Arie Doornenbal

Neurophysiological assessment of the sedative and analgesic effects of a constant rate infusion of dexmedetomidine in the dog

The sedative and analgesic effects of continuous rate infusion (CRI) of dexmedetomidine (DEX) were investigated in Beagle dogs (n=8) using auditory and somatosensory evoked potentials (AEPs and SEPs) recorded before, during and after a CRI of saline or DEX (1.0, 3.0, 5.0 μg/kg bolus, followed by 1.0, 3.0, 5.0 μg/kg/h CRI, respectively). The results showed a significant reduction in AEP at doses of 1.0 μg/kg/h and above and a significant reduction of the SEP at doses of 3.0 and 5.0 μg/kg/h. Neither the AEP nor the SEP was further reduced at 5.0 μg/kg/h when compared to 3.0 μg/kg/h, although a slower return towards baseline values was observed at 5.0 μg/kg/h. The mean plasma levels (±SEM) of DEX during infusion were 0.533±0.053 ng/mL for the 1.0 μg/kg/h dose, 1.869±0.063 ng/mL for the 3.0 μg/kg/h dose and 4.017±0.385 for the 5.0 μg/kg/dose. It was concluded that in adult dogs, a CRI of DEX had a sedative and analgesic effect that could be described quantitatively using neurophysiological parameters. Sedation was achieved at lower plasma levels than required for analgesia, and DEX had a longer (but not larger) effect with infusion rates above 3.0 μg/kg/h.

Evaluation of analgesic and sedative effects of continuous infusion of dexmedetomidine by measuring somatosensory- and auditory-evoked potentials in the rat

OBJECTIVE To study, the analgesic and sedative effects of different constant rate infusions (CRI) of dexmedetomidine, in the rat, by measurement of specific electroencephalographic parameters. The recorded parameters were somatosensory-evoked potentials (SEPs) and auditory-evoked potentials (AEPs), which have been shown to be related to analgesia and sedation respectively. ANIMALS Nine male Wistar rats (HsdCpb:Wu, Harlan Netherlands BV, body weight 300-350 g). METHODS Somatosensory-evoked potentials were recorded from the primary somatosensory cortex and the vertex location (SI/Vx-SEPs). Auditory-evoked potentials were recorded from the primary auditory cortex and vertex location (AI/Vx-AEPs). Primary somatosensory cortex and vertex location recorded SEPs and AI/Vx-AEPs were recorded alternately, during CRI of dexmedetomidine (4.0, 10.0, 20.0 microg kg(-1) hour(-1)) and a control (saline). RESULTS The primary somatosensory cortex-evoked potentials were not affected by the dexmedetomidine CRI, but the other three parameters were significantly affected; although the AI-SEP to a lesser extent than the Vx-SEP and Vx-AEP. A maximum effect on the Vx-AEP was reached at lower doses than on the Vx-SEP. CONCLUSIONS Based on the present findings, it is suggested that CRI of dexmedetomidine provided profound sedation at low doses, whereas higher doses are needed to provide concurrent analgesia. CLINICAL RELEVANCE A constant rate infusion of dexmedetomidine can be a valuable adjunct in the provision of sedation and/or analgesia. However, analgesia cannot be produced without sedation, and sedation is not necessarily accompanied by comparative degrees of analgesia.

Nociception and Conditioned Fear in Rats: Strains Matter

When using rats in pain research, strain-related differences in outcomes of tests for pain and nociception are acknowledged. However, very little is known about the specific characteristics of these strain differences. In this study four phylogenetically distant inbred rat strains, i.e. Wistar Kyoto (WKY), Fawn Hooded (FH), Brown Norway (BN) and Lewis (LE), were investigated in different tests related to pain and nociception. During Pavlovian fear conditioning, the LE and WKY showed a significantly longer duration of freezing behaviour than the FH and BN. Additionally, differences in c-Fos expression in subregions of the prefrontal cortex and amygdala between rat strains during retrieval and expression of conditioned fear were found. For example, the BN did not show recruitment of the basolateral amygdala, whereas the WKY, FH and LE did. During the hot plate test, the WKY and LE showed a lower thermal threshold compared to the BN and FH. In a follow-up experiment, the two most contrasting strains regarding behaviour during the hot plate test and Pavlovian fear conditioning (i.e. FH and WKY) were selected and the hot plate test, Von Frey test and somatosensory-evoked potential (SEP) were investigated. During the Von Frey test, the WKY showed a lower mechanical threshold compared to the FH. When measuring the SEP, the FH appeared to be less reactive to increasing stimulus intensities when considering both peak amplitudes and latencies. Altogether, the combined results indicate various differences between rat strains in Pavlovian fear conditioning, nociception related behaviours and nociceptive processing. These findings demonstrate the necessity of using multiple rat strains when using tests including noxious stimuli and suggest that the choice of rat strains should be considered. When selecting a strain for a particular study it should be considered how this strain behaves during the tests used in that study.

Treadmill but not wheel running improves fatigue resistance of isolated extensor digitorum longus muscle in mice

Aim:  The present study is the first to compare the physiological impact of either forced treadmill or voluntary wheel running exercise on hindlimb muscle in mice.

Forelimb and hindlimb ground reaction forces of walking cats: Assessment and comparison with walking dogs

The primary aim of this study was to assess the potential of force plate analysis for describing the stride cycle of the cat. The secondary aim was to define differences in feline and canine locomotion based on force plate characteristics. Ground reaction forces of 24 healthy cats were measured and compared with ground reaction forces of 24 healthy dogs. Force-time waveforms in cats generated by force plate analysis were consistent, as reflected by intra-class correlation coefficients for peak vertical force, peak propulsive force and peak braking force (0.94-0.95, 0.85-0.89 and 0.89-0.90, respectively). Compared with dogs, cats had a higher peak vertical force during the propulsion phase (cat, 3.89 ± 0.19 N/kg; dog, 3.03 ± 0.16 N/kg), and a higher hindlimb propulsive force (cat, -1.08 ± 0.13 N/kg; dog, (-0.87 ± 0.13 N/kg) and hindlimb impulse (cat, -0.18 ± 0.03 N/kg; dog, -0.14 ± 0.02 N/kg). Force plate analysis is a valuable tool for the assessment of locomotion in cats, because it can be applied in the clinical setting and provides a non-invasive and objective measurement of locomotion characteristics with high repeatability in cats, as well as information about kinetic characteristics. Differences in force-time waveforms between cats and dogs can be explained by the more crouched position of cats during stance and their more compliant gait compared with dogs. Feline waveforms of the medio-lateral ground reaction forces also differ between cats and dogs and this can be explained by differences in paw supination-pronation.

Reducing chloride conductance prevents hyperkalaemia‐induced loss of twitch force in rat slow‐twitch muscle

Exercise‐induced loss of skeletal muscle K+ can seriously impede muscle performance through membrane depolarization. Thus far, it has been assumed that the negative equilibrium potential and large membrane conductance of Cl− attenuate the loss of force during hyperkalaemia. We questioned this idea because there is some evidence that Cl− itself can exert a depolarizing influence on membrane potential (Vm). With this study we tried to identify the possible roles played by Cl− during hyperkalaemia. Isolated rat soleus muscles were kept at 25 °C and twitch contractions were evoked by current pulses. Reducing [Cl−]o to 5 mm, prior to introducing 12.5 mm Ko, prevented the otherwise occurring loss of force. Reversing the order of introducing these two solutions revealed an additional effect, i.e. the ongoing hyperkalaemia‐related loss of force was sped up tenfold after reducing [Cl−]o. However, hereafter twitch force recovered completely. The recovery of force was absent at [K+]o exceeding 14 mm. In addition, reducing [Cl−]o increased membrane excitability by 24%, as shown by a shift in the relationship between force and current level. Measurements of Vm indicated that the antagonistic effect of reducing [Cl−]o on hyperkalaemia‐induced loss of force was due to low‐Cl−‐induced membrane hyperpolarization. The involvement of specific Cl− conductance was established with 9‐anthracene carboxylic acid (9‐AC). At 100 μm, 9‐AC reduced the loss of force due to hyperkalaemia, while at 200 μm, 9‐AC completely prevented loss of force. To study the role of the Na+−K+−2Cl− cotransporter (NKCC1) in this matter, we added 400 μm of the NKCC inhibitor bumetanide to the incubation medium. This did not affect the hyperkalaemia‐induced loss of force. We conclude that Cl− exerts a permanent depolarizing influence on Vm. This influence of Cl− on Vm, in combination with a large membrane conductance, can apparently have two different effects on hyperkalaemia‐induced loss of force. It might exert a stabilizing influence on force production during short periods of hyperkalaemia, but it can add to the loss of force during prolonged periods of hyperkalaemia.

Bispectral index and the clinically evaluated anaesthetic depth in dogs

OBJECTIVE This study investigated whether the bispectral index (BIS monitor) corresponded with the clinical assessment of anaesthetic depth in dogs. STUDY DESIGN Prospective clinical study. ANIMALS Sixty-five dogs undergoing anaesthesia for surgery. METHODS Dogs were assigned to one of three different anaesthetic techniques. A three point scale was devised to determine the clinical depth of anaesthesia (CDA); CDA 1 represented light, CDA 2 surgical and CDA 3 excessive depth of anaesthesia. BIS values were recorded and CDA assessed at specific times and points throughout surgery. Data were statistically analysed using mixed model regression. RESULTS Clinical depth of anaesthesia was assessed as CDA 1 on 68, 2 on 748 and 3 on four occasions. The BIS recorded for CDA 1 differed significantly from that for CDA 2 (p<0.001). However, individual BIS values measured at light and surgical levels of anaesthesia overlapped considerably. The sensitivities and specificities calculated for BIS to diagnose CDA 1 compared to CDA 2 in the three anaesthetic protocols were 28-86% and 55-85%. The accompanying positive predictive value was 0.08-0.29 and the negative predictive value was 0.95-0.97. End-tidal isoflurane concentrations (anaesthetic techniques 1 and 3) and propofol infusion (technique 2) at CDA 1 was significantly lower than those at CDA 2 (p=0.001). CONCLUSIONS Although BIS values overall distinguished between CDA scores, the calculated specificities, sensitivities and predictive values were low, and there were anomalous results in individual cases. CLINICAL RELEVANCE The use of the BIS as the sole method to determine anaesthetic depth in dogs is imprudent.

Nociception-related somatosensory evoked potentials in awake dogs recorded after intra epidermal electrical stimulation

At present, the specific neurophysiologic methodology of recording pain‐related evoked potentials is considered a most promising approach to objectively quantify pain in man. This study was designed to characterise and evaluate the use of somatosensory evoked potentials to study nociception in a canine model. To this aim, somatosensory evoked potentials were evoked by intra‐epidermal electrical stimulation and recorded from the scalp in 8 beagle dogs. Characteristics determined were: (1) the conduction velocities of the peripheral nerve fibres involved, (2) the stimulus intensity response characteristics and (3) the evaluation of possible disturbance of the signals by muscular activity from the hind paw withdrawal reflex (EMG artefact). The results showed (1) the conduction velocities to be in the A‐delta fibre range (i.e. fibres involved in nociception), (2) an increase in amplitude and a decrease in latency of the evoked potential following increasing stimulus intensities and (3) the absence of EMG artefact in the signals. These data indicate that the evoked potentials recorded, are related to nociception and thus are suited to quantitatively characterise the perception of noxious stimuli making this model useful for pain‐ and analgesia‐related research.

Antinociceptive effects of low dose lumbosacral epidural ropivacaine in healthy ponies

The objective of this study was to evaluate the safety and efficacy of low dose lumbosacral epidural ropivacaine in ponies. Antinociceptive effects of epidural ropivacaine were evaluated by means of mechanical nociceptive thresholds (MNTs) at several spinal levels in conscious ponies. The effects of ropivacaine on nociceptive afferent transmission to the spinal cord were also assessed by measuring spinal cord somatosensory evoked potentials (SSEPs) in anaesthetised ponies. Ataxia scores were determined in conscious ponies to assess the effects on motor function. A randomised, placebo controlled, double blind cross-over design was used. Low dose lumbosacral epidural ropivacaine led to increases in MNTs at various anatomical locations with a maximum effect at the lumbosacral and sacrococcygeal regions, both with respect to increase in threshold and duration of effect. Analysis of SSEPs showed that epidural ropivacaine influenced both Aβ- and Aδ-mediated afferent transmission to the spinal cord at the level of the lumbosacral junction. Ponies showed mild ataxia after low dose lumbosacral epidural ropivacaine, but all ponies remained standing. Application of low dose lumbosacral epidural ropivacaine provided safe and efficacious antinociceptive effects in conscious and anaesthetised ponies, and could therefore be a valuable addition to multimodal analgesic protocols in Equidae.

The α2-adrenoceptor agonist dexmedetomidine suppresses memory formation only at doses attenuating the perception of sensory input

It was investigated whether continuous rate infusion of the alpha(2)-adrenoceptor agonist dexmedetomidine can suppress memory formation by mechanisms other than reducing perception of sensory input in a fear-conditioning paradigm. Different groups of rats infused with either saline or dexmedetomidine (2.0, 4.0 or 10.0microg/kg bolus, followed by 2.0, 4.0 or 10.0microg/kg/h continuous rate infusion respectively), were subjected to a somatosensory-evoked potential (SEP) fear-conditioning paradigm. This paradigm combined the pairing of an innoxious conditioned stimulus (CS) and a noxious unconditioned stimulus (US), of which the latter was used to generate the SEPs (training phase).The following day, the perception of the US during the training phase was assessed by presenting the CS only and subsequently scoring the resulting duration of freezing behaviour (testing phase). Freezing behaviour was reduced only in those groups which demonstrated reduced SEPs. Based on these findings, it is concluded that dexmedetomidine suppresses memory formation only at doses reducing central nervous system activity in response to sensory input.