Effects of a P-class CpG-ODN administered by intramuscular injection on plasma cytokines and on white blood cells of healthy horses.

Cytosine-phosphate-guanosine (CpG-ODN) has been described as a potent immunostimulatory agent in different species. No study reported the effect of a P-class CpG when administered systemically in healthy horses. The aim of this study was to evaluate the tolerance and the effect of an intramuscularly administered P-class CpG-ODN on hematology and on plasma cytokines (IFN-α, IL-10, TNF-α, IFN-γ) in 8 healthy horses. Intra-muscular CpG-ODN or placebo (PBS) was administered twice at a 7 days-interval. Groups were inversed after 2 months of washout period. A physical examination, complete blood count (CBC) and plasma cytokine measurements were performed from 2 days before injection up to 21 days after injection. P-class CpG-ODN injection was well tolerated with minor side effects. After the first injection a significant transient drop in circulating total leukocytes, lymphocytes and an increase in monocytes were observed. A transient drop in eosinophils was also noted after each CpG injection. P-class CpG-ODN at a dose of 5 mg did not create major side effects in 7 horses, one horse showed transient pyrexia. A redistribution of white blood cells was observed in horses receiving CpG, but no change in plasma cytokines was observed at the indicated dose, route of administration and sampling times.

Myeloid hypoxia-inducible factor 1α prevents airway allergy in mice through macrophage-mediated immunoregulation.

Hypoxia-inducible factor (HIF) has important roles in promoting pro-inflammatory and bactericidal functions in myeloid cells. Conditional genetic ablation of its major subunit Hif1α in the myeloid lineage consequently results in decreased inflammatory responses in classical models of acute inflammation in mice. By contrast, we report here that mice conditionally deficient for Hif1α in myeloid cells display enhanced sensitivity to the development of airway allergy to experimental allergens and house-dust mite antigens. We support that upon allergen exposure, MyD88-dependent upregulation of Hif1α boosts the expression of the immunosuppressive cytokine interleukin (IL)-10 by lung interstitial macrophages (IMs). Hif1α-dependent IL-10 secretion is required for IMs to block allergen-induced dendritic cell activation and consequently for preventing the development of allergen-specific T-helper cell responses upon allergen exposure. Thus, this study supports that, in addition to its known pro-inflammatory activities, myeloid Hif1α possesses immunoregulatory functions implicated in the prevention of airway allergy.

Detection of copy number variants in the horse genome and examination of their association with recurrent laryngeal neuropathy.

We used the data from a recently performed genome-wide association study using the Illumina Equine SNP50 beadchip for the detection of copy number variants (CNVs) and examined their association with recurrent laryngeal neuropathy (RLN), an important equine upper airway disease compromising performance. A total of 2797 CNVs were detected for 477 horses, covering 229 kb and seven SNPs on average. Overlapping CNVs were merged to define 478 CNV regions (CNVRs). CNVRs, particularly deletions, were shown to be significantly depleted in genes. Fifty-two of the 67 common CNVRs (frequency ≥ 1%) were validated by association mapping, Mendelian inheritance, and/or Mendelian inconsistencies. None of the 67 common CNVRs were significantly associated with RLN when accounting for multiple testing. However, a duplication on chromosome 10 was detected in 10 cases (representing three breeds) and two unphenotyped parents but in none of the controls. The duplication was embedded in an 8-Mb haplotype shared across breeds.

Serum concentration of surfactant protein D in horses with lower airway inflammation.

REASONS FOR PERFORMING STUDY: Surfactant protein D (SP-D), mainly synthesised by alveolar type II cells and nonciliated bronchiolar cells, is one important component of innate pulmonary immunity. In man, circulating concentrations of SP-D are routinely used as biomarkers for pulmonary injury. To date, serum SP-D levels have only been investigated in horses in an experimental model of bacterial airway infection. OBJECTIVES: To compare serum SP-D concentrations at rest and after exercise in horses with and without inflammatory airway disease (IAD). METHODS: Venous blood samples were collected from 42 Standardbred racehorses at rest and 60 min after performing a standardised treadmill exercise test. Tracheal wash and bronchoalveolar lavage fluid (BALF) samples were collected after exercise. Based on BALF cytology, 22 horses were defined as IAD-affected and 20 classified as controls. Serum SP-D concentrations were assessed using a commercially available ELISA kit and statistically compared between groups of horses and sampling times. RESULTS: Serum concentrations of SP-D in IAD-affected horses were significantly higher than those of control horses, both at rest and after exercise. Within the IAD-affected group, no significant correlation was found between serum SP-D concentrations and BALF cytology. Within each group of horses (IAD and control), no significant influence of exercise was found on serum SP-D levels. CONCLUSIONS: This is the first study determining serum SP-D concentrations in a noninfectious, naturally occurring form of lower airway inflammation in horses. The results highlight that IAD is associated with a detectable, though moderate, increase of circulating SP-D levels. POTENTIAL RELEVANCE: Serum concentration of surfactant protein D could represent a potentially valuable and readily accessible blood biomarker of equine lower airway inflammation.

Subclinical diseases underlying poor performance in endurance horses: diagnostic methods and predictive tests.

Thirty-eight endurance horses underwent clinical and ancillary examinations, including haematological and biochemical evaluation, standardised exercise tests both on a treadmill and in the field, Doppler echocardiography, impulse oscillometry, video endoscopy and collection of respiratory fluids. All of the examined poorly performing horses were affected by subclinical diseases, and most of them had multiple concomitant disorders. On the contrary, the well-performing horses were free of any subclinical disease. The most frequently diagnosed diseases were respiratory disorders, followed by musculoskeletal and cardiac problems. Poor performers exhibited lower speeds at blood lactate concentration of 4 mmol/l (VLA4) and at heart rates of 160 (V160) and 200 bpm (V200) on the treadmill and in the field, as well as slower recovery of heart rate.

Influence of subclinical inflammatory airway disease on equine respiratory function evaluated by impulse oscillometry.

REASONS FOR PERFORMING STUDY: Inflammatory airway disease (IAD) is a nonseptic condition of the lower respiratory tract. Its negative impact on respiratory function has previously been described using either forced expiration or forced oscillations techniques. However, sedation or drug-induced bronchoconstriction were usually required. The impulse oscillometry system (IOS) is a noninvasive and sensitive respiratory function test validated in horses, which could be useful to evaluate IAD-affected horses without further procedures. OBJECTIVES: To determine the sensitivity of IOS in detecting alterations of the respiratory function in subclinically IAD-affected horses without inducing bronchoprovocation and to characterise their respiratory impedance according to frequency for each respiratory phase. METHODS: Pulmonary function was evaluated at rest by IOS in 34 Standardbred trotters. According to the cytology of bronchoalveolar lavage fluid (BALF), 19 horses were defined as IAD-affected and 15 horses were used as control (CTL). Total respiratory resistance (Rrs) and reactance (Xrs) from 1-20 Hz as well as their inspiratory and expiratory components were compared between groups. RESULTS: A significant increase of Rrs at the lower frequencies (R1-10 Hz) as well as a significant decrease of Xrs beyond 5 Hz (X5-20 Hz) was observed in IAD compared to CTL horses. IOS-data was also significantly different between inspiration and expiration in IAD-affected horses. In the whole population, both BALF eosinophil and mast cell counts were significantly correlated with IOS measurements. CONCLUSIONS: Functional respiratory impairment may be measured, even in the absence of clinical signs of disease. In IAD-affected horses, the different parameters of respiratory function (Rrs or Xrs) may vary depending on the inflammatory cell profiles represented in BALF. POTENTIAL RELEVANCE: Impulse oscillometry could be used in a routine clinical setting as a noninvasive method for early detection of subclinical respiratory disease and of the results of treatment in horses.

Partial divergence of cytokine mRNA expression in bronchial tissues compared to bronchoalveolar lavage cells in horses with recurrent airway obstruction.

The aim of this study was to investigate mRNA levels of cytokines in bronchial epithelium in horses with recurrent airway obstruction (RAO) during acute crisis and remission. Additionally, cytokine mRNA levels in endobronchial biopsies and bronchoalveolar lavage (BAL) cells were compared. Seven RAO horses were examined while in respiratory crisis following provocation and again while in remission after 2 months on pasture, during which time six healthy horses on pasture were also examined. Quantitative real-time PCR (RT-PCR) was used to assess mRNA expression for cytokines IL-5, IL-6, IL-8, IL-10, IL-17 and transforming growth factor beta1 (TGF-beta1) in endobronchial biopsies and bronchoalveolar lavage. Expression of IL-8 mRNA was significantly upregulated during crisis in both endobronchial biopsies and BAL cells (p=0.036), while there was a similar trend for upregulation of IL-10 mRNA only in BAL cells that approached significance (p=0.059). Moreover, during crisis the expression of IL-8 mRNA in BAL cells was positively correlated to relative IL-6 mRNA expression (r(s)=0.971, p=0.001) and bronchial epithelial expression of IL-10 and TGF-beta1 mRNA were positively correlated (r(s)=0.943, p=0.005). In comparing the relationship of mRNA expression in BAL to biopsy in individual RAO horses, there was a positive correlation with IL-6 to IL-8 mRNA expression in BAL during respiratory crisis (r(s)=0.971, p=0.001) that also correlated positively with IL-8 expression in biopsies on pasture (r(s)=0.986, p<0.0001 for both). Regarding RAO horses at pasture versus controls neither the cytokine mRNA levels in endobronchial biopsy nor in BAL cells differed significantly. These results further support previous findings that IL-8 mRNA in both BAL cells and bronchial epithelium is upregulated in RAO horses during crisis. However, apart from IL-8, it appears that expression of other cytokines, including IL-5, IL-6, IL-10, IL-17 and TGF-beta1 in bronchial epithelium does not necessarily mirror cytokine expression in BAL cells in individual horses with RAO. Accordingly, examination of markers of inflammation in endobronchial tissue provides complementary but not necessarily identical information to that obtained in BAL cells. Given the potential for repeated sampling over time bronchial biopsy can serve as an invaluable additional tool for investigation of time-dependent changes in inflammatory process in this animal model of asthma.

Helenalin reduces Staphylococcus aureus infection in vitro and in vivo.

Staphylococcus (S.) aureus is a major udder pathogen causing bovine mastitis. Some pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha), enhance extracellular and intracellular growth of S. aureus, indicating that the inflammatory process favors S. aureus infection. Helenalin is a sesquiterpene lactone with potent anti-inflammatory properties. This study was designed to evaluate the effects of helenalin on S. aureus infection. First, in vitro experiments were conducted. These studies revealed that proliferation of S. aureus in bovine mammary epithelial MAC-T cells treated in the presence or absence of TNF-alpha was markedly reduced in the presence of helenalin. Secondly, in vivo effects of helenalin were investigated. Lactating mice treated in the presence or absence of helenalin were challenged by the intramammary route with S. aureus and the bacteria in the mammary glands were counted 12 h after infection. Significantly less numbers of bacteria were recovered from the infected glands of helenalin-treated mice compared with untreated mice. Moreover, histological examination of mammary tissue from helenalin-treated mice that were challenged with S. aureus indicated that helenalin is able to significantly reduce leukocyte infiltration in the mammary gland following S. aureus inoculation. Our results show that helenalin reduces S. aureus intracellular growth and experimental S. aureus infection. We conclude that helenalin may be of potential interest in the treatment of S. aureus-induced mastitis in the bovine species.

Prolactin-induced activation of nuclear factor kappaB in bovine mammary epithelial cells: role in chronic mastitis.

We sought to determine whether prolactin (PRL) could influence the neutrophilic inflammation that characterizes chronic mastitis. Most of the genes encoding inflammatory proteins depend on the nuclear factor kappaB (NF-kappaB) for their expression. We addressed the hypothesis that immunomodulatory activities of PRL might arise from an increase in NF-kappaB activity. MAC-T cells, a bovine mammary epithelial cell line, were stimulated with increasing concentrations of bovine PRL (1, 5, 25, 125, and 1,000 ng/mL). Level of NF-kappaB binding activity was measured and mRNA was evaluated for IL-1beta, IL-6, IL-8, granulocyte-macrophage colony-stimulating factor (GMCSF), IFN-gamma, and tumor necrosis factor (TNF)-alpha, cytokines known to require NF-kappaB for their maximal transcription. Prolactin activated NF-kappaB; maximal NF-kappaB activation was weaker with PRL than with TNF-alpha at 30 or 180 min poststimulation. In addition, PRL significantly amplified, in a dose-dependent manner, mRNA expression of IL-1beta, IL-6, IL-8, GMCSF, and TNF-alpha. We measured PRL concentrations in blood and milk from healthy and chronic mastitis-infected cows, and studied the relationship between the PRL concentration and the degree of inflammation in the mammary gland as indirectly assessed by somatic cell counts (SCC). Plasma PRL did not differ significantly between healthy and chronic mastitis-affected cows (63.7 and 67.5 ng/mL, respectively). Milk PRL concentration was significantly increased in chronic mastitis-affected quarters with the highest SCC, and had a positive significant correlation between SCC, as well as between the number of neutrophils present in milk samples. The present findings show that PRL promotes an inflammatory response in bovine mammary epithelial cells via NF-kappaB activation, and suggest a role for PRL in the pathogenesis of chronic mastitis.

Effects of exercise and oral antioxidant supplementation enriched in (n-3) fatty acids on blood oxidant markers and erythrocyte membrane fluidity in horses.

The aim of this study was to investigate in a placebo-controlled field study the effect of a (n-3)-vitamin supplementation on erythrocyte membrane fluidity (EMF), oxidant/antioxidant markers and plasmatic omega3/omega6 fatty acid ratio (FAR) in 12 eventing horses. Venous blood was sampled at rest before (PRE) and after (POST) a three week treatment period with either the supplement (group S, n=6) or a placebo (group P, n=6) as well as after 15min (POST E15') and 24h (POST E24h) after a standardised exercise test. The following markers were analysed: EMF, plasma antioxidant capacity of water and lipid soluble components, ascorbic acid, uric acid (UA), glutathione (reduced: GSH, oxidised: GSSG), vitamin E (Vit E), beta-carotene, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity, selenium, copper (Cu), zinc (Zn), oxidised proteins (Protox), lipid peroxides (Pool) and FAR. EMF did not differ between group S and P after treatment, but GPx remained unchanged in group S whereas it decreased in group P and plasma Cu/Zn ratio remained unchanged whereas it increased in group P. FAR were significantly increased in group S. Exercise induced a significant decrease of EMF (POST vs. E24h) in both groups, but which was significantly lower at E15' in group S than in group P. Exercise induced a significant increase of UA and ACW (POST vs. E15') and Protox (POST vs. E24h) in both groups. An exercise-related decrease in GSH and Pool (POST vs. E15') was found in group P, whereas Vit E and FAR (POST vs. E24h) significantly decreased in both groups. The study showed that exercise induced a decrease in EMF in horses associated with changes of blood oxidative balance. The (omega-3)-vitamin supplementation tested improved the oxidative balance poorly but delayed the exercise-induced decrease of EMF and increased the FAR.

Qualitative and quantitative evaluation of equine respiratory mechanics by impulse oscillometry.

REASONS FOR PERFORMING STUDY: The long-established conventional reference technique (CRT) for measuring respiratory mechanics in horses lacks sensitivity and there is a need for further refinement in new technology, such as the impulse oscillometry system (IOS). OBJECTIVES: To evaluate the potential use of the IOS as a clinical respiratory function test and compare it to the current CRT in horses suffering from common upper and lower airway dysfunctions. METHODS: Six healthy horses were tested before and after induction of a unilateral nasal obstruction (UNO) or transient left laryngeal hemiplegia (LLH). Six heaves-affected horses were tested in clinical remission and during a heaves crisis, before and after nebulisation of cumulative doses of a bronchodilator therapy (ipratropium bromide; IPB). RESULTS: As opposed to the CRT, the IOS was able to detect partial upper airway obstruction (UAO) caused by UNO or LLH in resting horses, without differentiating both conditions. Upper airway obstruction caused an upward shift of resistance (R(rs)) from 5 to 35 Hz without altering reactance (X(rs)). As for the CRT, IOS respiratory parameters measured in heaves-affected horses in crisis differed significantly from values measured during remission. The difference in frequency-dependent behaviour of R(rs) and X(rs) allowed discrimination between upper and lower airway obstructions. Bronchodilator treatment induced significant dose-dependent changes in X(rs) at 5 and 10 Hz, from the first dose. Total pulmonary resistance (RL) and R(rs) at 5 Hz were affected from the second dose and displayed similar sensitivity. Although post treatment RL values were comparable to remission, R(rs) and X(rs) remained significantly different, characterising persistent peripheral obstruction. CONCLUSIONS: The IOS was more sensitive than the CRT in detecting partial UAO in resting horses and persistent post treatment peripheral dysfunction in heaves-affected horses. The IOS is a sensitive test that provides graded quantitative and qualitative information on disease-induced respiratory dysfunctions as well as on treatment efficiency in horses. POTENTIAL RELEVANCE: The IOS could represent a practical and sensitive alternative respiratory function test for routine clinical investigations of common airway obstructive diseases and therapy in horses.

Investigation of blood oxidant/antioxidant markers in healthy competition horses of different breeds.

REASONS FOR PERFORMING STUDY: There is increasing evidence that the equine athlete is exposed to exercise-induced changes of its oxidant/antioxidant balance and antioxidant supplementation is frequently recommended. However, it is unknown whether there is a specific need for antioxidants according to performance, breed, gender or age. OBJECTIVES: To assess whether breed-, gender- and age-related differences of blood oxidant/antioxidant markers occur in competition horses. METHODS: Healthy horses (n = 493) underwent oxidant/ antioxidant blood marker determination. Vitamin E, lipophilic antioxidant capacity (ACL), ascorbic acid (AA), glutathione (GSH, GSSG), gluthione peroxidase (GPx), superoxide dismutase (SOD), selenium (Se), copper (Cu), zinc (Zn), lipid peroxides (Pool), oxidised proteins (Protox) were determined, as well as magnesium (Mg), creatine phosphokinase (CPK), lactate dehydrogenase (LDH), packed cell volume (PCV) and haemoglobin (Hb). A mixed linear model assessed the effect of breed, gender and age category. P<0.05 was considered significant. RESULTS: Thoroughbreds showed the highest values of vitamin E, ACL, GPx, PCV and Hb, whilst standardbreds had the highest values of AA and LDH. Jumping horses had the highest Protox values. Females had significantly higher SOD values, whereas most of the other markers were higher in stallions and geldings. Horses age 2-6 years had higher AA, SOD and LDH values than horses age >6 years. Correlation analyses were positive and significant between vitamin E and GPx, VitE and ACL, Se and GPx, Cu and Pool and negative between Pool and vitamin E, Pool and ACL, Protox and GPx, Protox and vitamin E. CONCLUSIONS: Blood oxidant/ antioxidant status of horses is influenced by breed, gender and age. The correlation analyses suggest synergistic relations between GPx, vitamin E and Se and an antagonistic relation between Protox-GPx, Protox-vitamin E, and Pool-vitamin E. POTENTIAL RELEVANCE: The results of this investigation provide definition of the specific need for antioxidants and vitamins in competition horses.

Effect of exercise on blood oxidant/antioxidant markers in standardbred horses: comparison between treadmill and race track tests.

REASONS FOR PERFORMING STUDY: Interest in establishing oxidant/ antioxidant profiles in competition horses is increasing. Earlier studies performed in horses have mainly been performed under laboratory conditions using a treadmill and it is not known to what extent laboratory results of oxidant/antioxidant studies might be transposed to field conditions. OBJECTIVE: To compare the impact on the blood oxidant/ antioxidant status of a standardised exercise test including a run up to fatigue performed on a treadmill (TM) and on a racetrack (RT) in healthy and trained Standardbred horses. MATERIAL AND METHODS: During TM and RT tests the following blood antioxidant markers were analysed in jugular venous blood at rest and 15 mins (E15) after an intense bout of exercise: uric acid (UA), ascorbic acid (AA), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (reduced: GSH and oxidised: GSSG), glutathione redox ratio (GRR) and protein thiol (PSH). Running time to fatigue (RTF), velocity during the last exercise stage (Vmax), final heart rate (HRfinal) and venous lactic acid (LA) were also recorded. RESULTS: Vmax was significantly (P<0.05) higher during the RT, whereas LA was significantly lower. HRfinal and RTF did not differ significantly between TM and RT. Exercise induced a significant increase (R vs. E15) of UA and AA in both tests, whereas GSH and PSH decreased significantly. GPx, SOD, GSSG and GRR remained unchanged. Differences between TM and RT were significant at E15 for UA, AA and PSH. CONCLUSIONS: Comparison of oxidant/antioxidant profiles from laboratory and field studies are difficult to standardise and should be interpreted with caution. POTENTIAL RELEVANCE: For the same RTF and final HR, the TM induced stronger changes in blood lactate and in blood oxidant/antioxidant balance than did RT.

The effects of dietary N-3 and antioxidant supplementation on erythrocyte membrane fatty acid composition and fluidity in exercising horses.

REASONS FOR PERFORMING STUDY: Fatty acid supplementation could modulate erythrocyte membrane fluidity in horses at rest and during exercise, but information is lacking on the effect of exercise. OBJECTIVES: To assess the effect of exercise with, and without, an oral antioxidant supplementation enriched with n-3 fatty acids on erythrocyte membrane fluidity (EMF) and fatty acid composition in eventing horses. METHODS: Twelve healthy and regularly trained horses were divided randomly into 2 groups: group S received an oral antioxidant cocktail enriched in n-3 fatty acid (alphatocopherol, eicosapentaenoic acid [EPA] and docosahexaenoic acid [DHA]) whereas group P was placebo-treated. At the end of 4 weeks, all horses performed a standardised exercise test (ET) under field conditions. Venous blood was sampled before starting treatment (TO), immediately before (T1) as well as 15 min (T2) and 24 h (T3) after ET. Spin labelled (16-DOXYL-stearic acid) red blood cell membranes were characterised using the relaxation correlation time (Tc in inverse proportion to EMF). Fatty acid composition (%) of the membrane was determined by gas-liquid chromatography. RESULTS: Supplementation did not induce changes in EMF (T1 vs. TO) but significant changes in membrane composition were observed and there were increases in n-3 polyunsaturated fatty acid PUFA, n-3/n-6 ratio, and total n-3 fatty acids. Exercise (T2 vs. T1) induced a significant decrease of EMF in group P (Tc: +19%, P<0.05) and nonsignificant decrease in group S (Tc: +5%), whereas membrane fatty acid composition did not change in either group. During the recovery period (T3 vs. T2), EMF decreased significantly in group S (Tc: +29%, P<0.05) and nonsignificantly in group P (Tc: +18%) without any significant changes in fatty acid composition. CONCLUSION AND POTENTIAL RELEVANCE: An enriched oral antioxidant supplementation induced changes in membrane composition, which modulated the decrease in EMF induced by exercise. Long chain n-3 fatty acid supplementation might therefore be beneficial.

The effect of inspired gas density on pulmonary artery transmural pressure and exercise induced pulmonary haemorrhage.

REASONS FOR PERFORMING STUDY: Pulmonary capillary stress failure, largely as a result of high pulmonary vascular pressures, has been implicated in the aetiology of EIPH. However, the role of the respiratory system in determining the magnitude of EIPH has received little attention. HYPOTHESIS: Horses breathing a gas of greater density than air will exhibit greater transmural pulmonary arterial pressures (TPAP) and more severe EIPH, and horses breathing a gas of lower density than air will exhibit lower TPAP and less severe EIPH, both compared with horses breathing air. METHODS: Following a warm-up, 8 Thoroughbred horses were exercised for 1 min at 10, 11 and 12 m/sec (5 degrees incline) breathing air or 21% oxygen/79% helium or 21% oxygen/79% argon in a randomised order. Heart rate, respiratory rate, pulmonary arterial pressure and oesophageal pressure were measured during exercise. Bronchoalveolar lavage fluid (BALF) was collected from the dorsocaudal regions of the left and right lungs 40 min post exercise and red blood cell (RBC) counts were performed. RESULTS: The exercise tests induced mild EIPH. Maximum changes in oesophageal pressure were lower on helium-oxygen compared to argon-oxygen (P<0.001). TPAP and median RBC counts did not differ between gas mixtures. BALF RBC counts from the left lung correlated with counts from the right lung (P<0.0001). However BALF RBC counts from the left lung were higher than those from the right lung (P = 0.004). CONCLUSION: As alterations in pulmonary arterial and oesophageal pressure caused by changes in inspired gas density were of similar magnitude, TPAP remained unchanged and there was no significant effect on EIPH severity. POTENTIAL RELEVANCE: Manipulations that decrease swings in intrapleural pressure may only decrease the degree of EIPH in horses severely affected by the condition.

Validation of a portable equine metabolic measurement system.

REASONS FOR PERFORMING STUDY: In equine sports medicine, VO2 has been measured exclusively with stationary systems, in laboratories equipped with a treadmill. Measurement during exercise in field conditions has not previously been reported because of the lack of portable equipment designed for horses. OBJECTIVES: A commercially available portable metabolic measurement system, based on breath-to-breath gas analysis and flow spirometry, was adapted to the horse's physiology and morphology (Cosmed K4b2 and Equimask) and its validity tested by (1) repeatability of the measures and (2) comparing metabolic data to those obtained by a reference method (RM). METHODS: To test the reproducibility of the measurements, 5 healthy saddle horses were subjected twice at 2 day intervals to a similar submaximal standardised incremental exercise test on a treadmill. The same horses performed twice at one week interval an incremental treadmill test to fatigue: the oxygen consumption and ventilation were measured once with the K4b2 system and once with the RM. The metabolic and ventilatory data obtained with both systems were compared. RESULTS: There was a good reproducibility of the metabolic measurements obtained by the K4b2 system at any workload. The VO2 obtained by both systems at any workload was not significantly different. However, the K4b2 expired fraction in CO2 (FETCO2) and carbon dioxide production (VCO2) were significantly lower at high and at maximal workloads. As a consequence, the values of the respiratory exchange ratio were too low and incompatible with normal physiological values. CONCLUSIONS: The good reproducibility of the metabolic and ventilatory measurements and the fact that the VO2 measurements at any workload were similar to the data obtained with the reference method suggested that this system may be used for comparison of repeated VO2 measurements in practical field conditions. POTENTIAL RELEVANCE: The K4b2 system could be used to improve knowledge of the energetic cost in different equine sports disciplines and offer the opportunity to undertake performance tests with genuine track conditions, on ridden or harnessed horses, rather than under laboratory conditions.

[Control of gene transcription as a new therapeutic target in asthma]. / Le contrôle de la transcription génique en tant que nouvelle cible thérapeutique dans le traitement de l'asthme.

The recent advances in the knowledge of the molecular mechanisms underlying asthma have lead to a significant improvement of the current treatments of the disease and opened new perspectives for the development of therapeutic alternatives to inhaled corticosteroids. The selective targeting of transcription factors controlling the expression of the genes implicated in the pathogenesis of asthma is one of these privileged strategies. This review aims at describing the most promising new therapeutic targets in the control of asthmatic inflammation at the gene transcription level.

Effect of carprofen treatment following experimentally induced Escherichia coli mastitis in primiparous cows.

Acute Escherichia coli mastitis is one of the major sources of economic loss in the dairy industry due to reduced milk production, treatment costs, discarded milk, and occasional fatal disease. Nonsteroidal anti-inflammatory drugs (NSAIDs) are frequently used as adjunctive therapy to antibiotics. The objective of the current study was to evaluate the effect of carprofen treatment following infusion of Escherichia coli into the mammary glands of primiparous cows during the periparturient period. Severity of mastitis was scored based on the average milk production in the uninfected quarters on d +2 postinoculation and a clinical severity score. Carprofen was administered intravenously at 9 h postchallenge, when clinical signs of mastitis appeared. In previous work, efficacy of NSAIDs was mainly evaluated using clinical symptoms. In the present study, the effect of carprofen on innate immune response was also assessed by quantification of inflammatory mediators. All primiparous cows reacted as moderate responders throughout the experimental period. Primiparous cows were intramammarily inoculated with 1 x 10(4) cfu of E. coli P4:O32 in 2 left quarters. Analysis of blood and milk parameters, including IL-8, complement component C5a, lipopolysaccharide-binding protein (LBP), soluble CD14, prostaglandin E2, and thromboxane B2 was performed from d 0 to d +6 relative to intramammary inoculation. Rectal temperature in carprofen-treated animals was lower than in control animals at 3 and 6 h posttreatment. Treatment also restored the decreased reticulorumen motility that occurs during E. coli mastitis to preinfection levels faster than in control animals. Carprofen treatment resulted in an earlier normalization of the clinical severity score. Eicosanoid (prostaglandin E2 and thromboxane B2) production in milk tended to be inhibited by carprofen. No significant differences in the kinetic patterns of somatic cell count, IL-8, complement component C5a, LBP, and soluble CD14 were observed. In conclusion, carprofen treatment improved general clinical condition by effective antipyrexia and restoration of reticulorumen motility but did not significantly inhibit eicosanoid production. Carprofen treatment did not result in a significant decrease of chemotactic inflammatory mediators, IL-8 and C5a, and early innate immune molecules, sCD14 and LBP. Therefore, major modulatory effects from NSAID administration were not observed in this mastitis model, although a larger study might confirm some apparent trends obtained in the present results.