Increased Plasma Nonesterified Fatty Acids Research Articles

Effect of exogenous insulin and fasting on growth hormone receptor and IGF-I expression in the pre-ovulatory follicle of ewes

The aim of this study was to investigate the effect of fasting and exogenous insulin administration on the expression of growth hormone receptor (GHR) and IGF-I mRNA in the pre-ovulatory follicle of ewes. Fifteen ewes received an intravaginal progesterone releasing device that was removed 6 days later (day of removal = day 0). On day −2, the ewes were divided into three groups: (i) fasting group (n = 5) that was fasted from day −2 to day 2; (ii) control group (n = 5) that received a maintenance diet; and (iii) insulin group (n = 5) that received insulin injections (0.25 IU/kg) every 12 h from day −2 to day 2 under the same diet as the control group. Follicular samples were obtained on day 2. Fasting increased plasma non-esterified fatty acids concentrations from day −1 to day 2 (P < 0.001). There was no difference (P > 0.05) in the number of follicles, although there was a tendency for an increase in the pre-ovulatory follicle diameter for the insulin group in comparison to the control group (P = 0.12). Thecal GHR mRNA expression was very low and was considered insignificant. Moreover, granulosa cells GHR mRNA expression increased (P < 0.05) in the insulin group. Expression of IGF-I mRNA was not different among groups in both tissues. In conclusion, insulin administration increases GHR mRNA but not IGF-I mRNA expression in granulosa cells of the pre-ovulatory follicle. However, fasting did not change the pattern of GHR/IGF-I mRNA expression in the pre-ovulatory follicle.

Biphasic adaptative responses in VLDL metabolism and lipoprotein homeostasis during Gram-negative endotoxemia

Dyslipidemia and hepatic overproduction of very low density lipoprotein (VLDL) are hallmarks of the septic response, yet the underlying mechanisms are not fully defined. We evaluated the lipoprotein subclasses profile and hepatic VLDL assembly machinery over 24 h in fasted LPS-treated rats. The response of serum non-esterified fatty acids (NEFA) and glucose to endotoxin was biphasic, with increased levels of NEFA and hypoglycemia in the first 12 h-phase, and low NEFA and high glucose in the second 12 h-phase. Hypertriglyceridemia was more marked in the first 12 h (6.8-fold), when triglyceride abundance increased in all lipoprotein subclasses, and preferentially in large VLDL. The abundance of medium-sized VLDL and the increase in the number of VLDL particles was higher in the second phase (10-fold vs 5-fold in the first phase); however, apoB gene transcript abundance increased only in the second phase. Analysis of putative pre-translational mechanisms revealed that neither increased Apob transcription rate nor increased transcript binding to mRNA stabilizing HuR (Hu antigen R) protein paralleled the increase in apoB transcripts. In conclusion, endotoxin challenge induces increases in plasma NEFA and large, triglyceride-rich VLDL. After approximately 12 h, the triglyceride-rich VLDLs are replaced by medium-sized, triglyceride-poor VLDL particles. Hepatic apoB mRNA abundance also increases during the second period, suggesting a role for apoB protein expression in the acute reaction against sepsis.

Adipose triacylglycerol lipase is a major regulator of hepatic lipid metabolism but not insulin sensitivity in mice

Hepatic steatosis is characterised by excessive triacylglycerol accumulation and is strongly associated with insulin resistance. An inability to efficiently mobilise liver triacylglycerol may be a key event mediating hepatic steatosis. Adipose triacylglycerol lipase (ATGL) is a key triacylglycerol lipase in the liver and we hypothesised that liver-specific overproduction of ATGL would reduce steatosis and enhance insulin action in obese rodents. Studies of fatty acid metabolism were conducted in primary hepatocytes isolated from wild-type and Atgl (also known as Pnpla2)⁻(/)⁻ mice. An ATGL adenovirus was utilised to overproduce ATGL in the livers of obese insulin-resistant C57Bl/6 mice (Ad-ATGL). Blood chemistry, hepatic lipid content and insulin sensitivity were assessed in mice. Triacylglycerol content was increased in Atgl⁻(/)⁻ hepatocytes and was associated with increased fatty acid uptake and impaired fatty acid oxidation. ATGL adenovirus administration in obese mice increased the production of hepatic ATGL protein and reduced triacylglycerol, diacylglycerol and ceramide content in the liver. Overproduction of ATGL improved insulin signal transduction in the liver but did not affect fasting glycaemia or insulinaemia. Inflammatory signalling was not suppressed by ATGL overproduction. While ATGL overproduction increased plasma non-esterified fatty acids, neither lipid deposition nor insulin-stimulated glucose uptake were affected in skeletal muscle. Liver ATGL overproduction decreases hepatic steatosis and mildly enhances liver insulin sensitivity. These effects are not sufficient to improve fasting glycaemia or insulinaemia in rodent obesity.

Increased Postprandial Nonesterified Fatty Acid Appearance and Oxidation in Type 2 Diabetes Is Not Fully Established in Offspring of Diabetic Subjects

BackgroundIt has been proposed that abnormal postprandial plasma nonesterified fatty acid (NEFA) metabolism may participate in the development of tissue lipotoxicity and type 2 diabetes (T2D). We previously found that non-diabetic offspring of two parents with T2D display increased plasma NEFA appearance and oxidation rates during intravenous administration of a fat emulsion. However, it is currently unknown whether plasma NEFA appearance and oxidation are abnormal during the postprandial state in these subjects at high-risk of developing T2D.MethodologyPalmitate appearance and oxidation rates and glycerol appearance rate were determined in eleven healthy offspring of two parents with T2D (positive family history, FH+), 13 healthy subjects without first-degree relatives with T2D (FH-) and 12 subjects with T2D at fasting, during normoglycemic hyperinsulinemic clamp and during continuous oral intake of a standard liquid meal to achieve steady postprandial NEFA and triacylglycerols (TG) without and with insulin infusion to maintain similar glycemia in all three groups.Principal FindingsPlasma palmitate appearance and oxidation were higher at fasting and during the clamp conditions in the T2D group (all P<0.05). In the postprandial state, palmitate appearance, oxidative and non oxidative rates were all elevated in T2D (all P<0.05) but not in FH+. Both T2D and FH+ displayed elevated postprandial TG vs. FH- (P<0.001). Acute correction of hyperglycemia during the postprandial state did not affect these group differences. Increased waist circumference and BMI were positively associated with elevated postprandial plasma palmitate appearance and oxidation.Conclusions/SignificancePostprandial plasma NEFA intolerance observed in subjects with T2D is not fully established in non-diabetic offspring of both parents with T2D, despite the presence of increased postprandial plasma TG in the later. Elevated postprandial plasma NEFA appearance and oxidation in T2D is observed despite acute correction of the exaggerated glycemic excursion in this group.

Effect of different dietary fats on hepatic gene expression in transition dairy goats

Eighteen second parity goats were selected for this study and divided in three homogenous groups of six animals each. Starting from week 2 before the expected kidding date to week 3 of lactation animals were fed the experimental diets: C, basal transition diet; PFISH, basal transition diet plus 47 g/day of rumen protected fish oil as source of n-3 fatty acids; HPALM, basal transition diet plus 47 g/day of hydrogenated palm oil (46.30% C16:0, 40.90% C18:0). Live body weight (LBW), body condition score (BCS), dry matter intake (DMI) and energy balance were assessed weekly. Milk yield and milk composition were recorded and analyzed on 4, 7, 15, and 21 days in milk (DIM). Alanine transaminase (ALAT), aspartate aminotrasferase (ASAT), non esterified fatty acids (NEFA), glucose, β-hydroxybutyrate (BHBA), and cholesterol plasma contents were measured weekly for all the experimental period. Liver mRNA levels for peroxisome proliferator-activated receptor-α (PPARA), acyl-coenzyme A dehydrogenase very long chain (ACADVL), acetyl coenzyme A carboxylase (ACACA), and carnitine palmitoyl-transferase 1A (CPT 1A) were determined 7 days (±1.43) before and 21 days after kidding by reverse transcription real time (RT) quantitative PCR (Q-PCR). Goats from the three experimental groups experienced negative energy balance after kidding with higher levels in both HPALM and PFISH than C ( P ≤ 0.01) while no differences were detected among fat supplemented goats. Performance, milk composition, plasma metabolites and liver enzymes were similar in C, HPALM and PFISH, except for a significant effect of treatment on milk yield ( P ≤ 0.01) in HPALM compared to C goats (3.65 kg/day vs. 2.42 kg/day, P = 0.01) and ALAT content ( P ≤ 0.01) in the first 21 DIM in HPALM than C and PFISH (14.25 IU vs. 17.54 IU and 16.80 IU respectively; P ≤ 0.05). Treatment ( P = 0.08), day of treatment ( P = 0.06), and the interaction among treatment and day of treatment ( P = 0.08) showed tendencies to increased plasma NEFA content in HPALM during the post kidding period respect to PFISH and C. An interaction effect between treatment and day of treatment was found for PPARA ( P = 0.02), CPT1A ( P = 0.05), and ACADVL ( P = 0.05) mRNA levels. At 21 days in milk HPALM showed higher values in PPARA mRNA levels than C. mRNA levels of PPARA, CPT 1A, ACADVL, and ACACA showed trends to increase from 7 days before to 21 days after kidding in HPALM, contrary to PFISH. The present trial shows the effectiveness of hydrogenated palm oil, as an oil mainly containing C16:0 and C18:0, in enhancing mRNA expression of PPARA and selected target genes in liver of transition dairy goat.

A Single Night of Partial Sleep Deprivation Induces Insulin Resistance in Multiple Metabolic Pathways in Healthy Subjects

Subsequent nights with partial sleep restriction result in impaired glucose tolerance, but the effects on insulin sensitivity have not been characterized. The aim of this study was to evaluate the effect of a single night of partial sleep restriction on parameters of insulin sensitivity. Nine healthy subjects (five men, four women) were studied once after a night of normal sleep duration (sleep allowed from 2300 to 0730 h), and once after a night of 4 h of sleep (sleep allowed from 0100 to 0500 h). Sleep characteristics were assessed by polysomnography. Insulin sensitivity was measured by hyperinsulinemic euglycemic clamp studies (from 1130 to 1430 h) with infusion of [6,6-(2)H(2)]glucose. Sleep duration was shorter in the night with sleep restriction than in the unrestricted night (226 +/- 11 vs. 454 +/- 9 min; P< 0.0001). Sleep restriction did not affect basal levels of glucose, nonesterified fatty acids, insulin, or endogenous glucose production. Sleep restriction resulted in increased endogenous glucose production during the hyperinsulinemic clamp study compared to the unrestricted night (4.4 +/- 0.3 vs. 3.6 +/- 0.2 micromol x kg lean body mass(-1) x min(-1); P = 0.017), indicating hepatic insulin resistance. In addition, sleep restriction decreased the glucose disposal rate during the clamp (32.5 +/- 3.6 vs. 40.7 +/- 5.1 micromol x kg lean body mass(-1) x min(-1); P = 0009), reflecting decreased peripheral insulin sensitivity. Accordingly, sleep restriction decreased the rate of glucose infusion by approximately 25% (P = 0.001). Sleep restriction increased plasma nonesterified fatty acid levels during the clamp study (68 +/- 5 vs. 57 +/- 4 micromol/liter; P = 0.005). Partial sleep deprivation during only a single night induces insulin resistance in multiple metabolic pathways in healthy subjects. This physiological observation may be of relevance for variations in glucoregulation in patients with type 1 and type 2 diabetes.

Fasting and sampling time affect liver gene expression of high-fat diet-fed mice

Several physiological and biological variables are known to affect peroxisome proliferator-activated receptor (PPAR)-α-dependent signaling pathway and plasma biochemical profiles. However, less is known about the effect of these variables on high-fat diet-fed mice. In a 5-week study, C57BL/6 mice were divided into control (C) and high-fat diet-fed (H) groups, whereby before dissection, each group was subdivided into non-fasted (nC and nH) and a 15-h fasted mice (fC and fH) killed in the early light cycle, and a 15-h fasted mice (eC and eH) killed in the late phase of the light cycle. Liver and blood from the vena cava were collected. Non-fasted nC and nH mice have a marginal difference in their body weight gain, whereas significant differences were found for fasted mice. In nH mice, PPAR-α, acyl-CoA oxidase and insulin-like growth factor-binding protein expressions were significantly elevated, in contrast to fatty acid synthase (Fasn), stearoyl CoA-desaturase (SCD)-1, and elongase (ELOVL)-6 expressions. Fasn was profoundly induced in fH mice, while decreased sterol regulatory-binding protein-1 and SCD-1 were found only in eH mice. Different from the gene expression profiles, plasma total cholesterol level of the eH mice was higher than controls, whereas nH mice have increased plasma non-esterified fatty acids. Only glucose level of the fH mice was higher than that observed for controls. Results showed that fasting and sampling time have significantly affected liver gene expression and plasma biochemical indices of the high-fat diet-treated mice. An overlook in these aspects can cause serious discrepancies in the experimental data and their interpretations.

Non-esterified fatty acids increase arterial pressure via central sympathetic activation in humans

Previous studies have shown that acute increases in plasma NEFAs (non-esterified fatty acids) raise SVR (systemic vascular resistance) and BP (blood pressure). However, these studies have failed to distinguish between CNS (central nervous system) mechanisms that raise sympathetic activity and paracrine mechanisms that increase SVR directly, independent of CNS involvement. The aim of the present study was to directly determine whether the sympathetic nervous system contributes to the pressor response to NEFAs. On 2 days separated by at least 2 weeks, 17 lean healthy volunteers (ten male/seven female; age, 22+/-1 years; body mass index, 23+/-1 kg/m2; values are means+/-S.E.M.) received a 4-h intravenous infusion of 20% Intralipid or placebo (in a single-blind randomized balanced order). MSNA (muscle sympathetic nerve activity), HR (heart rate), BP (oscillometric brachial measurement) and Q (cardiac output; acetylene rebreathing) were measured before and throughout infusion. The change in HR (+8.2+/-1.0 and +2.4+/-1.2 beats/min), systolic BP (+14.0+/-1.6 and +3.2+/-2.5 mmHg) and diastolic BP (+8.2+/-1.0 and -0.1+/-1.7 mmHg) were significantly greater after the 4-h infusion of Intralipid compared with placebo (P<0.001). The change in BP with Intralipid resulted from an increase in SVR (Q/mean arterial pressure; P<0.001) compared with baseline, without a change in Q. MSNA burst frequency increased during Intralipid infusion compared with baseline (+4.9+/-1.3 bursts/min; P<0.05), and total MSNA (frequencyxamplitude) was augmented 65% (P<0.001), with no change during placebo infusion. Lipid infusion increased insulin, aldosterone and F2-isoprostane, but not leptin, concentrations. On the basis of the concomitant increase in BP, MSNA and SVR, we conclude that central sympathetic activation contributes to the pressor response to NEFAs.

Changes of glycogen content in liver, skeletal muscle, and heart from fasted rats

Glycogen content of white and red skeletal muscles, cardiac muscle, and liver was investigated in conditions where changes in plasma levels of non-esterified fatty acids (NEFA) occur. The experiments were performed in fed and 12 and 48 h-fasted rats. The animals were also submitted to swimming for 10 and 30 min. Glycogen content was also investigated in both pharmacologically induced low plasma NEFA levels fasted rats and pharmacologically induced high plasma NEFA levels fed rats. The participation of Akt and glycogen synthase kinase-3 (GSK-3) in the changes observed was investigated. Plasma levels of NEFA, glucose, and insulin were determined in all conditions. Fasting increased plasma NEFA levels and reduced glycogen content in the liver and skeletal muscles. However, an increase of glycogen content was observed in the heart under this condition. Akt and GSK-3 phosphorylation was reduced during fasting in the liver and skeletal muscles but it remained unchanged in the heart. Our results suggest that in conditions of increased plasma NEFA levels, changes in insulin-stimulated phosphorylation of Akt and GSK-3 and glycogen content vary differently in liver, skeletal muscles, and heart. Akt and GSK-3 phosphorylation and glycogen content are decreased in liver and skeletal muscles, but in the heart it remain unchanged (Akt and GSK-3 phosphorylation) or increased (glycogen content) due to consistent increase of plasma NEFA levels.

Relationship between Total and High Molecular Weight Adiponectin Levels and Plasma Nonesterified Fatty Acid Tolerance during Enhanced Intravascular Triacylglycerol Lipolysis in Men

Increased plasma nonesterified fatty acid (NEFA) appearance during enhanced intravascular triacylglycerol (TG) lipolysis is a marker of metabolic adipose tissue dysfunction and may lead to the development of insulin resistance. The relationship between total and high molecular weight (HMW) adiponectin levels, NEFA appearance, and total TG lipolytic capacity has not been previously studied in humans. Our objective was to determine whether total and HMW adiponectin plasma levels are associated with plasma NEFA level and appearance, and with total TG lipolytic rate during enhanced intravascular TG lipolysis in men. This was a cross-sectional metabolic study. The study was performed at an academic clinical research center. There were 15 healthy men (mean +/- sd body mass index 25.5 +/- 4.7 kg/m(2)) aged 21-50 yr (mean +/- sd 31.1 +/- 10.2) without first-degree relatives with type 2 diabetes included in the study. Pancreatic clamps and iv infusion of stable isotopic tracers ([1,1,2,3,3-(2)H(5)]glycerol and [U-(13)C]palmitate) were performed, whereas intravascular TG lipolysis was clamped by iv infusion of heparin plus Intralipid at low (fasting) and high insulin levels. Total and HMW adiponectin levels were measured using an ELISA. Levels of total and HMW adiponectin, palmitate appearance (plasma palmitate appearance rate), and glycerol appearance (plasma glycerol appearance rate) were calculated. During heparin plus Intralipid infusion, total and HMW adiponectin was inversely correlated with plasma palmitate appearance rate (r = -0.65; P = 0.01), but this association was lost when expressed per nonlean weight. Adiponectin levels were positively associated with plasma glycerol appearance rate per nonlean weight (r = 0.71 and r = 0.66, respectively; P < or = 0.01). Increased adipose tissue mass likely explains the association between low adiponectin and reduced NEFA tolerance. Adiponectin level is a marker of total TG lipolytic rate per adipose tissue mass in men.

Short-term flexibility of myocardial triglycerides and diastolic function in patients with type 2 diabetes mellitus

Short-term caloric restriction increases plasma levels of nonesterified fatty acids (NEFAs) and is associated with increased myocardial triglyceride (TG) content and decreased myocardial function in healthy subjects. Whether this flexibility of myocardial TG stores and myocardial function is also present in patients with type 2 diabetes mellitus (T2DM) is yet unknown. Myocardial TG content and left ventricular (LV) ratio between the early (E) and atrial (A) diastolic filling phase (E/A) were determined using magnetic resonance (MR) spectroscopy and MR imaging, respectively, before and after a 3-day very low-calorie diet (VLCD) in 11 patients with T2DM. In addition, we studied patients after a 3-day VLCD combined with the antilipolytic drug acipimox. The VLCD induced myocardial TG accumulation [from 0.66 +/- 0.09% (mean +/- SE, baseline) to 0.98 +/- 0.16%, P = 0.028] and a decrease in E/A ratio [from 1.00 +/- 0.05 (baseline) to 0.90 +/- 0.06, P = 0.002]. This was associated with increased plasma NEFA levels (from 0.57 +/- 0.08 mmol/l at baseline to 0.92 +/- 0.12, P = 0.019). After the VLCD with acipimox, myocardial TG content, diastolic function, and plasma NEFA levels were similar to baseline values. In conclusion, in patients with T2DM, a VLCD increases myocardial TG content and is associated with a decrease in LV diastolic function. These effects were not observed when a VLCD was combined with acipimox, illustrating the physiological flexibility of myocardial TG stores and myocardial function in patients with T2DM.

Glycéronéogenèse et PEPCK-C

Obesity is a major risk factor for insulin resistance and type 2 diabetes. The link between hypertrophied adipose tissue and this pathology is thought to be non-esterified fatty acids (NEFA) arising from adipocyte lipolysis. Sustained increase in plasma NEFA induces insulin resistance. In adipocytes, a significant part of lipolytic NEFA is re-esterified to triacylglycerol. Re-esterification requires glycerol-3-phosphate which, during fasting, is synthesized from lactate, pyruvate or certain amino acids in a metabolic pathway named glyceroneogenesis. The key enzyme in this pathway is the cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C). In this review, we postulate that thiazolidinediones exert their hypolipidemic and antidiabetic effects in adipose tissue at least in part through a rapid and selective induction of PEPCK-C gene transcription leading to increased PEPCK-C and glyceroneogenesis. Subsequent fatty acid re-esterification participates in the reduction in blood NEFA and insulin resistance.

Lipid Oxidation According to Intensity and Exercise Duration in Overweight Men and Women

Our objective was to compare the effect of different exercise intensities on lipid oxidation in overweight men and women. Nine young, healthy, overweight men and women were studied (age, 31.4 +/- 2.3 and 26.7 +/- 2.1 years; BMI, 27.9 +/- 0.4 and 27.2 +/- 0.5; for men and women, respectively). On one study day, the subjects first performed 30 minutes of cycling exercise at 30% of their maximal oxygen uptake (Vo(2max); E1 session), followed by 30 minutes of exercise at 50% Vo(2max) (E2 session). On a second study day, a similar E1 session was followed by 30 minutes of exercise at 70% Vo(2max) (E3 session). From the gas exchange measurements, the respiratory exchange ratio (RER) and the fat oxidation rate (FOR) were calculated. Plasma concentrations of glycerol and non-esterified fatty acids (NEFAs) were assayed. RER was significantly lower for women during only the E1 session. For both sexes, RER decreased over time during the E2 and E3 sessions. During the E1 session, the FOR per kilogram of lean mass (LM) was higher among women, and it did not change over time despite an increase in plasma NEFAs. FOR per kilogram of LM was higher during the E2 exercise for both sexes. During E2 and E3 sessions, as the exercise time was prolonged, the FOR/kg LM increased simultaneously with the increase in the plasma glycerol. Lipid oxidation during exercise is optimized for moderate and lengthy exercise. The enhancement of lipid oxidation occurring over time during moderate- and high-intensity exercises could be, in part, linked to the improvement of lipid mobilization. This fact is discussed to shed light on exercise modalities as a tool for the management of overweight.

Short-Term Caloric Restriction Induces Accumulation of Myocardial Triglycerides and Decreases Left Ventricular Diastolic Function in Healthy Subjects

Diabetes and obesity are associated with increased plasma nonesterified fatty acid (NEFA) levels, myocardial triglyceride accumulation, and myocardial dysfunction. Because a very low-calorie diet (VLCD) also increases plasma NEFA levels, we studied the effect of a VLCD on myocardial triglyceride content and cardiac function in healthy subjects. Fourteen healthy nonobese men underwent (1)H-magnetic resonance spectroscopy (MRS) to determine myocardial and hepatic triglyceride content, (31)P-MRS to assess myocardial high-energy phosphate (HEP) metabolism (phosphocreatine/ATP), and magnetic resonance imaging of myocardial function at baseline and after a 3-day VLCD. After the dietary intervention, plasma NEFA levels increased compared with those at baseline (from 0.5 +/- 0.1 to 1.1 +/- 0.1 mmol/l, P < 0.05). Concomitantly, myocardial triglyceride content increased by approximately 55% compared with that at baseline (from 0.38 +/- 0.05 to 0.59 +/- 0.06%, P < 0.05), whereas liver triglyceride content decreased by approximately 32% (from 2.2 +/- 0.5 to 1.5 +/- 0.4%, P < 0.05). The VLCD did not change myocardial phosphocreatine-to-ATP ratio (2.33 +/- 0.15 vs. 2.33 +/- 0.08, P > 0.05) or systolic function. Interestingly, deceleration of the early diastolic flow across the mitral valve decreased after the VLCD (from 3.37 +/- 0.20 to 2.91 +/- 0.16 ml/s(2) x 10(-3), P < 0.05). This decrease in diastolic function was significantly correlated with the increase in myocardial triglyceride content. Short-term VLCD induces accumulation of myocardial triglycerides. In addition, VLCD decreases left ventricular diastolic function, without alterations in myocardial HEP metabolism. This study documents diet-dependent physiological variations in myocardial triglyceride content and diastolic function in healthy subjects.

Cytosolic Aspartate Aminotransferase, a New Partner in Adipocyte Glyceroneogenesis and an Atypical Target of Thiazolidinedione

We show that cytosolic aspartate aminotransferase (cAspAT) is involved in adipocyte glyceroneogenesis, a regulated pathway that controls fatty acid homeostasis by promoting glycerol 3-phosphate formation for fatty acid re-esterification during fasting. cAspAT activity, as well as the incorporation of [(14)C]aspartate into the neutral lipid fraction of 3T3-F442A adipocytes was stimulated by the thiazolidinedione (TZD) rosiglitazone. Conversely, the ratio of fatty acid to glycerol released into the medium decreased. Regulation of cAspAT gene expression was specific to differentiated adipocytes and did not require any peroxisome proliferator-activated receptor gamma (PPARgamma)/retinoid X receptor-alpha direct binding. Nevertheless, PPARgamma is indirectly necessary for both cAspAT basal expression and TZD responsiveness because they are, respectively, diminished and abolished by ectopic overexpression of a dominant negative PPARgamma. The cAspAT TZD-responsive site was restricted to a single AGGACA hexanucleotide located at -381 to -376 bp whose mutation impaired the specific RORalpha binding. RORalpha ectopic expression activated the cAspAT gene transcription in absence of rosiglitazone, and its protein amount in nuclear extracts is 1.8-fold increased by rosiglitazone treatment of adipocytes. Finally, the amounts of RORalpha and cAspAT mRNAs were similarly increased by TZD treatment of human adipose tissue explants, confirming coordinated regulation. Our data identify cAspAT as a new member of glyceroneogenesis, transcriptionally regulated by TZD via the control of RORalpha expression by PPARgamma in adipocytes.

Effects of Peripartum Propylene Glycol or Fats Differing in Fatty Acid Profiles on Feed Intake, Production, and Plasma Metabolites in Dairy Cows

Dry multiparous cows were used to investigate the effects on intake, production, and metabolism of either a supplement containing 55% dry propylene glycol (PGLY), a prilled fat supplement (PrFA) containing a low proportion of unsaturated fatty acids (FA), or calcium soaps of FA supplement (CaLFA) containing a high proportion of unsaturated FA. Fifty-three dry cows (256 d pregnant) were stratified into 4 groups and began one of the following dietary treatments: 1) control cows were fed a dry cow diet and at postpartum were fed a lactating cow diet; 2) diets of cows in the PGLY group were supplemented with 500 g/d per cow of dry PGLY until 21 d in milk (DIM); 3) diets of cows in the PrFA group were supplemented with 230 g/d per cow of PrFA until 100 DIM; 4) diets of cows in the CaLFA group were supplemented with 215 g/d per cow of CaLFA until 100 DIM. Prepartum DMI was lower in the PrFA and CaLFA groups than in the control and PGLY groups, whereas postpartum DMI in the PrFA group was higher than that in the control group. Milk production until 100 DIM in both fat-supplemented groups was 4.5% higher than that in the control group. Plasma glucose concentrations pre- and postpartum were higher in the PGLY group than in the PrFA and CaLFA groups, but were similar to those in the control group. Prepartum nonesterified FA (NEFA) concentrations in plasma were increased by 43 and 70% in the PrFA and CaLFA groups, respectively, as compared with the control and PGLY groups. Both fat supplements increased plasma β-hydroxybutyrate concentrations over those of the PGLY and control groups pre- and postpartum. Peripartum plasma insulin concentrations in the control group were 1.7-fold higher than in the PrFA group and 2.1-fold higher than in the CaFA group. Differences between the PrFA and CaLFA groups were observed: DMI was higher pre- and postpartum in the PrFA group than in the CaLFA group, and prepartum plasma NEFA concentrations were 19% higher and insulin concentrations were 21% lower in the CaLFA group than in the PrFA group. No significant differences were observed in DMI, plasma glucose, NEFA, and β-hydroxybutyrate concentrations between the control and PGLY groups. Feeding fat to cows during late pregnancy decreased the DMI and negatively affected the metabolic status of the cows, as reflected by plasma metabolites. Furthermore, protected fat with a high proportion of unsaturated FA (CaLFA) was more pronounced in increasing plasma NEFA concentrations and depressing plasma insulin concentrations than fat with a low proportion of unsaturated FA (PrFA).

Central administration of alpha-melanocyte-stimulating hormone changes lipid metabolism in chicks

Alpha-melanocyte-stimulating hormone (MSH) is well known as an anorexigenic peptide in the brain of mammals. In addition to this, brain alpha-MSH enhances heat production (HP), indicating that the peptide acts as a catabolic factor in the regulation of energy metabolism. The anorexigenic effect of alpha-MSH is also observed in chicks ( Gallus gallus), but no information has been available for its effect on HP. The present study was performed to examine whether intracerebroventricular (ICV) injection of alpha-MSH increases HP in chicks. The injection of alpha-MSH (10 and 100 pmol) did not affect oxygen consumption, carbon dioxide production and HP during the 1 h post-injection period. This result was supported by another result that ICV injection of alpha-MSH did not affect locomotion activity in chicks. In contrast, the respiratory quotient was significantly lowered by the ICV injection of MSH. We also found that alpha-MSH significantly increased plasma non-esterified fatty acid concentrations. In summary, brain alpha-MSH appears to exert generally catabolic effects on lipid metabolism in the chick, but does not appear to be involved in the regulation of HP.

Induction of Hyperlipidemia by Intravenous Infusion of Tallow Emulsion Causes Insulin Resistance in Holstein Cows

The objective was to test whether the induction of elevated blood nonesterified fatty acids (NEFA) by i.v. infusion of a tallow emulsion altered glucose tolerance and responsiveness to insulin in Holstein cows. Six non-lactating, nongestating Holstein cows were assigned to a crossover design. One cow was excluded before initiation of the experiment because of complications from mastitis. Treatments consisted of 11-h i.v. infusions of saline (control) or a 20% (wt/vol) triacylglycerol (TG) emulsion derived from tallow (tallow) to elevate plasma NEFA. Each period consisted of two 11-h infusions (INF1 and INF2), separated by 1 d in which cows were not infused. Intravenous glucose tolerance tests (IVGTT) and insulin challenges (IC) were performed 8h after initiation of INF1 and INF2, respectively. The infusion of treatments continued during the 3h of sampling for IVGTT and IC. Cows were fed every 4h at a rate to meet energy requirements for 5 d prior to each period, and every 2h during the first 8h of infusions. Infusion of tallow induced hyperlipidemia by increasing plasma NEFA (295±9 vs. 79±7μEq/L), serum TG (41.0±6 vs. 11.4±4.4mg/dL), and glycerol (0.81±0.09 vs. 0.23±0.1mg/dL) concentrations during INF1. During INF2, tallow treatment increased plasma NEFA (347 vs. 139±18μEq/L), serum TG (20.8±4.6 vs. 13.1±2.3mg/dL), and glycerol (0.88±0.04 vs. 0.31±0.02mg/dL) concentrations. Induction of hyperlipidemia impaired glucose clearance during IVGTT, despite the greater endogenous insulin response to the glucose infusion, leading to a lower insulin sensitivity index [0.29 vs. 1.88±0.31×10−4 min−1/(μIU/mL)]. Accordingly, hyperlipidemia impaired glucose clearance during IC (1.58 vs. 2.72%/min), reflecting lower responsiveness to insulin. These data show that induction of hyperlipidemia causes insulin resistance in Holstein cows by impairing both sensitivity and maximum responsiveness to insulin. The induction of insulin resistance by TG, NEFA, or both may increase the availability of glucogenic nutrients to the periparturient dairy cow. Yet excessive elevation of NEFA may potentially lead adipocytes to become more insulin resistant, further increasing plasma NEFA concentration and the risk of metabolic disorders.

Technical Note: Effect of Sampling Protocol on Plasma Nonesterified Fatty Acid Concentration in Dairy Cows

The objective of these experiments was to determine effects of sampling protocol on plasma nonesterified fatty acid (NEFA) concentration. In experiment 1, 8 nonlactating, nongestating dairy cows were blood sampled from a jugular vein catheter (basal, 0min), moved to an exercise lot for 15min, returned to stanchions, and sampled immediately and at 5, 15, 30, 60, and 120min following return to their stalls. Following 15min of exercise, plasma NEFA concentration increased, peaking at 5min (225μEq/L) and returning to basal (84μEq/L) by 30min (110μEq/L). Cows were then moved to box stalls overnight, and 24h after the basal sample, they were locked up and sampled again. Housing cows in a box stall overnight and locking them in headlocks increased plasma NEFA concentration (184μEq/L). In a second experiment at a large free-stall commercial dairy, 11 late-gestation nonlactating dairy cows were locked in headlocks at feeding, blood was sampled from the coccygeal artery or vein (0min), and cows were then released and allowed to finish eating and return to their stalls. Cows were then herded to headlocks and sampled immediately at 120min after initial sampling and at 135, 150, and 180min. Plasma NEFA concentration was highest at initial lockup (0min; 284μEq/L), lowest at 180min (178μEq/L), and intermediate at time points in between. A second group of 10 late-gestation nonlactating dairy cows were locked in headlocks at feeding, and blood was sampled immediately and at 5, 15, 30, and 60min. Plasma NEFA concentration was highest 15min after being placed in headlocks and lowest 60min after lockup (221 and 113μEq/L, respectively). At each time point in experiments 1 and 2, a behavior score was given (1 to 10; 1 = calm; 10 = extremely excited). In both experiments, there was a significant correlation between the plasma NEFA concentration and behavior score. In conclusion, plasma NEFA concentration was affected by sampling protocol.

Phlorizin Induces Lipolysis and Alters Meal Patterns in Both Early-and Late-Lactation Dairy Cows

Phlorizin is known to increase whole-body glucose demand, but it has also stimulated lipolysis in past studies in ruminants. Increased lipolysis complicates studies of dry matter intake (DMI) regulation by hepatic oxidation by providing the liver with additional oxidative substrate. Therefore, to assess whether increased glucose demand selectively increases DMI for cows in negative energy balance, phlorizin was administered to early- and late-lactation cows. Six Holstein cows in early lactation (19±6 DIM, 50.0±1.8kg/d of milk, mean±SD) and 6 Holstein cows in late lactation (228±18 DIM, 30.6±1.9kg/d of milk) were randomly assigned to treatment sequence in a crossover design. Periods were 14 d with 7-d adaptation periods and 7 d of treatment. Phlorizin (4g/d) and propylene glycol (carrier and control) were administered subcutaneously every 6h throughout the treatment periods. Feeding behavior and DMI data were collected for the final 4 d of each treatment period; blood samples and total urine output were collected on d 4 of each treatment period. Phlorizin caused urinary loss of glucose at 333g/d in early-lactation cows and 532g/d in late-lactation cows. Phlorizin increased plasma nonesterified fatty acid concentrations similarly in early- and late-lactation cows, but did not significantly alter plasma insulin concentrations. Treatment with phlorizin tended to decrease meal size, but also decreased intermeal interval, resulting in no effect on DMI. The effects of phlorizin on lipolysis, feeding behavior, and DMI are not dependent on relative energy balance.