Non-esterified Fatty Acid Response Research Articles

PMON244 Alterations in Nonesterified Free Fatty Acids (NEFA) Trafficking Rather Than Hyperandrogenism Contribute to Variations in Insulin Sensitivity Among Obese Women with Polycystic Ovary Syndrome

Abstract Objectives To compare the metabolic and hormonal features of metabolically unhealthy obese PCOS women (MU-PCOS) vs. metabolically healthy obese PCOS women (MH-PCOS) and determine whether alterations in non-esterified free fatty acids (NEFA) response to hyperinsulinemia contribute to variations in insulin sensitivity between MU-PCOS and MH-PCOS. Design Prospective cross-sectional study. Setting Tertiary-care academic center. Methods Insulin resistance (IR) in the basal state was assessed by HOMA-IR in 125 consecutive obese oligo-ovulatory PCOS women (BMI≥ 30 kg/m2; NIH 1990 or Rotterdam 2003 Phenotype A/B) undergoing an oral glucose tolerance test (oGTT); and IR in the dynamic state assessed by the modified frequently sampled intravenous glucose tolerance test (mFSIVGTT) in a subgroup of 16 non-diabetic obese PCOS women. Main Outcome Measures: Primary outcome measures were: 1) IR in AT was assessed in the basal state by the Adipose Insulin Resistance Index [Adipo-IR]) and dynamically by the mFSIVGTT-derived indices of insulin-mediated NEFA suppression [NEFAnadir, TIMEnadir, and %NEFAsupp]); 2) peak lipolysis rate [SNEFA]; and 3) peak NEFA uptake rate [KNEFA]. Secondary outcome measures were levels of insulin and glucose during oGTT, HOMA-β%, and degree of hyperandrogenism (i.e. mF-G score, free testosterone [T], total T, DHEAS). Results Eighty-five obese PCOS participants undergoing oGTT had a HOMA-IR index of ≥2.5 (MU-PCOS) and 40 had a HOMA-IR of <2.5 (MH-PCOS). MU-PCOS had higher mean HOMA-β%, levels of fasting, 1-hr and 2-hr. glucose and insulin, and peak insulin during the oGTT, and higher prevalence of prediabetes and elevated 1-hr. glucose and a trend towards higher prevalence of type 2 diabetes mellitus (T2DM), than MH-PCOS. Alternatively, measures of HA were similar between the groups. Assessing IR dynamically, subjects were subdivided into MU-PCOS by an insulin sensitivity index [Si] ≤1.7 [mU/L]−1·min−1 (n=8) or MH-PCOS by an Si ≥5.6 [mU/L]−1·min−1 (n=8), matched for age and BMI. MU-PCOS had lower Si and KNEFA, smaller %NEFAsupp, longer TIMEnidar, and higher NEFAnidar and Adipo-IR than MH-PCOS, although fasting plasma NEFA levels and SNEFA were similar. In bivalent analysis, Si correlated strongly and negatively with NEFAnadir, weakly and negatively with TIMEnadir, and positively and strongly with KNEFA and NEFAsupp, in MUO-PCOS only. Conclusion Independent of age and BMI, metabolically unhealthy obese PCOS women, compared with metabolically healthy obese PCOS women, had lower rates of NEFA uptake and altered insulin-mediated plasma NEFA suppression in response to mFSIVGTT-determined endogenous insulin secretion, rather than disparities in HA and rate of lipolysis, mechanism that may contribute to variations in insulin sensitivity among obese PCOS women. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Postprandial glycemic responses to a high-protein dairy snack and energy-enriched berry snacks in older adults.

We have previously shown that regular consumption of high-protein dairy- and energy-enriched berry-based products consumed as snacks improved nutritional status, functional status, and quality of life among vulnerable older people. These products contain protein, sugar and other components which may have acute effects on glycemic control. The aim of the present study was to determine the effects of these snack products on postprandial glycemic responses in older adults. In a randomized, single-blinded crossover design, 25 subjects aged 71.4±4.6 years consumed either a high-protein dairy drink or one of the two energy-enriched berry purées, as well as corresponding reference products, as mid-morning snacks, each containing 25g of available carbohydrates. Baseline and postprandial blood samples for measurements of glucose, insulin, and non-esterified fatty acids (NEFA) were collected at regular intervals up to 3h. In comparison with a protein-free reference, consumption of the dairy product resulted in a remarkably high insulin response, a fall in glucose concentration and suppression of late postprandial NEFA rebound. In comparison with a low-berry reference, both berry products led to significantly lower glucose, insulin and NEFA responses. The high-protein dairy product which induces short-term hyperinsulinemia, accompanied with reduced glycemia, may help to improve muscle protein and energy metabolism. The energy-enriched berry products maintain balanced postprandial glycemia. These are beneficial effects in older adults who may suffer from impaired muscle performance or glycemic control. NCT04175353.

162 Differential Late Gestation Metabolizable Energy Intake Alters Adipose-Specific Insulin Responsiveness in Antepartum Beef Cattle

Abstract We evaluated how metabolizable energy (ME) intake influences antepartum insulin sensitivity in beef cattle. Angus-Simmental cattle (primiparous: n = 17; multiparous: n = 21) were randomly assigned to treatments supplying 80 (LowME, n = 19) or 120% (HighME, n = 19) of predicted ME requirements for 53 d prepartum. At 7 d antepartum, intravenous glucose tolerance tests (IVGTT) were imposed by infusing 1.36 g glucose/kg BW0.75 and sampling from -3 to 90 min to measure plasma glucose and insulin and serum non-esterified fatty acids (NEFA) concentrations. Data were analyzed using PROC GLIMMIX with the fixed effects of treatment, parity, time (repeated), and treatment×time and considering cow(treatment) as random. Baseline glucose concentration increased (P = 0.03) and baseline insulin concentration tended to increase (P = 0.06) for HighME relative to LowME cattle irrespective (P ≥ 0.33) of parity, whereas baseline NEFA concentration was elevated (P < 0.01) for LowME vs. HighME. Plasma glucose-insulin concentrations during the IVGTT did not differ (P ≥ 0.25) by treatment, treatment×time, or parity. However, serum NEFA was elevated (P < 0.01) throughout the IVGTT for LowME vs. HighME cattle and tended to be affected (P = 0.09) by the treatment×time interaction as NEFA concentrations became similar at 90 min due to HighME cattle having a shallower (treatment×time: P < 0.01) decremental NEFA response than LowME cattle. Low ME cattle cleared NEFA more quickly (75% increase; P < 0.01) than HighME. The glucose positive incremental area-under-the-curve throughout the IVGTT tended to be lesser (P = 0.06) for heifers than cows but heifers had a blunted (P ≤ 0.05) glucose-stimulated reduction in NEFA and NEFA clearance rate compared to cows. These data demonstrate that ME provision may alter adipose insulin responsiveness in antepartum beef cattle and that heifers respond to excess glucose and NEFA differently than cows.

Exploration of robustness indicators using adaptive responses to short-term feed restriction in suckling primiparous beef cows

Animal robustness is a complex trait of importance for livestock production systems and genetic selection. Phenotyping is essential for evaluation of the adaptation of different genotypes to changing environments. This study tested an experimental framework to induce marked deviations in the adaptive responses of suckling beef cows and to identify relevant indicators of responses to characterise individual differences in the robustness of cows. The production and metabolic responses of primiparous suckling Charolais cows to two periods of feed restriction (FR, 50% of their net energy requirements) of different durations were monitored. After calving, 13 cows (aged 39 ± 2 months, BW of 680 ± 42 kg at calving) had ad libitum access to a diet composed of hay and supplemented with concentrate to meet their energy and protein requirements. Starting at 54 ± 6 days postcalving, the cows underwent two periods of FR: 4 days of FR (FR4), which was followed by 17 days of ad libitum intake to study the recovery from FR4, and 10 days of FR (FR10), which was followed by 18 days of ad libitum intake to study the recovery from FR10. The milk yield (MY), BW, body condition score and plasma non-esterified fatty acid (NEFA), β-hydroxybutyrate, glucose and urea concentrations were measured before, during and after each FR. Among all measured variables, the MY and NEFA concentrations showed the most significant changes in response to FR. A functional data analysis approach was applied to the MY and NEFA data to model the adaptive responses and extract quantifiable indicators of deviation and recovery. Linear correlations (P < 0.03–0.07) between FR4 and FR10 were found for some indicators describing MY and NEFA levels before and after FR. The overall repeatability of MY and NEFA responses between both FR accounted for 46% based on quartile analysis performed on average responses. Moreover, the variance in both the MY and NEFA variables did not differ significantly between FR4 and FR10, despite a trend for higher variances in FR10. Altogether, (1) the calculated variables derived from the functional data analysis of the time patterns of the MY and NEFA accounted for the differences in the cow responses to FR, and (2) the animal responses appeared to show concordance between FR4 and FR10. In conclusion, short-term FR is a relevant framework for studying productive and metabolic adaptive responses in suckling cows and allows the identification of potential robustness indicators.

Metabolic Responses to Epinephrine by Periparturient Dairy Cows Fed Prepartum Diets Differing in Predicted Metabolizable Protein Supply

We determined the effects of prepartum dietary metabolizable protein (MP) supply on lipolytic and glucose response to epinephrine stimulus during the periparturient period. Twelve non-lactating cows in second or greater pregnancies were assigned to a low MP (LMP) diet formulated to provide ~900 g/day MP or a high MP (HMP) diet to provide ~1,100 g/day MP. Cows received prepartum diets from 28 days before expected parturition to day of parturition and then received a common lactation diet until 35 days postpartum. Blood was sampled weekly to day −7, daily to day 7, and weekly through day 35 relative to parturition for analysis of glucose, non-esterified fatty acids (NEFAs), 3-hydroxybutyrate (BHB), urea N, and total protein concentrations. Epinephrine challenges were conducted on day −10 before expected parturition and days 7 and 14 postpartum. Epinephrine [1.4 μg/kg body weight (BW)] was administered via jugular vein and blood was collected from the opposite jugular vein from −45 min to +120 min relative to epinephrine administration. Plasma was analyzed for glucose and NEFA concentrations. Prepartum dry matter intake (DMI) (14.7 and 12.6 ± 1.5 kg/day for LMP and HMP, respectively), postpartum DMI (21.4 and 19.4 ± 1.3 kg/day for LMP and HMP, respectively), and 4% fat-corrected milk (FCM) (37.0 and 36.8 ± 1.5 kg/day for LMP and HMP, respectively) did not differ significantly between diets. Cows fed HMP had higher plasma concentrations of urea N prepartum than cows fed LMP (14.0 vs. 7.9 ± 0.8 mg/dl). Cows fed HMP tended to have greater prepartum concentrations of total protein in plasma than those fed LMP (7.5 vs. 7.0 ± 0.2 g/dl). The area under the curve (AUC) for NEFA response to epinephrine did not differ between diets, but differed by day relative to parturition [8,284, 29,018, and 18,219 ± 2,302 min × (μeq/l) for days −10, 7, and 14, respectively]. Maximal response of NEFA concentration to epinephrine was greater for HMP than for LMP (744 vs. 438 ± 72 μeq/l). The glucose AUC did not differ between diets or among days. Lipolytic response, but not glucose response, to epinephrine was enhanced during the early postpartum period relative to the late dry period. However, those responses were not affected by prepartum MP supply.

Heat stress affects some physiological and productive variables and alters metabolism in dairy ewes

Heat stress (HS) has a significant economic impact on the global dairy industry. However, the mechanisms by which HS negatively affects metabolism and milk synthesis in dairy ewes are not well defined. This study evaluated the production and metabolic variables in dairy ewes under controlled HS conditions. Eight Lacaune ewes (75.5 ± 3.2 kg of body weight; 165 ± 4 d of lactation; 2.31 ± 0.04 kg of milk per day) were submitted to thermoneutral (TN) or HS conditions in a crossover design (2 periods, 21 d each, 6-d transition). Conditions (day-night, 12-12 h; relative humidity; temperature-humidity index, THI) were: TN (15-20°C; 50 ± 5%; THI = 59-65) and HS (28-35°C; 45 ± 5%; THI = 75-83). Ewes were fed ad libitum and milked twice daily. Rectal temperature, respiratory rate, feed intake, water consumption, and milk yield were recorded daily. Milk and blood samples were collected weekly. Additionally, TN and HS ewes were exposed to glucose tolerance test, insulin tolerance test, and epinephrine challenge. Heat stress reduced feed intake (-11%), and increased rectal temperature (+0.77°C), respiratory rate (+90 breaths/min), and water consumption (+28%). Despite the reduced feed intake, HS ewes produced similar milk to TN ewes, but their milk contained lower fat (-1.7 points) and protein (-0.86 points). Further, HS milk tended to contain more somatic cells (+0.23 log points). Blood creatinine was greater in HS compared with TN, but no differences in blood glucose, nonesterified fatty acids, or urea were detected. When glucose was infused, TN and HS had similar insulin response, but higher glucose response (+85%) was detected in HS ewes. Epinephrine infusion resulted in lower nonesterified fatty acids response (-215%) in HS than TN ewes. Overall, HS decreased feed intake, but milk production was not affected. Heat stress caused metabolic adaptations that included increased body muscle degradation and reduced adipose tissue mobilization. These adaptations allowed ewes to spare glucose and to avoid reductions in milk yield.

Prior ingestion of a ketone monoester supplement reduces postprandial glycemic responses in young healthy-weight individuals.

The main objective of this study was to determine whether acute ingestion of a ketone monoester (KME) supplement impacted mixed-meal tolerance test (MMTT) glucose area under the curve (AUC). Nineteen healthy young volunteers (10 males/9 females; age, 24.7 ± 4.9 years; body mass index, 22.7 ± 2.4 kg/m2) participated in a double-blind, placebo-controlled crossover study. Following overnight fasting (≥10 h), participants consumed 0.45 mL/kg of a KME supplement or taste-matched placebo followed by an MMTT 15 min later. Blood samples were collected every 15-30 min over 2.5 h. KME supplementation acutely raised β-hydroxybutyrate AUC (590%, P < 0.0001, d = 2.4) and resulted in decreases in blood glucose AUC (-9.4%, P = 0.03, d = 0.56) and nonesterified fatty acid (NEFA) AUC (-27.3%, P = 0.023, d = 0.68) compared with placebo. No differences were found for plasma insulin AUC (P = 0.70) or gastric emptying estimated by co-ingested acetaminophen AUC (P = 0.96) between ketone and placebo. Overall, results indicate that KME supplementation attenuates postprandial glycemic and NEFA responses when taken 15 min prior to a mixed meal in young healthy individuals. Future studies are warranted to investigate whether KME supplementation may benefit individuals with impaired glycemic control. Novelty: Acute ketone monoester supplementation 15 min prior to a mixed meal decreased postprandial glucose and NEFA levels without significantly impacting postprandial insulin or estimates of gastric emptying. Glucose- and NEFA-lowering effects of ketone monoester supplementation are apparently not mediated by changes in insulin release or gastric emptying.

Administration of low and high doses of heparin causes changes in plasma non-esterified fatty acid concentration in merino and terminal sired lambs

Context The anticoagulant properties of heparin have led to the routine use of heparinised saline flushes to prevent thrombus formation and to prolong the patency of indwelling cannulas. However, limited work exists on very low-dose heparin used to retain cannula patency for the purpose of repeated blood sampling for metabolic studies. Of particular interest is whether low-dose heparin will cause increases in plasma non-esterified fatty acid (NEFA) concentration. This is most relevant in metabolic studies involving repeated sampling, as this may erroneously elevate apparent plasma NEFA concentrations. Aims The objective of the present study was to evaluate the impacts of low- and high-dose heparin administration on plasma NEFA response in lambs. Methods In total, 14 merino (3 female, 4 wether) and terminal (4 female, 3 wether) sired lambs were selected from the Katanning, Western Australia, site of the Meat and Livestock Australia genetic resource flock All lambs were subjected to the following three treatments: low heparin (0.25 mL, 250 IU), high heparin (1 mL, 1000 IU) or control (1 mL of 0.9% NaCl saline), with each challenge being randomly allocated over 1.5 days. Blood samples were collected at the following time points: –30, –15, –10, –5, 0, 2.5, 5, 10, 15, 20, 30, 45, 60, 90, 120, 125 and 130 min relative to the administration of the challenge (Time 0) and tested for NEFA concentration. A derived exponential function was fitted to the raw data, enabling the plasma NEFA concentration response curve to be modelled at different time pointspre- and post-challenge, using the area under curve (AUC), maximum concentration and return to basal concentration, to quantify the NEFA response. Results Heparin-challenge dose had a significant (P &amp;lt; 0.01) impact on peak NEFA response at 10 min following challenge administration (NEFA AUC10), with the values after high-heparin challenge (1.03 ± 0.086 mmol/L per 10 min) being ~25% higher (P &amp;lt; 0.05) than those after the low-heparin challenge (0.78 ± 0.086 mmol/L per 10 min). The NEFA AUC10 values with low-dose heparin and high-dose heparin were 0.76 units and 1.02 units higher than those with the saline treatment (0.02 ± 0.086 mmol/L per 10 min; P &amp;lt; 0.01). Heparin-challenge dose also had a significant impact on the maximum NEFA concentration (P &amp;lt; 0.05). The high-heparin challenge (0.32 ± 0.057 mmol/L) had 20% higher maximum NEFA concentrations than the low-heparin challenge (0.26 ± 0.057 mmol/L). Both high and low heparin-challenge groups had maximum NEFA concentrations that were 72% and 36% higher respectively, than for the saline-challenge (0.19 ± 0.057 mmol/L) group. NEFA returned to basal concentrations by 60 min for both challenges, although the high heparin-challenge group demonstrated a slower rate of return (P &amp;lt; 0.05). Conclusions High and low doses of heparin caused an increase in plasma NEFA response as measured by AUC10 and maximum NEFA concentration, but returned to basal concentrations within 1 h. Implications Results indicated that heparin as an anticoagulant should be avoided where frequent blood samples are required within intervals of less than 1 h. However, for repeated sampling at intervals greater than 1 h, judicious flushing with heparinised saline is unlikely to have an impact on plasma NEFA concentrations.

The Effect Of Fasted Cycling Exercise At Different Times Of The Day In Overweight Individuals

Intermittent fasting has become an increasingly popular intervention for metabolic health. Combining intermittent fasting with exercise may lead to benefits for weight management. PURPOSE: To investigate the effect of fasted exercise at different times of the day on metabolic responses and appetite regulation in overweight males. METHODS: Twelve healthy males (Mean ± SD; age 26 ± 4 y; body fat 23 ± 2%) completed four, 60 min cycle ergometry trials at 60% V̇O2peak in a randomised order; in the morning fasted (AMFAST), evening fasted (PMFAST) and after consuming a breakfast meal in the morning (AMFED) and evening (PMFED). Circulating levels of ghrelin, glucagon-like peptide-1 (GLP-1), pancreatic polypeptide (PP), peptide tyrosine tyrosine (PYY), insulin, triglycerides, non-esterified fatty acid (NEFA), glucose, and cholesterol were measured at baseline, post-breakfast, pre-exercise, post-exercise, pre-soup ingestion, then every 30 min post soup-ingestion for 2 h. Appetite was assessed at 15 min intervals throughout. Substrate utilisation was measured every 30 min and continuously throughout exercise. RESULTS: Area under the curve (AUC) values for NEFA were greater in PMFAST compared to all trials (186.5 ± 46.3 vs. AMFAST 120.5 ± 42.6, AMFED 80.8 ± 23.8, PMFED 91.1 ± 36.4 mmol/L 4.75 h; P<0.05). AMFAST NEFA was also greater than AMFED (P=0.029). AUC values were greater for GLP-1 in AMFED trial compared to PMFAST (8660.2 ± 12232.5 vs 5967.0 ± 12027.5 mmol/L 4.75 h, P<0.05), and PP values for PMFED greater compared to all trials (155411.9 ± 86064.1 vs. AMFAST 90165.4 ± 90145.6, AMFED 107162.5 ± 72846.2, PMFAST 105364.0 ± 81320.2 mmol/L 4.75 h, P<0.05). A time of day effect was seen for cholesterol with PM greater than AM (P<0.05). Fat oxidation was greater during AMFAST and PMFAST exercise compared to FED trials (P<0.05). Ratings of appetite did not change between trials once all participants consumed the lunch meal post exercise (P>0.05). No differences were seen in AUC between trials for ghrelin, PYY, glucose or insulin. CONCLUSION: Fasted exercise elicited greater NEFA responses, and some appetite hormones appear to respond differently to varying exercise conditions and time of day in overweight males. Regardless of the time of day, fasted exercise favours fat metabolism and may induce a short-term negative energy balance.

Nutritional Physiology and Biochemistry of Dairy Cattle under the Influence of Heat Stress: Consequences and Opportunities.

Simple SummaryModern dairy cows have elevated internal heat loads caused by high milk production, and the effects of accumulating incremental heat are exacerbated when temperature and humidity increases in the surroundings. To shed this additional heat, cows initiate a variety of adaptive mechanisms including increased respiration rate, panting, sweating, reduced milk yield, vasodilatation, and decreased reproductive performance. Hormonal changes based on reciprocal alterations to the energetic metabolism are particularly accountable for reduced efficiency of the dairy production under the heat stress. As animals experience negative energy balance; glucose, which is also a precursor of milk lactose, becomes the preferential energy fuel. In the absence of proper mitigations, heat stress possesses potential risk of economic losses to dairy sector. Besides physical measures for the timely prediction of the actual heat stress coupled with its proper amelioration, nutritional mitigation strategies should target modulating energetic metabolism and rumen environment.Higher milk yield and prolificacy of the modern dairy cattle requires high metabolism activities to support them. It causes high heat production by the body, which coupled with increasing environmental temperatures results in heat stress (HS). Production, health, and welfare of modern cattle are severely jeopardized due to their low adaptability to hot conditions. Animal activates a variety of physiological, endocrine, and behavioral mechanisms to cope with HS. Traditionally, decreased feed intake is considered as the major factor towards negative energy balance (NEBAL) leading to a decline in milk production. However, reciprocal changes related to insulin; glucose metabolism; failure of adipose mobilization; and skeletal muscle metabolism have appeared to be the major culprits behind HS specific NEBAL. There exists high insulin activity and glucose become preferential energy fuel. Physiological biochemistry of the heat stressed cows is characterized by low-fat reserves derived NEFA (non-esterified fatty acids) response, despite high energy demands. Besides these, physiological and gut-associated changes and poor feeding practices can further compromise the welfare and production of the heat-stressed cows. Better understanding of HS specific nutritional physiology and metabolic biochemistry of the dairy cattle will primarily help to devise practical interventions in this context. Proper assessment of the HS in cattle and thereby applying relevant cooling measures at dairy seems to be the basic mitigation approach. Score of the nutritional strategies be applied in the eve of HS should target supporting physiological responses of abatement and fulfilling the deficiencies possessed, such as water and minerals. Second line of abatement constitutes proper feeding, which could augment metabolic activities and synergizes energy support. The third line of supplemental supports should be directed towards modulating the metabolic (propionates, thiazolidinediones, dietary buffers, probiotics, and fermentates) and antioxidant responses (vitamins). Comprehensive understanding of the energetic metabolism dynamics under the impact of incremental heat load and complete outlook of pros and cons of the dietary ameliorating substances together with the discovery of the newer relevant supplementations constitutes the future avenues in this context.

Ovine prenatal growth-restriction and sex influence fetal adipose tissue phenotype and impact postnatal lipid metabolism and adiposity in vivo from birth until adulthood.

Adipose tissue development begins in utero and is a key target of developmental programming. Here the influence of nutritionally-mediated prenatal growth-restriction on perirenal adipose tissue (PAT) gene expression and adipocyte phenotype in late fetal life was investigated in both sexes in an ovine model. Likewise circulating leptin concentrations and non-esterified fatty acid (NEFA) and glycerol responses to glucose challenge were determined in relation to offspring adiposity at key stages from birth to mid-adult life. In both studies' singleton-bearing adolescent sheep were fed control or high nutrient intakes to induce normal or growth-restricted pregnancies, respectively. Fetal growth-restriction at day 130 of gestation (32% lighter) was characterised by greater body-weight-specific PAT mass and higher PAT expression of peroxisome proliferator-activated receptor gamma (PPARɤ), glycerol-3-phosphate dehydrogenase, hormone sensitive lipase (HSL), insulin-like growth factor 1 receptor, and uncoupling protein 1. Independent of prenatal growth, females had a greater body-weight-specific PAT mass, more multilocular adipocytes, higher leptin and lower insulin-like growth factor 1 mRNA than males. Growth-restricted offspring of both sexes (42% lighter at birth) were characterised by higher plasma NEFA concentrations across the life-course (post-fasting and after glucose challenge at 7, 32, 60, 85 and 106 weeks of age) consistent with reduced adipose tissue insulin sensitivity. Circulating plasma leptin correlated with body fat percentage (females>males) and restricted compared with normal females had more body fat and increased abundance of PPARɤ, HSL, leptin and adiponectin mRNA in PAT at necropsy (109 weeks). Therefore, prenatal nutrient supply and sex both influence adipose tissue development with consequences for lipid metabolism and body composition persisting throughout the life-course.

Responses to metabolic challenges in dairy cows with high or low milk yield during an extended lactation

Responses of dairy cows with high or low milk yield (MY) beyond 450 d in milk (DIM) to 3 metabolic challenges were investigated. Twelve multiparous Holstein-Friesian cows that calved in late winter in a pasture-based system were managed for a 670-d lactation by delaying re-breeding. Cows were selected for either high MY (18.9 ± 1.69 L/cow per d; n = 6) or low MY (12.3 ± 3.85 L/cow per d; n = 6) at 450 DIM. Cows were housed indoors for 2 periods of 12 d at approximately 460 and 580 DIM. Each cow was fed freshly cut pasture (460 DIM) or pasture silage (580 DIM) plus 6.0 kg of DM barley grain daily (approximately 200 MJ of total metabolizable energy/cow per day). At all other times, cows were managed as a single herd and grazed pasture supplemented with cereal grain to an estimated intake of 180 MJ of metabolizable energy/cow per d. Cows were fitted with a jugular catheter during the final week of each experimental period. Over a period of 3 d, each cow underwent an intravenous glucose tolerance test (0.3 g/kg of body weight), an insulin tolerance test (0.12 IU of insulin/kg of body weight), and a 2-dose epinephrine challenge (0.1 and 1.6 µg/kg of body weight). Cows selected for high MY had greater milk and milk solids yields between 450 and 580 DIM than low MY cows (17.3 vs. 10.8 ± 1.49 kg of milk/d and 2.4 vs. 1.5 ± 0.23 kg of milk solids/d). The results indicated that whole body and peripheral tissue responsiveness to insulin may vary between cows of high and low MY. Following the glucose tolerance test, high MY cows had a lower plasma insulin response with a greater glucose area under the curve than low MY cows. Further, high MY cows had slower plasma glucose clearance compared with low MY cows during an insulin tolerance test. The plasma nonesterified fatty acid (NEFA) responses to the IVGTT and the ITT were similar between cows of high and low MY, but the clearance of NEFA from the plasma following both the IVGTT and ITT were slower at 580 compared with 460 DIM. The sensitivity to epinephrine was greater in high MY cows compared with low MY cows as the glucose and NEFA area under the curve and the percentage change in NEFA were greater in high MY after the low dose epinephrine challenge. However, the lipolytic but not the glucose appearance in response to epinephrine was greater in high MY cows than low MY cows. Following the high dose of epinephrine, the glucose response was lower, but the NEFA response was greater in high MY compared with low MY cows. Cows able to sustain greater MY to 580 DIM had a greater propensity for lipid mobilization, possibly enhancing nutrient partitioning to the mammary gland during the late stages of an extended lactation.

Effects of continuous and increasing lipopolysaccharide infusion on basal and stimulated metabolism in lactating Holstein cows

Experimental objectives of this study were to characterize the systemic and intracellular metabolic response to continuous lipopolysaccharide (LPS) infusion in mid-lactation Holstein cows (169 ± 20 d in milk; 681 ± 16 kg of body weight). Following 3 d of acclimation, cows were enrolled in 2 experimental periods (P). During P1 (3 d), cows were fed ad libitum and baseline data were collected. In P2 (8 d), cows were assigned to 1 of 2 treatments: (1) saline-infused and pair-fed (CON-PF; i.v. sterile saline at 40 mL/h; n = 5) or (2) LPS-infused and fed ad libitum (LPS-AL; Escherichia coli O55:B5 at 0.017, 0.020, 0.026, 0.036, 0.055, 0.088, 0.148, and 0.148 µg/kg of body weight per hour for d 1 through 8, respectively; n = 6). During P2, CON-PF cows were pair-fed to LPS-AL cows to eliminate confounding effects of dissimilar nutrient intake. Blood samples were collected on d 1 and 2 of P1 and d 1, 3, 5, and 7 of P2. Following the P2 d 7 a.m. milking, adipose tissue, skeletal muscle, and liver biopsies were collected for reverse transcription quantitative PCR and Western blot analysis. To assess whole-body nutrient trafficking, an i.v. glucose tolerance test (GTT) was performed following the a.m. milking on P2 d 8; 4 h after the GTT, cows received an epinephrine challenge. During P2, there were no treatment differences in circulating glucose. Relative to P1, CON-PF cows had or tended to have decreased plasma β-hydroxybutyrate and insulin (29 and 47%, respectively) during P2, whereas neither variable changed in LPS-AL cows, leading to an overall increase in β-hydroxybutyrate and insulin (41 and 140%, respectively) relative to CON-PF cows. Circulating nonesterified fatty acids were increased from d 1 to 3 and subsequently decreased from d 3 to 7 in cows from both treatments. Blood urea nitrogen gradually decreased in CON-PF cows and increased in LPS-AL cows from d 1 to 5 of P2, resulting in an overall 25% increase in LPS-AL versus CON-PF cows. In response to the GTT, the glucose and insulin area under the curve were increased 33 and 56%, respectively, in LPS-AL compared with CON-PF cows; changes reflective of whole-body insulin resistance. However, protein abundance of insulin signaling markers within muscle, liver, and adipose tissue were similar between treatments. There were no observable treatment differences in the glucose or nonesterified fatty acids response to the epinephrine challenge. No treatment differences were observed in hepatic mRNA abundance of key gluconeogenic or lipid export enzymes. In conclusion, chronic LPS exposure altered multiple parameters of basal and stimulated metabolism, but did not appear to affect the molecular machinery evaluated herein.

Feed deprivation in Merino and Terminal sired lambs: (1) the metabolic response under resting conditions

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 (P < 0.05) by 25% from 4.04 ± 0.032 mmol/l to 3.04 ± 0.032 mmol/l. From 6 h NEFA concentration increased (P < 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 (P < 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 (P < 0.05). In Merino sired lambs, increasing PWT was also associated with an increase in glucose and decline in NEFA and BHOB concentration (P < 0.05). In Terminal sired lambs, increasing PFAT was associated with an increase in glucose and decline in NEFA concentration (P < 0.05). Contrary to the hypothesis, Merino sired lambs showed the greatest metabolic response to fasting especially in regards to fat metabolism.

Feed deprivation in Merino and Terminal sired lambs: (2) the metabolic response under pre-slaughter conditions and impact on meat quality and carcass yield

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 (P>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 acute effects of dietary carbohydrate reduction on postprandial responses of non-esterified fatty acids and triglycerides: a randomized trial

BackgroundPostprandial non-esterified fatty acid (NEFA) and triglyceride (TG) responses are increased in subjects with type 2 diabetes mellitus (T2DM) and may impair insulin action and increase risk of cardiovascular disease and death. Dietary carbohydrate reduction has been suggested as non-pharmacological therapy for T2DM, but the acute effects on NEFA and TG during subsequent meals remain to be investigated.MethodsPostprandial NEFA and TG responses were assessed in subjects with T2DM by comparing a carbohydrate-reduced high-protein (CRHP) diet with a conventional diabetes (CD) diet in an open-label, randomized, cross-over study. Each diet was consumed on two consecutive days, separated by a wash-out period. The iso-caloric CRHP/CD diets contained 31/54 E% from carbohydrate, 29/16 E% energy from protein and 40/30 E% from fat, respectively. Sixteen subjects with well-controlled T2DM (median HbA1c 47 mmol/mol, (37–67 mmol/mol) and BMI 30 ± 4.4 kg/m2) participated in the study. NEFA and TG were evaluated following breakfast and lunch.ResultsNEFA net area under curve (AUC) was increased by 97 ± 38 μmol/Lx270 min (p = 0.024) after breakfast but reduced by 141 ± 33 μmol/Lx180 min (p < 0.001) after lunch on the CRHP compared with CD diet. Likewise, TG net AUC was increased by 80 ± 28 μmol/Lx270 min (p = 0.012) after breakfast but reduced by 320 ± 60 μmol/Lx180 min (p < 0.001) after lunch on the CRHP compared with CD diet.ConclusionsIn well-controlled T2DM a modest reduction of dietary carbohydrate with a corresponding increase in protein and fat acutely reduced postprandial serum NEFA suppression and increased serum TG responses after a breakfast meal but had the opposite effect after a lunch meal. The mechanism behind this second-meal phenomenon of CRHP diet on important risk factors for aggravating T2DM and cardiovascular disease awaits further investigation.Trial registrationThe study was registered at clinicaltrials.gov ID: NCT02472951. https://clinicaltrials.gov/ct2/show/NCT02472951. Registered June 16, 2015.

Expression of deiodinase 2 (DIO2) and integrin alpha 9 (ITGA9) genes as indicators' of adaptability and their relationship with physio-biochemical parameters in Tharparkar and Karan Fries heifers during different seasons

The aim of the study was to observe the influence of ambient conditions (seasons) on expression pattern of deiodinase 2 (DIO2) and integrin alpha 9 (ITGA9) genes in peripheral blood mononuclear cells (PBMC) and their relationship with physiological and biochemical parameters of Tharparkar and Karan Fries heifers. Healthy heifers of Tharparkar and Karan Fries breed (6 each) were selected and blood samples were collected from these animals at weekly interval. These blood samples were centrifuged to separate the plasma (for biochemical parameters) and buffy coat (for total RNA isolation). The expression of DIO2 gene was down-regulated during hot humid season and up-regulated during winter season in both the breeds. The DIO2 gene showed a positive correlation with thyroid hormones (T3, T4) and negative correlation with physiological responses (RR, PR, RT, ST), cortisol hormone, antioxidant enzymes (SOD, GPx) and non-esterified fatty-acids (NEFA). The expression pattern of ITGA9 gene was opposite to DIO2 gene, i.e. up-regulated during hot humid season and down-regulated during winter season in both the breeds. ITGA9 gene expression showed a positive correlation with cortisol, antioxidant enzymes, NEFA and physiological responses, and negative correlation with thyroid hormones. The fold change in expression of DIO2 and ITGA9 genes was higher in Karan Fries than Tharparkar heifers during hot humid and winter season. The dry matter intake was lower during hot-humid season than autumn and winter season in both the breeds of heifer. The expression pattern of the DIO2 and ITGA9 genes and their relationship with physio-biochemical parameters revealed the essential role in adaptability of cattle as candidate gene. Based on the results of present study, it can be stated that Tharparkar heifers are better adapted to tropical climatic conditions than Karan Fries heifers.

Plasma glucose and nonesterified fatty acids response to epinephrine challenges in dairy cows during a 670-d lactation

This experiment investigated the metabolic response to a 2-dose epinephrine challenge of dairy cows undergoing an extended lactation. Twelve multiparous Holstein-Friesian cows that calved in late winter in a seasonally calving pasture-based dairying system were managed for a 670-d lactation by delaying rebreeding. In each of four 40-d experimental periods commencing at 73, 217, 422, and 520 (±9.1) d in milk (DIM), cows were offered a diet of perennial ryegrass (73 and 422 DIM) or pasture hay and silage (217 and 520 DIM), supplemented with 1 (CON; n = 6) or 6 kg of grain (GRN; n = 6) as a ration. Daily energy intake was approximately 160 and 215 MJ of metabolizable energy/cow for the CON and GRN treatments, respectively. At all other times, cows were managed as a single herd and grazed pasture supplemented with grain to an estimated daily total intake of 180 MJ of metabolizable energy/cow. Cows were fitted with a jugular catheter during the final week of each experimental period. Two doses of epinephrine (0.1 and 1.6 µg/kg of body weight) were infused via the catheter 2 h apart to each cow at approximately 100, 250, 460, and 560 DIM. Blood plasma concentrations of glucose and nonesterified fatty acids (NEFA) were measured before and after infusions. Cows in the GRN treatment had greater milk yield, milk fat and protein yields, and body weight than cows in the CON treatment. The maximum plasma glucose concentration was observed at 100 DIM for both the low and high doses of epinephrine. Thus, sensitivity and responsiveness to exogenous epinephrine were greater during early lactation, coinciding with increased priority of milk synthesis. Both the sensitivity and responsiveness to epinephrine decreased with decreasing milk yield, as measured by the acute appearance of NEFA in the plasma. Increased plasma glucose and NEFA clearance rates before 300 DIM indicated greater uptake of these substrates by the mammary gland for milk synthesis in early and mid lactation. These results support previous findings that major changes occur in terms of adipose tissue metabolism during extended lactations. Overall, sensitivity to epinephrine was not affected by diet, but responsiveness was greater in cows fed the GRN diet. The endocrine regulation of nutrient partitioning throughout traditional and extended lactations is complex, with many interactions between stage of lactation, diet, and milk yield potential.

Pre-Meal Effect of Whey Proteins on Metabolic Parameters in Subjects with and without Type 2 Diabetes: A Randomized, Crossover Trial.

Diabetic dyslipidemia with elevated postprandial triglyceride (TG) responses is characteristic in type 2 diabetes (T2D). Diet and meal timing can modify postprandial lipemia (PPL). The impact of a pre-meal of whey proteins (WP) on lipid metabolism is unidentified. We determined whether a WP pre-meal prior to a fat-rich meal influences TG and apolipoprotein B-48 (ApoB-48) responses differentially in patients with and without T2D. Two matched groups of 12 subjects with and without T2D accomplished an acute, randomized, cross-over trial. A pre-meal of WP (20 g) or water (control) was consumed 15 min before a fat-rich meal (supplemented with 20 g WP in case of water pre-meal). Postprandial responses were examined during a 360-min period. A WP pre-meal significantly increased postprandial concentrations of insulin (P < 0.0001), glucagon (P < 0.0001) and glucose-dependent insulinotropic peptide (GIP) (P < 0.0001) in subjects with and without T2D. We detected no effects of the WP pre-meal on TG, ApoB-48, or non-esterified fatty acids (NEFA) responses to the fat-rich meal in either group. Paracetamol absorption i.e., gastric emptying was delayed by the WP pre-meal (P = 0.039). In conclusion, the WP pre-meal induced similar hormone and lipid responses in subjects with and without T2D. Thus, the WP pre-meal enhanced insulin, glucagon and GIP responses but did not influence lipid or glucose responses. In addition, we demonstrated that a WP pre-meal reduced gastric emptying in both groups.

Effects of TNF receptor blockade on in vitro cell survival and response to negative energy balance in dairy cattle

BackgroundAssociative data and some controlled studies suggest that the inflammatory cytokine tumor necrosis factor (TNF) α can induce fatty liver in dairy cattle. However, research demonstrating that TNFα is a necessary component in the etiology of bovine fatty liver is lacking. The aim of this work was to evaluate whether blocking TNFα signaling with a synthetic cyclic peptide (TNF receptor loop peptide; TRLP) would improve liver metabolic function and reduce triglyceride accumulation during feed restriction.ResultsCapability of TRLP to inhibit TNFα signaling was confirmed on primary bovine hepatocytes treated with recombinant bovine TNFα and 4 doses of TRLP (0, 1, 10, 50 μmol/L) over 24 h. Next, 4 lactating Holstein cows (parity 1.4 ± 0.5, 433 ± 131 d in milk) in an incomplete Latin rectangle design (3 × 2) were subcutaneously administered with different TRLP doses (0, 1.5, 3.0 mg/kg BW) every 4 h for 24 h, followed by an intravenous injection of TNFα (5 μg/kg BW). Before and for 2 h after TNFα injection, TRLP decreased plasma non-esterified fatty acid (NEFA) concentration (P ≤ 0.05), suggesting an altered metabolic response to inflammation. Finally, 10 non-pregnant, non-lactating Holstein cows (3.9 ± 1.1 yr of age) were randomly assigned to treatments: control (carrier: 57% DMSO in PBS) or TRLP (1.75 mg TRLP /kg BW per day). Treatments were administrated every 4 h for 7 d by subcutaneous injection to feed-restricted cows fed 30% of maintenance energy requirements. Daily blood samples were analyzed for glucose, insulin, β-hydroxybutyrate, NEFA, and haptoglobin concentrations, with no treatment effects detected. On d 7, cows completed a glucose tolerance test (GTT) by i.v. administration of a dextrose bolus (300 mg glucose/kg BW). Glucose, insulin, and NEFA responses failed to demonstrate any significant effect of treatment during the GTT. However, plasma and liver analyses were not indicative of dramatic lipolysis or hepatic lipidosis, suggesting that the feed restriction protocol failed to induce the metabolic state of interest. Injection site inflammation, assessed by a scorer blinded to treatment, was enhanced by TRLP compared to control.ConclusionsAlthough the TRLP inhibited bovine TNFα signaling and altered responses to i.v. administration of TNFα, repeated use over 7 d caused apparent local allergic responses and it failed to alter metabolism during a feed restriction-induced negative energy balance. Although responses to feed restriction seemed atypical in this study, side effects of TRLP argue against its future use as a tool for investigating the role of inflammation in metabolic impacts of negative energy balance.