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See detailHigh concentrations of Myeloperoxidase in the equine uterus as an indicator of endometritis
Parrilla Hernandez, Sonia ULg; Ponthier, Jérôme ULg; Franck, Thierry ULg et al

in Theriogenology (in press)

Intra-luminal fluid and excessive abnormal hyper-edema are regularly used for the diagnosis of endometritis in the mare, which is routinely confirmed by the presence of neutrophils on endometrial smears ... [more ▼]

Intra-luminal fluid and excessive abnormal hyper-edema are regularly used for the diagnosis of endometritis in the mare, which is routinely confirmed by the presence of neutrophils on endometrial smears. Studies show a relation between neutrophils and myeloperoxidase (MPO), an enzyme contained in and released by neutrophils during degranulation or after cell lysis. This enzyme has been found in many fluids and tissues and associated with different inflammatory pathologies in the horse. The aims of this study were to assess the presence and concentration of MPO in the equine uterus, and to investigate its relation with neutrophils, and other clinical signs of endometritis. Mares (n=51) were evaluated for presence of intra-luminal fluid and excessive endometrial edema before breeding, and a small volume lavage and cytology samples were obtained. From 69 cycles, supernatant of the uterine flushes was analysed with a specific equine MPO ELISA assay to measure MPO concentration. Cytology samples were used for the diagnosis of endometritis. MPO was present in the uterus of all estrus mares in highly variable concentrations. MPO concentrations were significantly (p<0.05) higher in samples with positive cytologies and in presence of intra-luminal fluid. Occasionally, some samples with negative cytologies showed high MPO concentration, but the opposite was never observed. Cycles presenting hyper-edema weren’t associated to high concentration of MPO, intra-luminal fluid or positive cytology making it a poor diagnostic tool of endometritis. [less ▲]

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See detailConcentration, Activity and Biochemical Characterization of Myeloperoxidase in Fresh and Post-thaw equine semen and their Implication on Freezability
Ponthier, Jérôme ULg; Franck, Thierry ULg; Parrilla Hernandez, Sonia ULg et al

in Reproduction in Domestic Animals (in press)

Myeloperoxidase (MPO) is a pro-oxidant enzyme associated with decreased motility in thawed equine semen. This study aimed to describe MPO concentration, activity and subunits in raw and thawed semen and ... [more ▼]

Myeloperoxidase (MPO) is a pro-oxidant enzyme associated with decreased motility in thawed equine semen. This study aimed to describe MPO concentration, activity and subunits in raw and thawed semen and to correlate these data with motilities in raw and thawed semen. Semen samples from five stallions were collected four times. Motilities were assessed in raw and thawed semen. MPO assays were performed in raw seminal plasma, raw sperm-rich pellet and thawed semen. Total and active MPO concentrations were, respectively, assayed by enzyme-linked immunosorbent assay and specific immunological extraction followed by enzymatic detection. MPO subunits present in semen were characterized by Western 3 blot. Purified active MPO was added in PBS and freezing extender to control its activity during freezing procedure. Differences between medians were determined using Kruskal– Wallis test, and correlations were determined using Spearman’s test for nonparametric data. Active MPO concentration was low in seminal plasma and thawed semen, but high in pellet (p = 0.0058), as the opposite relation was observed for total MPO concentration (p < 0.0001). In seminal plasma and post- thaw semen, inactive 86-kDa MPO precursor was mainly observed. Purified MPO activity was decreased in the extender (p = 0.0286). MPO activity in pellet was highly correlated with thawed progressive motility (r = 􏰑0.5576, p = 0.0086). Inac- tive MPO precursor and unknown low molecular weight inactive MPO precursor subunits explain low MPO activity in semen. Major MPO activity was observed in pellet, and post- thaw loss of activity is partially explained by MPO inactiva- tion in extender. Thawed semen motility was negatively correlated with MPO activity in pellet, becoming a potential freezability predictor. [less ▲]

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See detailÉtude des contraintes mécaniques locomotrices chez le cheval
Noble, Prisca ULg; Collin, Bernard ULg; Denoix, Jean-Marie et al

in Annales de Médecine Vétérinaire (in press)

In locomotor biomechanics, three high groups of contraints are commonly encountared : pressure, traction and torsion. In supra-maximal conditions, all of these contraints would be responsible of some ... [more ▼]

In locomotor biomechanics, three high groups of contraints are commonly encountared : pressure, traction and torsion. In supra-maximal conditions, all of these contraints would be responsible of some equine diseased locomotor systems. In order to understand better the contraints in the equine locomotor dynamics, some investigations have been carried out. Moreover, some measurement methods, based on the mechanics of Newton, have been performed. This review shows the different measurement techniques and introduces the mechanical basis that are compulsory for the understanding of the equine locomotor apparatus functioning. [less ▲]

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See detailDevelopment and evaluation in vitro and in vivo of injectable hydrolipidic gels with sustained-release properties for the management of articular pathologies such as osteoarthritis.
Réeff, Jonathan; Oprenyeszk, Frédéric ULg; Franck, Thierry ULg et al

in International Journal of Pharmaceutics (2015), 490

This study aimed to evaluate glycerol monooleate (GMO) as a carrier to develop viscoelastic and injectable sustained-release drug delivery systems. The potential pro- and antioxidant activity of the ... [more ▼]

This study aimed to evaluate glycerol monooleate (GMO) as a carrier to develop viscoelastic and injectable sustained-release drug delivery systems. The potential pro- and antioxidant activity of the developed hydrolipidic gels were evaluated by measuring the production of ROS by polymorphonuclear leukocytes (PMNs). In addition, the biocompatibility and effectiveness of two selected gel candidates were evaluated in vivo by evaluating the benefit of a single intraarticular injection of these new treatments in a model of osteoarthritis in rabbits. The in vitro study demonstrated that the carrier F1 did not have a pro-oxidative effect and even protected PMNs against natural auto-activation, regardless of the incorporation of either clonidine chlorhydrate or betamethasone dipropionate. The in vivo study demonstrated that F1 and F1-BDP induced a loss of cartilage quality in comparison to the control and reference groups but that the lesions of cartilage observed were generally mild, with not much full-depth erosion. Moreover, no exacerbating inflammation was observed when considering the synovial membranes and the PGE2 and CRP levels. These results seemed to demonstrate that the sustained-release formulation based on GMO could be well-tolerated after intraarticular injection. Moreover, it could have the potential to prevent inflammatory conditions while sustaining drug activity locally over weeks. [less ▲]

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See detailAge-dependent expression of osteochondrosis-related genes in equine leukocytes
Mendoza García, Luis ULg; Piquemal, David; Lejeune, Jean-Philippe ULg et al

in Veterinary Record : Journal of the British Veterinary Association (2015), 2

Introduction: Osteochondrosis (OC) is a developmental disease in horses which has a significant impact on the horse’s welfare and performance. The early disturbance in the process of endochondral ... [more ▼]

Introduction: Osteochondrosis (OC) is a developmental disease in horses which has a significant impact on the horse’s welfare and performance. The early disturbance in the process of endochondral ossification progresses to inflammatory and repair processes in older horses. Previously, differentially expressed genes in leukocytes of OC-affected horses have been identified. The aim of the present study is to detect age-related changes in these differentially expressed genes. Materials and Methods: The expression of OC-related genes was analysed by real-time PCR and subsequent statistical analysis (ΔΔCT) in the leukocytes of 135 Belgian Warmblood horses divided into three different age groups: 30 months (n=38). Results: Relative expression of genes of horses less than 12 months of age showed significant induction of the genes MGAT4A, PRKCG, MHCI, ApoB, ApoB3G, B4GALT6 and a significantly lower expression of the genes OAS3. Horses of 18–24 months of age, showed a significantly higher expression of the genes TBC1D9, MGAT4A, IFIH1, MHCIIa and MMP1. Horses of more than 30 months of age showed a significantly higher expression of the genes MGAT4A, HP, SECTM1 compared with their age-matched control groups. Conclusions: The study demonstrates that OC-related genes are differentially expressed in horses of different ages compared with their age-matched controls. Some of the genes may be implicated in cell signalling and differentiation as well as carbohydrate and lipid metabolism and inflammation. However, the causal relationship between the differentially expressed genes and the development and progression of the OC lesions needs to be determined. [less ▲]

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See detailEffects of isoflurane and sevoflurane on the neutrophil myeloperoxidase system of horses
MINGUET, Grégory ULg; Franck, Thierry ULg; JORIS, Jean ULg et al

in Veterinary Immunology and Immunopathology (2015)

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See detailHigh plasma concentrations of sclerostin, an inhibitor of the Wnt signaling pathway in young horses affected by osteochondrosis
Serteyn, Didier ULg; Mendoza García, Luis ULg; Sandersen, Charlotte ULg et al

in Open journal of orthepedics (2014)

Osteochondrosis (OC) is a developmental disease in horses with a significant impact on the horse’s welfare and performance. The early disturbance of enchondral ossification progresses to inflammatory and ... [more ▼]

Osteochondrosis (OC) is a developmental disease in horses with a significant impact on the horse’s welfare and performance. The early disturbance of enchondral ossification progresses to inflammatory and healing process in older horses. Metabolic pathway analysis showed an obvious dysregulation of several signaling pathways related to cartilage formation and cartilage repair such as Wnt/β-catenin, Indian hedgehog and TGF-β signaling pathways. Other regulated genes appeared to be involved in high carbohydrate diet, abnormal insulin metabolism or inflammation. Sclerostin is an osteocyte-secreted soluble antagonist of the Wnt/β-catenin signaling pathway. It is crucial for osteoblast development and activity and is increased in naturally occurring lesions of equine osteochondrosis. The aim of this study is to compare the circulating sclerostin levels between OC-affected (n = 20) and healthy horses (n = 19). A significant linear regression between plasma sclerostin and age is observed especially in the healthy young horses. The mean plasma sclerostin concentration is significantly higher in young horses suffering from osteochondrosis compared to the control horses. These results reinforce the possible role of the Wnt/β-catenin signaling pathway in the OC pathogeny. The inhibition of this essential pathway could disturb the osteo-chondral differentiation. More studies are currently needed to define the eventual clinic interest of plasma sclerostin as future biomarker in bone and cartilage diseases. [less ▲]

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See detailMuscle Mitochondrial Dysfunction in Horses Affected by Acute Laminitis
Serteyn, Didier ULg; de la Rebière de Pouyade, Geoffroy ULg; Sandersen, Charlotte ULg et al

in Bioenergetics (2014)

Laminitis is a common and debilitating disease affecting horses and ponies. It often leads to the demise of the animal. Energy deficiency is suspected to entrain the disruption of the hemidesmosomes ... [more ▼]

Laminitis is a common and debilitating disease affecting horses and ponies. It often leads to the demise of the animal. Energy deficiency is suspected to entrain the disruption of the hemidesmosomes leading to the failure of the dermal-epidermal interface. The aim of this study was to measure the muscle mitochondrial function by high resolution respirometry. Muscle micro-biopsies were obtained from 11 horses affected by acute metabolic laminitis, 6 horses affected by acute laminitis resulting from a systemic inflammation response syndrome and 28 healthy horses distributed in 2 control groups: 17 horses with a body condition score [BSC, ranging from 0 (emaciated) to 5 (obese)] of 2 to 3 and 11 horses with a BSC of 4 to 5. During the acute phase of laminitis, a significant reduction of the muscle mitochondrial respiration was observed. The muscle mitochondrial dysfunction occurred independently of the etiology (metabolic disorder or systemic inflammation) leading to laminitis. The reduction of the oxidative phosphorylation and of the maximal respiratory capacity (after uncoupling) may induce depletion of the cell’s ATP content. If the same mitochondrial alteration occurs in the foot lamina, mitochondria targeting should be considered for the future, not only to better understand the physiopathology of the disease but also to maintain and to support the mitochondrial function before reaching the « mitochondrial dysfunction threshold » that may lead to the failure of the dermal-epidermal interface. [less ▲]

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See detailOsteochondrosis-Related Gene Expression in Equine Leukocytes Differs among Affected Joints in Foals
Serteyn, Didier ULg; Piquemal, David; Mendoza García, Luis ULg et al

in Journal of biomarkers and diagnosis (2014), 5

Osteochondrosis (OC) is a developmental disease in horses with a significant impact on the horse’s welfare and performance. Previously, differentially expressed genes in leukocytes of OC-affected have ... [more ▼]

Osteochondrosis (OC) is a developmental disease in horses with a significant impact on the horse’s welfare and performance. Previously, differentially expressed genes in leukocytes of OC-affected have been identified and were differentially expressed in horses of different ages when compared to their age-matched controls.As the time course of the development of OC lesions seems to be joint dependent,the aim of this study is to compare in young OCaffected horses (between 8 to 12 months), the different expression of selected genes depending the joints involved.The expression of OC-related genes were analysed by rt-PCR and subsequent statistical analysis (ΔΔCT) in the leukocytes of 30 Belgian Warmblood horses aged between 8 to 12 months divided in groups depending the affected joints (fetlock, hock and stifle).In the three groups, expression of ApoB-3G, MGAT4A, B4GALT6 and PRKCG genes were significantly higher in the OC-affected foals compared to the healthy foals. Based on the profiles of expression ofApoB-3G, Dsh1/Dvl1, Foxl1, Hp, ISG15, Mark2, PPR2A, RUSC2 and WASH1 genes,the localization of the disease can be determined: expression levels of ApoB3G, WASH1 and FOXl1 to identify fetlock, ApoB3G, PPR2A to identify OC-development in the hock and ApoB3G, Dsh1/Dvl1, WASH1, PPP2R1A and Mark2 geneto identify OC-development in the stifle. However at this moment, the rt-PCR analysis of the identified genes as biomarkers gives only diagnostic information. For the future, the profile of expression of these genes could give also some predictive information on the evolution of the disease such as remission or permanent OC-lesions. [less ▲]

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See detailAltered mitochondrial oxidative phosphorylation capacity in horses suffering from polysaccharide storage myopathy
Tosi, Irène ULg; Art, Tatiana ULg; Cassart, Dominique ULg et al

in Equine Veterinary Journal. Supplement (2014), 46(Suppl 46), 9

Introduction: Exertional myopathies are a common cause of exercise intolerance in the equine athlete, and Polysaccharide Storage Myopathy (PSSM) is a widely described muscular disease. It is characterized ... [more ▼]

Introduction: Exertional myopathies are a common cause of exercise intolerance in the equine athlete, and Polysaccharide Storage Myopathy (PSSM) is a widely described muscular disease. It is characterized by an accumulation of abnormal glycogen in myofibers due to a genetic defect in the skeletal muscle glycogen synthase (GYS1) enzyme. We hypothesized that energy production through oxidative phosphorylation (OXPHOS) in muscular mitochondria might be impaired in type-1 PSSM-affected horses. Methods: Eight horses with a history of exertional rhabdomyolysis were tested for the GYS1 mutation. Muscle biopsies were collected and used for histological analysis and high resolution respirometry (HRR). HRR values from 3 groups of horses (5 PSSM-positive horses, 3 horses with a history of myopathy but PSSM-negative and 16 healthy controls) were compared using a linear mixed model to take into account repeated (2–3 times) measurements made for each horse. Results: In 5/8 horses histology revealed an accumulation of abnormal glycogen in myofibers. These 5 horses also tested positive for the GYS1 mutation. A severe depression of maximal OXPHOS capacitywas observed by HRR in 7/8 horses with exertional rhabdomyolysis, with lower values in PSSM-positive cases (4/5). Conclusions: Our study shows a severely decreased OXPHOS capacity in PSSM-affected horses. PSSM is considered primarily a defect in glycogen synthesis but altered OXPHOS might play a central role in its pathogenesis. [less ▲]

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See detailNDS27 combines the effect of curcumin lysinate and hydroxypropyl-β-cyclodextrin to inhibit equine PKCδ and NADPH oxidase involved in the oxidative burst of neutrophils
Derochette, Sandrine ULg; Mouithys-Mickalad, Ange ULg; Franck, Thierry ULg et al

in FEBS Open Bio (2014), 4(0), 1021-1029

Polymorphonuclear neutrophils (PMNs) are involved in host defence against infections by the production of reactive oxygen species (ROS), but excessive PMN stimulation is associated with the development of ... [more ▼]

Polymorphonuclear neutrophils (PMNs) are involved in host defence against infections by the production of reactive oxygen species (ROS), but excessive PMN stimulation is associated with the development of inflammatory diseases. After appropriate stimuli, protein kinase C (PKC) triggers the assembly of NADPH oxidase (Nox2) which produces superoxide anion (O2●-), from which ROS derive. The therapeutic use of polyphenols is proposed to lower ROS production by limiting Nox2 and PKC activities. The purpose of this study was to compare the antioxidant effect of NDS27 and NDS28, two water-soluble forms of curcumin lysinate respectively complexed with hydroxypropyl-β-cyclodextrin (HPβCD) and γ-cyclodextrin (γ-CD), on the activity of Nox2 and PKCδ, involved in the Nox2 activation pathway. Our results, showed that NDS27 is the best inhibitor for Nox2 and PKCδ. This was illustrated by the combined effect of HPβCD and curcumin lysinate: HPβCD, but not γ-CD, improved the release of curcumin lysinate and its exchange against lipid or cholesterol as demonstrated by the lipid coloration with Oil red O, the extraction of radical lipophilic probes recorded by ESR and the HPLC measurements of curcumin. HPβCD not only solubilised and transported curcumin, but also indirectly enhanced its action on both PKC and Nox2 activities. The modulatory effect of NDS27 on the Nox2 activation pathway of neutrophils may open therapeutic perspectives for the control of pathologies with excessive inflammatory reactions. [less ▲]

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See detailEffect of inhaled hysrosoluble curcumin in lipopolysaccharide-induced lung neutrophilia in horses
Sandersen, Charlotte ULg; Cerri, Simona ULg; Franck, Thierry ULg et al

in Dorothy Russel Havemeyer Foundation IAD Workshop (2014)

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See detailEnhanced or Reduced Fetal Growth Induced by Embryo Transfer into Smaller or Larger Breeds Alters Post-Natal Growth and Metabolism in Pre-Weaning Horses
Peugnet, Pauline; Wimel, Laurence; Duchamp, Guy et al

in PLoS ONE (2014), 9(7), 102044

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See detailMyeloperoxidase and its products in synovial fluid of patients with treated or untreated rheumatoid arthritis.
Nzeusseu Toukap, A.; Delporte, C.; Noyon, C. et al

in Free radical research (2014), 48(4), 461-5

Abstract Objective. Plasma and synovial myeloperoxidase (MPO) and its products were strongly associated with osteoarthritis (OA) and rheumatoid arthritis (RA). In addition, it is well known that there is ... [more ▼]

Abstract Objective. Plasma and synovial myeloperoxidase (MPO) and its products were strongly associated with osteoarthritis (OA) and rheumatoid arthritis (RA). In addition, it is well known that there is a link between oxidative stress and cytokines. The present study aims at investigating the link between synovial MPO (and its products), interleukin (IL)-18, which is involved in the degradation of articular cartilage in RA, and IL-8, which is involved in recruitment and activation of neutrophils during inflammation. Effects of the treatment of RA on the biological parameters were also investigated. Methods. Patients (n = 105) were studied including 39 patients with OA, 33 with RA and 33 with RA receiving a specific treatment. Disease activity score (DAS-28) was calculated whereas MPO antigen/activity, neutrophils, chloro-tyrosine (Cl-Tyr), homocitrulline (Hcit), IL-8, and IL-18 were measured in synovial fluid (SF) and CRP was measured in serum. Results. DAS-28 and CRP levels were not significantly different between groups. MPO activity, and MPO, Cl-Tyr, and Hcit levels were significantly higher in SF of RA patients than OA patients. MPO specific activity (MPO activity/antigen ratio) was significantly lower in treated than in untreated RA patients as was IL-8. MPO activity and concentration were correlated with IL-8 and IL-18 in untreated but not in treated RA patients. Conclusions. MPO level is related to IL-8 and IL-18 levels in untreated RA patients. A link has been shown between treatment and decrease of IL-8, MPO specific activity and Hcit in SF. The causal role of MPO in SF inflammation and how treatment can affect MPO specific activity need further investigations. [less ▲]

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See detailEffect of different kinds of anoxia/reoxygenation on the mitochondrial function and the free radicals production of cultured primary equine skeletal myoblasts.
Ceusters, Justine ULg; Mouithys-Mickalad, Ange ULg; Franck, Thierry ULg et al

in Research in Veterinary Science (2013), 95

Horses are outstanding athletes, performing in many different disciplines involving different kinds of efforts and metabolic responses. Depending on exercise intensity, their skeletal muscle oxygenation ... [more ▼]

Horses are outstanding athletes, performing in many different disciplines involving different kinds of efforts and metabolic responses. Depending on exercise intensity, their skeletal muscle oxygenation decreases, and the reperfusion at cessation of the exercise can cause excessive production of free radicals. This study on cultured primary equine myoblasts investigated the effect of different kinds of anoxia/reoxygenation (A/R) on routine respiration, mitochondrial complex I specific activity and free radicals production. Our data revealed that short cycles of A/R caused a decrease of all the parameters, opposite to what a single long period of anoxia did. A preconditioning-like effect could explain our first pattern of results whereas mild uncoupling could be more appropriate for the second one. Anyway, it seems that mitochondrial complex I could play a major role in the regulation of the balance between metabolic and antioxidant protection of the muscular function of athletic horses. [less ▲]

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See detailEffect of non-sperm cells removal with single layer colloidal centrifugation on myeloperoxidase concentration in post-thaw equine semen
Ponthier, Jérôme ULg; Teague, Sheila; Franck, Thierry ULg et al

in Theriogenology (2013), 80(9), 1082-1087

Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in, and released by, neutrophils during degranulation or after lysis. Post-thaw semen contains MPO and its concentration is associated with ... [more ▼]

Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in, and released by, neutrophils during degranulation or after lysis. Post-thaw semen contains MPO and its concentration is associated with decreased sperm motility. Recently, MPO concentration in post-thaw semen was shown to be associated with presence of non-sperm cells. The aim of this study was to describe the effect of centrifugal fractionation of semen prior to cryopreservation on post-thaw concentrations of non-sperm cells and MPO. The experimental design consisted in freezing semen with or without previous centrifugation through two concentrations of single layer colloid media. Non-sperm cells and MPO concentrations were assessed in pellet and upper layer at each step of the procedure and MPO was detected in cells by immunocytochemistry. Single layer colloid centrifugation decreased non-sperm cells and MPO concentrations in post-thaw semen. The MPO concentration was correlated with concentration of non-sperm cells in the upper layer of the supernatant. In post-thaw semen, with or without previous single layer colloid centrifugation, MPO concentration was correlated with concentration of non-sperm cells. Overall, neutrophils were rarely observed and non-sperm cells were mainly epithelial cells or cellular debris, as demonstrated by MPO immunocytochemistry. Following single layer colloid centrifugation, MPO concentration was decreased, and correlations observed in different samples of the experiments revealed an association between concentrations of MPO and non-sperm cells. At all steps of the semen processing and cryopreservation, MPO immunostaining was clearly identified only on non-sperm cells. Our results demonstrate that non-sperm cells present in fresh semen release MPO during freezing. [less ▲]

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