Supplementation of oleic acid, stearic acid, palmitic acid and ß-hydroxybutyrate increase H3K9me3 in endometrial epithelial cells of cattle cultured in vitro.

There is growing evidence that greater than homeostatic blood concentrations of nonesterified fatty acids (NEFAs) and ß-hydroxybutyrate (BHBA) have negative consequences on dairy cow's fertility, but effects on cell homeostasis in the reproductive system is not completely understood. In this study, lipids accumulation, reactive oxygen species (ROS) concentrations, abundance of gene transcripts, and immunofluorescence signal of H3K4me3 and H3K9me3 were evaluated in endometrial epithelial cells of cattle cultured with NEFAs (Oleic (OA), Stearic (SA) and Palmitic (PA) acids), BHBA, NEFAs + BHBA or each of the three NEFAs alone. The cellular lipids were in greater concentrations as a result of NEFAs + BHBA, NEFAs, SA or OA supplementation, but not by BHBA or PA. The ROS concentrations were greater when there were treatments with NEFAs + BHBA, NEFAs or BHBA. The relative mRNA abundance for genes involved in the regulation of apoptosis (XIAP), glucose transport (GLUT3), and DNA methylation (DNMT1) were greater when there were NEFAs + BHBA, but not NEFAs, BHBA, OA, SA or PA treatments. The immunofluorescence signal for H3K9me3 was greater when there were NEFAs + BHBA, NEFAs or PA, but not by BHBA, OA or SA treatments. These findings indicate that NEFAs and BHBA have an additive effect on endometrial cells of cattle by altering epigenetic markers and the expression of genes controlling important cellular pathways. Furthermore, there was cellular lipid accumulation and increased H3K9me3 in cultured bovine endometrial cells that was mainly induced by OA and PA treatments, respectively.

Palmitic acid-rich oils with and without interesterification lower postprandial lipemia and increase atherogenic lipoproteins compared with a MUFA-rich oil: A randomized controlled trial.

BACKGROUND: Interesterified (IE) fats are widely used in place of trans fats; however, little is known about their metabolism. OBJECTIVES: To test the impact of a commonly consumed IE compared with a non-IE equivalent fat on in vivo postprandial and in vitro lipid metabolism, compared with a reference oil [rapeseed oil (RO)]. METHODS: A double-blinded, 3-phase crossover, randomized controlled trial was performed in healthy adults (n = 20) aged 45-75 y. Postprandial plasma triacylglycerol and lipoprotein responses (including stable isotope tracing) to a test meal (50 g fat) were evaluated over 8 h. The test fats were IE 80:20 palm stearin/palm kernel fat, an identical non-IE fat, and RO (control). In vitro, mechanisms of digestion were explored using a dynamic gastric model (DGM). RESULTS: Plasma triacylglycerol 8-h incremental area under the curves were lower following non-IE compared with RO [-1.7 mmol/L⋅h (95% CI: -3.3, -0.0)], but there were no differences between IE and RO or IE and non-IE. LDL particles were smaller following IE and non-IE compared with RO (P = 0.005). Extra extra large, extra large, and large VLDL particle concentrations were higher following IE and non-IE compared with RO at 6-8 h (P < 0.05). No differences in the appearance of [13C]palmitic acid in plasma triacylglycerol were observed between IE and non-IE fats. DGM revealed differences in phase separation of the IE and non-IE meals and delayed release of SFAs compared with RO. CONCLUSIONS: Interesterification did not modify fat digestion, postprandial lipemia, or lipid metabolism measured by stable isotope and DGM analysis. Despite the lower lipemia following the SFA-rich fats, increased proatherogenic large triacylglycerol-rich lipoprotein remnant and small LDL particles following the SFA-rich fats relative to RO adds a new postprandial dimension to the mechanistic evidence linking SFAs to cardiovascular disease risk.

Effect of supplementing palmitic acid and altering the dietary ratio of n-6:n-3 fatty acids in low-fibre diets on production responses of dairy cows.

Supplementing palmitic acid (C16 : 0) in combination with modifying the dietary n-6:n-3 fatty acid (FA) ratio may benefit energy metabolism and milk responses of dairy cows. Twelve Holstein cows (70 (sd 11) days in milk) were used in a replicated 4 × 4 Latin square and allocated to four low-fibre diets (18·5 % forage neutral-detergent fibre) supplemented with no FA (CON), or 2·4 % C16 : 0-enriched supplement (PAL), 2·4 % mixture (2:1) of C16 : 0 and n-6 FA (PW6), and mixture (2:1) of C16 : 0 and n-3 FA (PW3). The dietary ratio of n-6:n-3 was increased with PW6 (10:1) and decreased with PW3 (2·8:1), whereas PAL alone made no change in the ratio (about 7:1). Compared with CON, all FA-supplemented treatments increased milk yield. However, feed and energy intakes were higher in PAL than PW3 or PW6, resulting in greater feed efficiency for PW3 and PW6 than PAL. Dietary FA supplements decreased milk protein concentration but tended to increase protein yield. Compared with CON and FA mixtures, PAL increased milk fat content and tended to increase milk SFA and atherosclerotic index. The concentration of milk n-3 FA was similar between CON and PW3. Feeding PAL increased milk energy output and decreased energy partitioning towards body reserves (-4·2 %), while this measure was positive for other treatments. Blood TAG and NEFA concentrations, but not ß-hydroxybutyrate, were increased by FA-supplemented treatments. Feeding C16 : 0 combined with either n-6 or n-3 FA enhanced feed efficiency, alleviated the negative impacts on body energy reserves, but lowering the dietary n-6:n-3 ratio improved the FA profile of milk.

Effects of dietary deoiled soy lecithin supplementation on milk production and fatty acid digestibility in Holstein dairy cows.

Deoiled soy lecithin is a feed additive enriched in phospholipids. Our study evaluated the effects of dietary deoiled soy lecithin supplementation on (1) milk production and composition, (2) plasma and milk fatty acid (FA) content and yield, and (3) apparent FA digestibility and absorption in lactating dairy cows fed fractionated palm fat. In a split-plot Latin square design, 16 Holstein cows (160 ± 7 days in milk; 3.6 ± 1.2 parity) were randomly allocated to a main plot receiving a corn silage and alfalfa haylage-based diet with palm fat containing either moderate (MPA) or high palmitic acid (HPA) content at 1.75% of ration dry matter (72 or 99% palmitic acid, respectively; n = 8/palm fat diet). On each palm fat diet, deoiled soy lecithin was top-dressed at 0, 0.12, 0.24, or 0.36% of ration dry matter in a replicated 4 × 4 Latin square design. Following a 14-d covariate period, lecithin supplementation spanned 14 d, with milk and blood collected during the final 3 d. Milk composition and pooled plasma markers were measured. The statistical model included the fixed effects of palm fat type, lecithin dose, period, and the interaction between palm fat type and lecithin dose. The random effect of cow nested within palm fat group was also included. Lecithin linearly decreased dry matter intake. In cows fed HPA, lecithin feeding reduced milk fat content and tended to decrease milk fat yield. Although no changes in milk yield were observed, a quadratic reduction in 3.5% fat-corrected milk was observed with increasing lecithin dose. Lecithin linearly increased energy-corrected milk efficiency in cows fed MPA. Lecithin supplementation also decreased milk urea nitrogen, relative to unsupplemented cows. The proportion of 16-carbon FA in milk fat decreased linearly with lecithin dose, whereas 18-carbon FA increased linearly. Lecithin reduced de novo FA (<16-carbon) content and tended to increase preformed FA (>16-carbon) content in a linear manner. Compared with MPA, HPA diets reduced apparent total and 16-carbon FA digestibility and absorption. Deoiled soy lecithin feeding did not modify FA digestibility or absorption. Our observations suggest that soy lecithin feeding modifies rumen digestion to reduce dry matter intake and change milk composition.

Short communication: Effects of dietary deoiled soy lecithin supplementation on circulating choline and choline metabolites, and the plasma phospholipid profile in Holstein cows fed palm fat.

Dietary lecithin is a source of choline. Our objective was to evaluate the effects of dietary deoiled soy lecithin feeding on circulating choline, choline metabolites, and the plasma phospholipid profile in lactating dairy cows fed fractionated palm fatty acids. In a split-plot Latin square design, 16 Holstein cows (160 ± 7 d in milk; 3.6 ± 1.2 parity) were randomly allocated to a main plot receiving a corn silage and alfalfa haylage-based diet with palm fat containing either moderate or high palmitic acid content at 1.75% of ration dry matter (moderate and high palmitic acid containing 72 or 99% palmitic acid in fat supplement, respectively; n = 8/palm fat diet). Within each palm fat group, deoiled soy lecithin was top-dressed at 0, 0.12, 0.24, or 0.36% of ration dry matter in a replicated 4 × 4 Latin square design with 14-d experimental periods. A 14-d covariate period was used to acclimate cows to palm fat feeding without lecithin supplementation. Blood sampling occurred during the final 3 d of each experimental period. Plasma choline and choline metabolites were quantified using liquid chromatography and mass spectrometry. Plasma phospholipids were profiled using time-of-flight mass spectrometry. Whereas no effects of treatments were detected for plasma choline or methionine, lecithin feeding increased the plasma concentrations of choline metabolites trimethylamine N-oxide and dimethylglycine (24 and 11%, respectively). Plasma phosphatidylcholine (PC) and sphingomyelin (SM) concentrations increased with deoiled lecithin feeding (e.g., PC 16:0/22:6 and SM d18:1/18:3). Lecithin supplementation also increased plasma lysophosphatidylcholine (LPC) concentrations (e.g., LPC 18:0) while reducing plasma phosphatidylethanolamine (PE) concentrations (e.g., PE 16:0/20:5). Although increases in microbial-derived trimethylamine N-oxide suggest gastrointestinal lecithin degradation, elevations in plasma dimethylglycine, PC, LPC, and SM suggest that choline availability was improved by lecithin feeding in cows, thus supporting enhanced endogenous phospholipid synthesis.

Milk production responses to altering the dietary ratio of palmitic and oleic acids varies with production level in dairy cows.

We evaluated the effects of altering the dietary ratio of palmitic (C16:0; PA) and oleic (cis-9 C18:1; OA) acids on production responses of cows with a wide range of milk production (32 to 65 kg/d) in a crossover design experiment with a preliminary period. Thirty-two multiparous Holstein cows (144 ± 54 d in milk) were assigned randomly to a treatment sequence. Treatments were diets supplemented with fatty acid (FA) blends (1.5% of diet dry matter) that provided 80% C16:0 + 10% cis-9 C18:1 (PA) and 60% C16:0 + 30% cis-9 C18:1 (PA+OA). The corn silage and alfalfa-based diets contained 20.0% forage neutral detergent fiber (NDF), 28.5% starch, and 17.1% crude protein. Treatment periods were 21 d with the final 5 d used for data and sample collection. Treatment did not affect dry matter intake (DMI), milk yield, energy-corrected milk (ECM), body weight, or body weight change. The PA+OA diet increased total, 16-carbon, and 18-carbon FA digestibility compared with the PA diet. Compared with PA+OA, PA increased fat yield (1.97 vs. 1.91 kg/d) and protein yield (1.61 vs. 1.55 kg/d). The PA diet also increased the yield of de novo (448 vs. 428 g/d) and mixed (749 vs. 669 g/d) milk FA and decreased the yield of preformed FA (605 vs. 627 g/d) compared with PA+OA. Interactions were detected between treatment and preliminary milk yield for DMI, total FA intake, 16-carbon FA intake, ECM, 3.5% fat-corrected milk (linear interaction), and a tendency for milk yield (linear interaction); lower-producing cows (<45 kg/d) had increased DMI and ECM on the PA diet, whereas higher-producing cows (>55 kg/d) had increased DMI and ECM on the PA+OA diet. A linear interaction was detected between treatment and preliminary milk yield for mixed milk FA yield (linear interaction) and a tendency for de novo milk FA yield (linear interaction). Our results demonstrate that feeding a fat supplement containing more cis-9 C18:1 replacing C16:0 increased production responses (DMI, milk yield, and ECM) in higher-producing cows, but decreased production responses in lower-producing cows.

Effect of feeding a palmitic acid-enriched supplement on production responses and nitrogen metabolism of mid-lactating Holstein and Jersey cows.

This study evaluated the effect of feeding a palmitic acid-enriched supplement on production responses and nitrogen metabolism of mid-lactating Holstein and Jersey cows. Eighty mid-lactating dairy cows, 40 Holstein and 40 Jersey, were used in a randomized complete block design with a split-plot arrangement; the main plot was breed and the subplot was fatty acid treatment. Cows within each breed were assigned to 1 of 2 treatments: (1) control diet with no fat supplement or (2) control diet plus a palmitic acid-enriched supplement dosed at 1.5% of diet dry matter (PA treatment). The treatment period was 6 wk with the final 3 wk used for data and sample collection. There were no treatment × breed interactions for the variables analyzed. Compared with control, PA treatment increased milk fat yield (1.36 vs. 1.26 kg/d) and tended to increase 3.5% fat-corrected milk (35.6 vs. 34.0 kg/d) and energy-corrected milk (35.7 vs. 34.1 kg/d). There was no effect of PA treatment on dry matter intake, milk yield, milk protein yield, milk lactose yield, body condition score, body weight (BW) change, nitrogen intake, and variables related to nitrogen metabolism and excretion. Compared with Holstein cows, Jersey cows had greater dry matter intake as a percent of BW (4.90 vs. 3.37% of BW) and lower milk production (29.6 vs. 32.7 kg/d) and milk lactose yield (1.58 vs. 1.42 kg/d), but tended to have greater milk fat yield (1.36 vs. 1.26 kg/d). There was a breed effect on BW change; Holstein cows gained 0.385 kg/d during the experiment, and Jersey cows gained 0.145 kg/d. Jersey cows had lower nitrogen intake (636 vs. 694 g/d), blood urea nitrogen (12.6 vs. 13.8 mg/dL), urine total nitrogen (125 vs. 145 g/d), and urine total nitrogen as a percent of nitrogen intake (19.5 vs. 21.1%). Overall, feeding a palmitic acid-enriched supplement increased milk fat yield as well as dry matter and fiber digestibility in both Holstein and Jersey cows. The PA treatment did not have any major effects on nitrogen metabolism in both Holstein and Jersey cows. In addition, our results indicated that Jersey cows had lower urinary nitrogen excretion (g/d) than Holstein cows.

Palmitic acid triggers inflammatory responses in N42 cultured hypothalamic cells partially via ceramide synthesis but not via TLR4.

A high-fat diet induces hypothalamic inflammation in rodents which, in turn, contributes to the development of obesity by eliciting both insulin and leptin resistance. However, the mechanism by which long-chain saturated fatty acids trigger inflammation is still contentious. To elucidate this mechanism, the effect of fatty acids on the expression of the pro-inflammatory cytokines IL-6 and TNFα was investigated in the mHypoE-N42 hypothalamic cell line (N42). N42 cells were treated with lauric acid (LA) and palmitic acid (PA). PA challenge was carried out in the presence of either a TLR4 inhibitor, a ceramide synthesis inhibitor (L-cycloserine), oleic acid (OA) or eicosapentaenoic acid (EPA). Intracellular ceramide accumulation was quantified using LC-ESI-MS/MS. PA but not LA upregulated IL-6 and TNFα. L-cycloserine, OA and EPA all counteracted PA-induced intracellular ceramide accumulation leading to a downregulation of IL-6 and TNFα. However, a TLR4 inhibitor failed to inhibit PA-induced upregulation of pro-inflammatory cytokines.In conclusion, PA induced the expression of IL-6 and TNFα in N42 neuronal cells independently of TLR4 but, partially, via ceramide synthesis with OA and EPA being anti-inflammatory by decreasing PA-induced intracellular ceramide build-up. Thus, ceramide accumulation represents one on the mechanisms by which PA induces inflammation in neurons.

Palmitic acid damages gut epithelium integrity and initiates inflammatory cytokine production.

The mechanisms leading to the low-grade inflammation observed during obesity are not fully understood. Seeking the initiating events, we tested the hypothesis that the intestine could be damaged by repeated lipid supply and therefore participate in inflammation. In mice, 1-5 palm oil gavages increased intestinal permeability via decreased expression and mislocalization of junctional proteins at the cell-cell contacts; altered the intestinal bacterial species by decreasing the abundance of Akkermansia muciniphila, segmented filamentous bacteria, and Clostridium leptum; and increased inflammatory cytokine expression. This was further studied in human intestinal epithelial Caco-2/TC7 cells using the two main components of palm oil, i.e., palmitic and oleic acid. Saturated palmitic acid impaired paracellular permeability and junctional protein localization, and induced inflammatory cytokine expression in the cells, but unsaturated oleic acid did not. Inhibiting de novo ceramide synthesis prevented part of these effects. Altogether, our data show that short exposure to palm oil or palmitic acid induces intestinal dysfunctions targeting barrier integrity and inflammation. Excessive palm oil consumption could be an early player in the gut alterations observed in metabolic diseases.

Non-obesogenic doses of palmitate disrupt circadian metabolism in adipocytes.

Saturated fatty acids, such as palmitate, lead to circadian disruption. We aimed at studying the effect of low doses of palmitate on circadian metabolism and to decipher the mechanism by which fatty acids convey their effect in adipocytes. Mice were fed non-obesogenic doses of palm or olive oil and adipocytes were treated with palmitate and oleate. Cultured adipocytes treated with oleate showed increased AMPK activity and induced the expression of mitochondrial genes indicating increased fatty acid oxidation, while palmitate increased ACC activity and induced the expression of lipogenic genes, indicating increased fatty acid synthesis. Low doses of palmitate were sufficient to alter circadian rhythms, due to changes in the expression and/or activity of key metabolic proteins including GSK3ß and AKT. Palmitate-induced AKT and GSK3ß activation led to the phosphorylation of BMAL1 that resulted in low levels as well as high amplitude of circadian clock expression. In adipocytes, the detrimental metabolic alteration of palmitate manifests itself early on even at non-obesogenic levels. This is accompanied by modulating BMAL1 expression and phosphorylation levels, which lead to dampened clock gene expression.

Milk production and nutrient digestibility responses to triglyceride or fatty acid supplements enriched in palmitic acid.

The objective of our study was to evaluate the effects of feeding triglyceride and fatty acid (FA) supplements enriched in palmitic acid (PA; C16:0) on production and nutrient digestibility responses of mid-lactation dairy cows. Fifteen Holstein cows (137 ± 49 d in milk) were randomly assigned to a treatment sequence in a 3 × 3 Latin square design. Treatments consisted of a control diet (CON; no added PA) or 1.5% FA added as either a FA supplement (PA-FA) or a triglyceride supplement (PA-TG). The PA supplements replaced soyhulls, and diets were balanced for glycerol content. Periods were 21 d in length with sample and data collection occurring during the final 5 d. Compared with CON, PA treatments increased dry matter (66.5 vs. 63.9%) and neutral detergent fiber (NDF) apparent digestibility (42.0 vs. 38.2%). Although PA treatments tended to increase 18-carbon FA apparent digestibility (79.1 vs. 77.9%), PA treatments decreased 16-carbon (63.1 vs. 75.8%) and total FA (72.0 vs. 76.5%) apparent digestibilities compared with CON. The PA treatments increased milk fat content (3.60 vs. 3.41%), milk fat yield (1.70 vs. 1.60 kg/d), yield of 16-carbon milk FA (570 vs. 471 g/d), 3.5% fat-corrected milk (47.6 vs. 46.5 kg/d), and energy-corrected milk (47.4 vs. 46.6 kg/d) compared with CON. The PA treatments did not affect dry matter intake (28.5 vs. 29.2 kg/d), milk yield (47.0 vs. 47.4 kg/d), milk protein yield (1.42 vs. 1.45 kg/d), milk lactose yield (2.29 vs. 2.31 kg/d), yield of <16-carbon milk FA (360 vs. 370 g/d), yield of >16-carbon milk FA (642 vs. 630 g/d), body weight (720 vs. 723 kg), or body condition score (3.14 vs. 3.23). We did not observe differences in digestibilities of dry matter, NDF, and 18-carbon FA between PA-TG and PA-FA. In contrast, PA-FA increased 16-carbon (68.6 vs. 57.6%) and total FA apparent digestibility (73.8 vs. 70.1%) compared with PA-TG. This resulted in PA-FA supplementation increasing the apparent digestibility of the PA supplement by ∼10 percentage points compared with PA-TG. Compared with PA-TG, PA-FA increased 16-carbon FA intake by 60 g/d, absorbed 16-carbon FA by 86 g/d, and absorbed total FA by 85 g/d. Compared with PA-TG, PA-FA increased dry matter intake (29.1 vs. 27.8 kg/d), yield of 16-carbon milk FA (596 vs. 545 g/d), and tended to increase milk yield (47.6 vs. 46.4 kg/d), milk fat yield (1.70 vs. 1.66 kg/d), and 3.5% fat-corrected milk (48.1 vs. 47.2 kg/d). In conclusion, the production response of dairy cows to PA tended to be greater for a FA supplement compared with a triglyceride supplement. Overall, PA increased NDF digestibility, milk fat yield, energy-corrected milk, and feed efficiency in mid-lactation dairy cows.

The Differentiation of Spinal Cord Motor Neurons is Associated with Changes of the Mitochondrial Phospholipid Cardiolipin.

Motor neuron damage caused by diseases, traumatic insults or de-afferentation of the spinal cord is often incurable due to the poor intrinsic regenerative capacity. Moreover, regenerated peripheral nerves often do not reach normal functionality. Here, we investigated cardiolipin in the process of neuro-differentiation, since cardiolipin is closely linked to the mitochondrial energy supply in cells. The NSC-34 hybrid cell line, produced by fusing neuroblastoma cells with primary spinal cord motor neurons, was used, since it shares several morphological and physiological characteristics with mature primary motor neurons. Their neuro-differentiation was supported by switching from normal to differentiation medium or by fatty acid supplementation. Differentiation was evaluated by measuring neurite-sprouting parameters and PPARα expression. Cellular fatty acid distribution was analyzed to indicate changes in lipid metabolism during differentiation. Cardiolipin was characterized by acyl-chain composition and the distribution of molecular cardiolipin species. Both, the switch from normal to differentiation medium as well as the administration of palmitic and oleic acid promoted neuro-differentiation. Stimulated differentiation was accompanied by changes in cardiolipin content and composition. The positive correlation between neuro-differentiation and concentration of those molecular cardiolipin species containing palmitic and oleic acid implied a link between differentiation of NSC-34 cells and cardiolipin metabolism. We further demonstrated the impact of cellular lipid metabolism, and particularly cardiolipin metabolism, during and NSC-34 neuritogenesis. Thus, cardiolipin may represent a new therapeutic target for axon regeneration after peripheral nerve injuries or when axon sprouting is required to compensate for motor neuron loss in response to aging and/or disease.

Euodia daniellii Hemsl. (Bee-Bee Tree) Oil Attenuates Palmitate-Induced Lipid Accumulation and Apoptosis in Hepatocytes.

Hepatic lipid accumulation and apoptosis is elevated in patients with non-alcoholic steatohepatitis and is closely associated with severity. Saturated fatty acid palmitate stimulates lipid accumulation and apoptosis in hepatocytes. In the present study, we examined bee-bee tree oil (BO)-mediated protective effects on palmitate-induced lipid accumulation and apoptosis in mouse primary hepatocytes. Cells were cultured in a control media or the same media containing 150 or 300 µmol/L of albumin-bound palmitate for 24 h. BO concentrations used were 0, 0.1, 0.2, or 0.5%. Palmitate induced lipid accumulation and mRNA expression of lipogenic genes such as SREBP1c and SCD1. However, BO prevented these changes. Furthermore, palmitate stimulated caspase-3 activity and decreased cell viability in the absence of BO. BO reduced palmitate-induced activation of caspase-3 and cell death in a dose-dependent manner. AMP-activated protein kinase inhibitors abolished the effects of BO. Furthermore, BO suppressed palmitate-induced c-Jun N-terminal kinase (JNK) phosphorylation through the 5' adenosine monophosphate-activated protein kinase (AMPK)-dependent pathway. In conclusion, BO attenuated palmitate-induced hepatic steatosis and apoptosis through AMPK-mediated suppression of JNK signaling. These data suggest that BO is an important determinant of saturated fatty acid-induced lipid accumulation and apoptosis, and may be an effective therapeutic strategy for treatment of obesity-mediated liver diseases.

Long-term palmitic acid supplementation interacts with parity in lactating dairy cows: Production responses, nutrient digestibility, and energy partitioning.

The objective of our study was to evaluate the effects of long-term palmitic acid (C16:0) supplementation and parity on production, nutrient digestibility, and energy partitioning of mid-lactation dairy cows. Forty mid-lactation Holstein cows (18 primiparous and 22 multiparous) were used in a block design. Cows were assigned to receive either a control diet containing no supplemental fat (CON) or a C16:0-enriched supplemented diet (PA; 1.5% diet dry matter) fed for 10 wk. Compared with CON, PA increased dry matter intake, milk yield, cumulative milk yield, milk fat content, milk fat yield, 16-carbon milk fatty acid (FA) yield, 3.5% fat-corrected milk yield, and energy-corrected milk yield. Additionally, PA increased body weight change, but did not affect body condition score change compared with CON. A tendency for a treatment by parity interaction was observed for milk yield due to PA increasing milk yield in multiparous but not in primiparous cows. In addition, we observed interactions between treatment and parity for fat-corrected milk, energy-corrected milk, and milk fat yield due to PA increasing these variables to a greater extent in multiparous compared with primiparous cows. Interestingly, we observed an interaction between treatment and parity for body weight change, due to PA increasing body weight change in primiparous but not in multiparous cows. The PA treatment increased dry matter and neutral detergent fiber digestibilities compared with CON. Although PA did not affect 18-carbon FA digestibility, compared with CON, PA decreased 16-carbon and total FA digestibilities and increased total FA intake by 470 g/d and absorbed total FA by 316 g/d. We also observed an interaction between treatment and parity for total absorbed FA due to PA increasing it to a greater extent in multiparous than in primiparous cows. Compared with CON, PA increased apparent energy intake and milk energy output. We observed an interaction between treatment and parity for milk energy output due to PA increasing milk energy output to a greater extent in multiparous than primiparous cows. Additionally, an interaction between treatment and parity was observed for energy output in body reserves due to PA increasing energy output in body reserves in primiparous but not in multiparous cows. In conclusion, production responses of dairy cows to PA were consistent throughout the 10-wk treatment period. In addition, PA supplementation interacted with parity, with production responses increased to a greater extent in multiparous than primiparous cows and energy partitioned to body reserves only increased in primiparous cows.

Short communication: Comparison of a palmitic acid-enriched triglyceride supplement and calcium salts of palm fatty acids supplement on production responses of dairy cows.

The objective of our study was to evaluate the effects of feeding a palmitic acid-enriched triglyceride supplement or a calcium salts of palm fatty acid (FA) supplement on nutrient digestibility and production responses of mid-lactation dairy cows. Fifteen Holstein cows (139 ± 39 d in milk) were randomly assigned to treatment sequence in a 3 × 3 Latin square design. Treatments were a control diet (CON; no fat supplement) and 1.5% of FA added either as a palmitic acid-enriched triglyceride supplement (PA-TG) or as calcium salts of palm FA supplement (Ca-FA). Fat-supplemented treatments did not affect dry matter intake (DMI) compared with CON, but Ca-FA reduced DMI compared with PA-TG. Compared with CON, fat-supplemented treatments increased 18-carbon FA digestibility by 2.0 percentage units but did not affect digestibility of total FA or 16-carbon FA. Compared with Ca-FA, PA-TG reduced total FA digestibility by 8.7 percentage units due to a decrease in 16-carbon FA digestibility (21.7 percentage units). Both fat supplements increased neutral detergent fiber (NDF) digestibility compared with CON (3.90 percentage units), and PA-TG tended to increase NDF digestibility by 1.60 percentage units compared with Ca-FA. Compared with CON, fat-supplemented treatments increased milk yield (1.05 kg/d), 3.5% fat-corrected milk yield (2.20 kg/d), and energy-corrected milk yield (1.80 kg/d). Also, PA-TG increased milk fat yield (50 g/d) and milk energy output (1.0 Mcal/d) and tended to increase milk fat content (0.07 percentage units) and energy-corrected milk yield (1.0 kg/d) compared with Ca-FA. Fat-supplemented treatments reduced the yield of de novo milk FA (23 g/d) and increased the yields of mixed (43 g/d) and preformed (52 g/d) milk FA compared with CON. The PA-TG treatment increased the yield of 16-carbon (66 g/d) milk FA compared with Ca-FA, whereas Ca-FA increased the yield of preformed (60 g/d) milk FA. Fat-supplemented treatments increased intake of net energy for lactation by 1.80 Mcal/d, milk energy output by 1.30 Mcal/d, and energy in body reserves by 0.30 Mcal/d compared with CON. The Ca-FA treatment increased energy allocated to body reserves (0.60 Mcal/d), energy partitioning toward body reserves (1.20 percentage units), and body condition score change (0.06 units), and tended to increase body weight change (0.16 kg/d) and body condition score (0.08 units) compared with PA-TG. In conclusion, feeding a palmitic acid-enriched triglyceride supplement increased milk energy output due to increased yields of milk and milk fat, whereas feeding a calcium salts of palm FA supplement increased FA digestibility and energy partitioned to body reserves.

Effect of dietary palmitic and stearic acids on sucrose motivation and hypothalamic and striatal cell signals in the rat.

We have reported that motivation for sucrose is increased in rats fed a moderate (31%) mixed-fat diet for 4-6 wk. In this study, rats were fed diets containing 32% stearic (STEAR) or palmitic (PALM) acid, and behavior, metabolic profile, and cell signals were compared with those of rats fed a matched low-fat diet (LF; 11% fat) diet. Rats fed STEAR or PALM increased sucrose motivation relative to LF rats (one-way ANOVA for lever presses; P = 0.03). Diet did not change fasting glucose, insulin, total cholesterol, triglycerides, intravenous glucose tolerance test glucose profile, percent body fat, or total kilocalories, although kilocalories as fat were increased (ANOVA, P < 0.05). Cell signals were assessed in rats ranked from high to low sucrose motivation. Diet did not alter Thr and Ser phosphorylation of Akt in the medial hypothalamus (HYP) and striatum (STR). However, Ser phosphorylation of GSK3Β was decreased in HYP and STR from both high- and low-performer tertiles of STEAR and PALM rats (ANOVA within each brain region, P < 0.05). Two histone 3 (H3) modifications were also assessed. Although there was no effect of diet on the transcription-repressive H3 modification, H3K27me3, the transcription-permissive H3 modification, H3K4me3, was significantly decreased in the HYP of high performers fed PALM or STEAR (ANOVA, P = 0.013). There was no effect of diet on H3K4me3 levels in HYP of low performers, or in STR. Our findings suggest signal-specific and brain region-specific effects of PALM or STEAR diets and may link downstream signaling effects of GSK3Β activity and H3 modifications with enhanced motivational behavior.

Effect of palmitic acid on the mitigation of milk fat depression syndrome caused by trans-10, cis-12-conjugated linoleic acid in grazing dairy cows.

The objective of the study was to evaluate the effect of adding protected palmitic acid (PA) to the ration of grazing dairy cows supplemented with protected conjugated linoleic acid (CLA) on milk production, chemical composition and fat profile. Six cows were used, 3/4 American Swiss × Zebu, under a rotational grazing system in a mixed sward with Cynodon plectostachyus, Brachiaria decumbens and Brachiaria brizantha. Furthermore, each cow received daily 4 kg concentrates and 8 kg sorghum silage, which made up the basal diet. The cows were distributed into three two-cow groups. Three treatments were randomly assigned to the groups, using a cross design: (1) control (basal diet), (2) basal diet + CLA (50 g/d) and (3) basal diet + CLA (50 g/d) + PA (412 g/d). The following variables were evaluated: forage intake, milk production, protein, fat and lactose concentration in milk, and milk fatty acid (FA) profile. There were no differences in forage intake between treatments; however, there were differences in milk production, protein, fat and lactose yield and fat concentration, which increased significantly in group CLA + PA when compared with group CLA. The concentration of FA synthesised de novo was lower when PA was included in the diet. Adding PA to the diet of grazing cows mitigates the milk fat decline caused by including trans-10, cis-12 CLA in the diet.

Stearoyl-CoA desaturase-1, a novel target of omega-3 fatty acids for reducing breast cancer risk in obese postmenopausal women.

BACKGROUND/OBJECTIVES: Conversion of saturated fatty acids to monounsaturated fatty acids by the enzyme stearoyl-Co-A-desaturase (SCD-1) is emerging as a major factor in promoting carcinogenesis including breast cancer. The aim of our study was to explore the regulation of SCD-1 by Raloxifene and omega-3 fatty acids in women at increased risk of breast cancer based on high breast density. SUBJECTS/METHODS: As a reflection of SCD-1 activity, we measured the ratios of palmitoleic acid (C16:1n7) to palmitic acid (C16:0) (SCD-16) and oleic acid (C18:1n9) to steric acid (C18:0) (SCD-18) in plasma samples of postmenopausal women enrolled in our clinical trial (NCT00723398) designed to test the effects of the antiestrogen, Raloxifene and/or the omega-3 preparation Lovaza, on breast density, a validated biomarker of breast cancer risk. RESULTS: We report that Lovaza but not Raloxifene-reduced SCD-16 and SCD-18 for the 2-year duration of the trial. Importantly, decreasing levels of SCD-16 and SCD-18 were associated with a progressive reduction in breast density but only in obese women (body mass index ⩾30). CONCLUSIONS: Body mass index-related factors play an important role in the reduction of breast density and hence breast cancer risk by omega-3 fatty acids. SCD-1 may be a useful biomarker in future clinical trials testing the benefit of nutritional interventions in reducing obesity-associated breast cancer risk.

Nutrient digestibility and milk production responses to increasing levels of palmitic acid supplementation vary in cows receiving diets with or without whole cottonseed.

Our study evaluated the dose-dependent effects of a palmitic acid-enriched supplement in basal diets with or without the inclusion of whole cottonseed on nutrient digestibility and production responses of dairy cows. Sixteen Holstein cows (149 ± 56 days in milk) were used in a split plot Latin square design experiment. Cows were blocked by 3.5% fat-corrected milk (FCM) and allocated to a main plot receiving either a basal diet with soyhulls (SH, = 8) or a basal diet with whole cottonseed (CS, = 8) that was fed throughout the experiment. A palmitic acid-enriched supplement (PA 88.5% C16:0) was fed at 0, 0.75, 1.50, or 2.25% of ration DM in a replicated 4 × 4 Latin Square design within each basal diet group. Periods were 14 d with the final 4 d used for data collection. PA dose increased milk fat content linearly, and cubically affected yields of milk fat and 3.5% FCM. The PA dose did not affect milk protein and lactose contents, BW, and BCS, but tended to increase yields of milk, milk protein, and milk lactose. Also, PA dose reduced DMI and 16-carbon fatty acid digestibility quadratically, and increased 18-carbon fatty acid digestibility quadratically. There were no effects of basal diet on the yield of milk or milk components, but DMI tended to decrease in CS compared with SH, increasing feed efficiency (3.5% FCM/DMI). Compared with SH, CS diets increased yield of preformed milk fatty acids and 16-carbon fatty acid digestibility, and tended to decrease 18-carbon fatty acid digestibility. We observed basal diet × PA dose interactions for yields of milk and milk protein and for 16-carbon and total fatty acid digestibility, as well as tendency for yields of milk fat and 3.5% FCM. Also, there was a tendency for an interaction between basal diet and PA dose for NDF digestibility, which increased more for CS with increasing PA than for SH. PA dose linearly decreased digestibility of total fatty acids in SH diets but did not affect it in CS diets Results demonstrate that responses to PA dose are affected by the dietary basal diet. Additionally, the decrease in fatty acid digestibility only in the SH diets suggests that digestibility is impacted mainly by the profile of 16- and 18-carbon fatty acids reaching the duodenum. Under the dietary conditions evaluated, the yield of 3.5% FCM and milk fat were optimal when PA was fed at 1.5% of ration DM.

Methylation patterns of Vegfb promoter are associated with gene and protein expression levels: the effects of dietary fatty acids.

PURPOSE: We have investigated the epigenetic regulation by dietary fatty acids of Vegfb levels in rats' white adipose tissue and 3T3-L1 cells. METHODS: A group of rats were assigned to three diets, each one with a different composition of saturated, monounsaturated and polyunsaturated fatty acids. Samples of white adipose tissues were taken for the methylation and expression studies. Additionally, 3T3-L1 cells were treated with palmitic, oleic, and linoleic fatty acids. After treatment, cells were harvested and genetic material was extracted for the analysis of Vegfb levels. RESULTS: We report evidence of changes in the methylation levels of the CpG island at the Vegfb promoter and in the Vegfb expression levels in vivo and in vitro by dietary fatty acid, with the main contribution of the linoleic fatty acid. Vegfb promoter methylation levels were closely related to the Vegfb gene expression. CONCLUSION: According to our results, the regulation of Vegfb gene expression by dietary fatty acids may be mediated, at least in part, by epigenetic modifications on Vegfb promoter methylation. Considering the deep association between angiogenesis and tissue growth, we suggest the nutriepigenetic regulation of Vegfb as a key target in the control of the adipose tissue expansion.