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Funding Information: This study was supported by Academy of Finland Grant 309263 (H.Y.-J.) and Grant 138006 (J.P.), EU H2020 project ?Elucidating Pathways of Steatohepatitis? EPoS Grant 634413 (H.Y.-J.), H2020-JTI-IMI2 EU project 777377-2 Liver Investigation: Testing Marker Utility in Steatohepatitis (LITMUS) (H.Y.-J.), Government Funding (H.Y.-J.), Novo Nordisk Foundation (H.Y.-J., P.K.L., M.A.-K.), Ralph Gr?sbeck Scholarship of the Minerva Foundation (P.K.L.), Novo Nordisk Foundation (P.K.L.), Juho Vainio Foundation (J.P.), Finnish Medical Foundation (V.M.), British Heart Foundation Senior Research Fellowship in Basic Science (FS/15/56/31645) (L.H.) and Kuopio University Hospital Project grant (J.P., EVO/VTR grants 2005-2019). Funding Information: This study was supported by Academy of Finland Grant 309263 (H.Y.-J.) and Grant 138006 (J.P.), EU H2020 project ‘Elucidating Pathways of Steatohepatitis’ EPoS Grant 634413 (H.Y.-J.), H2020-JTI-IMI2 EU project 777377-2 Liver Investigation: Testing Marker Utility in Steatohepatitis (LITMUS) (H.Y.-J.), Government Funding (H.Y.-J.), Novo Nordisk Foundation (H.Y.-J., P.K.L., M.A.-K.), Ralph Gräsbeck Scholarship of the Minerva Foundation (P.K.L.), Novo Nordisk Foundation (P.K.L.), Juho Vainio Foundation (J.P.), Finnish Medical Foundation (V.M.), British Heart Foundation Senior Research Fellowship in Basic Science (FS/15/56/31645) (L.H.) and Kuopio University Hospital Project grant (J.P., EVO/VTR grants 2005-2019). Publisher Copyright: © 2021 The Author(s) Background & Aims: There is substantial inter-individual variability in the risk of non-alcoholic fatty liver disease (NAFLD). Part of which is explained by insulin resistance (IR) (‘MetComp’) and part by common modifiers of genetic risk (‘GenComp’). We examined how IR on the one hand and genetic risk on the other contribute to the pathogenesis of NAFLD. Methods: We studied 846 individuals: 492 were obese patients with liver histology and 354 were individuals who underwent intrahepatic triglyceride measurement by proton magnetic resonance spectroscopy. A genetic risk score was calculated using the number of risk alleles in PNPLA3, TM6SF2, MBOAT7, HSD17B13 and MARC1. Substrate concentrations were assessed by serum NMR metabolomics. In subsets of participants, non-esterified fatty acids (NEFAs) and their flux were assessed by D5-glycerol and hyperinsulinemic-euglycemic clamp (n = 41), and hepatic de novo lipogenesis (DNL) was measured by D2O (n = 61). Results: We found that substrate surplus (increased concentrations of 28 serum metabolites including glucose, glycolytic intermediates, and amino acids; increased NEFAs and their flux; increased DNL) characterized the ‘MetComp’. In contrast, the ‘GenComp’ was not accompanied by any substrate excess but was characterized by an increased hepatic mitochondrial redox state, as determined by serum β-hydroxybutyrate/acetoacetate ratio, and inhibition of hepatic pathways dependent on tricarboxylic acid cycle activity, such as DNL. Serum β-hydroxybutyrate/acetoacetate ratio correlated strongly with all histological features of NAFLD. IR and hepatic mitochondrial redox state conferred additive increases in histological features of NAFLD. Conclusions: These data show that the mechanisms underlying ‘Metabolic’ and ‘Genetic’ components of NAFLD are fundamentally different. These findings may have implications with respect to the diagnosis and treatment of NAFLD. Lay summary: The pathogenesis of non-alcoholic fatty liver disease can be explained in part by a metabolic component, including obesity, and in part by a genetic component. Herein, we demonstrate that the mechanisms underlying these components are fundamentally different: the metabolic component is characterized by hepatic oversupply of substrates, such as sugars, lipids and amino acids. In contrast, the genetic component is characterized by impaired hepatic mitochondrial function, making the liver less able to metabolize these substrates.

The endocannabinoid system (ECS) controls feed intake and energy balance in nonruminants. Recent studies suggested that dietary management alters the expression of members of the ECS in the liver and endometrium of dairy cows. The aim of this study was to determine the relationship between body condition score (BCS) loss and the mRNA abundance of genes related to fatty acid metabolism and the ECS in the subcutaneous adipose tissue (AT) of dairy cows. The BCS was determined in multiparous (3.2 ± 0.5 lactations) Holstein cows at -21 and 42 days relative to calving (designated as d = 0). Cows were grouped into three categories according to BCS loss between both assessments as follows: (1) lost ≤0.25 unit (n = 8, low BCS loss (LBL)), (2) lost between 0.5 and 0.75 units (n = 8, moderate BCS loss (MBL)) and (3) lost ≥1 unit (n = 8, high BCS loss (HBL)). Concentrations of haptoglobin and non-esterified fatty acids (NEFAs) were determined in plasma. Real-time PCR was used to determine mRNA abundance of key genes related to fatty acid metabolism, inflammation and ECS in AT. Milk yield (kg/day) between week 2 and 6 post-calving was greater in the LBL group (49.4 ± 0.75) compared to MBL (47.9 ± 0.56) and HBL (47.4 ± 0.62) groups (P < 0.05). The overall mean plasma haptoglobin and NEFA concentrations were greater in MBL and HBL groups compared with the LBL group (P < 0.05). The mRNA abundance of TNF-α, Interleukin-6 (IL-6) and IL-1β was greatest at 21 and 42 days post-calving in HBL, intermediate in MBL and lowest in LBL groups, respectively. Cows in the HBL group had the greatest AT gene expression for carnitine palmitoyltransferase 1A, hormone sensitive lipase and adipose triglyceride lipase at 21 and 42 days post-calving (P < 0.05). Overall, mRNA abundance for very long chain acyl-CoA dehydrogenase and peroxisome proliferator-activated receptor gamma, which are related to NEFA oxidation, were greater in MBL and HBL groups compared to the LBL group at 42 days post-calving. However, mRNA abundance of fatty acid amide hydrolase was lower at 21 and 42 days post-calving in HBL cows than in LBL cows (P < 0.05). In summary, results showed a positive association between increased degree of BCS loss, inflammation and activation of the ECS network in AT of dairy cows. Findings suggest that the ECS might play an important role in fatty acid metabolism, development of inflammation and cow's adaptation to onset of lactation.

Graphical abstract, Circulating non-esterified fatty acids (NEFA) are toxic to mammalian cells. They increase in diseases such as diabetes and sepsis. Herein we propose a serum albumin-fatty acid saturation test. • We based our test on three methodologies: isoelectric focusing (IF) of human plasma albumin, staining proteins after isoelectric focusing in gels with Coomassie Brilliant Blue, and serum albumin measurement with bromocresol green. • The test consists in the determination of albumin IF and staining with bromocresol green. If albumin is saturated with NEFA, it focuses on lower pH, meaning it is the threshold to bind to them. Excessive NEFA is free and toxic. Many other tests are available for NEFA quantification as NEFA kit assay. All colorimetric assays are used for quantification of NEFA and other tests need expensive equipment to read out the results, and they do not measure albumin levels. • Our method focused on albumin-NEFA saturation instead of just NEFA quantification. Critically ill patients have an alteration in both albumin and NEFA. Therefore, our test undergoes less daytime variation compared to assays that measure absolute NEFA values, allowing a more reliable use as an indicator of albumin-fatty acid saturation and NEFA toxicity.

Human leptospirosis is an acute infectious zoonosis presenting specific lipid disorders. Previous in vitro studies showed both leptospira glycolipoprotein endotoxin, and high oleic acid levels were associated with Na/K-ATPase inhibition that is amplified by the reduction of circulating albumin levels. In this study, we aimed to investigate the relationship of oleic acid/albumin (OA/A) molar ratio and clinical outcomes in Leptospirosis. Through a prospective observational cohort study employing high-performance liquid chromatography (HPLC) we sequentially determined serum concentrations of nonesterified fatty acids (NEFA) and albumin in twenty-eight patients with severe leptospirosis since their hospital admission. Twenty patients recovered, and eight died. Data was distributed in two groups according to clinical outcomes. Oleic acid/albumin molar ratios (OA/A), initial samples, were higher than those in healthy donors. The ratio OA/A, however, persisted high in dying patients, whereas patients who survived had a reduction matching to healthy donors. Biochemical alterations suggest that cure is correlated to the reestablishment of the OA/A molar ratio, while fatal outcomes related to persisting OA/A imbalances. Analysis by receiver operating characteristic (ROC) showed the area under the curve of 0.864 and the cutoff value of 0.715 being associated with a high odds ratio. Lipid analysis from patients with leptospirosis had an acute high serum OA/A molar ratio, and sustained imbalance has a high odds ratio and strong correlation with mortality., Leptospirosis clinical outcome; Acute lipotoxicity; Lipidome; Oleic acid/albumin; Na/K-ATPase.

In sheep production, economic efficiency strongly depends on the maternal health and feed efficiency status and on weaning performances of their offspring. Accordingly, an optimal level for the supply with macro- and microelements and the ewe energy status has impact on the fetal development during gestation and on maternal milk production during lactation. Furthermore, this study addressed intergenerational aspects, i.e., on associations between maternal energy metabolism profiles considering the macro- and microelement status, metabolic indicators (e.g. β-hydroxybutyrate (BHB)), body condition and methane (CH4) emissions with lamb BW (LBW) in two sheep breeds. Traits were recorded at the beginning of gestation (ewe traits), at lambing, three weeks postpartum, and at weaning (ewe and lamb traits). Trait recording included CH4 emissions (recorded via laser methane detector (LMD)), ewe BW (EBW), backfat thickness (BFT), and body condition score (BCS) from 46 ewes (24 Merinoland- (ML), 22 Rhonsheep (RH)), and LBW of their 87 (35 ML, 52 RH) purebred lambs. Serum levels of the following ewe blood parameters were determined: calcium (Ca), sodium (Na), potassium (K), phosphate (P), nonesterified fatty acids (NEFA), BHB, glutamate dehydrogenase (GLDH), selenium (Se), copper (Cu), iron (Fe), zinc (Zn), and magnesium (Mg). Mixed models were applied to infer associations between ewe blood parameters with EBW, BFT, BCS, and CH4 and with LBW recorded in offspring. At weaning, a maternal serum Mg level > 1.0 mmol/L was significantly associated with an increase of 13% in LBW in ML, compared to offspring from ML ewes with a serum Mg concentration within the lower reference range (0.8 mmol/L). Furthermore, higher Cu levels were favorably associated with ewe BCS and BFT at weaning in both breeds. In RH ewes, a Se level > 2.4 μmol/l was significantly associated with increased BCS. In the ML breed, high Zn levels during lactation were associated with reduced CH4 emissions. Ewe EBW was significantly larger for ML ewes representing low Ca levels. A low BHB level was associated with decreasing CH4 emissions in RH and ML. Serum levels for Na, K, P, GLDH, and Fe did not significantly affect the traits of interest. Trait associations from the present study indicate the importance of the mineral supply and metabolic status of the ewe with regard to body condition, CH4 emissions, and LBW development, but depending on the breed. Identified associations might contribute to energy efficiency in sheep production systems.

Optimal type and dietary inclusion rates of cereal grains for periparturient sheep are unknown. The objective was to determine effects of feeding diets with high (H) v. low (L) levels of ground corn grain (CN) v. combined ground wheat and barley grains (WB) on intake, rumen fermentation, colostrum and milk properties, and blood metabolites of periparturient sheep. Twenty Afshari×Merino ewes were used in a completely randomized design study from 24 days prepartum through 21 days postpartum. Ewes were kept indoors in individual boxes and received once daily at 0900 h total mixed rations. Treatments were mixed rations containing either (1) H or (2) L concentrate based on either (1) 100% CN or (2) 50 : 50 ratio of ground wheat : ground barley grains in a 2×2 factorial arrangement. Each treatment group had five ewes including two twin-lamb ewes and three single-lamb ewes. Postpartal dry matter intake (DMI) increased by feeding high CN v. high and low WB, while high v. low CN improved postpartum DMI. The DMI during lambing tended to increase with the high v. low WB. Feeding CN v. WB, and feeding both CN and WB at L v. H level increased fecal pH. Postpartal rumen pH was lower with the high v. low WB (5.7 v. 6.2). Rumen concentrations of propionate were lower and of acetate were higher with L v. H grain levels. Increased dietary grain reduced urine pH for WB (7.24 v. 7.83) but not for CN (7.63 v. 7.52) prepartum. Colostrum properties, postpartal urine pH, lamb weight at birth and 21 days of age, and placental weight and expulsion time were unaffected. Milk yield increased and milk fat yield tended to increase by H v. L grain diets. Plasma glucose was increased by feeding high v. low WB, whereas CN v. WB tended to reduce peripartal plasma non-esterified fatty acids (NEFA) and increased insulin to NEFA ratio. In conclusion, more cereal grains can be included in periparturient sheep diets and CN instead of WB may be fed to periparturient sheep to improve energy status. Findings suggest opportunities to optimize periparturient ewe physiology and performance through feeding certain cereals and avoiding high levels of WB.

Under current Australian industry pre-slaughter guidelines, lambs may be off feed for up to 48 h before slaughter. The purpose of this study was to examine what proportion of circulating metabolites at slaughter are due to stress and feed deprivation and if this response differs between Merino and Terminal genotypes. In addition the effect of feed deprivation on carcass weight and meat quality was examined. Jugular blood samples were collected from 88 Merino and Terminal sired lambs at rest and at slaughter following 24, 36 and 48 h of feed deprivation and plasma analysed for glucose, lactate, non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHOB). From the same carcasses hot carcass weight (HCWT) were measured as well as a suite of meat quality traits measured such as M. longissimus lumborum (loin) and M. semitendinosus pH at 24 h postmortem. Loin samples were also analysed for intramuscular fat content and Warner-Bratzer Shear Force. Merino sired lambs had a higher NEFA response compared to Terminal sired lambs at slaughter after 24, 36 and 48 h of feed deprivation, with NEFA levels up to 35% higher than previously reported in the same animals at rest in animal house conditions, whereas BHOB response to feed deprivation was not affected by sire type (Pgt;0.05) and similar to previously reported at rest. In addition to the metabolic effects, increasing feed deprivation from 36 h was associated with a 3% reduction in HCWT and dressing percentage as well as causing increased ultimate pH in the M. semitendinosus in Merino sired lambs. Findings from this study demonstrate that Merino and Terminal sired lambs differ in their metabolic response to feed deprivation under commercial slaughter conditions. In addition, commercial feed deprivation appears to have a negative effect on ultimate pH and carcass weight and warrants further investigation.

The aim of this study was to examine the metabolic response to feed deprivation up to 48 h in low and high yielding lamb genotypes. It was hypothesised that Terminal sired lambs would have decreased plasma glucose and increased plasma non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHOB) concentrations in response to feed deprivation compared to Merino sired lambs. In addition, it was hypothesised that the metabolic changes due to feed deprivation would also be greater in progeny of sires with breeding values for greater growth, muscling and leanness. Eighty nine lambs (45 ewes, 44 wethers) from Merino dams with Merino or Terminal sires with a range in Australian Sheep Breeding Values (ASBVs) for post-weaning weight (PWT), post-weaning eye muscle depth and post-weaning fat depth (PFAT) were used in this experiment. Blood samples were collected via jugular cannulas every 6 h from time 0 to 48 h of feed deprivation for the determination of plasma glucose, NEFA, BHOB and lactate concentration. From 12 to 48 h of feed deprivation plasma glucose concentration decreased (Plt; 0.05) by 25% from 4.04 ± 0.032 mmol/l to 3.04 ± 0.032 mmol/l. From 6 h NEFA concentration increased (Plt; 0.05) from 0.15 ± 0.021 mmol/l by almost 10-fold to 1.34 ± 0.021 mmol/l at 48 h of feed deprivation. Feed deprivation also influenced BHOB concentrations and from 12 to 48 h it increased (Plt; 0.05) from 0.15 ± 0.010 mmol/l to 0.52 ± 0.010 mmol/l. Merino sired lambs had a 8% greater reduction in glucose and 29% and 10% higher NEFA and BHOB response, respectively, compared to Terminal sired lambs (Plt; 0.05). In Merino sired lambs, increasing PWT was also associated with an increase in glucose and decline in NEFA and BHOB concentration (Plt; 0.05). In Terminal sired lambs, increasing PFAT was associated with an increase in glucose and decline in NEFA concentration (Plt; 0.05). Contrary to the hypothesis, Merino sired lambs showed the greatest metabolic response to fasting especially in regards to fat metabolism.

This experiment was conducted to evaluate the effects of Bupleurum extract (BE) on blood metabolites, antioxidant status, and immune function in dairy cows under heat stress. Forty lactating Holstein cows were randomly assigned to 1 of 4 treatments. The treatments consisted of 0, 0.25, 0.5, and 1.0 g of BE kg−1 dry matter. Supplementation with BE decreased (P0.05) on blood glucose (GLU), nonesterified fatty acid (NEFA), total triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C). Compared with control cows, cows fed BE had higher (P0.05) on total antioxidant capacity (T-AOC) or malondialdehyde (MDA) levels. The immunoglobulin (Ig) A and G contents increased (P0.05) on the CD4+ and CD8+ T lymphocyte ratios, CD4+/CD8+ ratio, or tumor necrosis factor-α (TNF-α) level among the groups. These findings suggest that BE supplementation may improve protein metabolism, in addition to enhancing antioxidant activity and immune function in heat-stressed dairy cows.

An exocrine pancreatic insufficient (EPI) pig model was used in the study. The effects of milk formula pre-digestion with immobilized microbial lipase, on the absorption and tissue accretion of long chain polyunsaturated fatty acids (LCPUFA) were assessed. Thirteen male EPI pigs, 10 weeks of age, were used in the study. Six healthy pigs, 6 weeks of age, were used as controls. The pigs were fed either a regular or a pre-digested milk formula. The formula pre-digestion resulted in the decreased faecal total fat and LCPUFA excretion (by 43% and 38% respectively), increased plasma and tissue LCPUFA content (up to 38%). In conclusion, we postulate that feeding a pre-digested milk formula, is an efficient method to develop a functional milk formula, the consumption of which will ensure an increase in total fat absorption (in particular LCPUFA) in human infants as was indicated in the present study in the EPI pig model.

International audience; Mitochondrial beta-oxidation is essential in fat metabolism and can be monitored with blood acylcarnitine profiling, as partly degraded fatty acids accumulate as their carnitine esters. To guarantee continuous energy supply during long-distance exercise, endurance horses oxidise considerable amounts of fat in the mitochondrion. In endurance races over 80 km, glycogen depletion is evident in equine slow-twitch high oxidative muscle fibres and as a consequence, horses participating in endurance races over 80 km rely almost entirely on beta-oxidation of fatty acids. This study investigated mitochondrial fatty acid beta-oxidation in endurance horses exposed to long-distance exercise. Electrospray tandem mass spectrometry analysis of serum acylcarnitine profiles from 10 Arab horses was performed before and after a 160 km endurance race. Results were analysed statistically using ANOVA.Mean speed over the entire race in finishing horses was 16.7 +/- 1.2 km/h. Endurance exercise increased mitochondrial beta-oxidation approximately eight-fold (pre-race, 5648.62 +/- 1508.52 nmol/L; post-race, 44,243.17 +/- 11,504.45 nmol/L; P = 0.001). In these horses, there was an approximately 17-fold increased lipolysis, as demonstrated by elevated serum concentrations of non-esterified fatty acids (NEFA; pre-race, 0.08 +/- 0.08 mmol/L; post-race, 132 +/- 0.36 mmol/L; P < 0.001). In comparison, four Arab horses with poor performance showed an approximately five-fold increase in mitochondrial p-oxidation (pre-race, 5286.17 +/- 3355.16 nmol/L; post-race, 26,660.57 +/- 10,064.27 nmol/L; P= 0.009); there was a 29-fold increase in NEFA (pre-race, 0.02 +/- 0.01 mmol/L; post-race, 0.58 +/- 0.07 mmol/L; P = 0.006) in these horses. Similar post-exercise free carnitine:acetylcarnitine ratios in both groups suggest that the availability of carnitine in long-distance endurance horses might limit performance.

We aimed to study the protein and gene expression of some hepatic enzymes of lipid metabolism along with plasma biomarkers in grazing dairy cattle during the transition period. Blood and liver biopsies from a group of eight multiparous cows were sampled at −28, −14, +4, +14, +28 and +56 days relative to parturition. Peak concentrations of NEFA and beta-hydroxybutyric acid with high triacylglycerol content in the liver were recorded on day 4 postpartum. Consistent with blood biomarkers, the gene expression of carnitine palmitoyltransferase 1A (CPT1A) and acyl-CoA oxidase 1 (ACOX1) increased, whereas that of diacylglycerol O-acyltransferase 1 (DGAT1) decreased. Nevertheless, CPT1A protein expression did not change during all the period evaluated and ACOX1 protein expression increased on day 56 postpartum. In addition, the protein expression of peroxisome proliferator-activated receptor alpha (PPAR-alpha) increased on day 28 postpartum. On the other hand, DGAT1 protein expression decreased on day 14 postpartum. As expected, the expression of genes associated with fatty acid oxidation increased on the first days postpartum but, notably, protein expression was highest after transition. Since most infectious diseases and metabolic disorders in dairy cattle occur particularly on the first days postpartum, it is not so clear whether an increase in the oxidation capacity of the liver at that time could help to prevent disease and improve dairy production. The valuable results about protein expression of enzymes involved in liver lipid metabolism could help to better characterize the metabolism of dairy cattle during the transition period. Fil: Angeli, Emmanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Trionfini, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina Fil: Gareis, Natalia Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Matiller, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Huber, Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Rey, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Salvetti, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Hein, Gustavo Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina

The potential of a mixture of glycosaminoglycans (GAGs) contained in an oral supplement for joint discomfort (OralviscTM) to prevent diet-induced obesity (DIO) and to serve as co-adjuvant in weight loss strategies was studied. Oral treatment with the GAG mix did not counteract the development of DIO in obesity-prone mice but reduced food intake under normal fat diet and led to increased insulin sensitivity under both normal and high fat diets. During reversal of DIO, GAG-treated obese mice showed higher and faster loss of body fat and displayed decreased endpoint adiposity, leptinaemia and hepatic steatosis and signs of increased insulin sensitivity compared with vehicle-treated controls. Expression of oxidative metabolism-related genes and mitochondrial DNA content were increased in visceral white fat depots of GAG-treated animals. The results sustain this GAG mix in functional foods/supplements for obesity and particularly for the (frequent) combination of obesity and joint dysfunction symptoms.

Objectives: The aim was investigate resveratrol, grape juice, or red wine effects on FNDC5 and UCP2 gene expression on muscle and adipose tissue of Wistar rats receiving a high-fat diet. Methods: Five groups of female Wistar rats formed with five animals in each (Control Group-CG, High-Fat Group-HFG-20%, Grape Juice Group-GJG; Red Wine Group-RWG and Resveratrol Group-RG) were treated for 2 months. At the end, adipose and muscle tissues were collected, FNDC5 and UCP2 mRNA expressions were obtained by using real-time polymerase chain reaction. Irisin levels were assessed through ELISA. For comparison: two way ANOVA, using Duncan as post-test. Results: FNDC5 mRNA expression was increased in RG and GJG muscle (p

Type 2 diabetes mellitus (T2DM) arises primarily due to lifestyle factors and genetics. A number of lifestyle factors are known to be important in the development of T2DM, including obesity. JTT-553, a novel Acyl CoA:diacylglycerol acyltransferase 1 inhibitor, reduced body weight depending on dietary fat in diet-induced obesity (DIO) rats in our previous study. Here, the effect of JTT-553 on glucose metabolism was evaluated using body weight reduction in T2DM mice. JTT-553 was repeatedly administered to DIO and KK-A(y) mice. JTT-553 reduced body weight gain and fat weight in both mouse models. In DIO mice, JTT-553 decreased insulin, non-esterified fatty acid (NEFA), total cholesterol (TC), and liver triglyceride (TG) plasma concentrations in non-fasting conditions. JTT-553 also improved insulin-dependent glucose uptake in adipose tissues and glucose intolerance in DIO mice. In KK-A(y) mice, JTT-553 decreased glucose, NEFA, TC and liver TG plasma concentrations in non-fasting conditions. JTT-553 also decreased glucose, insulin, and TC plasma concentrations in fasting conditions. In addition, JTT-553 decreased TNF-α mRNA levels and increased GLUT4 mRNA levels in adipose tissues in KK-A(y) mice. These results suggest that JTT-553 improves insulin resistance in adipose tissues and systemic glucose metabolism through reductions in body weight.

Background Caloric restriction and n-3 polyunsaturated fatty acid (PUFA) supplementation protect from some of the metabolic complications. The aim of this study was to assess the influence of a low calorie diet with or without n-3 PUFA supplementation on glucose dependent insulinotropic polypeptide (GIP) output and insulin sensitivity markers in obese subjects. Methods Obese, non-diabetic subjects (BMI 30–40 kg/m2) and aged 25–65 yr. were put on low calorie diet (1200–1500 kcal/day) supplemented with either 1.8 g/day n-3 PUFA (DHA/EPA, 5:1) (n = 24) or placebo capsules (n = 24) for three months in a randomized placebo controlled trial. Insulin resistance markers and GIP levels were analysed from samples obtained at fasting and during an oral glucose tolerance test (OGTT). Results Caloric restriction with n-3 PUFA led to a decrease of insulin resistance index (HOMA-IR) and a significant reduction of insulin output as well as decreased GIP secretion during the OGTT. These effects were not seen with caloric restriction alone. Changes in GIP output were inversely associated with changes in red blood cell EPA content whereas fasting GIP level positively correlated with HOMA-IR index. Blood triglyceride level was lowered by caloric restriction with a greater effect when n-3 PUFA were included and correlated positively with fasting GIP level. Conclusions Three months of caloric restriction with DHA + EPA supplementation exerts beneficial effects on insulin resistance, GIP and triglycerides. General significance Combining caloric restriction and n-3 PUFA improves insulin sensitivity, which may be related to a decrease of GIP levels., Highlights • Caloric restricted diet with n-3 PUFA improves insulin sensitivity in obese subjects • n-3 PUFA supplementation combined with low calorie diet decrease GIP output • Blood triglycerides level reduces after caloric restriction diet combined with n-3 PUFA • GIP level positively correlates with HOMA-IR index and triglycerides

Objective: Type 2 diabetes arises from insulin resistance of peripheral tissues followed by dysfunction of β-cells in the pancreas due to metabolic stress. Both depletion and supplementation of neutral amino acids have been discussed as strategies to improve insulin sensitivity. Here we characterise mice lacking the intestinal and renal neutral amino acid transporter B0AT1 (Slc6a19) as a model to study the consequences of selective depletion of neutral amino acids. Methods: Metabolic tests, analysis of metabolite levels and signalling pathways were used to characterise mice lacking the intestinal and renal neutral amino acid transporter B0AT1 (Slc6a19). Results: Reduced uptake of neutral amino acids in the intestine and loss of neutral amino acids in the urine causes an overload of amino acids in the lumen of the intestine and reduced systemic amino acid availability. As a result, higher levels of glucagon-like peptide 1 (GLP-1) are produced by the intestine after a meal, while the liver releases the starvation hormone fibroblast growth factor 21 (FGF21). The combination of these hormones generates a metabolic phenotype that is characterised by efficient removal of glucose, particularly by the heart, reduced adipose tissue mass, browning of subcutaneous white adipose tissue, enhanced production of ketone bodies and reduced hepatic glucose output. Conclusions: Reduced neutral amino acid availability improves glycaemic control. The epithelial neutral amino acid transporter B0AT1 could be a suitable target to treat type 2 diabetes.

Bile acid sequestrants (BASs) are cholesterollowering drugs that also affect hyperglycemia. The mechanism by which BASs exert these and other metabolic effects beyond cholesterol lowering remains poorly understood. The present study aimed to investigate the effects of a BAS, colestilan, on body weight, energy expenditure, and glucose and lipid metabolism and its mechanisms of action in high-fat-fed hyperlipidemic APOE*3 Leiden (E3L) transgenic mice. Mildly insulin-resistant E3L mice were fed a high-fat diet with or without 1.5% colestilan for 8 weeks. Colestilan treatment decreased body weight, visceral and subcutaneous fat, and plasma cholesterol and triglyceride levels but increased food intake. Blood glucose and plasma insulin levels were decreased, and hyperinsulinemic-euglycemic clamp analysis demonstrated improved insulin sensitivity, particularly in peripheral tissues. In addition, colestilan decreased energy expenditure and physical activity, whereas it increased the respiratory exchange ratio, indicating that colestilan induced carbohydrate catabolism. Moreover, kinetic analysis revealed that colestilan increased [ 3 H]NEFA incorporation in biliary cholesterol and phospholipids and increased fecal lipid excretion. Gene expression analysis in liver, fat, and muscle supported the above findings. In summary, colestilan decreases weight gain and improves peripheral insulin sensitivity in high-fat-fed E3L mice by enhanced NEFA incorporation in biliary lipids and increased fecal lipid excretion. Copyright © 2013 by the American Society for Biochemistry and Molecular Biology, Inc.

Directing protein and energy sources towards lactation is crucial to optimise milk production in sows but how this influences colostrum yield (CY) remains unknown. The aim of this study was to identify associations between CY and the sow’s use of nutrient resources. We included 37 sows in the study that were all housed, fed and managed similarly. Parity, back fat change (ΔBF), CY and performance parameters were measured. We obtained sow serum samples 3 to 4 days before farrowing and at D1 of lactation following overnight fasting. These were analysed for non-esterified fatty acids (NEFA), urea, creatinine, (iso) butyrylcarnitine (C4) and immunoglobulins G (IgG) and A (IgA). The colostrum samples collected 3, 6 and 24 h after the birth of the first piglet were analysed for their nutrient and immunoglobulins content. The technical parameters associated with CY were parity group (a; parities 1 to 3 =value 0 v. parities 4 to 7 =value 1) and ΔBF D85-D109 of gestation (mm) (b): CY (g) =4290–842a–113b. ( R2=0.41, P

Background and aims: Adipose tissue (AT) fatty acid (FA) composition is considered to be the gold standard long-term biomarker of dietary fatty acid intake. Typically this measurement is made directly from samples collected via large-needle-biopsy or incision. However, with growing interest in the role of AT in relation to health, ideally the fatty acid composition would be analysed along with other measurements, such as gene expression or histology, on a single AT sample. Here we assess alternative ways of obtaining AT for measuring FA composition, in some cases in conjunction with other measurements. Methods and results: The FA composition of tissue obtained via different methods was compared to that of tissue collected via large-needle or surgical biopsy. Fatty acid composition was not significantly different in AT collected by small-needle mini-biopsy (n=10), from an RNA 'lipid layer' (obtained during RNA extraction, 2 sites, n=6 for each), or from cryosectioned tissue prepared for histology (n=10). We also assessed the usefulness of the composition of plasma NEFA as a surrogate marker of subcutaneous AT (n=58-80). Most FAs in plasma NEFA correlated strongly with those in AT (P

Most athletic horses are fed a high-starch diet despite the risk of health problems. Replacing starch concentrate with high-energy forage would alleviate these health problems, but could result in a shift in major substrates for muscle energy supply from glucose to short-chain fatty acids (SCFA) due to more hindgut fermentation of fibre. Dietary fat inclusion has previously been shown to promote aerobic energy supply during exercise, but the contribution of SCFA to exercise metabolism has received little attention. This study compared metabolic response with exercise and lactate threshold (VLa4) in horses fed a forage-only diet (F) and a more traditional high-starch, low-energy forage diet (forage-concentrate diet - FC). The hypothesis was that diet F would increase plasma acetate concentration and increase VLa4 compared with diet FC. Six Standardbred geldings in race training were used in a 29-day change-over experiment. Plasma acetate, non-esterified fatty acids (NEFA), lactate, glucose and insulin concentrations and venous pH were measured in samples collected before, during and after a treadmill exercise test (ET, day 25) and muscle glycogen concentrations before and after ET. Plasma acetate concentration was higher before and after exercise in horses on diet F compared with diet FC, and there was a tendency (P = 0.09) for increased VLa4 on diet F. Venous pH and plasma glucose concentrations during exercise were higher in horses on diet F than diet FC, as was plasma NEFA on the day after ET. Plasma insulin and muscle glycogen concentrations were lower for diet F, but glycogen utilisation was similar for the two diets. The results show that a high-energy, forage-only diet alters the metabolic response to exercise and, with the exception of lowered glycogen stores, appears to have positive rather than negative effects on performance traits.

Electronegative LDL [LDL(-)] is a minor modified LDL subfraction present in blood with inflammatory effects. One of the antiatherogenic properties of HDL is the inhibition of the deleterious effects of in vitro modified LDL. However, the effect of HDL on the inflammatory activity of LDL(-) isolated from plasma is unknown. We aimed to assess the putative protective role of HDL against the cytokine released induced in monocytes by LDL(-). Our results showed that LDL(-) cytokine release was inhibited when LDL(-) was coincubated with HDL and human monocytes and also when LDL(-) was preincubated with HDL and reisolated prior to cell incubation. The addition of apoliprotein (apo) AI instead of HDL reproduced the protective behavior of HDL. HDL preincubated with LDL(-) promoted greater cytokine release than native HDL. Incubation of LDL(-) with HDL decreased the electronegative charge, phospholipase C-like activity, susceptibility to aggregation and nonesterified fatty acid (NEFA) content of LDL(-), whereas these properties increased in HDL. NEFA content in LDL appeared to be related to cytokine production because NEFA-enriched LDL induced cytokine release. HDL, at least in part through apoAI, inhibits phospholipase- C activity and cytokine release in monocytes, thereby counteracting the inflammatory effect of LDL(-). In turn, HDL acquires these properties and becomes inflammatory.-Bancells, C., J. L. Sanchez-Quesada, R. Birkelund, J. Ordoez-Llanos, and S. Benitez. HDL and electronegative LDL exchange anti-and pro-inflammatory properties. J. Lipid Res. 2010. 51: 2947-2956.

The current pilot study assessed the influence of N,N-dimethylglycine (DMG) on insulin sensitivity, glucose and fat metabolism, nutrient digestibility and reproductive performance of sows in the peripartal period. At day 105 of gestation, 25 sows were randomly assigned to the control (n = 13) or the DMG group (n = 12). Sows from the DMG group were supplemented with 1 g DMG/kg feed until day 3 of lactation. After an overnight fast 1 day after farrowing, a blood sample of each sow was drawn. The plasma was analyzed for insulin, glucose, fructosamine, leptin, thiobarbituric acid reactive substances (TBARS), ferric reducing ability of plasma (FRAP), non-esterified fatty acids (NEFA) and triglycerides (TG) and an oral glucose tolerance test was performed. A rectal feces sample was collected and the apparent fecal digestibility (AFD) of crude fat (CFAT), crude protein (CP) and nitrogen-free extract (NFE) was calculated after proximate analyses. Finally, a colostrum sample was collected from each sow and analyzed for the presence of DMG. Reproductive performance parameters were recorded. The results showed an improvement in the AFD of CFAT, CP and NFE when DMG was supplemented. This beneficial effect confirms the hypothesis that DMG acts as an emulsifying agent. The improvement in digestibility in the DMG group was accompanied by a numerical increase in plasma TG (P = 0.067). Plasma NEFA concentrations were not different between treatment groups. DMG supplementation neither affected glucose clearance nor influenced plasma insulin, glucose, fructosamine or leptin levels. TBARS and FRAP also remained unaffected, despite previously reported anti-oxidative properties of DMG. Furthermore, no significant impact on reproductive performance could be recorded. In conclusion, DMG supplementation significantly improved nutrient digestibility. Possible beneficial effects on energy metabolism and reproductive performance of sows should be tested when DMG is supplemented for a longer period of time or at a higher dose.

The aim of this experiment was to evaluate the effect of calf management during lactation on dam performance and nursing behaviour. Thirty-six multiparous beef cows (aged 7.5 6 0.52 years) of Parda de Montana breed (mean live-weight (LW) at calving 566 6 9.3 kg) with a moderate body condition (mean 2.6 6 0.02 on a 1 to 5 scale), were assigned to three nursing frequencies from the day after parturition: Once-daily restricted nursing during a 30 min period at 0800 h (RESTR1), twice-daily restricted nursing during two 30 min periods at 0800 and 1530 h (RESTR2) and ad libitum nursing (ADLIB). Cows were fed throughout the experiment 80 MJ/day of metabolizable energy. Cow-calf behaviour recordings were performed on days 30 and 66 of lactation in each treatment. Blood samples were collected fortnightly to analyse non-esterified fatty acids (NEFA) by an enzymatic colorimetric method, and twice weekly to determine progesterone concentrations by radioimmunoassay. Cow LW losses ( P , 0.05), fat-corrected milk yield ( P , 0.05) and calf average daily gain ( P , 0.01) during lactation were higher in ADLIB. Nursing frequency affected the mean serum concentrations of NEFA, being lower in RESTR1 (0.18 mmol/l; P , 0.05) than in RESTR2 and ADLIB cows (0.29 and 0.25 mmol/l, respectively). The highest occurrence of nursing bouts was observed in ADLIB, leading to greater nursing duration per day (not statistically compared) in this treatment (58.8 6 5.84 min) than in their restricted nursing counterparts (22.2 6 1.42 and 34.3 6 1.16 min, in RESTR1 and RESTR2, respectively). The nursing system did not affect the proportion of cows cycling within 3 months post partum (58%, 46% and 55% in RESTR1, RESTR2 and ADLIB, respectively; P. 0.05). The majority of cows in all treatments showed a short oestrus cycle after first ovulation (75%, 100% and 83%; P. 0.05). In conclusion, the post-partum luteal function in cows managed under continuous access to calves was similar to restricted nursing frequencies, despite the greater suckling intensity of the formers. Restricting calf presence does not enhance the resumption of ovarian activity when suckler cows are managed in moderate nutritional conditions.

We investigated the antidiabetic effects of E3030, which is a potent dual activator of peroxisome proliferator-activated receptor (PPAR) α and PPARγ, in an animal model of diabetes, C57BL/KsJ-db/db mice (db/db mice), and the lipidemic effects of E3030 in beagle dogs, whose PPARα and PPARγ transactivation responses to E3030 were similar to those of humans. E3030 activated human PPARα, mouse PPARα, dog PPARα, human PPARγ, mouse PPARγ, and dog PPARγ with EC50 values of 65, 920, 87, 34, 73, and 34 nM, respectively, in the chimeric GAL4-PPAR receptor transactivation reporter assay. In db/db mice orally administered E3030 decreased blood glucose, triglyceride (TG), non-esterified fatty acids (NEFA), and insulin levels and increased blood adiponectin levels during a 14-day experimental period. Significant effects on blood glucose and adiponectin levels were observed at a dose of 3 mg/kg or greater. Furthermore, significant effects on blood TG, NEFA, and insulin levels were observed at doses of 1 mg/kg or more. An oral glucose tolerance test (OGTT) performed on Day 15 showed that E3030 at 3 mg/kg improved glucose tolerance in this model. Fourteen days of oral treatment with E3030 at a dose of 0.03 mg/kg or greater showed remarkable TG- and non high-density lipoprotein (non-HDL) cholesterol–lowering effects in beagle dogs. These results were similar to those observed for the PPARα agonist fenofibrate. E3030 also reduced apo C-III levels on Days 7 and 14, and elevated lipoprotein lipase (LPL) levels on Day 15. These results indicate that the TG- and non-HDL cholesterol-lowering actions of E3030 involve combined effects on reduction of apo C-III and elevation of LPL, resulting in increased lipolysis. The experimental results in animals suggest that E3030 has potential for use in the treatment of various aspects of metabolic dysfunction in type 2 diabetes, including dyslipidemia, hyperglycemia, hyperinsulinemia, and impaired glucose disposal. Keywords:: E3030, peroxisome proliferator-activated receptor (PPAR) α, PPARγ, PPARα/γ agonist

Low-density lipoprotein (LDL) aggregation is central in triggering atherogenesis. A minor fraction of electronegative plasma LDL, termed LDL(-), plays a special role in atherogenesis. To better understand this role, we analyzed the kinetics of aggregation, fusion and disintegration of human LDL and its fractions, LDL(+) and LDL(-). Thermal denaturation of LDL was monitored by spectroscopy and electron microscopy. Initially, LDL(-) aggregated and fused faster than LDL(+), but later the order reversed. Most LDL(+) disintegrated and precipitated upon prolonged heating. In contrast, LDL(-) partially retained lipoprotein morphology and formed soluble aggregates. Biochemical analysis of all fractions showed no significant degradation of major lipids, mild phospholipid oxidation, and an increase in non-esterified fatty acid (NEFA) upon thermal denaturation. The main baseline difference between LDL subfractions was higher content of NEFA in LDL(-). Since NEFA promote lipoprotein fusion, increased NEFA content can explain rapid initial aggregation and fusion of LDL(-) but not its resistance to extensive disintegration. Partial hydrolysis of apoB upon heating was similar in LDL subfractions, suggesting that minor proteins importantly modulate LDL disintegration. Unlike LDL( +), LDL(-) contains small amounts of apoA-I and apoJ. Addition of exogenous apoA-I to LDL(+) hampered lipoprotein aggregation, fusion and precipitation, while depletion of endogenous apoj had an opposite effect. Therefore, the initial rapid aggregation of LDL(-) is apparently counterbalanced by the stabilizing effects of minor proteins such as apoA-I and apoj. These results help identify key determinants for LDL aggregation, fusion and coalescence into lipid droplets in vivo. (C) 2016 Elsevier B.V. All rights reserved.

We examined oxidative stress and metabolic characteristics of the spontaneously hypertensive hyperlipidemic rat (SHHR) when it was fed a high-fat diet and sucrose solution (HFDS) after NG-nitro-L-arginine methyl ester ingestion to develop a rat model of metabolic syndrome. This study was carried out to assess the effects of pioglitazone on levels of lipid peroxide (LPO), Cu,Zn superoxide dismutase (Cu,Zn-SOD), catalase (CAT), glutathione peroxidase (GPx), and non-esterified fatty acids (NEFA) in the plasma and liver tissue in HFDS-SHHR compared with Sprague-Dawley rats (SD). In the HFDS-treated groups, levels of LPO, CAT, GPx, and NEFA were elevated and levels of Cu,Zn-SOD were reduced in the plasma and liver tissue, with a marked accumulation of visceral fat. The changes induced by HFDS feeding were severe in the SHHR model that had essential hypertension and hyperlipidemia, when compared with SD that did not have these essential risk factors. Subcutaneous injection of 10 mg/kg per day of pioglitazone for 2 months significantly restored levels of LPO, CAT, GPx, Cu,Zn-SOD, and NEFA in the HFDS-SHHR group, and visceral fat accumulation was reduced. These results suggest that HFDS-SHHR is a suitable model of metabolic syndrome and that pioglitazone treatment can improve oxidative dysregulation in this rat model. Keywords:: visceral fat accumulation, metabolic syndrome model, oxidative stress, pioglitazone, anti-oxidant enzyme

The purpose of this experiment was to investigate how early lactating cows adjust their metabolism and production to acute, but moderate changes in the energy density of the diet. Sixty dairy cows were randomly assigned to one of four treatments: two change-over groups (HNH and NHN) and two control groups (HHH and NNN), where H and N refer to a high and normal energy density in the total mixed ration (TMR), respectively. The experimental period covered the first 9 weeks post calving, which was split up in three 3-week periods. Thus, cows assigned to HNH or NHN shifted TMR in weeks 4 and 7 after calving while cows assigned to HHH or NNN were fed the same TMR for all 9 weeks. Results from cows on treatment HNH were compared with group HHH while cows on treatment NHN were compared with group NNN. When the diet changed from N to H and H to N, cows increased and decreased their dry-matter intake (DMI), respectively compared with control groups. Cows adjusted milk yield accordingly to changes in DMI, although not always significantly. Energy-corrected milk yield was not significantly affected by any of the changes in the energy density of the diet but generally showed same tendencies as milk yield. Non-esterified fatty acids (NEFA), beta-hydroxybutyrate in blood and milk and triacylglycerol and glycogen content in the liver were not significantly affected by changes in the energy density of the diet, except from NEFA at one change. Glucose increased more when the diet changed from N to H and increased less when the diet changed from H to N, compared with control groups, although not always significantly. Collectively, these results suggest that cows adjust their DMI and partly milk yield according to the energy density of the diet and therefore only limited effects were observed in physiological parameters.

The aim of this paper is to develop a mathematical model capable of simulating the metabolic response to a variety of mixed meals in fed and fasted conditions with particular emphasis placed on the hepatic triglyceride element of the model. Model validation is carried out using experimental data for the ingestion of three mixed composition meals over a 24-hour period. Comparison with experimental data suggests the model predicts key plasma lipids accurately given a prescribed insulin profile. One counter-intuitive observation to arise from simulations is that liver triglyceride initially decreases when a high fat meal is ingested, a phenomenon potentially explained by the carbohydrate portion of the meal raising plasma insulin.

Postprandial plasma triglyceride (ppTG) and NEFA clearance were stratified by plasma acylation-stimulating protein (ASP) and gender to determine the contribution of fasting ASP in a normal population (70 men; 71 women). In the highest ASP tertile only, ASP decreased over 8 h (90 ± 9.7 nM to 70 ± 5.9 nM, P < 0.05 males; 61.9 ± 4.0 nM to 45.6 ± 6.2 nM, P < 0.01 females). Fasting ASP correlated positively with ppTG response. ppTG (P < 0.0001, 2-way ANOVA, both genders) and NEFA levels progressively increased from lowest to highest ASP tertile, with the greatest differences in males. By stepwise multiple regression, the best prediction of ppTG was: (fasting ASP + apolipoprotein B + insulin + TG; r = 0.806) for men and (fasting ASP + total cholesterol; r = 0.574) for women. Leptin, body mass index, and other fasting variables did not improve the prediction. Thus, in men and women, ASP significantly predicted ppTG and NEFA clearance and, based on lower ASP, women may be more ASP sensitive than men.Plasma ASP may be useful as a fasting variable that will provide additional information regarding ppTG and NEFA clearance.

Phospholipid transfer protein (PLTP) activity is elevated in obese and diabetic subjects. No prospective studies have examined the effect of weight loss on PLTP activity and assessed whether the resultant changes in activity are related to changes in body weight, insulin resistance, or both. PLTP activity was measured at baseline in 46 subjects (body mass index = 19-64 kg/m2) and after diet-induced weight loss in 19 of the obese subjects. Total body fat mass (FM) by dual-energy X-ray absorptiometry, intraabdominal fat (IAF), and abdominal subcutaneous fat (SQF) by CT scan, insulin sensitivity (SI) by frequently sampled intravenous glucose tolerance test, leptin, and lipids were determined. At baseline, PLTP activity correlated with FM (r = 0.36, P = 0.02) and SQF (r = 0.31, P = 0.045), but not with IAF (r = 0.16, P = 0.32) or SI (r = 0.10, P = 0.52). With diet-induced weight loss (16 +/- 7.3 kg), PLTP activity significantly decreased 9.1% (P = 0.002). The change in PLTP activity correlated with the change in SQF (r = 0.55, P = 0.014) (33.6% decrease), but not with IAF (r = 0.09, P = 0.73) or SI (r = 0.18, P = 0.44), and was highly correlated with the change in nonesterified fatty acid (NEFA) (r = 0.71, P < 0.001). In conclusion, elevated PLTP activity in obese subjects is likely a result of increased body fat, reflected by SQF, and is influenced by NEFAs but is not directly related to insulin resistance.

Graphical abstract In situ derivatization to methyl esters (FAME) and fractionation of EFA and NEFA., Despite their important role in tissues, fluids and foods, the analysis of non-esterified fatty acids (NEFA) as methyl esters (NEFAME) is performed using expensive, cumbersome and time-consuming procedures that needs of isolation, fractionation and derivatization steps. However, Yi et al. [1] proposed a promising in situ, single-step procedure to analyze esterified fatty acids (EFA) and NEFA from a same sample on the basis that acylglycerols and free fatty acids can be derivatized using specific reactions. However, according to the data presented in this research work, some modifications need to be performed to increase the reliability of the method:•Increment of the transesterification performance by adding hexane to the reaction mixture, decreasing the time for the derivatization of acylglycerols from 10 min to 3–4 min and stopping the reaction with sulfuric acid.•Avoid cross-contamination of the NEFAME extract by adding 500 μL of water after collection of EFA methyl esters (EFAME).•Samples are spiked with three internal standards: a triacylglycerol (to calculate the concentration of EFA), a free fatty acid (to calculate NEFA) and a FAME (to control isolation of FAME and cross-contamination).

Fibrates and other lipid regulator drugs are widespread in the aquatic environment including estuaries and coastal zones, but little is known on their chronic effects on non-target organisms as marine fish. In the present study, turbot juveniles were exposed to the PPARα model agonist WY-14,643 for 21 days by repeated injections at the concentrations of 5 mg/kg (lo-WY) and 50 mg/kg (hi-WY), and samples taken after 7 and 21 days. Enzyme activity and mRNA expression of palmitoyl-CoA oxidase and catalase in the liver were analyzed as first response, which validated the experiment by demonstrating interactions with the peroxisomal fatty acid oxidation and oxidative stress pathways in the hi-WY treatment. In order to get mechanistic insights, alterations of plasma lipids (free cholesterol, FC; HDL associated cholesterol, C-HDL; triglycerides, TG; non-esterified fatty acids, NEFA) and hepatic mRNA expression of 17 genes involved in fatty acid and lipid metabolism were studied. The exposure to hi-WY reduced the quantity of plasma FC, C-HDL, and NEFA. Microsomal triglyceride transfer protein and apolipoprotein E mRNA expression were higher in hi-WY, and indicated an increased formation of VLDL particles and energy mobilization from liver. It is speculated that energy depletion by PPARα agonists may contribute to a higher susceptibility to environmental stressors.

The effects of a shortage of dietary N and of a rumen protected CLA (rpCLA) supplement on DMI, rumination, rumen fluid characteristics, milk yield (MY), and milk N/N intake ratio (ENU) were studied in mid-late lactating cows. Twenty cows housed in 4 pens in groups of 5, homogeneous for parity, days in milk (DIM) and MY, were fed on 4 different diets: with 150 g (CP15) or 123 g (CP12) of CP/kg DM, with or without an rpCLA supplement (containing 6.34 g/d of C18:2c9,t11 and 6.14 g/d of C18:2t10,c12). A 4 × 4 Latin Square experimental design was used with periods of 3 wks, although the sequence of the 4 treatments (CP15 or CP12, with or without rpCLA) was such that each group received CP15 or CP12 for 6 consecutive wks. The CP12 diet was formulated from CP15 by replacing soybean meal with barley grain to maintain similar energy, fiber content and feed particle size. Rumination activity, DMI, and MY were recorded daily. Rumen fluid was analyzed for VFA and ammonia N content, and milk for quality traits. Nutrient digestibility was estimated using Lignin(sa) as a marker. Period, treatment and group (random) were included as sources of variation in the statistical analysis. Dietary CP restriction tended to reduce DMI (−7.7%; P = 0.09) and digestibility, but increased time spent in rumination (+10%; P = 0.009), decreased rumen fluid ammonia N (−36%; P

The objective of this study was to examine the postprandial response to an exogenous fat source in eight weight-stable postobese subjects (2-3 years after gastric by- pass) and eight matched control women, using a stable iso- tope, ( 13 C)oleate. After a high fat breakfast meal (1,062 cal, 67% fat), ( 13 C)oleate in triglyceride (TG)-rich lipoproteins (S f . 400 and S f 20-400) and nonesterified fatty acids (NEFA), and 13 C in breath CO 2 , were monitored over 8 h. There were no differences in resting energy expenditure, thermic effect of food, carbohydrate/fat oxidation ratio, breath 13 CO 2 enrichment, or fecal fat content between post- obese and control subjects. Postprandially, there was no difference in S f 20-400 TG or NEFA, but postobese sub- jects had lower S f . 400 incremental area under the curve (AUC) (-33%, P , 0.0025) and glucose ( P , 0.01 by re- peated measures analysis of variance (RM ANOVA)). Post- prandial 13 C in S f . 400 TG returned to fasting levels 4 h earlier in postobese subjects and was lower than in control subjects at 4 and 6 h ( P , 0.05 by RM ANOVA). The great- est difference was in the ( 13 C)NEFA profiles. In control sub- jects ( 13 C)NEFA increased markedly over 8 h; postobese subject ( 13 C)NEFA remained close to fasting nonenriched values, and was strikingly lower than in control subjects (72% lower by AUC, P , 0.0001 by RM ANOVA). Finally, postobese subjects tended to have lower postprandial insu- lin ( P , 0.01, 4 h), lower postprandial acylation-stimulating protein, and lower fasting leptin ( 2 46%, P , 0.02). This study demonstrates clear metabolic differences in exoge- nous dietary fat partitioning in postobese women. These findings are compatible with an increased efficiency of di- etary fat storage and suggest one possible mechanism for promotion of weight regain in postobese individuals. — Faraj, M., P. Jones, A. D. Sniderman, and K. Cianflone. En- hanced dietary fat clearance in postobese women. J. Lipid Res. 2001. 42 : 571-580.

A polymorphism in FABP2 that results in an alanine-to-threonine substitution at amino acid 54 of the intestinal fatty acid-binding protein (IFABP) is associated with insulin resistance in Pima Indians. In vitro, the threonine form (Thr54) has a higher binding affinity for long-chain fatty acids than does the alanine form (Ala54). We tested whether this polymorphism affected metabolic responses to dietary fat, in vivo. Eighteen healthy Pima Indians, half homozygous for the Thr54 form of IFABP and half homozygous for the Ala54 form, were studied. The groups were matched for sex, age, and body mass index. Plasma triglyceride, nonesterified fatty acid (NEFA), glucose, and insulin responses were measured after a mixed meal (35% of daily energy requirements, 50 g of fat) and after a high fat challenge (1362 kcal, 129 g of fat). NEFA concentrations were ∼15% higher after the mixed meal and peaked earlier and were ∼20% higher at 7 h in response to the high fat test meal in Thr54 homozygotes compared with Ala54 homozygotes. Insulin responses to the test meals tended to be higher in Thr54 homozygotes, but glucose and triglyceride responses were not different. The results of this study suggest that the Thr54 form of IFABP is associated with higher and prolonged NEFA responses to dietary fat in vivo. Higher NEFA concentrations may contribute to insulin resistance and hyperinsulinemia in individuals with this allele.—Pratley, R. E., L. Baier, D. A. Pan, A. D. Salbe, L. Storlien, E. Ravussin, and C. Bogardus. Effects of an Ala54Thr polymorphism in the intestinal fatty acid-binding protein on responses to dietary fat in humans. J. Lipid Res. 2000. 41: 2002–2008.

There is evidence to indicate that the high rates of coronary heart disease and myocardial infarction amongst Indians of Asian descent may be partly related to circulating non-esterified fatty acids (NEFA). As docosahexaenoic acid (DHA,22:6n–3) in NEFA form has been found to exhibit anti-platelet aggregatory and anti-arrythmic potential in vitro, the effect of supplementary DHA was examined in healthy subjects of Asian Indian background. Furthermore, time- and dose-dependent changes in absolute levels of DHA as NEFA or phospholipid (PL) were compared. The subjects consumed 8 capsules daily of placebo (DHA-free) or low DHA (0.75 g/day) or high DHA (1.50 g/day) over 6 wks. Fasting blood samples were drawn at days 0, 21, and 42 for analysis of serum lipid/lipoprotein composition. No significant effect of DHA supplementation on the levels of serum lipid/lipoproteins (including Lp[a]) or blood pressure was found. However, the DHA level in serum phospholipid rose by 167% overall with low-dose supplementation (from 2.4–6.4 mol%) but only by an additional 23% upon doubling the dose from 0.75 g to 1.50 g/day. Furthermore, after 6 weeks of supplementation with 0.75 g or 1.5 g DHA/day, absolute concentrations of DHA as PL were not significantly different from the corresponding 3-week values. Interestingly, the absolute concentrations of serum DHA as NEFA showed a marked rise with low-dose supplementation (by 212% overall, from 2.4 to 7.5 μm) and a further 70% rise (to 12.7 μm) upon doubling the supplementation from 0.75 to 1.50 g/day. As well, the 6-week concentrations (DHA-NEFA) were significantly different than the corresponding 3-week values at both dose levels. Elevation of circulating DHA-NEFA levels via DHA supplementation, as shown herein, to concentrations that exhibit anti-thrombotic and anti-arrhythmic potential in vitro needs to be extended to trials where clinical end-points are determined.—Conquer, J. A., and B. J. Holub. Effects of supplementation with different doses of DHA on the levels of circulating DHA as non-esterified fatty acid in subjects of Asian Indian background.

Docosahexaenoic acid (22:6) decreases blood platelet function and is highly concentrated in the brain where its depletion leads to functional impairments. Because the platelets and blood brain barrier capillary endothelium cannot hydrolyze the complex lipids for fatty acid (FA) uptake, nonesterified FA (NEFA) bound to albumin are assumed to be the delivery route of FA to these cells. The supply of 13C-labeled 22:6 to blood cells by plasma albumin was studied in humans after a single ingestion of this FA esterified in a triglyceride (TG). The 22:6 13C/12C ratio, measured by gas chromatography combustion-isotope ratio mass spectrometry was measured in lipid classes from albumin, platelets, leukocytes, and erythrocytes (taken as a tentative index of the brain uptake). Nonesterified [13C]22:6 bound to albumin was rapidly produced after ingestion, as a result of the hydrolysis of very low density lipoprotein (VLDL) plus chylomicron TG. We found that albumin carried another source of 22:6, lyso-phosphatidylcholines (lyso-PC), in which [13C]22:6 accumulated while the nonesterified [13C]22:6 reached its minimal plasma concentrations. Computation of the relative contribution of NEFA and lyso-PC for the [13C]22:6 delivery to platelets and erythrocytes showed that the [13C]22:6 supply to platelets occurred uniquely through NEFA, whereas this pool was weakly involved in the delivery to erythrocytes. In contrast, lyso-PC was uniquely concerned with the 22:6 delivery to erythrocytes and represented the major part of this supply. We conclude that plasma albumin carries 22:6 in two lipid forms that are involved differently in the delivery of this FA to target cells.

The objectives of this study were to assess the clinical and metabolic risk factors for clinical endometritis, the likelihood for having a normal vaginal discharge during postpartum, and the effects of endometritis on milk yield, reproductive efficiency, and metabolic status in Holstein cows. The study was conducted in a commercial dairy herd (Cordoba, Argentina) where 303 Holstein cows were enrolled. Cows were body condition scored (1 to 5) and tail bled on -14, 7, 21, 31, 41, and 50. d relative to parturition. Cows having a vaginal discharge with presence of pus between 21 and 41. d postpartum (dpp) were diagnosed as having clinical endometritis. Plasma blood samples were analyzed for nonesterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), and blood urea nitrogen using commercial kits and insulin-like growth factor 1, insulin, and leptin by RIA. Data were analyzed with PROC MIXED, PROC GENMOD, and PROC PHREG of SAS (SAS Institute Inc., Cary, NC). Abnormal calving and puerperal metritis increased the risk for endometritis [adjusted odds ratio (AOR)=2.21 for both]. High prepartum NEFA and high postpartum BHBA increased the risk for endometritis (AOR=1.003 and 1.001, respectively), whereas high prepartum blood urea nitrogen reduced it (AOR=0.853). Cut-offs of 456.6 μ M NEFA and 402.5 μM BHBA had sensitivities of 0.69 and 0.58, and specificities of 0.88 and 0.86, respectively. The likelihood for having normal vaginal discharge increased with time (∼1% × dpp) and with normal calving. Cows with endometritis had higher milk yield than normal herdmates (27.8±0.9 vs. 25.7±0.4. kg/d), lower risk for pregnancy by 100. dpp (AOR=0.10), higher nonpregnancy risk by 200. dpp (AOR=2.87), and higher risk for culling than normal cows (AOR=2.28). Cows with endometritis had a lower hazard rate (0.44) for pregnancy and had approximately 70. d longer calving-to-conception intervals. Finally, endometritis had no effect on metabolic hormones. In conclusion, the risk for clinical endometritis increases with abnormal calving and puerperal metritis, as prepartum NEFA and postpartum BHBA concentrations increase. Prepartum NEFA and postpartum BHBA could be useful for the prediction of endometritis. Last, clinical endometritis has detrimental effects on reproductive efficiency, as affected cows take longer to get pregnant and are at higher risk for culling., Facultad de Ciencias Veterinarias