Cyclic and constant hyperoxia cause inflammation, apoptosis and cell death in human umbilical vein endothelial cells.

BACKGROUND: Perioperative high-dose oxygen (O2 ) exposure can cause hyperoxia. While the effect of constant hyperoxia on the vascular endothelium has been investigated to some extent, the impact of cyclic hyperoxia largely remains unknown. We hypothesized that cyclic hyperoxia would induce more injury than constant hyperoxia to human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were exposed to cyclic hyperoxia (5-95% O2 ) or constant hyperoxia (95% O2 ), normoxia (21% O2 ), and hypoxia (5% O2 ). Cell growth, viability (Annexin V/propidium iodide and 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide, MTT) lactate dehydrogenase (LDH), release, cytokine (interleukin, IL and macrophage migration inhibitory factor, MIF) release, total antioxidant capacity (TAC), and superoxide dismutase activity (SOD) of cell lysate were assessed at baseline and 8, 24, and 72 h. A signal transduction pathway finder array for gene expression analysis was performed after 8 h. RESULTS: Constant and cyclic hyperoxia-induced gradually detrimental effects on HUVECs. After 72 h, constant or cyclic hyperoxia exposure induced change in cytotoxic (LDH +12%, P = 0.026; apoptosis +121/61%, P < 0.01; alive cells -15%, P < 0.01; MTT -16/15%, P < 0.01), inflammatory (IL-6 +142/190%, P < 0.01; IL-8 +72/43%, P < 0.01; MIF +147/93%, P < 0.01), or redox-sensitive (SOD +278%, TAC-25% P < 0.01) markers. Gene expression analysis revealed that constant and cyclic hyperoxia exposure differently activates oxidative stress, nuclear factor kappa B, Notch, and peroxisome proliferator-activated receptor pathways. CONCLUSIONS: Extreme hyperoxia exposure induces inflammation, apoptosis and cell death in HUVECs. Although our findings cannot be transferred to clinical settings, results suggest that hyperoxia exposure may cause vascular injury that could play a role in determining perioperative outcome.

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.

In vitro and in vivo evaluation of a new large animal spirometry device using mainstream CO2 flow sensors.

REASONS FOR PERFORMING STUDY: A spirometry device equipped with mainstream CO2 flow sensor is not available for large animal anaesthesia. OBJECTIVES: To measure the resistance of a new large animal spirometry device and assess its agreement with reference methods for volume measurements. STUDY DESIGN: In vitro experiment and crossover study using anaesthetised horses. METHODS: A flow partitioning device (FPD) equipped with 4 human CO2 flow sensors was tested. Pressure differences were measured across the whole FPD and across each sensor separately using air flows (range: 90-720 l/min). One sensor was connected to a spirometry monitor for in vitro volume (3, 5 and 7 l) measurements. These measurements were compared with a reference method. Five anaesthetised horses were used for tidal volume (VT) measurements using the FPD and a horse-lite sensor (reference method). Bland-Altman analysis, ANOVA and linear regression analysis were used for data analysis. RESULTS: Pressure differences across each sensor were similar suggesting equal flow partitioning. The resistance of the device increased with flow (range: 0.3-1.5 cmH2 O s/l) and was higher than that of the horse-lite. The limits of agreement for volume measurements were within -1 and 2% in vitro and -12 and 0% in vivo. Nine of 147 VT measurements in horses were outside of the ± 10% limits of acceptance but most of these erroneous measurements occurred with VTs lower than 4 l. The determined correction factor for volume measurements was 3.97 ± 0.03. CONCLUSIONS: The limits of agreement for volume measurements by the new device were within ± 10% using clinically relevant range of volumes. The new spirometry device can be recommended for measurement of VT in adult Warmblood horses.

Breathing pattern and thoracoabdominal asynchrony in horses with chronic obstructive and inflammatory lung disease.

The aim of the study was to show that changes in thoracoabdominal asynchrony (TAA) between quiet breathing and CO2-induced hyperpnoea can be used to differentiate between horses with healthy airways and those suffering from inflammatory airway disease (IAD) or recurrent airway obstruction (RAO). The level of TAA was displayed by the Pearson's correlation coefficient (PCC) of thoracic and abdominal signals, generated by respiratory ultrasonic plethysmography (RUP) during quiet breathing and hyperpnoea. Changes in TAA were expressed as the quotient of the PCCs (PCCQ) during normal breathing and hyperpnoea. Horses with RAO and IAD showed significant higher median PCCQ than healthy horses. Median PCCQ of horses with RAO and IAD was not significantly different. Horses affected by a pulmonary disorder showed lower TAA compared to the control group. This study suggests that TAA provides a useful parameter to differentiate horses with RAO and IAD from healthy horses.

The effect of airflow on thermographically determined temperature of the distal forelimb of the horse.

REASONS FOR PERFORMING STUDY: Current literature suggests that thermographic imaging of horses should be performed in a draught-free room. However, studies on the effect of airflow on determined temperature have not been published. OBJECTIVES: To investigate effects of airflow on thermographically determined temperature of horses' forelimbs; to assess the relationship of wind velocity, rectal temperature, ambient temperature and humidity. METHODS: Thermographic images were obtained for the forelimbs of 6 horses in a draught-free room. Three replicates (R) with defined wind velocities (R1, 0.5-1.0 m/s; R2, 1.3-2.6 m/s; and R3, 3.0-4.0 m/s) were conducted. Each replicate consisted of a baseline image, a 15 min phase with the wind on and a 15 min phase with the wind off. We exposed only the right leg to airflow and determined the temperature by thermography with the wind on and wind off. Temperature differences between baseline and wind on, between wind on and wind off and between different wind velocities were analysed by a general linear model, Student's paired t test and ANOVA. RESULTS: After the onset of wind, the temperature on the right forelimb decreased within 1-3 min (by approximately 0.6°C at R1, 1.5°C at R2 and 2.1°C at R3). With the wind off, the temperature increased within 3 min (by approximately 1.2°C at R1, 1.7°C at R2 and 2.1°C at R3). With increasing wind velocity, the temperature differences between baseline and wind on and between wind on and wind off increased significantly. CONCLUSIONS: Barely noticeable wind velocities caused a decrease in thermographically determined temperatures of the forelimbs of the horse. Further research is required to assess the influence of airflow on other parts of the body and at different ambient temperatures, as well as the effect on horses with inflammatory lesions, especially of the distal limbs. POTENTIAL RELEVANCE: It is essential for practitioners to perform thermography on horses in a draught-free environment in order to avoid false-positive or -negative diagnoses.