Katarina Nostell

Resting concentrations of cardiac troponin I in fit horses and effect of racing.

OBJECTIVES To determine normal resting values for cardiac troponin I (cTnI) in healthy Standardbred, Thoroughbred and Warmblood horses and investigate if racing has an influence on cTnI concentrations. BACKGROUND Measuring cTnI concentrations in plasma is the gold standard for detecting myocardial injury in humans. Cardiac troponin I is highly conserved between species and has gained interest as a marker for cardiac injury in horses. Increased levels of cTnI have been reported in association with endurance and short-term strenuous exercise on a treadmill in horses. However, the effect of true racing conditions has not yet been reported. ANIMALS, MATERIALS AND METHODS Blood samples for analysis of cTnI concentrations in plasma were collected from 67 Standardbred racehorses, 34 Thoroughbred racehorses and 35 Warmblood dressage horses at rest. Blood samples were also collected prior to and after racing in 22 Standardbred racehorses and 6 Thoroughbred racehorses. RESULTS All horses except one had resting plasma cTnI concentrations <0.022 microg/L. Mild increases in cTnI concentrations were seen in some horses 1-2h after the race (1/17 Standardbreds and 2/6 Thoroughbreds) as well as 10-14 h after the race (4/21 Standardbreds and 1/6 Thoroughbreds). CONCLUSIONS Resting cTnI concentrations in horses are low but mildly elevated cTnI concentrations may be detected in some horses 1-14 h after racing. These findings could be of importance when evaluating horses with suspected cardiac disease that recently have performed hard exercise.

Cardiac troponin I and the occurrence of cardiac arrhythmias in horses with experimentally induced endotoxaemia.

The aim of this study was to determine whether experimentally-induced endotoxaemia induced elevations in plasma cardiac troponin I (cTnI) concentrations in horses and how this might affect the incidence of cardiac arrhythmias. Eight Standardbred horses received an intravenous continuous rate infusion of endotoxin (total dose 500 ng/kg) for 6 h while being monitored using electrocardiography (ECG). Blood samples were collected before the start of the endotoxin infusion, every 60 min during the infusion, then 1, 2, 3, 8, 10 and 24 h post-infusion, and analysed for cTnI concentrations. One horse was excluded from the study owing to a high initial cTnI concentration. Endotoxin infusion induced an increase in cTnI concentrations in all horses, reaching mean peak concentration of 0.135±0.094 μg/L by 1 h post-infusion. The cTnI concentrations then decreased and were no longer significantly different from pre-infusion concentrations at 6, 10 and 24 h post-infusion. The number of ventricular events was generally low during the infusion period, but increased during the first 3 h post-infusion in 6/7 horses. In conclusion, elevated cTnI concentrations could be detected early after an endotoxaemic insult using an ultrasensitive cTnI assay, with peak cTnI concentrations preceding the occurrence of ventricular events on ECG.

Lipopolysaccharide-induced inhibition of transcription of tlr4 in vitro is reversed by dexamethasone and correlates with presence of conserved NFκB binding sites.

Engagement of Toll-like receptor 4 (TLR4) by lipopolysaccharide (LPS) is a master trigger of the deleterious effects of septic shock. Horses and humans are considered the most sensitive species to septic shock, but the mechanisms explaining these phenomena remain elusive. Analysis of tlr4 promoters revealed high similarity among LPS-sensitive species (human, chimpanzee, and horse) and low similarity with LPS-resistant species (mouse and rat). Four conserved nuclear factor kappa B (NFκB) binding sites were found in the tlr4 promoter and two in the md2 promoter sequences that are likely to be targets for dexamethasone regulation. In vitro treatment of equine peripheral blood mononuclear cells (eqPBMC) with LPS decreased transcripts of tlr4 and increased transcription of md2 (myeloid differentiation factor 2) and cd14 (cluster of differentiation 14). Treatment with dexamethasone rescued transcription of tlr4 after LPS inhibition. LPS-induced transcription of md2 was inhibited in the presence of dexamethasone. Dexamethasone alone did not affect transcription of tlr4 and md2.

Interday variation and effect of transportation on indirect blood pressure measurements, plasma endothelin-1 and serum cortisol in Standardbred and Icelandic horses.

BackgroundSystemic hypertension is a prominent feature in humans with metabolic syndrome (MS) and this is partly caused by an enhanced endothelin-1 (ET-1) mediated vasoconstriction. There are indications that systemic hypertension might be a feature in equine metabolic syndrome (EMS) but if ET-1 is involved in the development of hypertension in horses is not known. Increased levels of cortisol have also been found in humans with MS but there are no reports of this in horses. Before blood pressure, plasma ET-1 and serum cortisol can be evaluated in horses with EMS, it is necessary to investigate the interday variation of these parameters on clinically healthy horses. The aims of the present study were therefore to evaluate the interday variation and influence of transportation on systemic blood pressure, plasma ET-1 and serum cortisol in healthy Standardbred and Icelandic horses, and to detect potential breed differences.MethodsNine horses of each breed were included in the study. Blood pressure was measured and blood samples were collected between 6 and 9 am on two separate days. Eight of the horses (four of each breed) were transported to a new stable were they stayed overnight. The next morning, the sampling procedure was repeated.ResultsThe interday variation was higher for plasma ET-1 (37%) than for indirect pressure measurements (8-21%) and serum cortisol (18%). There were no differences in systemic blood pressure between the two breeds. The Icelandic horses had significantly lower serum cortisol and significantly higher plasma ET-1 concentrations compared to the Standardbred horses. Plasma ET-1 was significantly elevated after transportation, but systemic blood pressure and serum cortisol did not differ from the values obtained in the home environment.ConclusionsIndirect blood pressure, plasma ET-1 and serum cortisol are of interest as markers for cardiovascular dysfunction in horses with EMS. The elevated plasma ET-1 concentrations recorded after transportation was likely caused by a stress response. This needs to be considered when evaluating plasma ET-1 in horses after transportation. The differences detected in plasma ET-1 and serum cortisol between the two breeds might be related to differences in genetic setup, training status as well as management conditions.

Effect of repeated oral administration of glucose and leucine immediately after exercise on plasma insulin concentration and glycogen synthesis in horses

OBJECTIVE To determine whether repeated oral administration of glucose and leucine during the period immediately after intense exercise would increase the release of insulin and thereby enhance glycogen synthesis in horses. ANIMALS 12 Standardbred horses. PROCEDURES In a crossover study design, after glycogen-depleting exercise, horses received oral boluses of glucose (1 g/kg at 0, 2, and 4 hours) and leucine (0.1 g/kg at 0 and 4 hours) or boluses of water (10 mL/kg at 0, 2, and 4 hours; control treatment). Blood samples for determination of glucose, insulin, and leucine concentrations were collected prior to and during a 6-hour period immediately after exercise. Biopsy specimens of a gluteus muscle were obtained before and immediately after exercise and at 3, 6, and 24 hours after exercise for measurement of glycogen concentration. RESULTS When glucose and leucine were administered to the horses, plasma insulin concentration was significantly higher during the 6 hours immediately after exercise than it was when water was administered to the horses. Serum glucose concentration during the 4 hours immediately after exercise was significantly higher when glucose and leucine were administered than the serum glucose concentration when water was administered. Muscle glycogen concentrations did not differ between the 2 treatments during the 24 hours after exercise. CONCLUSIONS AND CLINICAL RELEVANCE Synthesis of muscle glycogen after intense intermittent exercise was not enhanced by oral boluses of glucose and leucine after exercise despite pronounced increases in plasma insulin and serum glucose concentrations.

Repeated post-exercise administration with a mixture of leucine and glucose alters the plasma amino acid profile in Standardbred trotters

BackgroundThe branched chain amino acid leucine is a potent stimulator of insulin secretion. Used in combination with glucose it can increase the insulin response and the post exercise re-synthesis of glycogen in man. Decreased plasma amino acid concentrations have been reported after intravenous or per oral administration of leucine in man as well as after a single per oral dose in horses. In man, a negative correlation between the insulin response and the concentrations of isoleucine, valine and methionine have been shown but results from horses are lacking. This study aims to determine the effect of repeated per oral administration with a mixture of glucose and leucine on the free amino acid profile and the insulin response in horses after glycogen-depleting exercise.MethodsIn a crossover design, after a glycogen depleting exercise, twelve Standardbred trotters received either repeated oral boluses of glucose, 1 g/kg body weight (BW) at 0, 2 and 4 h with addition of leucine 0.1 g/kg BW at 0 and 4 h (GLU+LEU), or repeated boluses of water at 0, 2 and 4 h (CON). Blood samples for analysis of glucose, insulin and amino acid concentrations were collected prior to exercise and over a 6 h post-exercise period. A mixed model approach was used for the statistical analyses.ResultsPlasma leucine, isoleucine, valine, tyrosine and phenylalanine concentrations increased after exercise. Post-exercise serum glucose and plasma insulin response were significantly higher in the GLU+LEU treatment compared to the CON treatment. Plasma leucine concentrations increased after supplementation. During the post-exercise period isoleucine, valine and methionine concentrations decreased in both treatments but were significantly lower in the GLU+LEU treatment. There was no correlation between the insulin response and the response in plasma leucine, isoleucine, valine and methionine.ConclusionsRepeated post-exercise administration with a mixture of leucine and glucose caused a marked insulin response and altered the plasma amino acid profile in horses in a similar manner as described in man. However, the decreases seen in plasma amino acids in horses seem to be related more to an effect of leucine and not to the insulin response as seen in man.

Effects of diet-induced weight gain and turnout to pasture on insulin sensitivity in moderately insulin resistant horses

OBJECTIVE To quantify insulin sensitivity and monitor glucose, insulin, and lipid concentrations in a group of moderately insulin-resistant horses during induction of obesity by use of a forage diet supplemented with fat and during subsequent turnout to pasture. ANIMALS 9 adult Standardbred mares (11 to 20 years old). PROCEDURES Weight gain of horses was induced during 22 weeks by use of a forage diet supplemented with fat fed in gradually increasing amounts, followed by feeding of that fat-supplemented diet at 2.5 times the daily maintenance requirements. Horses were then turned out to pasture. Insulin sensitivity was measured with the euglycemic hyperinsulinemic clamp method before and after weight gain and after 4 weeks at pasture. Body weight, body condition score, and cresty neck score as well as fasting and postprandial concentrations of plasma insulin, plasma glucose, serum triglyceride, and serum nonesterified fatty acids were measured during the study. RESULTS Body weight typically increased by 10%, and body condition score (scale, 1 to 9) increased by > 1.5 from the start to the end of the weight-gain period. There was no difference in insulin sensitivity or metabolic clearance rate of insulin during the weight-gain period. Four weeks at pasture generally improved insulin sensitivity and metabolic clearance rate of insulin by 54% and 32%, respectively, but there was no change in body weight or body condition score. CONCLUSIONS AND CLINICAL RELEVANCE Findings indicated that dietary composition played a more important role than did short-term weight gain on alterations in insulin sensitivity of horses.

Serum thymidine kinase activity in clinically healthy and diseased horses: a potential marker for lymphoma.

Serum thymidine kinase (sTK) activity is a tumour marker used as a prognostic indicator for lymphoma in humans, dogs and cats. The aim of this study was to evaluate the clinical utility of sTK as a biomarker for lymphoma in horses. Serum samples were collected from clinically normal horses (n = 37), horses with lymphoma (n = 23), horses with non-haematopoietic neoplasia (n = 9) and horses with inflammatory disease (n = 14). sTK was measured using a radioenzyme assay. A reference cut-off value of <2.7 U/L (mean + 2 standard deviations, SDs) was established using data from clinically normal horses. sTK activity (mean ± SD) was 26.3 ± 91.5 U/L (range 0.8-443 U/L) for horses with lymphoma, 2.3 ± 1.4 U/L (range 0.6-5.7 U/L) for horses with non-haematopoietic neoplasia and 1.5 ± 0.6 U/L (range 0.6-2.8 U/L) for horses with inflammatory disease. Horses with lymphoma had significantly higher sTK activity than horses without clinical signs of disease (P <0.01), horses with inflammatory disease (P <0.01) and horses with non-haematopoietic neoplasia (P <0.05). sTK activity is a potentially useful biomarker for equine lymphoma.

Relationship Between β-cell Response and Insulin Sensitivity in Horses based on the Oral Sugar Test and the Euglycemic Hyperinsulinemic Clamp

Background A hyperbolic relationship between β‐cell response and insulin sensitivity (IS) has been described in several species including rodents, dogs, and humans. This relationship has not been elucidated in the horse. Hypothesis/Objectives To determine whether the hyperbolic relationship between β‐cell response and IS exists in horses by using indices of β‐cell response from the oral sugar test (OST) and IS measurements from the euglycemic hyperinsulinemic clamp (EHC). A second aim was to compare how well IS estimates from the OST and EHC correlate. Animals Forty‐nine horses with different degrees of insulin regulation (normal‐to‐severe insulin dysregulation). Methods Cross‐sectional study. Horses were examined with an OST and an EHC. Results Decreased IS was associated with increased β‐cell response in the horses. Nine of 12 comparisons between indices of β‐cell response and IS measures fulfilled the criteria for a hyperbolic relationship. Indices of IS calculated from the OST correlated highly with the insulin‐dependent glucose disposal rate (M) and the insulin‐dependent glucose disposal rate per unit of insulin (M/I) determined from the EHC (r = 0.81–0.87). Conclusions and Clinical Importance A hyperbolic relationship between β‐cell response and IS exists in horses, which suggest that horses with insulin dysregulation respond not only with postprandial hyperinsulinemia but are also insulin resistant. The OST is primarily a test for β‐cell response rather than a test for IS, but calculated indices of IS from the OST may be useful to estimate IS in horses, especially when the horse is insulin resistant.

Comparison between the euglycemic-hyperinsulinemic clamp and the use of proxies for determination of insulin sensitivity in horses.

Background Accurate quantification of insulin resistance is important for diagnosing and determining the efficacy of treatment in patients with equine metabolic syndrome (EMS). The euglycemic-hyperinsulinemic clamp (EHC) is a goldstandard method for measuring insulin sensitivity (IS) but the complexity of the technique limits its use in research. The use of proxies based on fasting values offers an attractive alternative.