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See detailIMPACT OF THE TEMPERAMENT OF YOUNG STALLIONS ON THEIR STRESS REACTIONS WHEN SUBJECTED TO A STANDARDISED VETERINARY EXAMINATION
Peeters, Marie ULg; Verwilghen, Denis ULg; Serteyn, Didier ULg et al

in Journal of Veterinary Behavior: Clinical Applications and Research (2011, September), 6

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See detailHorseshoeing styles comparison and detection of subclinic equine digit discomfort during movement
Noble, Prisca ULg; Lejeune, Jean-Philippe ULg; Serteyn, Didier ULg et al

Conference (2011, July 07)

Introduction: For the sound equine forelimb, ground reaction force (GRF) peaks to ~0.8-1 BodyWeight (BW) during the trot (4m/s). Little work investigates the effect of different horsehoeing styles ... [more ▼]

Introduction: For the sound equine forelimb, ground reaction force (GRF) peaks to ~0.8-1 BodyWeight (BW) during the trot (4m/s). Little work investigates the effect of different horsehoeing styles, directly on the limb GRF distribution and indirectly on the equine digit comfort during movement. This study investigates GRF distribution after application of different aluminium horseshoeing styles during movement. Methods: Two horses (H1,H2; mean 575kg) were used. However H2 had an old healed flexor tendons lesion, the horses were judged to be sound on locomotor examination (without lameness). According to the rules that respect the foot biomechanical balance, they were trimmed and shod with a non-broken foot-pastern axis. On the day of the tests, they were led on a treadmill at a trot (4m/s). Kinetics were collected, using a F-Scan system, during 3 following sessions : to both forehooves after application of classic roller, eggbarr and equi+ horseshoes. For each session, data from 3 strides for each left forelimb of each horse were averaged and kinetics (GRF) were obtained. Results: For the classic roller, eggbarr and equi+ horseshoes, GRF peaked respectively to 0.57+-0.006;0.36+-0.005;0.83+-0.009BW for H1 and 0.61+-0.007;0.84+-0.007;0.51+-0.009BW for H2. Theses horseshoes loading differences show a dynamic load transfer from the forelimbs to the hindlimbs, that is the result of a subclinic (without lameness) equine digit discomfort during movement. Conclusions: Subclinic equine digit discomfort after application of horshoes has been detected using the F-Scan system during movement. These results confirm the interest of the equi+ horseshoes on horses without flexor tendon lesion. [less ▲]

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See detailTime trends of blood leucocytes, neutrophils and plasmatic myeloperoxidase in the perioperative period of horses undergoing colic surgery.
Salciccia, Alexandra ULg; Grulke, Sigrid ULg; Detilleux, Johann ULg et al

Poster (2011, July)

Background: Despite the recent advances in this area, colic remains a major cause of morbidity and death in horses. Neutrophilic activation and degranulation may play a key role in the postoperative ... [more ▼]

Background: Despite the recent advances in this area, colic remains a major cause of morbidity and death in horses. Neutrophilic activation and degranulation may play a key role in the postoperative complications. Activated neutrophils release enzymes like proteases and myeloperoxidase (MPO). MPO concentrations in plasma and tissue are considered as a marker of neutrophil activation. (McConnico et al. 1999; Hoy et al. 2002). When freed in the tissue, active MPO is able to oxidize, nitrate and chlorate most organic molecules (Klebanoff 2005). Objectives: The aim of this study was 1) to determine the time trends of blood leukocyte and neutrophil counts as well as of plasmatic MPO concentrations in the perioperative period of horses undergoing colic surgery and 2) to relate these time trends to the location of the pathology, the severity of postoperative complications and to the outcome of the patients. Methods: Fifty two horses undergoing colic surgery at the Equine Teaching Hospital of the University of Liege were included in this study. The location of the predominant lesion of the intestine, the severity of the postoperative complications and the outcome were recorded for each horse. Total leukocyte and neutrophil counts were performed in all of the horses while plasmatic myeloperoxidase levels were determined in 16 of them. The blood samplings were realized before and during the surgery (after correction of the intestinal lesion), during the recovery and every 4 hours during the first 4 days (from day 0 until day 4) and then every 12 hours until day 6 (150th hour after the first blood sampling) or until euthanasia. Hematologic analyses were performed at the time of sampling by use of the Medonic CA 530 (Menarini, Zaventem, Belgium). The blood was then centrifuged and the plasma was aliquoted and frozen at -20° C until assayed. MPO was assayed with a specific enzyme-linked immunosorbent assay (Equine MPO-ELISA kit, BiopTis, Liège, Belgium). A mixed model was used to analyze the time trends of leukocytes, neutrophils and MPO. All computations were done with the SAS (Statistical Analysis System) procedure Proc Mixed, with Satterthwaite degrees of freedom. The significance level was set at p=0.05. Results: The main pathology was found in the large intestine in 30 horses (58%) and the small intestine in 22 horses (42%). Forty horses (77%) survived to discharge from the clinic. Twelve horses were euthanized during the postoperative period. Their survival time varied from 0.5 day to 20 days with a mean of 7.8 days. Twenty-two horses (42%) showed none or mild complications, 12 horses (23%) showed moderate complications and 18 horses (35%) suffered from severe complications. Time trends for leukocytes and neutrophils were similar to each other (p= 0.7205) and significantly different (p< 0.0001) from the MPO time trend, which increased during the first hours, while the neutrophil time trend decreased immediately after the admission. The time trend of neutrophils was higher in large intestinal than in small intestinal pathologies and the time trend of MPO was lower in large intestinal than in small intestinal pathologies. The time trends of neutrophils were significantly different between the degrees of complications (no/mild vs moderate vs severe). For the first part of the curve, the more severe the complication, the lower is the time trend. The time trend of MPO was lower in survivors. The time trend of neutrophils in survivors was higher during the first 4 days thereafter it becomes lower than in non survivors. Conclusions: These results confirm that neutrophil counts and MPO levels undergo timely changes and that they are related to the severity of the inflammatory reaction in surgical colic cases. Knowing the kinetics of these parameters is an essential step to further determine cut-off values (with a larger group of horses) for the prognosis of horses after colic surgery. References Hoy, A., Leininger-Muller, B., Kutter, D., Siest, G. and Visvikis, S. (2002) Growing significance of myeloperoxidase in non-infectious diseases. Clin. Chem. Lab. Med. 40, 2-8. Klebanoff, S.J. (2005) Myeloperoxidase: friend and foe. J. Leukoc. Biol. 77, 598-625. McConnico, R.S., Weinstock, D., Poston, M.E. and Roberts, M.C. (1999) Myeloperoxidase activity of the large intestine in an equine model of acute colitis. Am. J. Vet. Res. 60, 807-813. [less ▲]

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See detailEvaluation of the effect of general anaesthesia on ultrasonographic images of the small intestine in horses.
Salciccia, Alexandra ULg; Gougnard, Alexandra ULg; de la Rebière de Pouyade, Geoffroy ULg et al

Poster (2011, July)

ultrasonography is currently more and more used in equine acute abdominal disease as well as in the follow-up of surgical colic patients because of its sensibility for the detection of small intestinal ... [more ▼]

ultrasonography is currently more and more used in equine acute abdominal disease as well as in the follow-up of surgical colic patients because of its sensibility for the detection of small intestinal distension. General anaesthesia is known to diminish gastrointestinal motility even if there are no clinical signs associated with it. The aim of this study was to evaluate the effect of general anaesthesia on transabdominal ultrasonographic images of the small intestine in horses undergoing routine non abdominal surgeries. The ultrasonographic exams were performed in 19 horses before and after the general anaesthesia (immediately after the recovery and then 2h, 12h and 24h after it). Each ultrasonographic exam focused on the duodenum, the jejunum and the presence of peritoneal fluid. For the jejunum, 5 locations were used: on the ventral midline just caudally to the sternum, on the left and right cranial ventral parts of the abdomen, on the left and right inguinal regions. Other parameters such as the gut sounds and the postoperative fecal output were also recorded. Anova and Chi-square tests were used for the statistical analysis. No horse showed colic signs. No significant difference was found between the pre and post anaesthetic period considering the maximal diameter of the duodenum, the maximal diameter of the jejunum on the 5 locations and the peritoneal fluid. The contractions of the duodenum were increased at the recovery compared to before the anaesthesia (p= 0,0299). The small intestine was most visible at the recovery (57,5%) and then at 2 hours after it (38,3%). It seemed that the ventral midline just caudally to the sternum and the right and left inguinal regions were the best locations to observe the jejunum (with a mean of respectively 44,6%, 39,8% and 38,5% of visualization of the jejunum).The gut sounds were very significantly decreased at the recovery (p < 0,0001) and at 2h after it (p = 0,0006). The postoperative fecal output was not decreased. In conclusion, even if general anaesthesia seems to reduce temporarily the intestinal activity (decreased gut sounds in the early post anaesthetic period), it does not cause significant distension of the small intestine (almost the small intestinal diameters were in the normal range). If an increased diameter of small intestine is observed by ultrasonography after surgery it should therefore be attributed to a pathological process and not to the anaesthesia. [less ▲]

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See detailMesure de la respiration musculaire par respirométrie à haute résolution
Votion, Dominique ULg; Fraipont, Audrey ULg; Robert, Céline et al

in Pratique Vétérinaire Equine (2011), 43(170), 35-37

This technical note describes the reference method for screening the mitochondrial function in horses.

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See detailSpecific Immuno Extraction Followed by Enzymatic Detection (SIEFED), a new tool to measure active myeloperoxidase in complex media and to screen potential inhibitors of this enzyme.
Serteyn, Didier ULg; Mouithys-Mickalad, Ange ULg; Deby-Dupont, Ginette et al

in The 7th International Human Peroxidase Meeting, 22-25 May 2011, Brussels (2011, May 25)

A method called SIEFED (“Specific Immuno Extraction Followed by Enzymatic Detection”) was developed for the specific measurement of myeloperoxidase (MPO) activity without interference of the sample medium ... [more ▼]

A method called SIEFED (“Specific Immuno Extraction Followed by Enzymatic Detection”) was developed for the specific measurement of myeloperoxidase (MPO) activity without interference of the sample medium. In this method, MPO is first extracted out of aqueous or biological samples by its capture by immobilized anti-MPO antibodies. A washing step then allows to eliminate the solution or biological fluid with interfering materials (proteins, lipids, reducing or oxidizing molecules, ...) and in a last step the activity of MPO bound to its antibodies is measured by using a sensitive detection system containing a fluorogenic substrate, hydrogen peroxide and nitrite as reaction enhancer. SIEFED is a method of choice to measure easily, quantitatively, and specifically the active part of MPO present in biological samples or complex media. Results obtained with this technique applied to biological samples emphasize the importance to distinguish the total MPO concentration of a sample obtained by ELISA from the active part of MPO, which is the real witness of the oxidant potential of the enzyme. SIEFED is also a powerful tool to study compounds or natural extracts that could have an inhibitory effect on the activity of MPO. Since the potential inhibitor is first incubated with the enzyme solution, and further eliminated by washing after the immunocapture of MPO, the tested compound or extract cannot interfere with the chromogenic substrate used to measure the activity of MPO or with products derived from the enzyme activity. Thus, an inhibitory effect could only be attributed to a direct interaction of the tested compound with the enzyme, either on the active site or another key position of the structure. In conclusion, the SIEFED technique opens new perspectives to study pathologies in which the release of active MPO is relevant and to select interesting compounds or extracts able to modulate the MPO activity. [less ▲]

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See detailA pendulum test as a tool to evaluate viscous friction parameters in the equine fetlock joint
Noble, Prisca ULg; Lumay, Geoffroy ULg; Coninx, Marc ULg et al

in Veterinary Journal (2011), 188(2), 204-209

An equine fetlock joint pendulum test was studied and the influence of post mortem time and intra-articular lipid solvent on the viscous frictional response examined. Fresh equine digits (group 1, n=6 ... [more ▼]

An equine fetlock joint pendulum test was studied and the influence of post mortem time and intra-articular lipid solvent on the viscous frictional response examined. Fresh equine digits (group 1, n=6 controls; group 2, n=6 lipid solvent) were mounted on a pendulum tribometer. Assuming that pendular joint damping could be modelled by a harmonic oscillator fluid damping (HOFD), damping time (τ), viscous damping coefficient (c) and friction coefficient (μ) were monitored for 5h under experimental conditions (400N; 20°C). In all experiments, pendular joint damping was found to follow an exponential decay function (R(2)=0.99714), which confirmed that joint damping was fluid. The evolution of τ, c and μ was found to be significantly (P<0.05) different in the two groups, with a decrease in τ and an increase in c and μ that was faster and more prominent in digits from group 2. It was concluded that pendular joint damping could be modelled by a HOFD model. The influence of post mortem time on results suggested that, ideally, joint mechanical properties should only be tested on fresh cadavers at the same post mortem time. Moreover, the addition of lipid solvent was found to be responsible for upper viscous friction parameters and for a reduced damping time, which suggested that articular lubricating ability was compromised. This equine pendulum test could be used to test the efficacy of various bio-lubricant treatments. [less ▲]

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See detailEffect of benzoic acid analogs on myeloperoxidase activity measured by a new technique to study their direct interaction with the enzyme.
Franck, Thierry ULg; Mazloum, Ali; Mouithys-Mickalad, Ange ULg et al

Poster (2011, May)

Myeloperoxidase (MPO) plays a key role in inflammatory response and constitutes a target for new drug development. The effects of some benzoic acid analogs were studied on the specific activity of human ... [more ▼]

Myeloperoxidase (MPO) plays a key role in inflammatory response and constitutes a target for new drug development. The effects of some benzoic acid analogs were studied on the specific activity of human MPO measured by SIEFED (“Specific Immunologic Extraction Followed by Enzymatic Detection”), an original method that consists of incubation of the compound with MPO, followed by capture of the enzyme by specific antibodies, washing (elimination of the compounds) and enzymatic detection of the immunocaptured enzyme. The compounds tested at 10-4, 10-5 and 10-6 M were studied in terms of structure activity relationship. Gallic acid (3,4,5-trihydroxybenzoic acid) with 3 hydroxyl groups had an important dose dependent inhibitory effect on MPO activity. Other molecules with less or without hydroxyl groups [3,4-dihydroxybenzoic acid, 2-hydroxybenzoic acid (salicylic acid) and benzoic acid] had rather an activator effect at 10-5 and 10-6 M. 2,4,6-Trihydroxybenzoic acid, with two hydroxyl groups adjacent to the carboxyl group, had a less efficient inhibitory effect. Caffeic acid (3,4-dihydroxycinnamic acid) with a propenoic acid group presented a dose dependent inhibitory effect on MPO activity contrary to its analog 3,4-dihydroxybenzoic acid. The esterification of the carboxyl group of gallic acid to obtain propyl gallate induced an activation of MPO at 10-5 and 10-6 M. Finally, the substitution of one or two hydroxyl groups by methoxyl ones (ferulic and syringic acids) decreased the efficiency of the molecules on the enzyme inhibition. The SIEFED technique appears as an innovative pharmacological tool to study the direct interaction of compounds with MPO. Number and position of hydroxyl groups and the extension of the carboxyl group of benzoic acid play a crucial role in the inhibition of MPO activity probably by facilitating the interaction with the active site or another elements of the enzyme structure. [less ▲]

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See detailFat analysis in muscle samples from horses affected with atypical myopathy
Van Galen, Gaby ULg; Deby-Dupont, G; Serteyn, Didier ULg et al

Poster (2011, April 15)

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See detailReversal of acepromazine-induced hemodynamic alterations by norepinephrine in standing horses
pequito, Manuel; Amory, Hélène ULg; Peters, Fabrice et al

Poster (2011, March 18)

Acepromazine (ACP) is a phenothiazine commonly used to sedate horses. Additionally, ACP exerts strong anti-inflammatory effects, which might have a therapeutic potential in horses suffering from systemic ... [more ▼]

Acepromazine (ACP) is a phenothiazine commonly used to sedate horses. Additionally, ACP exerts strong anti-inflammatory effects, which might have a therapeutic potential in horses suffering from systemic inflammatory response syndrome (SIRS). However, the ACP-induced vasodilation precludes its use in horses with SIRS-related cardiovascular compromise. The objective of this study was to test if the hemodynamic effects of the administration of 0,1 mg/kg of ACP could be counteracted by an intravenous infusion of norepinephrine (NOR) at 1ug/kg/min in healthy horses. In 5 healthy adult horses, a 15 minutes NOR IV infusion was administered 45 minutes after an injection of 0,1 mg/kg of ACP IV. The systolic arterial blood pressure (SAP) was non-invasively measured by Doppler sphingometry at the tail. Hemodynamics of the median artery of the left forelimb were studied using Doppler ultrasonography, through calculation of the vessel’s surface (SURF), diameter (DIAM), circumference (CIRC), and peak systolic velocity (PSV), end diastolic velocity (EDV), mean velocity (MV), volumetric flow (VF) and resistivity index (RI) of the flow. Both SAP and Doppler parameters were determined at regular intervals during the entire study. ACP induced a hypotension and a vasodilatation, that were evidenced by a significant rise of the SURF, DIAM, CIRC, PSV, EDV, MV and VF and reduction of the SAP and RI. During NOR infusion, all these ACP-induced hemodynamic changes were reversed. These findings suggest that a continuous IV NOR infusion at 1ug/kg/min is able to revert ACP-induced hypotension and vasodilation in healthy adult horses. [less ▲]

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See detailIncidences pratiques tirées des dernières séries cliniques de myopathie atypique sur le territoire français
Patarin, Florence; Van Galen, Gaby ULg; Marcillaud-Pitel, Christel et al

in Institut français du cheval et de l'équitation (Ed.) Actes de colloque: 37ème Journée de la Recherche Equine (2011, February 24)

Epidemiological data of atypical myopathy cases reported in France in autumn 2006 to spring 2010 were compared with those of European cases. During the observation period, 38.9% of European cases were ... [more ▼]

Epidemiological data of atypical myopathy cases reported in France in autumn 2006 to spring 2010 were compared with those of European cases. During the observation period, 38.9% of European cases were French. The demographics of the French case were identical to the European cases but the survival rate was higher (32% vs. 26%). Differences in the management of horses and pastures have emerged. From these elements, techniques, case management and the environment are proposed [less ▲]

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See detailThe use of vacuum assisted closure therapy for wound healing in horses
Lazzaretti, Sara; Grulke, Sigrid ULg; Serteyn, Didier ULg et al

Conference (2011, February 05)

Traumatic wounds and particularly limb wounds in horses can be difficult to treat. Horses show protracted and weak inflammatory reaction in the first stages of the wound healing causing delays[1]. Several ... [more ▼]

Traumatic wounds and particularly limb wounds in horses can be difficult to treat. Horses show protracted and weak inflammatory reaction in the first stages of the wound healing causing delays[1]. Several types of treatments have been tested in order to promote wound healing in horses, with variable success[2]. Vacuum-assisted closure(VAC) has been used in human medicine for over a decade now, but only one regarding a horse is available [3]. This presentation descrebis the use and the experiences of the authors with VAC in horses. [less ▲]

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See detailTHE USE OF VAC® THERAPY IN HORSES
Lazzaretti, Sarah; Grulke, Sigrid ULg; Serteyn, Didier ULg et al

Poster (2011, January 22)

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See detailModulating effects of acepromazine on the reactive oxygen species production by stimulated equine neutrophils
Sandersen, Charlotte ULg; Mouithys-Mickalad, Ange ULg; de la Rebière de Pouyade, Geoffroy ULg et al

in Veterinary Anaesthesia & Analgesia (2011), 38

To investigate the effect of acepromazine (ACP) on reactive oxygen species (ROS) production by stimulated equine neutrophils.

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See detailFlow cytometric detection of myeloperoxidase in horse neutrophils: a novel technique in equine diagnostic research.
Wauters, Jella; Franck, Thierry ULg; Pille, Frederik et al

in Veterinary Immunology and Immunopathology (2011), 144(3-4), 417-22

Myeloperoxidase (MPO) is a protein of interest due to its involvement in equine pathologies. Until now, results in equine diagnostic research were achieved through extracellular MPO detection. However ... [more ▼]

Myeloperoxidase (MPO) is a protein of interest due to its involvement in equine pathologies. Until now, results in equine diagnostic research were achieved through extracellular MPO detection. However, studying the cellular MPO content in neutrophils has revealed important insights in human diseases. This study aimed to develop a technique for the specific detection of MPO on the single cell level defining a flow cytometric protocol for the detection of both equine surface-bound and cellular MPO. Both indirect and direct labeling techniques are described which include the comparison of two secondary antibodies and two linking-fluorochromes, respectively. [less ▲]

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See detailClinical significance of active myeloperoxidase in carotid atherosclerotic plaques
GACH, Olivier ULg; Magne, Julien ULg; Franck, Thierry ULg et al

in International Journal of Cardiology (2011), 152(1), 149-151

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See detailProduction of free radicals and oxygen consumption by primary equine endothelial cells during anoxia-reoxygenation.
de la Rebière de Pouyade, Geoffroy ULg; Salciccia, Alexandra ULg; Ceusters, Justine ULg et al

in Open Biochemistry Journal (The) (2011), 5

The endothelium plays an active role in ischemia/reperfusion injuries. Herein, we report the effect of a single or successive cycles of anoxia/reoxygenation (A/R) on the mitochondrial respiratory function ... [more ▼]

The endothelium plays an active role in ischemia/reperfusion injuries. Herein, we report the effect of a single or successive cycles of anoxia/reoxygenation (A/R) on the mitochondrial respiratory function of equine endothelial cells (cultured from carotids) monitored by high resolution oxymetry, and on their production of reactive oxygen species (ROS). ROS were measured by electron paramagnetic resonance (ESR) using POBN and DMPO spin traps, and by gas chromatography (GC) of ethylene released by ROS-induced alpha-keto-gamma-(methylthio)butyric acid (KMB) oxidation. The oxygen consumption significantly decreased with the number of A/R cycles, and POBN-ESR spectra were specific of adducts formed in the cells from superoxide anion. After a one-hour A/R cycle, high intensity DMPO-ESR spectra were observed and assigned to superoxide anion trapping; the GC results confirmed an important production of ROS compared to normoxic cells. These results show that A/R induces mitochondrial alterations in endothelial cells, and strongly stimulates their oxidative activity as demonstrated by ESR and GC methods. [less ▲]

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See detailThe role of activated neutrophils in the early stage of equine laminitis.
de la Rebière de Pouyade, Geoffroy ULg; Serteyn, Didier ULg

in Veterinary Journal (2011), 189(1), 27-33

Despite ongoing research and a widening range of treatment options, laminitis remains a severely damaging condition with poorly understood pathophysiology. Results obtained from cytokine regulation ... [more ▼]

Despite ongoing research and a widening range of treatment options, laminitis remains a severely damaging condition with poorly understood pathophysiology. Results obtained from cytokine regulation studies during the last decade have highlighted the inflammatory nature of laminitis. This review will describe the role of systemic activation and local infiltration of neutrophils in laminar tissues in the induction of laminitis. Particular emphasis is placed on the role of neutrophil activation in subsequent vascular dysfunction and oxidative and proteolysis imbalances that are pathways previously implicated in laminitis. Neutrophils, by the way of their interdependent relationship with endothelial cells and keratinocytes, dramatically increase the inflammatory response culminating in the failure of the laminar dermal-epidermal interface. [less ▲]

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See detailColl2-1, Coll2-1NO2 and myeloperoxidase concentrations in the synovial fluid of equine tarsocrural joints affected with osteochondrosis.
Verwilghen, Denis ULg; Martens, Ann; Busschers, Evita et al

in Veterinary Research Communications (2011), 35(7), 401-8

The measurement of biomarkers that reflect cartilage breakdown is a powerful tool for investigating joint damage caused by disease or injury. Particularly in cases of osteochondrosis, synovial ... [more ▼]

The measurement of biomarkers that reflect cartilage breakdown is a powerful tool for investigating joint damage caused by disease or injury. Particularly in cases of osteochondrosis, synovial concentrations of these biomarkers may reveal the presence of osteoarthritic changes. Coll2-1, Coll2-1 NO2 and myeloperoxidase have recently been introduced in equine osteoarticular research but comparison between the concentrations of these markers in OCD affected and healthy joints has not been made. Therefore, this study aimed at reporting the synovial concentrations of these biomarkers in joints affected with osteochondral fragments in the tarsocrural joint compared to unaffected joints. Myeloperoxidase and Coll2-1NO2 revealed to have similar levels between affected joints and controls. However, in contrast to previous studies using C2C the present study demonstrated that synovial levels of Coll2-1 were significantly elevated in tarsocrural joints affected with osteochondrosis. Thus, Coll2-1 may be an earlier marker of cartilage degeneration than other cartilage degradation markers that have been previously used in equine medicine. [less ▲]

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