Tamas D Ambrisko

Vitamin D Is a Regulator of Endothelial Nitric Oxide Synthase and Arterial Stiffness in Mice

The vitamin D hormone 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3] is essential for the preservation of serum calcium and phosphate levels but may also be important for the regulation of cardiovascular function. Epidemiological data in humans have shown that vitamin D insufficiency is associated with hypertension, left ventricular hypertrophy, increased arterial stiffness, and endothelial dysfunction in normal subjects and in patients with chronic kidney disease and type 2 diabetes. However, the pathophysiological mechanisms underlying these associations remain largely unexplained. In this study, we aimed to decipher the mechanisms by which 1,25(OH)2D3 may regulate systemic vascular tone and cardiac function, using mice carrying a mutant, functionally inactive vitamin D receptor (VDR). To normalize calcium homeostasis in VDR mutant mice, we fed the mice lifelong with the so-called rescue diet enriched with calcium, phosphate, and lactose. Here, we report that VDR mutant mice are characterized by lower bioavailability of the vasodilator nitric oxide (NO) due to reduced expression of the key NO synthesizing enzyme, endothelial NO synthase, leading to endothelial dysfunction, increased arterial stiffness, increased aortic impedance, structural remodeling of the aorta, and impaired systolic and diastolic heart function at later ages, independent of changes in the renin-angiotensin system. We further demonstrate that 1,25(OH)2D3 is a direct transcriptional regulator of endothelial NO synthase. Our data demonstrate the importance of intact VDR signaling in the preservation of vascular function and may provide a mechanistic explanation for epidemiological data in humans showing that vitamin D insufficiency is associated with hypertension and endothelial dysfunction.

Influence of drugs on the response characteristics of the LiDCO sensor: an in vitro study

BACKGROUND In a previous study, the authors found a large bias (50%) for lithium (LiDCO) compared with thermodilution cardiac output measurement methods in ponies receiving i.v. infusions of xylazine, ketamine, and midazolam. This prompted the authors to examine the effect of drugs on the LiDCO sensor. METHODS Drugs and lithium were dissolved in 0.9% saline to produce the following solutions: saline, saline-lithium, saline-drug, and saline-drug-lithium. The drug concentrations were overlapping the range of clinical interest as estimated from the published literature. These 38°C solutions were pumped through the LiDCO sensor in predetermined order. Sensor voltages were measured. Differences between lithium-induced voltage changes in the absence and presence of drugs indicated erroneous lithium detections that, if they occurred in vivo, may cause biases in LiDCO measurements. RESULTS Clonidine, detomidine, dexmedetomidine, medetomidine, romifidine, xylazine, ketamine, S-ketamine, lidocaine, and rocuronium caused concentration-dependent increases in sensor voltages and negative biases in lithium detection that were mathematically equivalent to greater than +10% biases in LiDCO. The drug-induced voltage changes correlated with calculated biases in LiDCO (r(2)=0.91). Atipamezole, acepromazine, butorphanol, diazepam, midazolam, and guaifenesin caused minimal or no interaction in this study. CONCLUSIONS A number of drugs influenced the accuracy of the LiDCO sensor in vitro but, based on published pharmacokinetic data, only xylazine, ketamine, lidocaine, and rocuronium may cause biases at clinically relevant concentrations. These findings need to be confirmed in vivo. Relevant (>3 mV) changes in sensor voltages due to the presence of drugs may indicate possible interactions with the LiDCO sensor.

Lithium dilution, pulse power analysis, and continuous thermodilution cardiac output measurements compared with bolus thermodilution in anaesthetized ponies

BACKGROUND This study compares cardiac output (CO) measurements obtained by lithium dilution (LiDCO), pulse power analysis (PulseCO), and continuous thermodilution (CTD) with bolus thermodilution (BTD) in ponies. METHODS Eight isoflurane-anaesthetized Shetland ponies received xylazine, ketamine, and midazolam infusions (0.3, 1.2, and 0.018 mg kg(-1) h(-1), respectively). CO was measured with BTD, CTD, LiDCO, and PulseCO. Lithium was injected into the jugular vein and blood was sampled from the facial artery for lithium detection and this artery was also used for PulseCO. Measurements were obtained during four stable haemodynamic conditions in the following order: isoflurane 1% (end-tidal concentration), isoflurane 2%, isoflurane 1%, and isoflurane 1%+dobutamine 5 µg kg(-1) min(-1). RESULTS The bias (2 sd) was 2.5 (2.1) and 0.5 (2.9) litre min(-1) for LiDCO-BTD and for CTD-BTD comparisons, respectively. The limits of agreement were wider than ±30%; therefore, interchangeability was rejected for both comparisons. A possible error in LiDCO might explain the bias observed because CTD-BTD comparison showed less bias. Changes in PulseCO did not correlate with those of BTD and a weak correlation (r(2)=0.23; P=0.018) and concordance (Pc=0.42) was found between CTD and BTD. CONCLUSIONS This is the first study to show a large bias for LiDCO-BTD comparison in animals receiving xylazine, ketamine, and midazolam infusions. The trending abilities of neither PulseCO nor CTD were reliable. Further studies are needed to elucidate possible influences of drugs on the accuracy of the LiDCOplus system.

Measurement of tidal volume using Respiratory Ultrasonic Plethysmography in anaesthetized, mechanically ventilated horses

OBJECTIVE To compare tidal volume estimations obtained from Respiratory Ultrasonic Plethysmography (RUP) with simultaneous spirometric measurements in anaesthetized, mechanically ventilated horses. STUDY DESIGN Prospective randomized experimental study. ANIMALS Five experimental horses. METHODS Five horses were anaesthetized twice (1 week apart) in random order in lateral and in dorsal recumbency. Nine ventilation modes (treatments) were scheduled in random order (each lasting 4 minutes) applying combinations of different tidal volumes (8, 10, 12 mL kg(-1)) and positive end-expiratory pressures (PEEP) (0, 10, 20 cm H(2)O). Baseline ventilation mode (tidal volume=15 mL kg(-1), PEEP=0 cm H(2)O) was applied for 4 minutes between all treatments. Spirometry and RUP data were downloaded to personal computers. Linear regression analyses (RUP versus spirometric tidal volume) were performed using different subsets of data. Additonally RUP was calibrated against spirometry using a regression equation for all RUP signal values (thoracic, abdominal and combined) with all data collectively and also by an individually determined best regression equation (highest R(2)) for each experiment (horse versus recumbency) separately. Agreement between methods was assessed with Bland-Altman analyses. RESULTS The highest correlation of RUP and spirometric tidal volume (R(2)=0.81) was found with the combined RUP signal in horses in lateral recumbency and ventilated without PEEP. The bias ±2 SD was 0±2.66 L when RUP was calibrated for collective data, but decreased to 0±0.87 L when RUP was calibrated with individual data. CONCLUSIONS AND CLINICAL RELEVANCE A possible use of RUP for tidal volume measurement during IPPV needs individual calibration to obtain limits of agreement within ±20%.

Voltage changes in the lithium dilution cardiac output sensor after exposure to blood from horses given xylazine

BACKGROUND In a previous in vitro study using saline medium, the authors showed that certain drugs changed the voltages of lithium dilution cardiac output (LiDCO) sensors and also influenced their accuracy in measuring lithium concentrations. These two parameters correlated and so we examined whether such drug-sensor interaction exists when LiDCO sensor was exposed to xylazine in blood. METHODS Five healthy adult warm-blood horses were injected with 0.5 mg kg(-1) xylazine i.v. Physiological saline solution and venous blood were consecutively sampled through the same LiDCO sensor at 60, 45, 30, 15, and 0 min before and then 5, 15, 30, 45, and 60 min after xylazine injection. Sensor voltages were recorded and the differences between saline- and blood-exposed sensor voltages were compared at each time point. RESULTS Saline-exposed sensor voltages continuously increased in a non-linear pattern during the experiment. Blood-exposed sensor voltages also increased in a similar pattern, but it was interrupted by an abrupt increase in voltage after xylazine injection. The differences between saline- and blood-exposed sensor voltages were 7 (6.1-8) mV [median (range)] before xylazine but decreased significantly at 5 and 15 min after xylazine treatment. The highest drug-induced voltage change was 3.4 (1.6-7) mV. CONCLUSIONS This study showed that exposure of a LiDCO sensor to blood after a single clinically relevant dose of xylazine in horses changed the voltages of the sensors for 15 min. Comparison of saline- and blood-exposed sensor voltages could become a tool to detect drug-sensor interactions.

Assessment of distribution of ventilation and regional lung compliance by electrical impedance tomography in anaesthetized horses undergoing alveolar recruitment manoeuvres

OBJECTIVE To examine changes in the distribution of ventilation and regional lung compliances in anaesthetized horses during the alveolar recruitment manoeuvre (ARM). STUDY DESIGN Experimental study in which a series of treatments were administered in a fixed order on one occasion. ANIMALS Five adult Warmblood horses. METHODS Animals were anaesthetized (xylazine, midazolam-ketamine, isoflurane), placed in dorsal recumbency and ventilated with 100% oxygen using peak inspiratory pressure (PIP) and positive end-expiratory pressure (PEEP) of 20 cmH2O and 0 cmH2O, respectively. Thoracic electrical impedance tomography (EIT), spirometry and routine anaesthesia monitoring were performed. At 90 minutes after induction of anaesthesia, PIP and PEEP were increased in steps of 5 cmH2O to 50 cmH2O and 30 cmH2O, respectively, and then decreased to baseline values. Each step lasted 10 minutes. Data were recorded and functional EIT images were created using three breaths at the end of each step. Arterial blood samples were analysed. Values for left-to-right and sternal-to-dorsal centre of ventilation (COV), lung compliances and Bohr dead space were calculated. RESULTS Distribution of ventilation drifted leftward and dorsally during recruitment. Mean±standard deviation (SD) values at baseline and highest airway pressures, respectively, were 49.9±0.7% and 48.0±0.6% for left-to-right COV (p=0.009), and 46.3±2.0% and 54.6±2.0% for sternal-to-dorsal COV (p=0.0001). Compliance of dependent lung regions and PaO2 increased, whereas compliance of non-dependent lung regions decreased during ARM and then returned to baseline (p<0.001). Bohr dead space decreased after ARM (p=0.007). Interestingly, PaO2 correlated to the compliance of the dependent lung (r2=0.71, p<0.001). CONCLUSIONS AND CLINICAL RELEVANCE The proportion of tidal volume distributed to dependent and left lung regions increased during ARM, presumably as a result of opening atelectasis. Monitoring compliance of the dependent lung with EIT may substitute PaO2 measurements during ARM to identify an optimal PEEP.

A novel flow partition device for spirometry during large animal anaesthesia

OBJECTIVE We describe and test a novel device for large animal anaesthesia monitoring that uses standard human medicine spirometry sensors. STUDY DESIGN In-vitro study. METHODS The device consists of two adapters that enable the flow to be split evenly into four tubes in parallel, each tube containing a D-lite sensor. The performance of this flow partitioning device (FPD) over a range of flows from 100 to 700 L minute⁻¹ was determined and the pressure versus flow relation, resistance and dead space was compared with a Horse-lite (Moens 2010). RESULTS Equipped with four D-lite sensors, and a flow of 700 L minute⁻¹ the pressure drop of the FPD was 13.5 cm H₂O, resistance 1.17 cm H₂O second L⁻¹ and volume (potential dead space) 182 mL, compared to 2.8 cm H₂O, 0.24 cm H₂O second L⁻¹ and 54 mL respectively for the Horse-lite. The predicted value of the flow partition of ¼ could be confirmed. Limits of agreement were found to be 4.2% in inspiratory direction and 7.1% in expiratory direction. CONCLUSIONS AND CLINICAL RELEVANCE The FPD is an affordable device that extends the specification of any commercially available human spirometry sensors to large animal applications. However, an increase in total resistance and dead space has to be taken into account. Therefore, the new device could be useful during equine anaesthesia.

Occupational exposure to isoflurane during anaesthesia induction with standard and scavenging double masks in dogs, pigs and ponies

Induction of anaesthesia using a face mask may cause workplace pollution with anaesthetics. The aim of this study was to compare the effect of the use of a standard versus a scavenging double face mask on isoflurane pollution during induction of anaesthesia in experimental animals: six dogs, 12 pigs and five ponies. Pigs were anaesthetized only once using either mask type randomly (n = 6). Dogs and ponies were anaesthetized twice, using different mask types for each occasion in a random order with at least 14 days between experiments. The masks were attached to a Bain breathing system (dogs and pigs) or to a circle system (ponies) using a fresh gas flow of 300 or 50 mL/kg/min, respectively, with 5% vaporizer dial setting. Isoflurane concentrations were measured in the anaesthetists breathing zone using an infrared photoacoustic spectrometer. The peak isoflurane concentrations (pollution) during baseline and induction periods were compared with Wilcoxon test in all species, and values between the mask types were compared with either Wilcoxon (ponies and dogs) or Mann–Whitney tests (pigs) (P < 0.05). Pollution was higher during induction when compared with baseline regardless of the mask type used but it was only statistically significant in dogs and pigs. Pollution was lower during induction with double versus single masks but it was only significant in pigs. Despite the lack of statistical significance, large and consistent differences were noted in all species, hence using scavenging masks is recommended to reduce isoflurane workplace pollution.

Assessment of distribution of ventilation by electrical impedance tomography in standing horses

The aim was to evaluate the feasibility of using electrical impedance tomography (EIT) in horses. Thoracic EIT was used in nine horses. Thoracic and abdominal circumference changes were also measured with respiratory ultrasound plethysmography (RUP). Data were recorded during baseline, rebreathing of CO2 and sedation. Three breaths were selected for analysis from each recording. During baseline breathing, horses regularly took single large breaths (sighs), which were also analysed. Functional EIT images were created using standard deviations (SD) of pixel signals and correlation coefficients (R) of each pixel signal with a reference respiratory signal. Left-to-right ratio, centre-of-ventilation and global-inhomogeneity-index were calculated. RM-ANOVA and Bonferroni tests were used (P < 0.05). Distribution of ventilation shifted towards right during sighs and towards dependent regions during sighs, rebreathing and sedation. Global-inhomogeneity-index did not change for SD but increased for R images during sedation. The sum of SDs for the respiratory EIT signals correlated well with thoracic (r(2) = 0.78) and abdominal (r(2) = 0.82) tidal circumferential changes. Inverse respiratory signals were identified on the images at sternal location and based on reviewing CT images, seemed to correspond to location of gas filled intestines. Application of EIT in standing non-sedated horses is feasible. EIT images may provide physiologically useful information even in situations, such as sighs, that cannot easily be tested by other methods.

Comparison of an infrared anaesthetic agent analyser (Datex-Ohmeda) with refractometry for measurement of isoflurane, sevoflurane and desflurane concentrations

OBJECTIVE To assess agreement between infrared (IR) analysers and a refractometer for measurements of isoflurane, sevoflurane and desflurane concentrations and to demonstrate the effect of customized calibration of IR analysers. STUDY DESIGN In vitro experiment. SUBJECTS Six IR anaesthetic monitors (Datex-Ohmeda) and a single portable refractometer (Riken). METHODS Both devices were calibrated following the manufacturers recommendations. Gas samples were collected at common gas outlets of anaesthesia machines. A range of agent concentrations was produced by stepwise changes in dial settings: isoflurane (0-5% in 0.5% increments), sevoflurane (0-8% in 1% increments), or desflurane (0-18% in 2% increments). Oxygen flow was 2 L minute(-1) . The orders of testing IR analysers, agents and dial settings were randomized. Duplicate measurements were performed at each setting. The entire procedure was repeated 24 hours later. Bland-Altman analysis was performed. Measurements on day-1 were used to yield calibration equations (IR measurements as dependent and refractometry measurements as independent variables), which were used to modify the IR measurements on day-2. RESULTS Bias ± limits of agreement for isoflurane, sevoflurane and desflurane were 0.2 ± 0.3, 0.1 ± 0.4 and 0.7 ± 0.9 volume%, respectively. There were significant linear relationships between differences and means for all agents. The IR analysers became less accurate at higher gas concentrations. After customized calibration, the bias became almost zero and the limits of agreement became narrower. CONCLUSIONS AND CLINICAL RELEVANCE If similar IR analysers are used in research studies, they need to be calibrated against a reference method using the agent in question at multiple calibration points overlapping the range of interest.