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See detailDevelopment of an enzyme-linked immunosorbent assay for equine neutrophil elastase measurement in blood: Preliminary application to colic cases.
de la Rebière de Pouyade, Geoffroy ULg; Franck, Thierry ULg; Salciccia, Alexandra ULg et al

in Veterinary immunology and immunopathology (2010)

Equine neutrophil elastase (NE) is a protease released in inflammatory diseases and participating in tissue destruction. To measure NE in horse plasma to assess its role in pathological conditions, we ... [more ▼]

Equine neutrophil elastase (NE) is a protease released in inflammatory diseases and participating in tissue destruction. To measure NE in horse plasma to assess its role in pathological conditions, we purified elastase from equine neutrophils by a double step chromatography and obtained a pure protein of 27kDa, 4kDa smaller than the NE 2A previously purified (Scudamore et al., 1993; Dagleish et al., 1999), which was likely to be NE 2B. We developed an ELISA by using two specific polyclonal antibodies obtained from rabbit and guinea pig. The sandwich complex was detected using a secondary antibody conjugated to alkaline phosphatase. The ELISA showed good precision and accuracy, with intra- and inter-assay coefficients of variation below 10% for equine NE concentrations ranging from 1.875 to 60ng/ml. A stable plasma NE value, unaffected by the delay of centrifugation (over 4h), was obtained with plasma from EDTA anticoagulated blood. The mean value (+/-SEM) measured in 37 healthy horses was 32.53+/-4.6ng/ml. NE level in plasma of horses with colic at the time of admission was significantly higher than in healthy horses. Our results indicate that the ELISA technique we developed to measure plasmatic NE is a powerful tool for studying the role of elastase in equine inflammatory disease. In future, the application will be extended to other equine biological fluids. [less ▲]

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See detailLes tendinopathies et desmopathies des régions métacarpienne et –tarsienne: revue des thérapies actuelles. Seconde partie : les traitements
Verwilghen, Denis ULg; Caudron, Isabelle; Van Galen, Gaby ULg et al

in Annales de Médecine Vétérinaire (2010), 154(1), 1-15

Lesions to the horse’s tendon are very frequent and are a non negligible reason for early retirement of sport and leisure horses. For decades man has been searching for “the” treatment but unfortunately ... [more ▼]

Lesions to the horse’s tendon are very frequent and are a non negligible reason for early retirement of sport and leisure horses. For decades man has been searching for “the” treatment but unfortunately most of the known treatments have empiric bases and very few have actually proven their real efficiency. Diagnostic techniques like ultrasound allows to better evaluate the improvements made in tendonitis therapies. Stem cells and gene therapy are probably the most innovating of them, but their long-term effectiveness still has to stand the test of times. This article gives an overview of the possible treatments of tendonitis today. [less ▲]

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See detailPrevalence of radiographic findings in candidate sires (2001-2008)
Verwilghen, Denis ULg; Serteyn, Didier ULg; Pille, Frederik et al

in Vlaams Diergeneeskundig Tijdschrift (2009), 78

Radiographic screening of horses prior to purchase or admission to studbooks has become an important tool for evaluating the osteo-articular status of the animals. Developmental orthopedic disorders (DOD ... [more ▼]

Radiographic screening of horses prior to purchase or admission to studbooks has become an important tool for evaluating the osteo-articular status of the animals. Developmental orthopedic disorders (DOD) are the main cause of potential lameness in youngsters. The genetic factor associated with DOD has been established, and specific selection schemes have been developed to diminish the prevalence of lesions. Increasing knowledge of prevalence in different breeds may help in developing specific breeding strategies. This article presents the results of the radiographic screening of 676 stallions presented for sire admission to the Royal Belgian Sports Horse Society (Studbook sBs). On average, two radiographic findings were noted per horse. The prevalence of DOD in this population was 38.9%. A high percentage of horses showed radiographic signs of effusion of the distal interphalangeal joint. An uncommon fragment location at the proximal tubercle of the talus was found in 7 horses. The present paper presents a review and discussion of all the radiographic findings. [less ▲]

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See detailLes tendinopathies et desmopathies de la région métacarpienne et –tarsienne: une revue des thérapies actuelles. Première partie : la structure, les lésions du tendon et le diagnostic.
Verwilghen, Denis ULg; Gabriel, Annick ULg; Busoni, Valeria ULg et al

in Annales de Médecine Vétérinaire (2009), 3(153), 145-155

Lesions to the horse’s tendon are very frequent and are a non negligible reason for early retirement of sport and leisure horses. For decades man has been searching for “the” treatment but unfortunately ... [more ▼]

Lesions to the horse’s tendon are very frequent and are a non negligible reason for early retirement of sport and leisure horses. For decades man has been searching for “the” treatment but unfortunately most of the known treatments have empiric bases and very few have actually proven their real efficiency. Better knowledge of tendon anatomy and pathophysiology of tendonitis together with evolutions in diagnostic techniques like ultrasonography allows us to better evaluate the improvements made in tendonitis therapies today. The first part of this article gives a review on tendon structure, tendon healing and ultrasonographic evaluation of the tendon. In the second part an overview of therapeutic options for tendonitis will be discussed. [less ▲]

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See detailEvolution of blood myeloperoxidase in the perioperative period of horses undergoing emergency celiotomy.
Salciccia, Alexandra ULg; Grulke, Sigrid ULg; de la Rebière de Pouyade, Geoffroy ULg et al

Poster (2009, July)

Colic can cause an activation of neutrophils with release in the blood flow of myeloperoxidase (MPO), a specific enzyme with strong oxidative activity. The aim of this study was to describe the evolution ... [more ▼]

Colic can cause an activation of neutrophils with release in the blood flow of myeloperoxidase (MPO), a specific enzyme with strong oxidative activity. The aim of this study was to describe the evolution of plasma MPO after colic surgery. Materials included 13 adult horses that underwent an emergency celiotomy for acute intestinal obstruction. Venous blood samples were collected into EDTA anticoagulated tubes before surgery, during surgery after correction of the intestinal lesion and during the recovery of anaesthesia. In the postoperative period samples were taken every 4 hours during the first 4 days (from day 0 until day 4), every 12 hours during the days 4 and 5 and every 24 hours until day 10. MPO was assayed with a specific enzyme-linked immunosorbent assay. Statistical analysis was performed by one way Anova with student- Newman-Keuls post test on data obtained for each time point. Significance was set at p < 0.05. The horses were operated for an obstruction strangulated or not of the small or the large intestine. In six cases the postoperative period was uneventful, the 7 remaining developed one or 2 severe complications. Eight horses were discharged and 5 died during the hospitalization.The general aspect of the curve of mean plasmatic MPO can be described as follow: An increase was observed from the admission on until a peak of concentration occurring generally during the time of recovery from the anaesthesia with the highest mean value reaching 740.84 +/- 507.61ng/ml. This was followed by a progressive decrease until the lowest value, usually near to day 2 after the recovery from anaesthesia corresponding to 171.79 +/- 76.21 ng/ml of MPO. Afterwards, the mean concentrations increased slowly until postoperative day 10. In the majority of cases a stable and low MPO value (plateau) was observed during approximately 2 days (from day 1 to day 3 postoperatively).The initial peak of MPO after surgery could be associated to the neutrophil activation consequent to the intestinal disorder and the intense stimulation of the coeliotomy. The following significant reduction in concentration could be attributed to MPO infiltration into the tissues with a critical point at approximately 2 days after surgery. This study may contribute to a better understanding of the role of the MPO and neutrophils in the pathophysiology of horses in the postoperative period after colic surgery. [less ▲]

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See detailEvaluation of low-molecular-weight heparin for the prevention of equine laminitis after colic surgery
de la Rebière de Pouyade, Geoffroy ULg; Grulke, Sigrid ULg; Detilleux, Johann ULg et al

in Journal of Veterinary Emergency and Critical Care (2009), 19(1), 113-119

Objectives – The aim of this study is to describe the prevalence of postoperative laminitis in colic cases and to determine if low-molecular-weight heparin (LMWH) is effective in preventing this ... [more ▼]

Objectives – The aim of this study is to describe the prevalence of postoperative laminitis in colic cases and to determine if low-molecular-weight heparin (LMWH) is effective in preventing this complication. Design – Retrospective clinical study. Animals – Client-owned horses. Interventions – SC administration of enoxaparin during the postoperative period. Measurements and Main Results – Medical records of 360 horses undergoing surgery for colic and surviving at least 3 days were evaluated. Fifty-six horses admitted before 1995 did not receive LMWH (control group) and 304 admitted after 1995 received LMWH as a prophylaxis for laminitis (treatment group). Three grades of severity were defined for laminitis. Prevalence and severity of laminitis were compared between the 2 groups. Several parameters recorded on admission (sex, age, breed, site and nature of the disease, heart rate, PCV, gravity score, and shock score) and the administration of LMWH were tested as risk factors in the development of laminitis in a logistic regression procedure. Prevalence and grade of laminitis were significantly lower in the treatment group. Only the absence of LMWH was recognized as a significant risk factor in the logistic regression model. Conclusions – The administration of LMWH appears to be effective in the prophylaxis of laminitis following colic surgery and may be useful in the postoperative management of these horses. [less ▲]

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See detailRelationship between biochemical markers and radiographic scores in the evaluation of the osteoarticular status of Warmblood stallions
Verwilghen, Denis ULg; Busoni, Valeria ULg; Gangl, Monika et al

in Research in Veterinary Science (2009), 87(2), 319-28

Establishing the osteoarticular status of the horse is often performed by means of radiological screening of the animals. Widespread blood sampling could reveal to be an interesting alternative to this ... [more ▼]

Establishing the osteoarticular status of the horse is often performed by means of radiological screening of the animals. Widespread blood sampling could reveal to be an interesting alternative to this procedure which is time consuming and sometimes technically difficult. The aim of this study was to investigate the relationship between the radiological status of the horses and the levels of biochemical markers of cartilage degradation and synovial inflammation. A specific radiological scoring and classification system was therefore developed and applied on 63 stallions presented for studbook admission. Additionally, groups of horses were established according to the occurrence of osteochondrosis, degenerative joint disease and distal interphalangeal joint effusion. Insulin growth factor-I, myeloperoxidases, Coll2-1 and Coll2-1NO(2) were used as blood markers. The combination of the blood parameters did not seem to correlate with the used scoring system. Coll2-1NO(2) levels however tended to increase with poorer radiological class and this could therefore potentially be a useful predictor of the osteoarticular status in the horse. Coll2-1 levels were significantly higher in the degenerative joint disease group. A high percentage of horses with distal interphalangeal joint effusion was present in this study and was associated with decreased IGF-I and increased Coll2-1 levels. [less ▲]

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See detailDie exokrine Pankreasfunktion beim kranken Pferd
Sandersen, Charlotte ULg; Serteyn, Didier ULg; Grulke, Sigrid ULg

in Vervuert; Achenbach; Gäbel (Eds.) et al LBH, Proccedings 5. Leipziger Tieraerzte Tag (2009)

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See detailBiochemical markers and radiographic scores as an evaluation for the osteoarticular status of Warmblood stallions.
Verwilghen, Denis ULg; Busoni, Valeria ULg; Salciccia, Alexandra ULg et al

(2009)

Establishing the osteoarticular status of the horse is often performed by means of radiological screening of the animals. Widespread blood sampling could potentially be an alternative to this procedure ... [more ▼]

Establishing the osteoarticular status of the horse is often performed by means of radiological screening of the animals. Widespread blood sampling could potentially be an alternative to this procedure which is time consuming and sometimes technically difficult. The aim of this study was to investigate the relationship between the radiological status of the horses and the levels of biochemical markers (BM) of cartilage degradation and synovial inflammation. Materials and Methods: A specific radiological scoring (RS) system was developed and applied on 63 stallions presented for studbook admission. Additionally, groups of horses were established according to the occurrence of osteochondrosis (OC), degenerative joint disease (DJD) and distal interphalangeal joint (DIPJ) effusion. Insulin growth factor-I, myeloperoxidases, Coll2-1 and Coll2-1NO2 were used as BM. The effects of age and weight on the BM and of the BM on the RS were measured. Mean values of BM between OC positive versus negative, DJD positive versus negative and DIPJ effusion positive versus negative were compared using SAS statistical program. Results: No effect of the combined BM was found on the RS. No significant differences between the values of the BM were found within the different radiological classes (RC). However, considering only the Coll2-1NO2 marker, the probability to belong to class A was given by the equation 9.63 -0.31 Coll2-1NO2 + 0.04 (Coll2-1NO2)². Equally a tendency (p= 0.06) towards an increase in RC by 0.45 for each increasing unit of Coll2-1NO2 was present. Significant positive correlations were found between Coll2-1 values and Coll2-1 NO2 values. MPO values were found to be significantly higher in OC negative horses (357.86  64.43 ng/ml) than in OC positive horses (231.17  27.15 ng/ml). IGF-I levels were found significantly lower in the positive DIPJ effusion group (416.05  17.57 ng/ml) compared to the negative DIPJ effusion group (477.1  22.40 ng/ml). Coll2-1 values were significantly higher in the positive DIPJ effusion group (911.04  41.13 nM) compared to the negative DIPJ effusion group (769.04  37.92 nM). Mean values of Coll2-1 were significantly higher in the DJD positive group (901.24  36.75 nM) compared to the DJD negative group (790.590  39.397 nM) Discussion: The combination of the blood parameters did not seem to correlate with the used RS system. Coll2-1NO2 levels however tend to increase with poorer RC and could therefore be used as a useful predictor of the osteoarticular status of the horse. Coll2-1 levels were significantly higher in the degenerative joint disease group. A high percentage of horses with DIPJ effusion was present in this study and was associated with decreased IGF-I and increased Coll2-1 levels. [less ▲]

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See detailVariations of plasmatic concentrations of Insulin-like Growth Factor-I in post-pubescent horses affected with developmental osteochondral lesions.
Verwilghen, Denis ULg; Vander Heyden, Laurent ULg; Franck, Thierry ULg et al

in Veterinary Research Communications (2009)

Developmental osteochondral lesions are often encountered in the equine population and are a major cause of lameness. Different growth factors that act systemically as well as locally regulate the growth ... [more ▼]

Developmental osteochondral lesions are often encountered in the equine population and are a major cause of lameness. Different growth factors that act systemically as well as locally regulate the growth of cartilage. Among them is Insulin-like Growth Factor I that has been demonstrated to promote chondrocyte growth and differentiation and that has been shown to influence cartilage repair. The aims of this study were to investigate differences in circulating plasma levels of Insulin-like Growth Factor-I in post-pubescent horses affected with developmental osteochondral lesions compared to unaffected ones. Significantly higher values of circulating Insulin-like Growth Factor-I levels were found in the affected group (n = 82) compared to controls (n = 86). This result may still reflect an earlier imbalance in IGF-I levels from horses with developmental osteochondral lesions considering the aetiopathological link which has been made between IGF-I and the occurrence of osteochondrosis. However, other studies have shown increased expression of IGF-I after cartilage damage. The higher levels found in this study could be due to a healing response of the cartilage to the damage caused by the osteochondral lesions. [less ▲]

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See detailIn vivo administration of acepromazine or promethazine to horse decreases the reactive oxygen species production response of subsequently isolated neutrophils to stimulation with phorbol myristate acetate
Péters, Fabrice; Franck, Thierry ULg; Pequito, Manuel et al

in Journal of Veterinary Pharmacology & Therapeutics (2009)

The previous experiments have shown that some phenothiazines have antioxidant and anti-inflammatory properties in vitro. In this study the inhibition of the production of reactive oxygen species (ROS) by ... [more ▼]

The previous experiments have shown that some phenothiazines have antioxidant and anti-inflammatory properties in vitro. In this study the inhibition of the production of reactive oxygen species (ROS) by neutrophils was studied in two groups of horses, which received a dose of 0.1 mg⁄ kg of either acepromazine or promethazine intravenously. Blood samples were collected before (T0) and 0.5, 1, 3 and 5 h after drug administration. The chemiluminescence (CML) response of neutrophils was measured ex vivo in the presence of luminol for a period of 10 min and the maximum CML value (peak value) recorded. There was a significant inhibition of the ROS production in the acepromazine treated group (49% inhibition) at 5 h after administration and in the promethazine group (24% inhibition) at 3 h after administration (P < 0.05 vs. T0). These findings are of therapeutic relevance in the use of phenothiazines in equine patients with inflammatory diseases where neutrophil activation and ROS production are implicated. [less ▲]

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See detailEvaluation of sedation depth and quality of performed procedures following administration of α2-agonists combined or not with butorphanol in horses.
Barbry, Stephanie; Verwilghen, Denis ULg; Van Galen, Gaby ULg et al

Poster (2008, October)

Several molecules available in equine medicine provide profound sedation and good analgesia when performing diagnostic or surgical procedures. They differ slightly in degree of action, as well in time as ... [more ▼]

Several molecules available in equine medicine provide profound sedation and good analgesia when performing diagnostic or surgical procedures. They differ slightly in degree of action, as well in time as in depth. The aim of this study was to evaluate the depth of sedation and quality of performing a standardized procedure after injection of an 2-agonist alone or associated with butorphanol. Horses presented to the Equine Clinic of Liege University from December 2007 to June 2008 for an endoscopic, dental, radiological or ophthalmological examination of which owner and clinician agreed to include the horse in the study were retained. All horses (n=133) were sedated in a randomized double blind fashion using one of the six following protocols: detomidine (D) (0.01mg/kg IV), romifidine (R) (0.04mg/kg IV), or xylazine (X) (0.5mg/kg IV) alone or associated with butorphanol (B) (0.02mg/kg IV). Stability, head position and the reaction to a stimulus (auditive and visual) were evaluated before the start of the procedure. An overall subjective grade using a 10 cm line scale was given to evaluate sedation depth (ranging from no effect to maximal depth on line) and the quality of the performed procedure (ranging from not realised to ideal examination). For each specific procedure, the same clinician experienced in his particular field evaluated the parameters. Data were related by a variance analysis using SAS system and considered significant if P<0.01. The used protocols significantly influenced both sedation depth and examination quality. Sedation depth increased significantly when B was added in all the three 2-agonists used (mean ± SD): D: 6.46±0.33; D+B: 7.06±0.37; R: 4.93±0.46; R+B: 7.04±0.45; X: 4.94±0.43; X+B: 7.75±0.40. Detomidine provided a significantly better quality of the examination compared to romifidine and xylazine. The addiction of butorphanol significantly decreased the examination quality when added to detomidine and increased the quality when combined with romifidine and xylazine. Mean (± SD) of the quality of performed procedures: D: 8.41±0.54; D+B: 6.06±0.64; R: 6.68±0.71; R+B: 7.16±0.67; X: 4.47±0.67; X+B: 4.52±0.64. Comparison of sedation depth and quality of performed examination revealed an increase in quality of examination scores with increasing depth of sedation until a certain level. As from sedation depths with scores of 8 or more, a decrease in quality of examination was encountered. This report shows the important effect of butorphanol addition on sedation. However, increasing sedation depth was not necessarily linked to a better quality of procedure, notably due to difficulties in handling strongly sedated horses for the procedures performed in this study. [less ▲]

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See detailMandibular osteodistraction for correction of deep bite class II malocclusion in a horse
Verwilghen, Denis ULg; Van Galen, Gaby ULg; Vander Heyden, Laurent ULg et al

in Veterinary Surgery : The Official Journal of the American College of Veterinary Surgeons (2008), 37(6), 571-579

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See detailMandibular osteodistraction as a corrective method for class II malocclusions in the horse.
Verwilghen, Denis; Van Galen, Gaby; Busoni, Valeria ULg et al

Poster (2008, July)

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See detailMyeloperoxidase Assay in Plasma and Peritoneal Fluid of Horses with Gastrointestinal Disease
Grulke, Sigrid ULg; Franck, Thierry ULg; Gangl, Monika et al

in Canadian Journal of Veterinary Research = Revue Canadienne de Recherche Vétérinaire (2008), 72(1), 37-42

Gastrointestinal disorders, especially strangulating intestinal obstructions, are still a major cause of illness and death in the horse. Circulating lipopolysaccharides may activate both neutrophils and ... [more ▼]

Gastrointestinal disorders, especially strangulating intestinal obstructions, are still a major cause of illness and death in the horse. Circulating lipopolysaccharides may activate both neutrophils and monocytes. The activated neutrophils release myeloperoxidase (MPO), a specific enzyme with strong oxidative activity. The aim of this study was to evaluate MPO concentrations in the plasma and peritoneal fluid (PF) of horses with colic and to check the hypothesis that these concentrations would be higher in a case of strangulating obstruction than in cases of nonstrangulating disease. By using a specific enzyme-linked immunosorbent assay for equine MPO, we determined the MPO concentrations in horses admitted to a clinic for colic. Horses with nonstrangulating or strangulating obstruction of the large intestine (NSLI or SLI), strangulating obstruction of the small intestine (SSI), or inflammatory bowel disease (IBD) were compared with healthy horses. The horses with SLI, SSI, or IBD had significantly higher MPO levels in plasma and PF than did those in the other 2 groups. The mean plasma level was significantly higher in the horses with NSLI than in the healthy horses. High MPO values in PF indicated necrotic bowel. These results show that neutrophil activation occurs during nonstrangulating and strangulating intestinal obstruction in horses and that the plasma and PF MPO concentrations may be a marker of the severity of the disease. [less ▲]

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See detailInhibitory effect of curcuminoids and tetrahydrocurcuminoids on equine activated neutrophils and myeloperoxidase activity
Franck, Thierry ULg; Kohnen, Stephan ULg; Grulke, Sigrid ULg et al

in Physiological Research (2008), 57(4), 577-587

In the horse, the inflammation response to various pathologies (intestinal strangulations, laminitis, etc.) involves an excessive stimulation of the polymorphonuclear neutrophils releasing reactive oxygen ... [more ▼]

In the horse, the inflammation response to various pathologies (intestinal strangulations, laminitis, etc.) involves an excessive stimulation of the polymorphonuclear neutrophils releasing reactive oxygen species (ROS) and myeloperoxidase (MPO). The aim of the present work was to study the effect of natural polyphenols, curcuminoids and tetrahydrocurcuminoids (THC) on isolated stimulated equine neutrophils and on the activity of purified MPO. The ROS production and the release of MPO by activated neutrophils were measured by chemiluminescence and ELISA techniques, respectively. The activity of purified MPO was measured by studying its nitration, chlorination or oxidation capacity and by using an original method called SIEFED allowing the study of drug interaction with the enzyme without interferences of the medium. Curcuminoids and THC had dose-dependent inhibitory effects on ROS production and MPO release by activated neutrophils and on purified MPO activity. We suggest that the higher efficacy of curcuminoids versus THC could be explained, at least partially, by its chemical structure: the conjugated double bounds and the plane structure of curcuminoids made easier the neutralization of the radical species generated by activated neutrophils and the interaction of the drug with the active site of MPO. These inhibitory effects of curcuminoids on the oxidant activity of equine neutrophils and on MPO activity open therapeutic perspectives in equine pathologies with excessive inflammatory reactions. [less ▲]

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