potentially benefiting patients during breast cancer NAC.

Abstract Lymphedema is a chronic lymphatic disorder characterized by persistent tissue swelling, pain, and recurrent infections, often secondary to cancer treatment, surgery, or obesity. Obesity-associated increases in saturated fatty acids (SFAs) have been linked to lipotoxicity. In this study, patients with secondary lymphedema showed a significantly lower plasma polyunsaturated to saturated fatty acid (PUFA/SFA) ratio compared to BMI-matched controls. Stearic acid, a common dietary SFA, induced apoptosis, oxidative stress, and endoplasmic reticulum (ER) stress in human lymphatic endothelial cells. In a mouse model, a short-term high-SFA diet was used to lower the plasma PUFA/SFA ratio, which worsened tail swelling, oxidative stress, ER stress, and tissue damage following lymphatic injury. Switching to a standard chow diet after surgery prevented these effects. Patients with lymphedema also exhibited elevated levels of fatty acid-binding protein 4 (FABP4), a lipid chaperone associated with metabolic stress. FABP4 inhibition reduced stearic acid-induced cell death in vitro and mitigated tissue damage in vivo. These findings suggest a pathogenic role for SFAs and support dietary modulation and FABP4 inhibition as potential therapeutic strategies for lymphedema.

Background: Omega-3 long-chain-polyunsaturated fatty acids (LCPUFAs) are important dietary components for maternal and infant health during pregnancy and lactation. Objectives: This study investigated determinants of maternal and infant LCPUFAs status at 3 mo postpartum and the relationship between maternal serum, maternal milk, and infant LCPUFAs. Methods: This cross-sectional study included mothers (n = 1481) and their offspring (n = 526) at 3 mo postpartum from the Alberta Pregnancy Outcomes and Nutrition (APrON) cohort. Maternal dietary intake (24-h recall), blood samples from mothers and infants, and maternal milk were collected. Fatty acid composition (relative % of total fatty acids) was determined by gas-liquid chromatography. Linear regression analyses explored associations between diet, sociodemographic factors, and fatty acid status. Results: In a multivariable-adjusted analysis, maternal total dietary intake (supplement + food) was positively associated with the percentage of DHA (standardized B [S beta] = 0.158; B = 0.394; 95% [confidence interval] CI: 0.192, 0.558; P < 0.001) in maternal serum phospholipids. Similar associations were found for DHA and eicosapentaenoic acid in maternal milk and plasma phospholipids of infants. Prepregnancy body mass index (BMI) was negatively associated with DHA (SB =-0.073; B =-0.003; 95% CI:-0.006,-0.001; P = 0.008) and positively associated with total saturated fatty acids (SB = 0.086; B = 0.111; 95% CI: 0.042, 0.180; P = 0.002) in maternal milk. Infants receiving formula combination with maternal milk had lower percentage of DHA (SB =-0.177; B =-0.390; 95% CI:-0.604,-0.175; P < 0.001) and arachidonic acid (SB =-0.106; B =-0.595; 95% CI:-1.122,-0.067; P = 0.027) in their plasma phospholipids compared with those who fed exclusively maternal milk. Conclusions: Maternal total dietary intake and prepregnancy BMI are independently associated with their serum fatty acid status during lactation, whereas maternal diet, milk fatty acid composition, and lactation status are important determinants of infant n-3 LCPUFAs fatty acid status. Future research should investigate the impact of these differences in fatty acid status on infant health outcomes.

Supplementation of omega-3 (n-3) polyunsaturated fatty acids has been associated with reduced side effects and improved quality of life (QoL) in breast cancer patients receiving chemotherapy. The current study reports secondary outcomes from the DHA WIN randomized controlled trial which was designed to evaluate docosahexaenoic acid (DHA) supplementation (4.4 g/day) in conjunction with six cycles of neoadjuvant chemotherapy (NAC) (3 weeks/cycle) in women with non-metastatic breast cancer (n = 49). The objective of the current study was to assess the effects of DHA supplementation on QoL and exercise behaviour in women undergoing NAC for breast cancer. Self-administered questionnaires were used to measure QoL and exercise behaviour before starting chemotherapy (baseline), before each chemotherapy cycle (exercise), and after completing chemotherapy. DHA supplementation did not significantly affect QoL, aerobic exercise volume or resistance training frequency during treatment. However, mean aerobic exercise volume was significantly lower at week 12 (-53.5 minutes/week; 95% CI, -100.5 to -6.3; p = 0.02) and week 18 (-70.8 minutes/week; 95% CI, -123.0 to -18.6; p = 0.01) compared to baseline. Mean resistance training frequency was lower at week 12 (-0.57 times/week; 95% CI, -1.0 to -0.13; p = 0.02) compared to baseline. Meeting exercise guidelines during chemotherapy was not associated with better QoL. In the current exploratory study, QoL and exercise decreased during treatment regardless of DHA supplementation, highlighting the need for supportive care and potential therapies that may mitigate these declines in breast cancer patients receiving NAC. Adequately powered studies are needed to determine if DHA supplementation improves these two indices of health. The trial is registered at ClinicalTrials.gov (NCT03831178).

Breast cancer neoadjuvant therapy may negatively impact the immune system. As a secondary outcome of the docosahexaenoic acid (DHA) for women with breast cancer in the neoadjuvant setting (DHA-WIN trial), we sought to assess the effects of an intervention with DHA on parameters of immune function of women undergoing neoadjuvant therapy. Women with early-stage breast cancer in the neoadjuvant setting were recruited for the DHA-WIN trial and randomly assigned to receive either 4.4 g/day of DHA or a placebo for 18 weeks in conjunction with their neoadjuvant chemotherapy for breast cancer. Venous blood was collected to isolate peripheral blood mononuclear cells. Immune parameters were assessed by measuring white blood cell concentration, flow cytometry, and cytokines concentration after mitogen-stimulated immune response. In the placebo group the proportion of T cells (CD3 +), and functionally active monocytes (CD14 + HLA-DR +) was reduced at the last cycle of chemotherapy (15 weeks) but remained constant in the DHA group (P interaction < 0.05). The neutrophil-to-lymphocyte ratio (NLR) was maintained in the DHA group but increased in the placebo at the end of chemotherapy (P-interaction = 0.02). An increase in this ratio was associated with lower chance of achieving pathological complete response (OR = 0.32, 95

There is limited clinical evidence of docosahexaenoic acid (DHA) efficacy during breast cancer neoadjuvant chemotherapy (NAC). This randomized, double‐blind, placebo‐controlled trial aimed to investigate the safety and efficacy of DHA supplementation in breast cancer patients undergoing NAC. Participants ( n = 49) were assigned to receive either DHA 4.4 g/day orally (algae triacylglycerol) or a placebo (corn/soy oil) over six cycles (18 weeks) of NAC. The primary outcome was the evaluation of changes in the percentage of Ki‐67 expression, assessed by immunohistochemistry analysis from pre‐ to post‐treatment. Secondary outcomes included pathological complete response, incidence of adverse effects, and 3‐year survival analysis. Compliance was evaluated by fatty acid analysis of plasma phospholipids and erythrocyte total lipids quantified by gas–liquid chromatography. The expression of Ki‐67 significantly decreased in both groups, with no significant effects of the DHA intervention ( p = 0.38). When stratified by breast cancer subtype, there was a trend of greater reduction in Ki‐67 expression in the human epidermal growth receptor 2 (HER2+++) subtype in the DHA group compared to placebo ( p = 0.1). The % of DHA in erythrocytes and plasma phospholipids was increased by two‐fold at 9 and 15 weeks of therapy in the DHA group, while it remained unchanged in the placebo group ( p ‐interaction <0.001). There was no reported incidence of adverse effects related to the intervention, and no significant effects were found in the other secondary outcomes. NAC significantly decreased the expression of Ki‐67, with no additional beneficial effects observed by DHA supplementation. Further research is necessary to confirm these findings.

BACKGROUND:Suckling and weaning arachidonic acid (ARA) + docosahexaenoic acid (DHA) supplementation promoted oral tolerance (OT) development in pups, however, the effect of it on the intestine to promote OT development remains unknown. OBJECTIVE:We aimed to explore the impact of this supplementation on intestinal fatty acid composition, structure, and indicators that are supportive of OT development. METHODS:Allergy-prone Brown Norway dams were randomly assigned to a control (0% ARA, 0% DHA) or ARA + DHA diet (0.45% ARA, 0.8% DHA) during suckling (0-3 wk). At weaning (3-8 wk), offspring were randomly assigned to a control (0% ARA, 0% DHA) or ARA + DHA diet (0.5% ARA, 0.5% DHA). At 3 wk, offspring in each group received an oral gavage of sucrose or ovalbumin (OVA) solution for five consecutive days. At 7 wk, all offspring received an intraperitoneal OVA injection. At 8 wk, offspring were terminated to evaluate jejunum morphology and measure mucosal food allergy-related secretory immunoglobulin A (sIgA) and cytokines, ileum phospholipid and triglyceride fatty acid compositions, and fecal calprotectin. RESULTS:Weaning ARA + DHA resulted in a higher percentage of DHA in ileum phospholipids and triglycerides (both P < 0.001), without affecting the percentage of ARA. Despite no lasting effect of suckling ARA + DHA on the DHA content in ileum phospholipids, a programming effect was found on the allergy-related intestinal immune profile [higher concentrations of mucosal IL-2 (P = 0.049) and sIgA (P = 0.033)]. OVA treatment resulted in a lower concentration of mucosal IL-6 (P = 0.026) regardless of dietary interventions. Offspring fed ARA + DHA during suckling and/or weaning had a higher concentration of mucosal transforming growth factor-beta (TGF-β) after OVA treatment but this was not observed in offspring fed control diets during suckling and weaning (P = 0.04). CONCLUSIONS:Early life dietary ARA + DHA supplementation to allergy-prone rats enhanced the DHA concentration in intestinal phospholipids (weaning period) and increased the mucosal sIgA, IL-2, and TGF-β levels (suckling and weaning period), indicating its ability to create a tolerogenic intestinal environment to support OT development.

BACKGROUND:Developmental responses to nutrient deprivation may differ by fetal sex. Despite this, relationships between maternal prenatal iron biomarkers and birth outcomes when stratifying by offspring sex are poorly described, especially in healthy cohorts. OBJECTIVES:This study aimed to determine associations between maternal iron biomarkers and birth weights (BWs) and birth head circumferences (BHCs) among female and male newborns to assess whether the potential predictive ability of iron biomarkers on birth outcomes differs by offspring sex. METHODS:The Alberta Pregnancy Outcomes and Nutrition (APrON) cohort study recruited 2189 pregnant individuals from Calgary and Edmonton, Canada. Maternal blood was drawn at each trimester and 3 mo postpartum. Maternal serum ferritin (SF) concentrations were measured using chemiluminescent immunoassays and erythropoietin (EPO), hepcidin, and soluble transferrin receptor (sTfR) using enzyme-linked immunosorbent assays. Ratios of sTfR:SF and hepcidin:EPO were calculated and birth outcomes accessed through delivery records. Directed acyclic graphs informed multivariate regression models. RESULTS:The risk of maternal iron deficiency increased throughout pregnancy because ∼61% showed depleted iron stores (SF < 15 μg/L) by the third trimester. Maternal hepcidin, SF, sTfR, and sTfR:SF concentrations changed across time (P < 0.01), and participants carrying female fetuses consistently (across 6 biomarkers) showed a lower iron status during the third trimester compared with those with male fetuses (P < 0.05). Higher maternal SF and hepcidin:EPO during the third trimester was associated with lower BWs in males (P = 0.006 for SF; P = 0.03 for hepcidin:EPO) and females (P = 0.02 for SF; P = 0.02 for hepcidin:EPO). There were additional inverse associations between BWs and third trimester maternal hepcidin (P = 0.03) and hemoglobin (P = 0.004) and between BHCs and maternal SF (second trimester; P < 0.05) and Hb (third trimester P = 0.02) but only in males. CONCLUSIONS:Relationships between maternal iron biomarkers and BWs and BHCs may depend on the timing of pregnancy and offpsring sex. There was a high risk of third trimester iron storage depletion among generally healthy pregnant individuals.

To study the effects of feeding docosahexaenoic acid (DHA, derived from novel canola oil), with same amount of arachidonic acid (ARA), supplemented diet to lactating dams on the immune system development of suckled offspring using a T helper type-2 (Th2)-dominant BALB/c mouse. Dams received nutritionally complete control (no ARA or DHA) or DHA + ARA diet (1

To determine the effect of feeding buttermilk-derived choline metabolites on the immune system development in Sprague–Dawley rat pups. Sprague–Dawley dams were randomized to one of the three diets containing 1.7 g/kg choline: 1-Control (100% free choline (FC)), 2-Buttermilk (BM, 37% phosphatidylcholine (PC), 34% sphingomyelin (SM), 17% glycerophosphocholine (GPC), 7% FC, 5% phosphocholine), and 3-Placebo (PB, 50% PC, 25% FC, 25% GPC) until the end of the lactation period. At weaning, pups continued on the same diet as their mom. Cell phenotypes and cytokine production by mitogen-stimulated splenocytes isolated from 3- and 10-week-old pups were measured. At 3 weeks, BM-pups had a higher proportion of cytotoxic T cells (CTL; CD3 + CD8 +) while both BM- and PB-pups had an increased proportion of cells expressing CD28 + , CD86 + and CD27 + (all p > 0.05). Following ConA stimulation, splenocytes from BM- and PB-pups produced more TNF-α and IFN-γ and after LPS stimulation produced more IL-10 and TNF-α (all p > 0.05). Starting at week 6 of age, BM-pups had a higher body weight. At 10 weeks, both the BM- and PB-pups had a higher proportion of CTL expressing CD27 + . After ConA stimulation, splenocytes from BM- and PB-pups produced more IL-2, IFN-γ and IL-6 and more IL-10 after LPS stimulation (all p > 0.05). The proportion of lipid soluble forms of choline in the diet during lactation and weaning periods influence the immune system development in rat offspring.

BACKGROUND:Docosahexaenoic acid (DHA) and arachidonic acid (AA) on oral tolerance (OT) development in allergy-prone infants is less known. OBJECTIVES:We aim to determine the effects of early life DHA supplementation (1% of total fat, from novel canola oil), along with AA, on OT toward ovalbumin (ova, egg protein) in allergy-prone BALB/c pups at 6-wk. METHODS:Breastfeeding dams (n ≥ 10/diet) were fed DHA+AA (1% DHA, 1% AA wt/wt of total fat) or control (0% DHA, 0% AA) suckling period diet (SPD) during which pups consumed dam's milk. At 3-wk, pups from each SPD group were assigned to either the control or DHA+AA weaning diet. For OT, pups from each diet group were either orally fed ova or placebo daily from 21-25 d. Systemic immunization to ova was induced through intraperitoneal injections before euthanizing 6-wk pups. Ova-specific immunoglobulin (ova-Ig) and splenocytes ex-vivo cytokine response to different stimuli were analyzed using a 3-factor analysis of variance. RESULTS:OT-induced suppression was seen in ova-stimulated splenocyte ex-vivo response, where ova-tolerized pups showed significantly lower total immunoglobulin (Ig)G, IgG1, interleukin (IL)-2 and IL-6 production than sucrose (placebo) pups. DHA+AA SPD was associated with 3 times lower plasma concentrations of ova-IgE (P = 0.03) than controls. DHA+AA weaning diet resulted in lower T helper type-2 cytokines (IL-4 and IL-6) with ova stimulation than controls, which may benefit OT. DHA+AA SPD resulted in significantly higher T cell cytokine response [IL-2, interferon-gamma, (IFNγ) and IL-1β] to anti-CD3/CD28 stimulation than controls. The splenocytes stimulated with lipopolysaccharide produced lower inflammatory cytokines (IFNγ, tumor necrosis factor-alpha, IL-6, and C-X-C motif ligand 1), which may be because of lower CD11b+CD68+ splenocytes proportion in pups from DHA+AA SPD than control (all P < 0.05). CONCLUSIONS:DHA and AA in early life may influence OT in allergy-prone BALB/c mouse offspring, as they effectively promote T helper type-1 immune responses.

Docosahexaenoic acid (DHA) reduces breast cancer tumor growth in preclinical models. To better understand how DHA amplifies the actions of docetaxel (TXT) chemotherapy, we examined the effects of two doses of dietary DHA on tumor size, membrane DHA content and necroptosis using a drug resistant triple negative breast cancer (TNBC) patient derived xenograft (PDX) model. Female NOD.Cb-PrkdcscidIl2rg mice bearing TNBC PDXs were randomized to one of three nutritionally complete diets (20% w/w fat): control (0% DHA), high DHA (3.8% HDHA), or low DHA (1.6% LDHA) with or without intraperitoneal injections of 5 mg/kg TXT, twice weekly for 6 weeks (n=8 per group). Tumors from mice fed either HDHA+TXT or LDHA+TXT were similar in size to each other, but were 36% and 32% smaller than tumors from mice fed control+TXT, respectively (P<.05). A dose effect of DHA incorporation was observed in plasma total phospholipids and in phosphatidylethanolamine and phosphatidylinositol. Both doses of DHA resulted in similarly increased necrotic tissue and decreased NFκB protein expression compared to control tumors, however only the HDHA+TXT had increased expression of necroptosis related proteins: RIPK1, RIPK3 and MLKL (P<.05). Increased MLKL was observed in the lipid raft portion of HDHA+TXT tumor extracts. This work confirms the efficacy of a combination therapy consisting of DHA supplementation and TXT chemotherapy using two doses of DHA as indicated by reduced tumor growth in a TNBC PDX model. Moreover, the results suggest that decreased growth may occur through increased DHA incorporation into tumor phospholipid membranes and necroptosis.

Background In humans, the development of gut-associated lymphoid tissue (GALT) occurs in the first years of life and can be influenced by diet. Objectives The objective of this study was to determine the effect of dietary choline on the development of gut-associated lymphoid tissue (GALT). Methods Three feeding trials were conducted in female Sprague-Dawley rats. Beginning 3 d before parturition (studies 1 and 3) or at day 10 of gestation (study 2), control dams consumed a 100% free choline (FC) diet until the end of the lactation period. In studies 1 and 3, test dams consumed a high-glycerophosphocholine (HGPC) diet [75% glycerophosphocholine (GPC), 12.5% phosphatidylcholine (PC), 12.5% FC] and a 100% PC diet, respectively (both 1 g of choline/kg diet). In study 2, test dams consumed a high-sphingomyelin (SM) and PC (SMPC) diet (34% SM, 37% PC, 17% GPC, 7% FC, 5% phosphocholine) or a 50% PC diet (50% PC, 25% FC, 25% GPC), both 1.7 g of choline/kg diet. Immune cell phenotypes and ex vivo cytokine production by mitogen-stimulated immune cells were measured. Results Feeding of the HGPC diet lowered T-cell IL-2 (44%), IFN-gamma (34%), and TNF-alpha (55%) production in mesenteric lymph nodes (MLNs) compared with control. Feeding both SMPC and 50% PC diets during the lactation and weaning periods increased IL-2 (54%) and TNF-alpha (46%) production after T-cell stimulation compared with control. There was a lower production of IL-2 (46%), IL-6 (66%), and TNF-alpha (45%), and a higher production of IL-10 (44%) in both SMPC and 50% PC groups following ovalbumin stimulation compared with control in MLNs. Feeding a diet containing 100% PC increased the production of IFN-gamma by 52% after T-cell stimulation compared with control. Conclusion Feeding a diet containing a mixture of choline forms with a high content of lipid-soluble forms during both the lactation and weaning periods enhances ex vivo immune responses from the GALT in female Sprague-Dawley offspring.