The horse-human dyad: can we align horse training and handling activities with the equid social ethogram?

This article examines the recently completed equid ethogram and shows how analogues of social interactions between horses may occur in various human-horse interactions. It discusses how some specific horse-horse interactions have a corresponding horse-human interaction - some of which may be directly beneficial for the horse while others may be unusual or even abnormal. It also shows how correspondent behaviours sometimes become inappropriate because of their duration, consistency or context. One analogue is unlikely to hold true for all horse-human contexts, so when applying any model from horse-horse interactions to human-horse interactions, the limitations of the model may eclipse the intended outcome of the intervention. These limitations are especially likely when the horse is being ridden. Such analyses may help to determine the validity of extrapolating intra-specific interactions to the inter-specific setting, as is advocated by some popular horse-training methods, and highlight the subsequent limitations where humans play the role of the 'alpha mare' or leader in horse handling and training. This examination provides a constructive framework for further informed debate and empirical investigation of the critical features of successful intra-specific interactions.

Energy replacement attenuates the effects of prior moderate exercise on postprandial metabolism in overweight/obese men.

BACKGROUND: The extent to which exercise-induced changes to postprandial metabolism are dependant on the associated energy deficit is not known. OBJECTIVE: To determine the effects of exercise, with and without energy replacement, on postprandial metabolism. DESIGN: Each subject underwent three 2-day trials in random order. On day 1 of each trial subjects rested (control), walked at 50% maximal oxygen uptake to induce a net energy expenditure of 27 kJ kg(-1) body mass (energy-deficit) or completed the same walk with the net energy expended replaced (energy-replacement). On day 2 subjects completed an 8.5-h metabolic assessment. For 3 days prior to day 2, subjects consumed an isocaloric diet, avoided planned exercise (apart from exercise interventions) and alcohol. SUBJECTS: A total of 13 overweight/obese men (age: 40+/-8 years, body mass index: 31.1+/-3.0 kg m(-2)). MEASUREMENTS: Postprandial triglyceride, insulin, glucose, non-esterified fatty acid and 3-hydroxybutyrate concentrations and substrate utilization rates were determined. RESULTS: Energy-deficit lowered postprandial triglyceride concentrations by 14 and 10% compared with control and energy-replacement (P<0.05 for both). Energy-deficit increased postprandial 3-hydroxybutyrate concentrations by 40 and 19% compared with control and energy-replacement (P<0.05 for both). Postprandial insulin concentrations were 18 and 10% lower for energy-deficit and energy-replacement compared with control and 10% lower for energy-deficit than energy-replacement (P<0.05 for all). Postprandial fat oxidation increased by 30 and 14% for energy-deficit and energy-replacement compared to control and was 12% higher for energy-deficit than energy-replacement (P<0.05 for all). CONCLUSION: Exercise with energy replacement lowered postprandial insulinaemia and increased fat oxidation. However an exercise-induced energy deficit augmented these effects and was necessary to lower postprandial lipaemia.

Spontaneous Ca2+ transients in rat pulmonary vein cardiomyocytes are increased in frequency and become more synchronous following electrical stimulation.

The pulmonary veins have an external sleeve of cardiomyocytes that are a widely recognised source of ectopic electrical activity that can lead to atrial fibrillation. Although the mechanisms behind this activity are currently unknown, changes in intracellular calcium (Ca2+) signalling are purported to play a role. Therefore, the intracellular Ca2+ concentration was monitored in the pulmonary vein using fluo-4 and epifluorescence microscopy. Electrical field stimulation evoked a synchronous rise in Ca2+ in neighbouring cardiomyocytes; asynchronous spontaneous Ca2+ transients between electrical stimuli were also present. Immediately following termination of electrical field stimulation at 3 Hz or greater, the frequency of the spontaneous Ca2+ transients was increased from 0.45 ± 0.06 Hz under basal conditions to between 0.59 ± 0.05 and 0.65 ± 0.06 Hz (P < 0.001). Increasing the extracellular Ca2+ concentration enhanced this effect, with the frequency of spontaneous Ca2+ transients increasing from 0.45 ± 0.05 Hz to between 0.75 ± 0.06 and 0.94 ± 0.09 Hz after electrical stimulation at 3 to 9 Hz (P < 0.001), and this was accompanied by a significant increase in the velocity of Ca2+ transients that manifested as waves. Moreover, in the presence of high extracellular Ca2+, the spontaneous Ca2+ transients occurred more synchronously in the initial few seconds following electrical stimulation. The ryanodine receptors, which are the source of spontaneous Ca2+ transients in pulmonary vein cardiomyocytes, were found to be arranged in a striated pattern in the cell interior, as well as along the periphery of cell. Furthermore, labelling the sarcolemma with di-4-ANEPPS showed that over 90% of pulmonary vein cardiomyocytes possessed T-tubules. These findings demonstrate that the frequency of spontaneous Ca2+ transients in the rat pulmonary vein are increased following higher rates of electrical stimulation and increasing the extracellular Ca2+ concentration.

The Use of Ratiometric Fluorescence Measurements of the Voltage Sensitive Dye Di-4-ANEPPS to Examine Action Potential Characteristics and Drug Effects on Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) and higher throughput platforms have emerged as potential tools to advance cardiac drug safety screening. This study evaluated the use of high bandwidth photometry applied to voltage-sensitive fluorescent dyes (VSDs) to assess drug-induced changes in action potential characteristics of spontaneously active hiPSC-CM. Human iPSC-CM from 2 commercial sources (Cor.4U and iCell Cardiomyocytes) were stained with the VSD di-4-ANEPPS and placed in a specialized photometry system that simultaneously monitors 2 wavebands of emitted fluorescence, allowing ratiometric measurement of membrane voltage. Signals were acquired at 10 kHz and analyzed using custom software. Action potential duration (APD) values were normally distributed in cardiomyocytes (CMC) from both sources though the mean and variance differed significantly (APD90: 229 ± 15 ms vs 427 ± 49 ms [mean ± SD, P < 0.01]; average spontaneous cycle length: 0.99 ± 0.02 s vs 1.47 ± 0.35 s [mean ± SD, P < 0.01], Cor.4U vs iCell CMC, respectively). The 10-90% rise time of the AP (Trise) was ∼6 ms and was normally distributed when expressed as 1/[Formula: see text] in both cell preparations. Both cell types showed a rate dependence analogous to that of adult human cardiac cells. Furthermore, nifedipine, ranolazine, and E4031 had similar effects on cardiomyocyte electrophysiology in both cell types. However, ranolazine and E4031 induced early after depolarization-like events and high intrinsic firing rates at lower concentrations in iCell CMC. These data show that VSDs provide a minimally invasive, quantitative, and accurate method to assess hiPSC-CM electrophysiology and detect subtle drug-induced effects for drug safety screening while highlighting a need to standardize experimental protocols across preparations.

Effect of sustained stretch on dispersion of ventricular fibrillation intervals in normal rabbit hearts.

OBJECTIVE: To determine the effect of acute left ventricular dilatation on refractoriness in normal hearts. METHODS: During sustained ventricular fibrillation (VF) in isolated perfused hearts, recording of local activation time yields VF intervals which provide an index of local refractoriness. Simultaneous measurement from multiple sites enables study of spatial aspects of changes in refractoriness. We studied the effects of stretch on the magnitude and dispersion of changes in VF interval in 10 isolated, Langendorff-perfused rabbit hearts using a flexible epicardial array containing 240 unipolar electrodes. The left ventricular pressure was increased from 0 to 40 mmHg by inflation of an intraventricular balloon during sustained VF. RESULTS: The current threshold for VF induction fell from 64 +/- 11 mA to 43 +/- 11 mA (mean +/- SE, P < 0.01) following ventricular dilatation. Mean VF interval at 0 mmHg was 79.8 +/- 1.3 ms and fell to 70.2 +/- 1.7 ms (P < 0.01) at 40 mmHg. There was a corresponding increase in dispersion of VF interval (coefficient of variation) from 8.13 +/- 0.8 to 13.3 +/- 0.8 (P < 0.01). There was regional heterogeneity in the areas of greatest reduction in VF interval, which varied between hearts. Following balloon inflation there was an increase in the number of activation waves. CONCLUSIONS: Acute ventricular dilatation produces spatially heterogeneous changes in refractoriness which would predispose to the maintenance of reentrant arrhythmias.

Observational study of "urine testing" in the horse and donkey stallion.

Although "urine testing" is said to enable the male equid to assess the sexual status of the mare, there are no reports in the literature of any detailed study of this behavioural response of the stallion. Behavioural response to conspecific urine was studied in two horse stallions and one donkey stallion. The relevant nasopalatine anatomy is described. Events observed during urine testing included head, neck, lip, jaw, tongue movements, penile changes and nasal secretion. Nasal endoscopy indicated that the source of part of the nasal secretion was the secretory glands of the vomeronasal organ complex. The significance and probable function of these events in urine testing is discussed.

The effect of cromakalim on intracellular [Ca2+] in isolated rat skeletal muscle during fatigue and metabolic blockade.

The effects of the ATP-sensitive K+ channel (KATP channel) opener cromakalim on excitation-contraction (E-C) coupling were studied in skeletal muscle during fatiguing and non-fatiguing activity. Intracellular calcium concentration ([Ca2+]i) was monitored using the fluorescent indicator fura-2 in isolated single skeletal muscle fibres enzymatically dissociated from rat flexor digitorum brevis. A protocol of tetanic stimulation (50 Hz for 300 ms) with progressively shorter durations between tetani was used to induce E-C coupling failure in these cells. Cromakalim (100-800 microM) had little effect on peak [Ca2+]i during twitch and non-fatiguing tetanic stimulation. However, with 0.4 s between tetani, 100 microM cromakalim decreased peak tetanic [Ca2+]i from 1.47 +/- 0.11 microM to 8.35 +/- 55 nM, but did not affect resting [Ca2+]i (control, 220 +/- 40 nM; with cromakalim, 171 +/- 33 nM). Cyanide (2 mM) decreased tetanic [Ca2+]i and increased resting [Ca2+]i during the stimulus protocol; with 0.4 s between tetani, peak [Ca2+]i was 820 +/- 50 nM and resting [Ca2+]i was 443 +/- 32 nM. The ability of cromakalim to inhibit E-C coupling was enhanced by the presence of cyanide. Complete blockade of metabolism by cyanide and iodoactetate (0.1 mM) caused a marked rise in resting [Ca2+]i and inhibition of the tetanic rise of [Ca2+]i. With cromakalim (100 microM) present, E-C coupling failed during metabolic blockade but without a significant increase in resting [Ca2+]i. These results are consistent with a role for the KATP channel in the failure of Ca2+ release during fatigue.

Sensitivity to flow of intrinsic gating in inwardly rectifying potassium channel from mammalian skeletal muscle.

1. Current through inwardly rectifying K+ channels was measured in inside-out patches from rat and human sarcolemmal vesicles and from dispersed rat flexor digitorum brevis muscle fibres. The patches were positioned so as to face the aperture of a large-diameter pipette from which solution of the same composition as the bath solution could be ejected. The solution within the patch pipette and the bath solution both contained principally 140 mM-KCl. 2. The kinetic behaviour of the inwardly rectifying channel was found to vary according to whether the patch was in static or flowing solution. At negative holding potentials, when the channel is open most of the time in static solution, flow produced a reversible and repeatable decrease in open probability. 3. In Mg2(+)-free solution the inwardly rectifying channel allows outward current to pass at positive holding potentials. This allows the kinetic behaviour of the channel in static and flowing solution to be compared over a wider voltage range. 4. In both static and flowing solution, the open probability-voltage relation is sigmoidal and can be fitted by a Boltzmann curve. As a result of flow, the maximum open probability at negative potentials is decreased and the mid-point of the relation is shifted to the right by more than 20 mV. 5. No evidence could be found for the existence of a local concentration gradient sensitive to flow. Application of suction to the patch pipette showed the inwardly rectifying channels not to be sensitive to membrane stretch.(ABSTRACT TRUNCATED AT 250 WORDS)

Ventricular fibrillation threshold and local dispersion of refractoriness in isolated rabbit hearts with left ventricular dysfunction.

Patients with congestive heart failure or left ventricular dysfunction (LVD) have a high incidence of ventricular tachyarrhythmias and sudden cardiac death. In addition to structural, metabolic and neuroendocrine changes, mechanoelectrical feedback may play a role in arrhythmogenesis in heart failure. Three groups of rabbits (n = 10 for each) were studied: chronic coronary ligation with ejection fraction (EF) > or = 0.45 or < 0.45, and sham-operated controls. Ventricular fibrillation (VF) thresholds were measured at LV pressures of 0 and 40 mm Hg during modified Langendorff perfusion. Intervals between local activations during VF (VFI) were used as an index of refractoriness. Global dispersion was expressed as coefficient of variation of VFI; local dispersion by maximum difference in VFI between adjacent sites. Median VF threshold was lower at 0 mm Hg in the lower EF group compared to controls (30 vs. 67.5 mA, P<0.05). VF threshold in control hearts was lower at 40 mm Hg than at 0 mm Hg (P<0.01), but there was no further reduction in threshold in LVD hearts at 40 mm Hg. Global dispersion of VFI did not differ significantly between groups. Local dispersion of VFI in the lower EF group was greater than in controls at 0 mm Hg in the infarct border zone (P<0.05). At 40 mm Hg, local dispersion of VFI in zones bordering and remote from the infarct were greater in both LVD groups than in controls (P<0.05). Local inhomogeneity of refractoriness is more marked in the infarct border zone, but latent abnormalities are evident in normal myocardium of rabbits with left ventricular dysfunction and are revealed by left ventricular distension.

Mechanical properties of normal and mdx mouse sarcolemma: bearing on function of dystrophin.

The tensile strength of the muscle fibre surface membrane was estimated (1) from the suction required to burst membrane patches and (2) by aspiration of sarcolemmal vesicles into micropipettes of uniform bore. Each method gave an average value close to 60 microN cm-1 for the maximum tension sustainable by normal mouse sarcolemma and only slightly lower values for sarcolemma from mdx mice which lack dystrophin. The elastic modulus of area expansion, as measurable by pipette aspiration of sarcolemmal vesicles, was found to have an average value of 3160 microN cm-1 for normal and 2770 microN cm-1 for mdx mouse sarcolemma. The tensile strength of the sarcolemma is much too small for any differences in it to be the basis for the different osmotic behaviour of normal and mdx muscle fibres reported recently (Menke & Jockusch, 1991). By analogy with the better understood origin of the osmotic fragility of different types of red blood cells, the higher osmotic fragility of mdx muscle fibres is suggested to be of morphological origin. We postulate that dystrophin functions as an element of the submembrane cytoskeleton so as to maintain the normal folding which safeguards the sarcolemma against mechanical damage.

The electrophysiological and mechanical effects of 2,3-butane-dione monoxime and cytochalasin-D in the Langendorff perfused rabbit heart.

UNLABELLED: Procedures that reduce contraction are used to facilitate optical measurements of membrane potential, but it is unclear to what extent they affect the excitability of the heart. This study has examined the electrophysiological consequences of a range of extracellular [Ca2+] (0.7-2.5 mmol l(-1)), 2,3-butane-dione monoxime (BDM; 1-20 mmol l(-1)) and cytochalasin-D (Cyto-D; 1-5 micromol l(-1)). METHODS: Monophasic action potentials (MAPs) were recorded from the basal epicardial surface of the left ventricle of isolated rabbit hearts. Conduction delay (CD) and time to 90% repolarisation of the monophasic action potential (MAPD90) were measured. The effects of BDM and Cyto-D on restitution were studied at a [Ca2+] of 1.9 mmol l(-1). Restitution curves for MAPD90 were generated using a standard S1-S2 protocol. RESULTS: All manoeuvres decreased left ventricular developed pressure (LVDP): 0.7 mmol l(-1) Ca2+ to 74.0 +/- 6.1%, 20 mmol l(-1) BDM to 4.5 +/- 1.0%, and 5 micromol l(-1) Cyto-D to 12.8 +/- 3.5% of control value. CD decreased from a control value (33.3 +/- 1.0 ms, n= 16) to 93.0 +/- 2.2% in 0.7 mmol l(-1) Ca2+, but increased to 133.7 +/- 10.5% in 20 mmol l(-1) BDM and 127.4 +/- 10.6% in 5 micromol l(-1) Cyto-D. At 350 ms pacing cycle length, MAPD90 (control = 119.6 +/- 1.7 ms n= 16) was prolonged by reduced extracellular [Ca2+]. BDM had no effects on MAPD90 at control pacing rates. Cyto-D caused a significant prolongation (to 115.0 +/- 3.0% of control, n= 6) at the highest concentration studied (5 micromol l(-1)). Both BDM (20 mmol l(-1)) and Cyto-D (3 micromol l(-1)) flattened the restitution curves but neither agent altered maximum MAPD90. CONCLUSIONS: Extracellular [Ca2+] of 1.9 mmol l(-1) in conjunction with a moderate dose of Cyto-D (3 micromol l(-1)) reduced contractility with minimal effects on action potential duration and conduction at a fixed pacing cycle length. However, both BDM and Cyto-D had pronounced effects on electrical restitution.

P(i) inhibits the SR Ca(2+) pump and stimulates pump-mediated Ca(2+) leak in rabbit cardiac myocytes.

Measurements of sarcoplasmic reticulum (SR) Ca(2+) uptake were made from aliquots of dissociated permeabilized ventricular myocytes using fura 2. Equilibration with 10 mM oxalate ensured a reproducible exponential decline of [Ca(2+)] from 600 nM to a steady state of 100-200 nM after addition of Ca(2+). In the presence of 5 microM ruthenium red, which blocks the ryanodine receptor, the time course of the decline of [Ca(2+)] can be modeled by a Ca(2+)-dependent uptake process and a fixed Ca(2+) leak. Partial inhibition of the Ca(2+) pump with 1 microM cyclopiazonic acid or 50 nM thapsigargin reduced the time constant for Ca(2+) uptake but did not affect the SR Ca(2+) leak. Addition of 10 mM inorganic phosphate (P(i)) decreased the rate of Ca(2+) accumulation by the SR and increased the Ca(2+) leak rate. This effect was reversed on addition of 10 mM phosphocreatine. 10 mM P(i) had no effect on Ca(2+) leak from the SR after complete inhibition of the Ca(2+) pump. In conclusion, P(i) decreases the Ca(2+) uptake capacity of cardiac SR via a decrease in pump rate and an increase in Ca(2+) pump-dependent Ca(2+) leak.