Effectiveness of furosemide in attenuating exercise-induced pulmonary haemorrhage in horses when administered at 4- and 24-h prior to high-speed training.

BACKGROUND: Due to the high prevalence of EIPH in racehorses and its potential impact on the horse's health, furosemide administration is permitted up to 4-h prior to post time in most North American racing jurisdictions. Anecdotal reports suggest that administration of furosemide 24-h prior to strenuous exercise may be equally effective in decreasing the severity of EIPH. OBJECTIVES: To 1) compare the efficacy of furosemide in reducing the presence and severity of EIPH when administered 4- or 24-h prior to strenuous exercise 2) characterise electrolyte and blood parameters following administration of furosemide at 4- and 24-h prior to exercise. STUDY DESIGN: 3-way crossover. METHODS: Fifteen Thoroughbred racehorses received 5 mL of 0.9% NaCl or 250 mg of furosemide either 4- or 24-h prior to a 5-furlong simulated race. Blood samples were collected prior to and post-run for determination of furosemide, lactate, haemoglobin and electrolyte concentrations. One-hour post-race, an endoscopic exam and bronchoalveolar lavage (BAL) were performed. Horses were assigned an EIPH score based on predetermined criteria and the number of red blood cells in BAL fluid was determined. RESULTS: Endoscopic EIPH scores were lower in the 4-h vs. the 24-h (P = 0.03) furosemide groups. RBC counts in BAL fluid were lower in the 4-h furosemide vs. saline treatment groups (P = 0.01) but no difference was noted between the saline and 24-h furosemide groups (P = 0.3), nor between the 4- and 24-h groups (P = 0.5). MAIN LIMITATIONS: Small sample size and large range of running times for the 5-furlong work. CONCLUSIONS: While none of the treatments prevented EIPH, endoscopic scores and RBC counts in BAL fluid support the efficacy of furosemide in reducing the severity of EIPH. Endoscopic scores were lower in the 4-h furosemide group compared with 24-h administration. Red blood cell counts were lower in the 4-h furosemide group compared with saline treatment.

Pharmacokinetics of furosemide administered 4 and 24 hours prior to high-speed exercise in horses.

Furosemide is a diuretic agent used commonly in racehorses to attenuate the bleeding associated with exercise-induced pulmonary hemorrhage (EIPH). The current study describes serum and urine concentrations and the pharmacokinetics of furosemide following administration at 4 and 24 hrs prior to maximal exercise. Eight exercised adult Thoroughbred horses received a single IV administration of 250 mg of furosemide at 4 and 24 hrs prior to maximal exercise on a high-speed treadmill. Blood and urine samples were collected at time 0 and at various times for up to 72 hrs and furosemide concentrations determined using liquid chromatography-tandem mass spectrometry. Serum furosemide concentrations remained above the LOQ (0.05 ng/ml) for 36 hrs in 3/8 and 1/8 horses in the 4- and 24-hrs groups, respectively. Serum concentration data were best fit by a two-compartment model. There was not a significant difference in the volume of distribution at steady-state (0.594 ± 0.178 [4 hrs] and 0.648 ± 0.147 [24 hrs] L/kg) or systemic clearance (0.541 ± 0.094 [4 hrs] and 0.617 ± 0.114 [24 hrs] L/hrs/kg) between horses that were exercised at 4- and 24 hrs postdrug administration. The mean ± SD elimination half-life was 3.12 ± 0.387 and 3.23 ± 0.407 hrs following administration at 4 and 24 hrs prior to exercise, respectively.

Disease ecology, health and the environment: a framework to account for ecological and socio-economic drivers in the control of neglected tropical diseases.

Reducing the burden of neglected tropical diseases (NTDs) is one of the key strategic targets advanced by the Sustainable Development Goals. Despite the unprecedented effort deployed for NTD elimination in the past decade, their control, mainly through drug administration, remains particularly challenging: persistent poverty and repeated exposure to pathogens embedded in the environment limit the efficacy of strategies focused exclusively on human treatment or medical care. Here, we present a simple modelling framework to illustrate the relative role of ecological and socio-economic drivers of environmentally transmitted parasites and pathogens. Through the analysis of system dynamics, we show that periodic drug treatments that lead to the elimination of directly transmitted diseases may fail to do so in the case of human pathogens with an environmental reservoir. Control of environmentally transmitted diseases can be more effective when human treatment is complemented with interventions targeting the environmental reservoir of the pathogen. We present mechanisms through which the environment can influence the dynamics of poverty via disease feedbacks. For illustration, we present the case studies of Buruli ulcer and schistosomiasis, two devastating waterborne NTDs for which control is particularly challenging.This article is part of the themed issue 'Conservation, biodiversity and infectious disease: scientific evidence and policy implications'.

The evolutionary dynamics of timing of maternal immunity: evaluating the role of age-specific mortality.

If a female survives an infection, she can transfer antibodies against that particular pathogen to any future offspring she produces. The resulting protection of offspring for a period after their birth is termed maternal immunity. Because infection in newborns is associated with high mortality, the duration of this protection is expected to be under strong selection. Evolutionary modelling structured around a trade-off between fertility and duration of maternal immunity has indicated selection for longer duration of maternal immunity for hosts with longer lifespans. Here, we use a new modelling framework to extend this analysis to consider characteristics of pathogens (and hosts) in further detail. Importantly, given the challenges in characterizing trade-offs linked to immune function empirically, our model makes no assumptions about costs of longer lasting maternal immunity. Rather, a key component of this analysis is variation in mortality over age. We found that the optimal duration of maternal immunity is shaped by the shifting balance of the burden of infection between young and old individuals. As age of infection depends on characteristics of both the host and the pathogen, both affect the evolution of duration of maternal immunity. Our analysis provides additional support for selection for longer duration of maternal immunity in long-lived hosts, even in the absence of explicit costs linked to duration of maternal immunity. Further, the scope of our results provides explanations for exceptions to the general correlation between duration of maternal immunity and lifespan, as we found that both pathogen characteristics and trans-generational effects can lead to important shifts in fitness linked to maternal immunity. Finally, our analysis points to new directions for quantifying the trade-offs that drive the development of the immune system.

Mars atmosphere. The imprint of atmospheric evolution in the D/H of Hesperian clay minerals on Mars.

The deuterium-to-hydrogen (D/H) ratio in strongly bound water or hydroxyl groups in ancient martian clays retains the imprint of the water of formation of these minerals. Curiosity's Sample Analysis at Mars (SAM) experiment measured thermally evolved water and hydrogen gas released between 550° and 950°C from samples of Hesperian-era Gale crater smectite to determine this isotope ratio. The D/H value is 3.0 (±0.2) times the ratio in standard mean ocean water. The D/H ratio in this ~3-billion-year-old mudstone, which is half that of the present martian atmosphere but substantially higher than that expected in very early Mars, indicates an extended history of hydrogen escape and desiccation of the planet.

Effects of three warm-up regimens of equal distance on VO2 kinetics during supramaximal exercise in Thoroughbred horses.

REASONS FOR PERFORMING STUDY: Several studies have indicated that even low-intensity warm-up increases O(2) transport kinetics and that high-intensity warm-up may not be needed in horses. However, conventional warm-up exercise for Thoroughbred races is more intense than those utilised in previous studies of equine warm-up responses. OBJECTIVES: To test the hypothesis that warm-up exercise at different intensities alters the kinetics and total contribution of aerobic power to total metabolic power in subsequent supramaximal (sprint) exercise in Thoroughbred horses. METHODS: Nine well-trained Thoroughbreds ran until fatigue at 115% of maximal oxygen consumption (VO2max) 10 min after warming-up under each of 3 protocols of equal running distance: 400 s at 30% VO2max (LoWU), 200 s at 60% VO2max (MoWU) and 120 s at 100% VO2max (HiWU). Variables measured during exercise were rates of O(2) and CO(2) consumption/production (VO2,VO2), respiratory exchange ratio (RER), heart rate, blood lactate concentration and accumulation rate and blood gas variables. RESULTS: VO2 was significantly higher in HiWU than in LoWU at the onset of the sprint exercise and HR was significantly higher in HiWU than in LoWU throughout the sprint. Accumulation of blood lactate, RER, P(a)CO(2) and PvCO2 in the first 60 s were significantly lower in HiWU than in LoWU and MoWU. There were no significant differences in stroke volume, run time or arterial-mixed venous O(2) concentration. CONCLUSIONS: These results suggest HiWU accelerates kinetics and reduces reliance on net anaerobic power compared with LoWU at the onset of the subsequent sprint.

Comparison of net anaerobic energy utilisation estimated by plasma lactate accumulation rate and accumulated oxygen deficit in Thoroughbred horses.

REASONS FOR PERFORMING STUDY: Accumulated O(2) deficit (AOD) and plasma lactate accumulation rate (PLAR) are alternative methods for estimating net anaerobic energy utilisation (NAEU) in exercising horses. How they compare or their accuracy is unknown. OBJECTIVES: We hypothesised net anaerobic energy utilisation calculated by PLAR (NAUE(PLAR)) is equivalent to NAUE estimated by AOD (NAUE(AOD)). METHODS: Six Thoroughbred horses ran at identical supramaximal speeds (118% aerobic capacity) until exhaustion for 2 runs while breathing normoxic (NO, 21% O(2)) or hyperoxic (HO, 26% O(2)) gas. Jugular blood was sampled at 15 s intervals to measure plasma lactate concentration. Horses also ran at incremental submaximal speeds from 1.7-11.0 m/s to determine the linear relationship between speed and O(2) consumption to estimate O(2) demand for AOD calculations. RESULTS: Maximum O(2) consumption of horses increased 11.6 ± 2.3% in HO and NAEU(PLAR) and NAUE(AOD) decreased 38.5 ± 8.0% and 46.2 ± 17.7%, respectively. The NAEU(PLAR) in NO was 114.5 ± 27.4 mlO(2) (STPD) equivalent/kg bwt contributing 23.5 ± 3.7% to total energy turnover and in HO was 70.9 ± 19.8 mlO(2) (STPD) equivalent/kg bwt contributing 14.6 ± 3.8% to total energy turnover. The NAUE(AOD) in NO was 88.6 ± 24.3 mlO(2) (STPD) equivalent/kg bwt contributing 19.9 ± 2.1% to total energy turnover and in HO was 56.2 ± 19.1 mlO(2) (STPD) equivalent/kg bwt contributing 10.9 ± 4.3% to total energy turnover. Overall, NAEU(AOD) was systematically biased -23.5 ± 16.8 mlO(2) (STPD) equivalent/kg bwt below NAEU(PLAR). Total energy demand estimated by PLAR was 11.1 ± 5.4% greater than that estimated by AOD and was higher in every horse. CONCLUSIONS: The NAUE(PLAR) estimates average 40.0 ± 29.6% higher than NAUE(AOD) and are highly correlated (r(2) = 0.734), indicating both indices are sensitive to similar changes in NAEU. Accuracy of the estimates remains to be determined. Multiple considerations suggest NAUE(AOD) may underestimate total energy cost during high-speed galloping, thus biasing low the AOD estimate of NAEU.

The effect of sildenafil citrate administration on selected physiological parameters of exercising Thoroughbred horses.

REASONS FOR PERFORMING STUDY: Sildenafil, a phosphodiesterase-5 inhibitor vasodilator, increases cGMP concentrations by inhibiting enzymatic degradation. Marketed to treat erectile dysfunction in men, it also reduces pulmonary arterial pressure (PAP). Because it reduces PAP, sildenafil may enhance performance and/or prevent exercise induced-pulmonary haemorrhage (EIPH). OBJECTIVE: To determine if sildenafil citrate administration altered commonly measured indices of performance or reduced EIPH in exercised horses. METHODS: Thirteen athletically conditioned Thoroughbred horses (2 mares and 11 geldings, age 3-12 years) were administered sildenafil citrate or placebo in 2 crossover design exercise testing studies. In a step-wise test to exhaustion, inspired/expired gas analysis, blood lactate, heart rate, runtime and bronchoalveolar lavage (BAL) cytology were measured. In a 13 m/s test to exhaustion, blood lactate, heart rate, runtime, BAL cytology and pulmonary arterial pressure were measured. Data were analysed with paired and unpaired t tests, one-way ANOVA and Tukey's pair-wise multiple comparison and Friedman repeated measure analysis of variance on ranks. RESULTS: The administration of sildenafil did not alter mean inspired/expired gas measurements, plasma lactate concentrations or acute pulmonary haemorrhage in either exercise test or pulmonary arterial pressure measurement in the 13 m/s trial. Heart rates in both stress tests were significantly different at submaximal speeds and during the early recovery period. Run time was not affected by sildenafil administration in the step-wise trial (P = 0.622) or in the 13 m/s trial (P = 0.059). CONCLUSIONS: Sildenafil did not alleviate pulmonary haemorrhage or enhance performance-related indices in these trials. Sildenafil administration altered cardiovascular adaptation to intense exercise as evidenced by altered heart rates at submaximal speeds and post exercise. The effect of these alterations on other performance perimeters was not evident.

The "fire stick farming" hypothesis: Australian Aboriginal foraging strategies, biodiversity, and anthropogenic fire mosaics.

Aboriginal burning in Australia has long been assumed to be a "resource management" strategy, but no quantitative tests of this hypothesis have ever been conducted. We combine ethnographic observations of contemporary Aboriginal hunting and burning with satellite image analysis of anthropogenic and natural landscape structure to demonstrate the processes through which Aboriginal burning shapes arid-zone vegetational diversity. Anthropogenic landscapes contain a greater diversity of successional stages than landscapes under a lightning fire regime, and differences are of scale, not of kind. Landscape scale is directly linked to foraging for small, burrowed prey (monitor lizards), which is a specialty of Aboriginal women. The maintenance of small-scale habitat mosaics increases small-animal hunting productivity. These results have implications for understanding the unique biodiversity of the Australian continent, through time and space. In particular, anthropogenic influences on the habitat structure of paleolandscapes are likely to be spatially localized and linked to less mobile, "broad-spectrum" foraging economies.

Effect of high versus low initial doses of L-thyroxine for congenital hypothyroidism on thyroid function and somatic growth.

BACKGROUND AND AIMS: The optimal dose of thyroxine (T4) in congenital hypothyroidism (CH) during infancy is controversial. Higher doses lead to improvement in cognitive scores, but have been linked to later behavioural difficulties. We have examined the effects of initial T4 dosage on somatic growth--a putative surrogate marker of overtreatment. METHODS: 314 CH children (214 girls, 100 boys) were analysed according to initial daily dose of T4: Group 1 (25 mug, n = 152), Group 2 (30-40 mug, n = 63) and Group 3 (50 mug, n = 99). Thyroid function and weight, length and occipito-frontal head circumference (OFC) standard deviation score (SDS) were compared at 3, 6, 12, 18, 24 and 36 months of age. Linear growth SDS was compared between the three groups using a regression adjustment model at 12 and 18 months of age using birth weight and 3-month data as baselines. Thyroid function was also compared at diagnosis (T 0), and 7-21 days after the start of treatment (T1). RESULTS: At T1 median thyroid stimulating hormone (TSH) for Groups 1, 2 and 3 was 58, 29 and 4.1 mU/l, respectively (p<0.001), Group 3 values remaining significantly lower at 3 and 6 months. Median free T4 (fT4) was within or just above the reference range in all groups at T1, but 7.4% of Group 1 had values <9 pmol/l compared with 5.1% and 0% for Groups 2 and 3, respectively. At 3 months weight, length and OFC SDS values were -0.39, -0.35, 0.09; -0.30, -0.47, 0.32; and -0.03, -0.13, 0.18 for Groups 1, 2 and 3, respectively, indicating relatively large OFC in all infants. A regression adjustment model showed no significant difference in growth rate from baseline and 12 or 18 months of age, between the three groups. CONCLUSION: An initial T4 dose of 50 mug daily, normalises thyroid function several months earlier than lower-dose regimes, with no evidence of sustained somatic overgrowth between 3 months and 3 years.

Combined ultrasound and isotope scanning is more informative in the diagnosis of congenital hypothyroidism than single scanning.

BACKGROUND: Thyroid imaging is helpful in confirming the diagnosis of congenital hypothyroidism and in establishing the aetiology. Although isotope scanning is the standard method of imaging, ultrasound assessment may be complementary. AIM: To determine the strengths and weaknesses of thyroid ultrasound and isotope scanning in neonates with thyroid stimulating hormone (TSH) elevation. METHODS: Babies from the West of Scotland with raised capillary TSH (>15 mU/l) on neonatal screening between January 1999 and 2004 were recruited. Thyroid dimensions were measured using ultrasonography, and volumes were calculated. Isotope scanning was carried out with a pinhole collimator after an intravenous injection of 99m-technetium pertechnetate. RESULTS: 40 infants (29 female) underwent scanning at a median of 17 days (range 12 days to 15 months). The final diagnosis was athyreosis (n = 11), ectopia (n = 12), hypoplasia (n = 8; 3 cases of hemi-agenesis), dyshormonogenesis (n = 5), transient hypothyroidism (n = 2), transient hyperthyrotropinaemia (n = 1) and uncertain status with gland in situ (n = 1). 6 infants had discordant scans with no isotope uptake but visualisation of thyroid tissue on ultrasound. This was attributed to TSH suppression from thyroxine (n = 3); maternal blocking antibodies (n = 1); cystic degeneration of the thyroid (n = 1); and possible TSH receptor defect (n = 1). CONCLUSIONS: Isotope scanning was superior to ultrasound in the detection of ectopic tissue. However, ultrasound detected tissue that was not visualised on isotope scanning, and showed abnormalities of thyroid volume and morphology. We would therefore advocate dual scanning in newborns with TSH elevation as each modality provides different information.

Improvement in screening performance and diagnosis of congenital hypothyroidism in Scotland 1979-2003.

AIM: To assess the Scottish newborn screening programme for congenital hypothyroidism from 1994 to 2003 (period 2) for performance and compare with an initial audit covering 1979 to 1993 (period 1). DESIGN: Performance data-age at blood spot sampling, notification by screening laboratory, start of treatment, and the prevalence of late testing, notification or treatment-were compared, together with the incidence of congenital hypothyroidism. RESULTS: Comparing data for period 2 with period 1, the mean annual incidence of true congenital hypothyroidism was 1:3655 live births v 1:4363. Median age for Guthrie sampling (all referrals) was 6 v 7 days (p<0.0001). Late sampling (>10 days) had fallen from 10.7% to 7%. For infants requiring repeat sampling before notification, the median (range) interval between initial and final repeat samples was 11 (1 to 52) compared with 14 (3 to 73) days. Median age at notification for true congenital hypothyroidism was 10 v 12 days (p <0.0001). Late notification (>15 days) was justifiable (mild TSH elevation) in 10 of 13 patients in period 2. Median age at start of treatment for true congenital hypothyroidism had improved to 11 days from 13.5 days. For true congenital hypothyroidism, late treatment (>16 days) occurred in 7% of patients compared with 19% (p<0.0001). CONCLUSIONS: There has been an improvement in performance measures for the congenital hypothyroidism screening programme in Scotland. However, late sampling, occurring primarily in inpatients and which is never justified, remains a problem, while the interval between initial and recall sampling is a further source of delay.

Effect of detraining on cardiorespiratory variables in young thoroughbred horses.

REASONS FOR PERFORMING STUDY: Thoroughbred racehorses often experience interruptions to their training. Identifying the effects of these changes and how they alter athletic performance might provide an insight on to how to prevent these changes from occurring. HYPOTHESIS: Training and detraining young Thoroughbreds alters their aerobic capacities with correlated changes in circulatory capacities; if horses remained spontaneously active in a pasture during their detraining period, their decreases in aerobic capacity during detraining would be reduced. METHODS: We trained 6 Thoroughbred yearlings for 6 months using a conventional yearling race training programme. They were then detrained for 10 weeks with free range on pasture for 8 h/day and stall rest at night. Treadmill measurements of O2 transport variables were made before training (PRE), after training (TR) and after detraining (DT). A step-test protocol identified each horse's aerobic capacity (VO2max) and speed to attain it, and a steady-state run at VO2max was used to quantify 02 transport variables at each time period. RESULTS: The mass-specific and whole-body VO2max, cardiac output (Q) and stroke volume (Vs) increased from PRE to TR. All mass-specific values decreased significantly from TR to DT; however, because body mass increased by 8.3% from TR to DT, none of the variables changed significantly from TR to DT on a whole-body basis. CONCLUSIONS: Changes in aerobic capacity are highly correlated with changes in Vs and circulatory capacity during training and detraining. Exercise activity of trained young horses free at pasture for 8 h/day is sufficient to maintain VO2max, (Q and Vs during 10 weeks of DT. POTENTIAL RELEVANCE: Aerobic and cardiovascular fitness may be maintained in young Thoroughbred horses during at least 10 weeks of detraining by maintaining modest spontaneous exercise activity.

Energetic cost of locomotion on different equine treadmills.

REASON FOR PERFORMING STUDY: Human athletes run faster and experience fewer injuries when running on surfaces with a stiffness 'tuned' to their bodies. We questioned if the same might be true for horses, and if so, would running on surfaces of different stiffness cause a measurable change in the amount of energy required to move at a given speed? HYPOTHESIS: Different brands of commercial treadmills have pans of unequal stiffness, and this difference would result in different metabolic power requirements to locomote at a given speed. METHODS: We tested for differences in stiffness between a Mustang 2200 and a Säto I commercial treadmill by incrementally loading each treadmill near the centre of the pan with fixed weights and measuring the displacement of the pan as weights were added or removed from the pan. We trained six 3-year-old Thoroughbreds to run on the 2 treadmills. After 4 months the horses ran with reproducible specific maximum rates of O2 consumption (VO2max/kg bwt, 2.62 +/- 0.23 (s.d.) mlO2 STPD/sec/kg) at 14.2 +/- 0.7 (s.d.) m/sec. They were alternately run on the 2 treadmills at identical grade (0.40 +/- 0.02%) and speeds (1.83 (walk), 4.0 (trot) and 8.0 (canter) m/sec, all +/- 0.03 m/sec) while wearing an open-flow mask for measurement of VO2. RESULTS: The Mustang treadmill was over 6 times stiffer than the Säto. The VO2/kg bwt increased by approximately 4-fold over the range of speeds studied on both treadmills. Oxygen consumption was significantly lower at all speeds for the Mustang treadmill compared to the Säto. The fractional difference in energy cost decreased by a factor of 6 with increasing speed, although absolute difference in cost was relatively constant. CONCLUSIONS: We suggest it costs less energy for horses to walk, trot or canter on a stiffer treadmill than on a more compliant treadmill, at least within the ranges of stiffness evaluated. POTENTIAL RELEVANCE: It may be possible to define a substrate stiffness 'tuned' to a horse's body enabling maximal energetic economy when running. The differences between treadmills allows more accurate comparisons between physiological studies conducted on treadmills of different stiffness, and might help to identify an ideal track stiffness to reduce locomotor injuries in equine athletes.

Method for quantifying net anaerobic power in exercising horses.

REASON FOR PERFORMING STUDY: There is no good method for measuring net anaerobic power in exercising horses to allow accurate estimates of total metabolic power. HYPOTHESIS: The increase in VO2max when breathing hyperoxic (HO) gas should be accompanied by a stoichiometrically equal (in terms of ATP turnover, i.e. energy equivalents) decrease in plasma lactate accumulation rate (Mlactate). METHODS: Six 3-year-old Thoroughbreds were trained on an equine treadmill wearing a semi-open flow mask for measurement of VO2. After 4 months the horses ran with reproducible specific VO2max (VO2max/kg bwt). The mask design allowed mixing of O2 or N2 with the inward bias flow of gas so that inspired O2 concentration of the horse could be controlled. While the horse breathed either HO (25.1% O2), normoxic (NO, 21% O2) or hypoxic (LO, 19.5% O2) gas, it ran at a speed sufficient to elicit VO2max in NO while jugular venous blood was drawn at 15 sec intervals over a period of 2 min to determine Mlactate. RESULTS: VO2max/kg bwt was not significantly different between LO and NO conditions, and LO data could not be used in the comparison. The VO2max/kg bwt increased from 2.59 +/- 0.24 (s.d.) to 2.86 +/- 0.24 mlO2 (STPD)/sec/kg in NO and HO, respectively, while Mlactate decreased from 11.5 +/- 4.2 to 9.0 +/- 3.9 mmol/min as VO2 increased. CONCLUSIONS: The ratio of delta Mlactate to delta VO2max/kg bwt suggests that Mlactate of approx 11.1 +/- 6.7 mmol/min is associated with net anaerobic power approximately equivalent to 1.0 mlO2 (STPD)/sec/kg of aerobic power (20.1 W/kg(-1)). The high variability in VO2max/kg bwt observed in data from some runs, particularly in LO, suggests that caution must be used when comparing data from the same horse during different runs. POTENTIAL RELEVANCE: This study provides a tool for estimating net anaerobic power and, more accurately, evaluating total metabolic power of horses exercising at or above their aerobic capacities.

Mucociliary transport determined by in vivo microdialysis in the airways of normal and CF mice.

We report a novel method to measure mucociliary transport (MCT) in both the upper and lower airways of normal and CF mice. The in vivo microdialysis technique involves placing a small quantity of dye on the airway surface and a microdialysis probe a defined distance from the site of dye deposition. The dye is transported toward the probe by ciliary transport and, upon reaching the microdialysis probe, diffuses across the dialysis membrane and is collected in the dialysate leaving the probe. The rate of MCT is calculated from the length of time from dye deposition to recovery. The rate of tracheal MCT in normal mice was 2.2 +/- 0.45 (SE) mm/min (n = 6), a value similar to that in reports using other techniques. MCT in CF mice was not different (2.3 +/- 0.29, n = 6), consistent with previous observations suggesting that tracheal ion transport properties are not different between CF and normal mice. The rate of MCT in the nasal cavity of normal mice was slower than in the trachea (1.3 +/- 0.26, n = 4). MCT in the CF mouse nasal cavity (1.4 +/- 0.31, n = 8), a region in which the CF mouse exhibits bioelectric properties similar to the human CF patient, was, again, not different from the normal mouse, perhaps reflecting copious gland secretion offsetting Na(+) and liquid hyperabsorption. In conclusion, we have developed a versatile, simple in vivo method to measure MCT in both upper and lower airways of mice and larger animals.

Physiological responses of young Thoroughbreds during their first year of race training.

Yearling horses are typically trained for more than a year before they begin racing; therefore, we questioned how relevant analyses of the initial responses to training are compared to physiological responses that occur over a year of training, and whether young horses with no history of training would respond the same as older horses that had been trained previously. We hypothesised that changes in O2 transport over the last months of a year of training would be different than at the beginning. We trained 5 yearling Thoroughbreds and evaluated metabolism, O2 transport and echocardiograms. Measurements were made before breaking (T1), after 6 months of training (T2) and following an additional 4 months of training (T3). We compared 5 trained horses (TR) with 5 untrained (UT) sex-, size- and age-matched yearlings kept at pasture and in boxes. Satellite telemetry indicated UT moved less total daily distance than TR during winter and more during summer, but UT walked for 80% of their distance, TR only 25%. The UT increased body mass (Mb) after T1 by 13% and were significantly heavier and fatter than TR. Specific aerobic capacity (VO2max/Mb) increased by 16% in both groups at T2, but by T3 was not different from T1 in UT, but was higher in TR (19%>T1, 15%>UT). In TR, specific cardiac output (Q/Mb) increased by 13% at T2, and specific stroke volume (V(S)/Mb) were larger at T2 and T3 than T1 and UT at the same times both by physiological (15-16%) and echocardiographical (22-23%) estimates. Increased Vs was a primary correlate of the sustained increase in VO2max/Mb in TR. The large increases in V(S) and VO2max had occurred by T2 and changed only slightly by T3.

Does training affect mRNA transciption for cytokine production in circulating leucocytes?

Physical exertion is a stimulus for the upregulation of cytokine production including IL-1beta, IL-1ra, IL-2, IL-4, IL-6, IL-10 and TNF-alpha in horses. To investigate that hypothesis, we initiated training of 5 stall-rested Thoroughbreds. Blood samples were drawn before and weekly during training. The relative transcription of mRNA within the leucocytes was measured using real time TaqMan quantitative PCR. The training protocol was walking (3 min), trotting (3 min) and cantering/galloping (6 min) increasing in intensity weekly (6 to 12 m/s) and culminating in an intense exercise period. Comparisons of mRNA concentrations were made using a repeated measures ANOVA on ranks and a Student-Newman-Keuls pair-wise multiple comparison (P<0.05). The training programme or intense exercise bout did not affect IL-2, IL-4, IL-6 and IL-10. IL1-beta and TNF-alpha transcription increased on Day 23. TNF-alpha peaked on Day 23 and IL1-beta on Day 30. Neither demonstrated a response to intense exercise. IL-1ra decreased significantly on Day 9; rose significantly from Day 9 to Days 16 and 23; remained significantly elevated through Days 30 and 37 over Day 9, and rose very slightly after intense exercise on Day 56. Alterations in leukocyte cytokine responses may influence susceptibility to infectious disease, metabolic responses to exercise or exercised induced syndromes.

Heterogeneity of intrapleural pressures during exercise.

We determined whether the caudodorsal region of the intrapleural space in exercising horses experiences larger pressure fluctuations than other regions and whether systematic phase-shifting of peak intrapleural pressures along the length of the thorax suggests the existence of locomotor-induced intrapleural pressure waves. We utilised percutaneous introducers and solid-state pressure-tip transducers implanted along the dorsal aspect of the thorax, mid-thorax or oesophagus to measure regional intrapleural pressures while 3 horses galloped on a flat treadmill at 13-14 m/s, then recorded pressures from the same catheters when horses exercised intensely (heart rate 170-190 beats/min) while swimming with no ground concussion. Pressure excursions in the caudodorsal region did not vary systematically from other regions during galloping or swimming, nor more than a few torr between different locations. During swimming, peak expiratory pressures were higher than during galloping (68-79 vs. 26-32 torr), and horses breathed explosively at frequencies 5 times slower than while galloping (28 vs. 120/min). During galloping, individual catheter locations registered locomotor concussion; however, this was variable and did not indicate a systematic pressure wave passing through the lung or intrapleural space.