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Selected AbstractsSusceptibility of isolated myofibrils to in vitro glutathionylation: Potential relevance to muscle functions,CYTOSKELETON, Issue 2 2010Chiara Passarelli Abstract In this study we investigated the molecular mechanism of glutathionylation on isolated human cardiac myofibrils using several pro-glutathionylating agents. Total glutathionylated proteins appeared significantly enhanced with all the pro-oxidants used. The increase was completely reversed by the addition of a reducing agent, demonstrating that glutathione binding occurs by a disulfide and that the process is reversible. A sensitive target of glutathionylation was ,-actin, showing a different reactivity to the several pro-glutathionylating agents by ELISA. Noteworthy, myosin although highly sensitive to the in vitro glutathionylation does not represent the primary glutathionylation target in isolated myofibrils. Light scattering measurements of the glutathionylated ,-actin showed a slower polymerisation compared to the non-glutathionylated protein and force development was depressed after glutathionylation, when the myofibrils were mounted in a force recording apparatus. Interestingly, confocal laser scanning microscopy of cardiac cryosections indicated, for the first time, the constitutive glutathionylation of ,-cardiac actin in human heart. Due to the critical location of ,-actin in the contractile machinery and to its susceptibility to the oxidative modifications, glutathionylation may represent a mechanism for modulating sarcomere assembly and muscle functionality under patho-physiological conditions in vivo. © 2009 Wiley-Liss, Inc. [source] Pathology of lethal peripartum broad ligament haematoma in 31 Thoroughbred maresEQUINE VETERINARY JOURNAL, Issue 6 2010T. UENO Summary Reasons for performing study: Broad ligament haemorrhage in peripartum mares is a life-threatening disease and there are few reports on the aetiology and pathogenesis of broad ligament haematoma. Objectives: To obtain information regarding the sites for the early diagnosis and pathogenesis of broad ligament haematoma of mares. Methods: Thirty-one mares that died of broad ligament haematoma peripartum were examined pathologically for bleeding sites. The arterial distribution of 5 young mares with several parities served as negative controls. Results: Age and/or multiparity were the predisposing factors for the disease. Arterial injuries were most commonly observed in the uterine artery (24 of 31 mares). Among these, the proximal uterine artery that lies within 15 cm of the bifurcation of the iliac artery was the most frequent site for rupture (18 mares). The lesions occurred preferentially at the bifurcations, lateral part of curvatures and abrupt flexures of the artery. The morphology of the injuries was classified into 3 types: ruptures with and without longitudinal fissures, and transections. Histologically, the arterial wall adjacent to the rupture showed atrophy of smooth muscle cells with fibrosis of the tunica media and disruption and/or calcification of the internal elastic lamina. Conclusions: Arterial injuries that led to broad ligament haematoma in peripartum mares occurred most frequently in the proximal uterine artery, and atrophy of smooth muscle cells with fibrosis of the arterial wall was as one of the predisposing factors in aged and multiparous mares. Potential relevance: Monitoring small aneurysms, mural tearing, medial fibrosis at the proximal uterine artery by transrectal echography could provide useful information for the early diagnosis and possible prevention of broad ligament haematoma of peripartum mares. [source] Lateralised motor behaviour leads to increased unevenness in front feet and asymmetry in athletic performance in young mature Warmblood horsesEQUINE VETERINARY JOURNAL, Issue 5 2010M. C. V. Van HEEL Summary Reason for performing study: Foot stance in grazing significantly influences hoof conformation and development from foal to yearling age. Objectives: To conduct a longitudinal study to establish if the relationship between motor laterality and uneven front feet persisted in 3-year-old horses at the time of studbook selection and to investigate if such laterality and unevenness might influence the horses' ability to perform symmetrically while trotting, cantering and free jumping. Methods: Seventeen clinically sound but untrained (with only minimal experience of handling) and sound Warmblood horses that had participated in a previous study were assessed as per the protocol reported. Laterality was tested in a preference test (PT) and z -values were calculated for analysis purposes. Laterality and hoof unevenness were related to both relative limb length and relative head size, while the ability to perform symmetrically was tested in free trot-canter transitions and free jumping exercises. Differences in performance between horses with and without a limb preference in the PT and those with ,uneven' and ,even' feet were tested for differences in performance metrics using Students' t test, while linearity was tested using a regression analysis (P<0.05). Results: Significant laterality was still present in 24% of the 3-year-old horses and the relationship between laterality and uneven feet pairs was stronger than at foal and yearling stages. Horses with significant motor laterality had almost 4 times more unevenness, a smaller head and longer limbs and the relationship between body conformation and laterality was still present. There was a strong linear relation between unevenness, laterality and a bias or side preference for trot-canter transitions. However, this relationship was not significant during the free jumping exercise. Conclusion: Motor laterality and uneven feet pairs were still present and significantly related in the 3-year-old horses and both variables were also strongly related to sidedness in trot-canter transitions. Potential relevance: Warmblood studbooks should include quantitative data on laterality at the time of studbook admission as part of the selection criteria. [source] Intralesional bovine papillomavirus DNA loads reflect severity of equine sarcoid diseaseEQUINE VETERINARY JOURNAL, Issue 4 2010R. HARALAMBUS Summary Reasons for performing study: Sarcoids are nonmetastasising, yet locally aggressive skin tumours that constitute the most frequent neoplasm in equids. Infection by bovine papillomaviruses types 1 and 2 (BPV-1, BPV-2) has been recognised as major causative factor in sarcoid pathogenesis, but a possible correlation of intralesional virus load with disease severity has not been established thus far. Hypothesis: Given the pathogenic role of BPV-1 and BPV-2 in sarcoid disease, we suggest that intralesional viral DNA concentration may reflect the degree of affection. Methods: Severity of disease was addressed by recording the tumour growth kinetics, lesion number and tumour type for 37 sarcoid-bearing horses and one donkey. Viral load was estimated via quantitative real-time PCR (qPCR) of the E2, E5, L1 and L2 genes from the BPV-1/-2 genome for one randomly selected lesion per horse and correlated with disease severity. Results: Quantitative PCR against E2 identified viral DNA concentrations ranging from 0,556 copies/tumour cell. Of 16 horses affected by quiescent, slowly growing single tumours or multiple mild-type lesions, 15 showed a viral load up to 1.4 copies per cell. In stark contrast, all equids (22/22) bearing rapidly growing and/or multiple aggressive sarcoids had a viral load between 3 and 569 copies per cell. Consistent results were obtained with qPCR against E5, L1 and L2. Conclusions: While tumours of the same clinical type carried variable virus load, confirming that viral titre does not determine clinical appearance, we identified a highly significant correlation between intralesional viral load and disease severity. Potential relevance: The rapid determination of BPV viral load will give a reliable marker for disease severity and may also be considered when establishing a therapeutic strategy. [source] Comparisons of overground endoscopy and treadmill endoscopy in UK Thoroughbred racehorsesEQUINE VETERINARY JOURNAL, Issue 3 2010K. J. ALLEN Summary Reasons for performing study: To date there is no information on the comparison of the more recently documented technique of performing endoscopy during ridden exercise in the field, with the more traditional method of endoscopy during high-speed treadmill exercise. Objectives: To compare the results of upper respiratory tract endoscopy in UK Thoroughbred racehorses performed during ridden exercise in the field with those obtained during exercise on the treadmill. Methods: A direct comparison was undertaken in 4 horses whereby both procedures were performed in the same horse within 10 days of each other. An indirect comparison was also undertaken whereby the results of overground endoscopy performed in 50 racehorses was compared to the results obtained during treadmill endoscopy in a further 50 racehorses. Horses were matched for age, gender, use (National Hunt vs. Flat) and presenting complaint (abnormal respiratory noise vs. poor performance). Results: Dorsal displacement of the soft palate was diagnosed less frequently during overground endoscopy than during treadmill endoscopy. There was no significant difference in the diagnosis of dynamic laryngeal collapse between the 2 techniques. The treadmill exercise test was performed over longer distances at higher inclines, albeit at lower speeds than the overground test. In contrast to the treadmill test, the overground test was frequently performed in intervals. Conclusions: The results of both the direct and indirect comparisons suggest that dorsal displacement of the soft palate is diagnosed less often during overground endoscopy than during treadmill endoscopy. Strenuous exercise tests may be more easily performed on a treadmill than by performing multiple exercise intervals in the field. Potential relevance: Care should be taken in interpreting negative findings during both procedures, but particularly during overground endoscopy if racing conditions have not been appropriately replicated. [source] Effect of intravenous lidocaine administration on laminar inflammation in the black walnut extract model of laminitisEQUINE VETERINARY JOURNAL, Issue 3 2010J. M. WILLIAMS Summary Reasons for performing study: Laminitis is a serious complication of horses suffering from sepsis/endotoxaemia-related events. Laminitis in horses and organ injury in human sepsis are both reported to involve inflammatory injury to the laminae/organs including early activation of endothelium and leucocytes leading to emigration of neutrophils into the tissue interstitium. In the black walnut extract (BWE) model, systemic inflammatory events coincide with marked increase in laminar mRNA concentrations of inflammatory genes including proinflammatory cytokines (i.e. IL-1,, IL-6), COX-2, chemokines (i.e. IL-8) and endothelial adhesion molecules (i.e. ICAM-1 and E-selectin). In models of human sepsis, i.v. lidocaine has been reported to decrease leucocyte and endothelial activation, and the expression of proinflammatory cytokines and chemokines. Objectives: To evaluate the effect of i.v. lidocaine therapy on the inflammatory processes documented to occur in the BWE model of laminitis. Methods: Twelve horses were administered BWE and treated immediately with either lidocaine (1.3 mg/kg bwt bolus, followed by 0.05 mg/kg bwt/min CRI, n = 6) or saline (n = 6) for 10 h. At 10 h post BWE administration, laminar samples were obtained under general anaesthesia for assessment of proinflammatory gene expression (using RT-qPCR) and leucocyte emigration (via CD13 immunohistochemistry). At 0, 3 and 10 h post BWE administration, skin samples were obtained for assessment of leucocyte emigration (via calprotectin immunohistochemistry). Results: No significant differences between groups were noted for inflammatory gene mRNA concentrations (IL-1,, IL-6, IL-8, COX-2) or for number of leucocytes present within the laminar interstitium or skin dermis. Increased (P<0.05) laminar E-selectin mRNA concentrations were present in the LD group (vs. SAL group). Conclusions: Continuous administration of i.v. lidocaine does not inhibit inflammatory events in either the laminae or skin in the horse administered black walnut extract. Potential relevance: This work questions the use of continuous i.v. administration of lidocaine as an effective anti-inflammatory therapy for systemic inflammation. [source] Repeatability of subjective evaluation of lameness in horsesEQUINE VETERINARY JOURNAL, Issue 2 2010K. G. KEEGAN Summary Reasons for performing study: Previous studies have suggested that agreement between equine veterinarians subjectively evaluating lameness in horses is low. These studies were limited to small numbers of horses, evaluating movement on the treadmill or to evaluating previously-recorded videotape. Objectives: To estimate agreement between equine practitioners performing lameness evaluations in horses in the live, over ground setting. Methods: 131 mature horses were evaluated for lameness by 2,5 clinicians (mean 3.2) with a weighted-average of 18.7 years of experience. Clinicians graded each limb using the AAEP lameness scale by first watching the horse trot in a straight line only and then after full lameness evaluation. Agreement was estimated by calculation of Fleiss' (,). Evaluators agreed if they picked the same limb as lame or not lame regardless of the severity of perceived lameness. Results: After only evaluating the horse trot in a straight line clinicians agreed whether a limb was lame or not 76.6% of the time (,= 0.44). After full lameness evaluation clinicians agreed whether a limb was lame or not 72.9% of the time (,= 0.45). Agreement on forelimb lameness was slightly higher than on hindlimb lameness. When the mean AAEP lameness score was >1.5 clinicians agreed whether or not a limb was lame 93.1% of the time (,= 0.86), but when the mean score was ,1.5 they agreed 61.9% (,= 0.23) of the time. When given the task of picking whether or not the horse was lame and picking the worst limb after full lameness evaluation, clinicians agreed 51.6% (,= 0.37) of the time. Conclusions: For horses with mild lameness subjective evaluation of lameness is not very reliable. Potential relevance: A search for and the development of more objective and reliable methods of lameness evaluation is justified and should be encouraged and supported. [source] In vitro effects of lidocaine on the contractility of equine jejunal smooth muscle challenged by ischaemia-reperfusion injuryEQUINE VETERINARY JOURNAL, Issue 1 2010M. GUSCHLBAUER Summary Reasons for performing study: Post operative ileus (POI) in horses is a severe complication after colic surgery. A commonly used prokinetic drug is lidocaine, which has been shown to have stimulatory effects on intestinal motility. The cellular mechanisms through which lidocaine affects smooth muscle activity are not yet known. Objectives: To examine the effects of lidocaine on smooth muscle in vitro and identify mechanisms by which it may affect the contractility of intestinal smooth muscle. Hypothesis: Ischaemia and reperfusion associated with intestinal strangulation can cause smooth muscle injury. Consequently, muscle cell functionality and contractile performance is decreased. Lidocaine can improve basic cell functions and thereby muscle cell contractility especially in ischaemia-reperfusion-challenged smooth muscle. Methods: To examine the effects of lidocaine on smooth muscle function directly, isometric force performance was measured in vitro in noninjured and in vivo ischaemia-reperfusion injured smooth muscle tissues. Dose-dependent response of lidocaine was measured in both samples. To assess membrane permeability as a marker of basic cell function, release of creatine kinase (CK) was measured by in vitro incubations. Results: Lidocaine-stimulated contractility of ischaemia-reperfusion injured smooth muscle was more pronounced than that of noninjured smooth muscle. A 3-phasic dose-dependency was observed with an initial recovery of contractility especially in ischaemia-reperfusion injured smooth muscle followed by a plateau phase where contractility was maintained over a broad concentration range. CK release was decreased by lidocaine. Conclusion: Lidocaine may improve smooth muscle contractility and basic cell function by cellular repair mechanisms which are still unknown. Improving contractility of smooth muscle after ischaemia-reperfusion injury is essential in recovery of propulsive intestinal motility. Potential relevance: Characterisation of the cellular mechanisms of effects of lidocaine, especially on ischaemia-reperfusion injured smooth muscle, may lead to improved treatment strategies for horses with POI. [source] Equine laminitis: Ultrastructural lesions detected in ponies following hyperinsulinaemiaEQUINE VETERINARY JOURNAL, Issue 7 2009A. R. NOURIAN Summary Reasons for performing study: Anatomical changes in the hoof lamellar tissue induced by prolonged hyperinsulinaemia have not been described previously. Analysis of the induced lesions may promote understanding of hyperinsulinaemic laminitis pathogenesis and produce clinical benefit. Objectives: To use light and transmission electron microscopy (TEM) to document hoof lamellar lesions in ponies clinically lame after prolonged hyperinsulinaemia. Methods: Nine clinically normal, mature ponies were allocated randomly to either a treatment group (n = 5) or control group (n = 4). The treatment group received insulin via a modified, prolonged euglycaemic hyperinsulinaemic clamp technique (EHCT) and were subjected to euthanasia when clinical signs of Obel grade II laminitis occurred. The control group was sham treated with an equivalent volume of 0.9% saline and killed at 72 h. Lamellar tissues of the right front feet were harvested and processed for TEM. Results: Lamellae from insulin treated ponies were attenuated and elongated with many epidermal basal cells (EBC) in mitosis. Unlike carbohydrate induced laminitis in horses there was no global separation at the lamellar dermal/epidermal interface among ponies. Sporadic EBC basement membrane (BM) separation was associated with the proximity of infiltrating leucocytes. In 2 ponies, the lamellar BM was thickened. The number of hemidesmosomes/,m of BM was decreased in all insulin treated ponies. Conclusions: Prolonged hyperinsulinaemia causes unique lamellar lesions normally characteristic of acute and chronic laminitis. Lamellar proliferation may be an insulin effect through its mitogenic pathway. Aberrant lamellar mitosis may lengthen and weaken the lamellar, distal phalanx attachment apparatus and contribute to the clinical signs that developed. Potential relevance: The study shows that insulin alone, in higher than normal circulating concentrations, induces profound, changes in lamellar anatomy. Medical control of insulin resistance and hyperinsulinaemia may ameliorate lesions and produce clinical benefit. [source] Regional distribution of collagen and haemosiderin in the lungs of horses with exercise-induced pulmonary haemorrhageEQUINE VETERINARY JOURNAL, Issue 6 2009F. J. Derksen Summary Reasons for performing study: Regional veno-occlusive remodelling of pulmonary veins in EIPH-affected horses, suggests that pulmonary veins may be central to pathogenesis. The current study quantified site-specific changes in vein walls, collagen and haemosiderin accumulation, and pleural vascular profiles in the lungs of horses suffering EIPH. Hypothesis: In the caudodorsal lung regions of EIPH-affected horses, there is veno-occlusive remodelling with haemosiderosis, angiogenesis and fibrosis of the interstitium, interlobular septa and pleura. Methods: Morphometric methods were used to analyse the distribution and accumulation of pulmonary collagen and haemosiderin, and to count pleural vascular profiles in the lungs of 5 EIPH-affected and 2 control horses. Results: Vein wall thickness was greatest in the dorsocaudal lung and significantly correlated with haemosiderin accumulation. Increased venous, interstitial, pleural and septal collagen; lung haemosiderin; and pleural vascular profiles occurred together and changes were most pronounced in the dorsocaudal lung. Further, haemosiderin accumulation colocalised with decreased pulmonary vein lumen size. Vein wall thickening, haemosiderin accumulation and histological score were highly correlated and these changes occurred only in the caudodorsal part of the lung. Conclusion: The colocalisation of these changes suggests that regional (caudodorsal) venous remodelling plays an important role in the pathogenesis of EIPH. Potential relevance: The results support the hypothesis that repeated bouts of venous hypertension during strenuous exercise cause regional vein wall remodelling and collagen accumulation, venous occlusion and pulmonary capillary hypertension. Subjected to these high pressures, there is capillary stress failure, bleeding, haemosiderin accumulation and, subsequently, lung fibrosis. [source] Faecal shedding and serological cross-sectional study of Lawsonia intracellularis in horses in the state of Minas Gerais, BrazilEQUINE VETERINARY JOURNAL, Issue 6 2009C. V. Guimarăes-Ladeira Summary Reason for performing the study: Proliferative enteropathy, caused by the intracellular bacterium Lawsonia intracellularis, has been described in horses in Australia, the USA, Canada and European countries but has not been reported in Latin America. The prevalence of the disease in horses worldwide is unknown. Objective: To evaluate the presence of subclinical L. intracellularis infection in horses in the state of Minas Gerais, Brazil. Methods: A longitudinal study using serology and PCR for detecting antibodies (IgG) and shedding of L. intracellularis in faecal samples, respectively, was conducted using a total of 223 horses from 14 different horse farms in Minas Gerais, and from the Veterinary School of UFMG equine herds in Minas Gerais. The immunoperoxidase technique in glass slides was used as the serological test. Results: Twenty-one horse sera had immunoglobulin G titres of 1:60 and were considered positive. The PCR technique in faeces for L. intracellularis DNA identified 7 horses as faecal shedders. Horses shedding the organism appeared healthy, indicating that subclinical infection of L. intracellularis occurred in the horses. Conclusion: Seropositivity and detection of faecal shedding of L. intracellularis indicates the presence of the agent in the equine population in Minas Gerais. Potential relevance: Results of this study should alert clinicians in countries where proliferative enteropthy in horses has not been reported to consider this disease as a possible cause of enteric disease. [source] The effect of training on stride parameters in a cohort of National Hunt racing Thoroughbreds: A preliminary studyEQUINE VETERINARY JOURNAL, Issue 5 2009M. FERRARI Summary Reasons for performing study: The influence of training on stride parameters is controversial and to date there is no information on how training influences stride parameters during high-speed locomotion in the field. Objective: To determine the influence of training on stride variables during high-speed locomotion in Thoroughbred racehorses. Methods: Speed, stride frequency, stance and protraction times were quantified in 8 Thoroughbreds with foot mounted accelerometers and GPS sensors during their first week of canter after the summer break and 6 months into training. Results: At a speed of 11 m/s, stride frequency was (mean ± s.d.) 2.160 ± 0.120 strides/s pre- and 2.167 ± 0.083 strides/s post training; mean stance time was 125.3 ± 9 ms pre- and 125.9 ± 7 ms post training; protraction time was 340.7 ± 20.4 ms pre- and 337.2 ± 14.3 ms post training. The increase in stride frequency and the decrease in protraction time after training were significant. There was no statistically significant difference in the maximum speed reached by each horse pre- and post training. Conclusions: Stance time stayed constant throughout the training season in the tested horses. A significant decrease in protraction time and a corresponding significant increase in stride frequency were observed after training. Potential relevance: Training of racehorses could be adapted to maximise the effect on modifiable parameters and reduce the risk of training-induced pathologies. Further research will be conducted to investigate the effect of different training protocols on a large number of horses. [source] Influence of the position of the foot on MRI signal in the deep digital flexor tendon and collateral ligaments of the distal interphalangeal joint in the standing horseEQUINE VETERINARY JOURNAL, Issue 5 2009M. SPRIET Summary Reasons for performing study: Hyperintense signal is sometimes observed in ligaments and tendons of the equine foot on standing magnetic resonance examination without associated changes in size and shape. In such cases, the presence of a true lesion or an artifact should be considered. A change in position of a ligament or tendon relative to the magnetic field can induce increased signal intensity due to the magic angle effect. Objectives: To assess if positional rotation of the foot in the solar plane could be responsible for artifactual changes in signal intensity in the collateral ligaments of the distal interphalangeal joint and in the deep digital flexor tendon. Methods: Six isolated equine feet were imaged with a standing equine magnetic resonance system in 9 different positions with different degrees of rotation in the solar plane. Results: Rotation of the limb induced a linear hyperintense signal on all feet at the palmar aspect of one of the lobes of the deep digital flexor tendon and at the dorsal aspect of the other lobe. Changes in signal intensity in the collateral ligaments of the distal interphalangeal joint occurred with rotation of the limb only in those feet where mediolateral hoof imbalance was present. Conclusions: The position and conformation of the foot influence the signal intensity in the deep digital flexor tendon and in the collateral ligaments of the distal interphalangeal joint. Potential relevance: The significance of increased signal intensity in the deep digital flexor tendon and in the collateral ligaments of the distal interphalangeal joint should be interpreted with regard to the position and the conformation of the foot. [source] Effect of magnitude and direction of force on laryngeal abduction: Implications for the nerve-muscle pedicle graft techniqueEQUINE VETERINARY JOURNAL, Issue 4 2009P. CRAMP Summary Reasons for performing study: The nerve-muscle pedicle graft technique is a treatment for recurrent laryngeal neuropathy (RLN), but the optimal placement of the pedicles within the cricoarytenoideus dorsalis (CAD) muscle is unknown. Hypothesis: The magnitude and direction of force placed on the muscular process of the left arytenoid cartilage affects the magnitude of laryngeal abduction. Methods: Five larynges were harvested from cadavers. Using increments of 0.98 N, a dead-weight force generator applied a force of 0,14.7 N for 1 min each to the left muscular process at 0, 10, 20, 30, 40, 50, 60 and 70° angles. The rima glottis was photographed digitally 1 min after each force had been applied. Distances between biomarkers (Lines 1,4) and right to left angle quotient (RLQ) were used to assess the degree of left arytenoid abduction. Results: Increasing force from 0,14.7 N progressively and significantly increased the length of all lines and RLQ, indicating abduction. Furthermore, there was a significant interaction between force and angles. Applying forces of 7.84 N or greater (Lines 2,4 and RLQ) or 11.76 N or greater (Line 1) at angles 0, 10, 20 and 30° resulted in significantly greater abduction than applying the same forces at 40, 50, 60 and 70°. Angles of 0,30° correspond with the direction of pull exerted by the lateral compartment of the CAD muscle. Conclusion: In RLN, nerve-muscle pedicle grafts should be placed preferentially in the lateral rather than in the medial compartment of the CAD muscle. Potential relevance: The information presented can be used to assist surgeons in the planning and application of the nerve-muscle pedicle graft procedure. [source] Oral endoscopy as an aid to diagnosis of equine cheek tooth infections in the absence of gross oral pathological changes: 17 casesEQUINE VETERINARY JOURNAL, Issue 2 2009P. H. L. RAMZAN Summary Reasons for performing study: Removal of cheek teeth in all but the aged horse or pony is a serious undertaking with potentially deleterious sequellae. Rigid endoscopy permits detailed examination of the oral cavity and erupted dental tissues and has the potential to assist in the correct identification of the diseased tooth. Objectives: To document oral endoscopic findings associated with infected equine cheek teeth in cases without gross oral pathological changes and thereby determine the usefulness of rigid oral endoscopy as an aid to diagnosis of such infections. Methods: Records of all cases of equine cheek tooth removal attempted under standing sedation over a 38 month period were examined. Cases were excluded from the study if apical infection was associated with gross dental fracture, malalignment, diastema/periodontal pocketing or supernumerary teeth. Endoscopic and radiographic findings were analysed and correlated to diseased tooth location. Results: Seventeen cases of apical dental infection fitting the inclusion criteria (nonresponsive to antibiotics and with no gross oral abnormality of the affected arcade) were identified in which oral endoscopy was used as an aid to diagnosis. In 15 (88%) of the 17 cases, oral endoscopy revealed abnormalities specific to the infected tooth. Focal gingival recession (10/17 cases) was the most common visible abnormality associated with infected teeth. Conclusions: In the majority of cases of apical infection of equine cheek teeth there is visible intraoral evidence implicating the affected tooth. Potential relevance: Oral endoscopy facilitates detailed examination of the mouth and should be considered along with radiography as an important aid to diagnosis in cases of equine dental infection. [source] Prediction of incipient pasture-associated laminitis from hyperinsulinaemia, hyperleptinaemia and generalised and localised obesity in a cohort of poniesEQUINE VETERINARY JOURNAL, Issue 2 2009R. A. CARTER Summary Reasons for performing study: The ability to predict ponies at increased risk of laminitic episodes, when exposed to nutrient dense pasture, would facilitate management to avoid disease. Objectives: To identify variables and clinically useful cut-off values with reproducible diagnostic accuracy for the prediction of ponies that subsequently developed laminitis when exposed to nutrient dense pasture. Methods: A cohort of predominantly Welsh and Dartmoor ponies from a closed herd was evaluated in March 2006 (n = 74) and March 2007 (n = 57). Ponies were categorised as never laminitic or previously laminitic according to reported laminitic history and as clinically laminitic (CL) if laminitis was observed within 3 months following evaluation. Body condition score (BCS), cresty neck score (CNS), girth and neck circumferences (NC), withers height, blood pressure and hoof surface temperature, and plasma insulin, glucose, triglyceride, leptin, cortisol, ACTH, uric acid and TNF-, concentrations were measured. Analysis of sensitivity, specificity and receiver operating characteristic curves was used to evaluate the diagnostic accuracy for a variable to predict CL ponies. Results: Variables with diagnostic accuracy for the prediction of CL ponies included insulin, leptin, BCS, CNS, and NC:height ratio. Specific cut-off values of insulin (>32 mu/l), leptin (>7.3 ng/ml), BCS (,7), CNS (,4) and NC:height ratio (>0.71) had reproducible diagnostic accuracy for the prediction of laminitis. Combining tests did not result in higher diagnostic accuracy than individual tests of insulin or leptin during either evaluation. Conclusions: Tests of insulin and leptin concentrations and measures of generalised (BCS) and localised (CNS or NC:height ratio) obesity were beneficial in the prediction of laminitic episodes. Potential relevance: These results highlight the importance of monitoring and reducing insulin concentration, and generalised and regional obesity in ponies to reduce risk of laminitis. [source] Prevalence of various radiographic manifestations of osteochondrosis and their correlations between and within joints in Dutch Warmblood horsesEQUINE VETERINARY JOURNAL, Issue 1 2009E. M. Van Grevenhof Summary Reasons for performing study: Osteochondrosis (OC) is the most important orthopaedic developmental disorder in horses and may manifest in several different forms. No detailed study on the prevalence and/or interrelation of these forms is available, even though these data are a prerequisite for conclusive genetic studies. Objectives: To assess the prevalence of the various manifestations of OC as detected radiographically and to evaluate possible relationships between their occurrence within the same joint and between different joints. Methods: The FP (femoropatellar), TC (tarsocrural) and MCP/MTP (metacarpophalangeal/metatarsophalangeal) joints of 811 yearlings selected randomly, descending from 32 representative stallions, were radiographed and scored for the presence and grade of osteochondrotic lesions. Results were compared at the sire, animal, joint and predilection site levels. Results: In the FP joint, the percentage of animals showing normal joint contours in all sites was 60.7%. For the TC joint and the combined MCP/MTP joints, these figures were 68.6 and 64.6%, respectively. For all joints combined, the percentage dropped to 30.5%. Sedation improved detection of OC lesions in the FP joint. There was a high correlation between the right and left joints. The correlation between flattened bone contours and fragments was considerably less. Conclusions: Scoring on a detailed scale is necessary to achieve good insight into the prevalence of OC. Observations on the right and left joints can be combined in further analyses, whereas flattened bone contours and fragments should be evaluated as statistically different disorders. Potential relevance: This study provides insight into the prevalences of various manifestations of OC and their relationships, within and between joints. These results form the basis for detailed quantitative and/or molecular genetic studies that should lead to the establishment of breeding indices and/or genetic marker sets for OC. [source] Diagnostic accuracy of digital photography and image analysis for the measurement of foot conformation in the horseEQUINE VETERINARY JOURNAL, Issue 7 2008J. M. WHITE Summary Reasons for performing study: Rigorous evaluation of practicable methods for the objective assessment of foot conformation has not been performed. Objectives: To assess the practicability, precision and accuracy of the process of obtaining measurements of horses' feet using photography and image processing software. Methods: Precision study: Lateral photographs of horses' feet were obtained twice by 2 veterinary surgeons (image acquisition - IAc). Photographs were analysed by 2 masked veterinary surgeons on 2 occasions (image analysis - IAn). Measurements were compared within and between operators for self and non-self acquired photographs. Agreement indices (AIs) and 95% limits of agreement (LOA) were calculated for the IAn process alone and for the combined IAc + IAn processes. Accuracy study: Measurements obtained from lateral photographs were compared with those obtained from lateromedial radiographs. AIs and 95% LOA were calculated for each measurement. Results: Precision study: Mean intra- and interoperator AIs for the IAn process alone were ,0.90 while those for the combined IAc + IAn processes were ,0.89 for all measurements. Similar mean AIs and 95% LOA were calculated regardless of image origin. The 95% LOA for hoof angle, heel height/toe height% and coronary band angle for all comparisons were within target values. Accuracy study: Mean AIs were ,0.89 for all measurements. The 95% LOA for heel height/toe height% and coronary band angle were within target values. Conclusions: Excellent precision was identified within and between operators regardless of image origin. High levels of accuracy were also identified, especially for heel height/toe height% and coronary band angle, indicating that photography and radiography may be used interchangeably. Potential relevance: Acquisition and analysis of photographic images is an appropriate method for the objective measurement of foot conformation, both in clinical and research settings. [source] Effects of a MAPK p38 inhibitor on lung function and airway inflammation in equine recurrent airway obstructionEQUINE VETERINARY JOURNAL, Issue 6 2008J.-P. LAVOIE Summary Reasons for performing study: It has been suggested that many of the beneficial effects of corticosteroids are mediated through mitogen-activated protein kinase (MAPK) p38 inhibition. Objective: To investigate the efficacy of the MAPK p38 inhibitor compound MRL-EQ1 to either prevent (Phase 1) or treat (Phase 2) recurrent airway obstruction (RAO) in horses. Methods: MRL-EQ1 was administered i.v. at a dosage of 0.75-1.5 mg/kg bwt q. 12 h. In Phase 1, susceptible horses in clinical remission were divided into 2 groups (n = 5/group), based on historical values of respiratory mechanics. All horses were entered in the study in pairs (one control, one treated horse) and exposed to the same environmental challenge (stabling, mouldy hay and dusty conditions). The treatment group received MRL-EQ1 for 14 days while the control horses were untreated during the same period. In Phase 2, affected horses were ranked by severity of respiratory dysfunction and split randomly into either dexamethasone or MRL-EQ1 treatment groups (n = 5/group). Bronchoalveolar lavage fluid, respiratory mechanic measurements, MRL-EQ1 plasma concentration and tumour necrosis factor (TNF) whole blood activity were evaluated sequentially. Results: In Phase 1, MRL-EQ1 did not prevent the occurrence of clinical signs and pulmonary inflammation. However, treatment was associated with a reduction in severity and a delay in the onset of signs and a reduction in pulmonary neutrophilia. In Phase 2, plasma concentrations achieved resulted in ex vivo suppression of lipopolysaccharide-induced TNF production in equine blood. MRL-EQ1 did not improve airway inflammation or lung function and was associated in a dose dependent manner with behavioural (depression, excitability) and blood changes (neutrophilia, increased serum muscle enzyme concentrations). Conclusions: Inhibition of p38 in the horse was partially effective in reducing clinical signs and airway inflammation when administered prior to, but not during clinical exacerbation in RAO. Potential relevance: Inhibitors of p38 MAPK with a better toxicity profile may be effective in the prevention or treatment of RAO. [source] Intra-articular stabilisation of the equine cricoarytenoid jointEQUINE VETERINARY JOURNAL, Issue 6 2008J. CHEETHAM Summary Reasons for performing study: The success of laryngoplasty is limited by abduction loss in the early post operative period. Objective: To determine the efficacy of polymethylmethacrylate (PMMA) in stabilising the cricoarytenoid joint (CAJ) and reducing the force on the laryngoplasty suture. Hypothesis: Injection into the cricoarytenoid joint resists the forces produced by physiological laryngeal air flows and pressures thereby reducing the force experienced by the laryngoplasty suture. Methods: Ten cadaver larynges were collected at necropsy and PMMA was injected into one CAJ at selected random. Each larynx was subjected to physiological conditions with with constant (static) or cycling (dynamic) flow. The specimens were tested sequentially in each of 4 conditions: 1) bilateral full abduction (Control 1); 2) transection of the suture on the side without PMMA; 3) bilateral abduction achieved by replacing the suture (Control 2); and 4) cutting the suture on the PMMA side. Tracheal pressure and flow and pressure in the flow chamber were recorded using pressure and flow transducers. The strain experienced by each suture during bilateral abduction (Controls 1 and 2) was measured. Statistical comparison of the 4 conditions was performed using a mixed effect model with Tukey's post hoc test for multiple comparisons. The strain gauge data were analysed by paired comparison of the regression slopes. Results: In the static and dynamic states, tracheal pressure increased and tracheal flow decreased when the suture on the non-cement side was cut (P<0.05). There was no significant difference in any outcome measure between PMMA injected into the CAJ and bilaterally abducted specimens (Controls 1 and 2) for either condition. The rate of increase in strain with increasing translaryngeal pressure was significantly less on the suture with PMMA placed in the CAJ (P = 0.03). Conclusions: These data provide strong evidence that injecting PMMA into the CAJ resists the collapsing effect of physiological airflows and pressures in vitro and reduces the force experienced by the laryngoplasty suture during maximal abduction. Potential relevance: Augmentation of prosthetic laryngoplasty with this technique may reduce arytenoid abduction loss in the early post operative period. [source] The effect of metformin on measurements of insulin sensitivity and , cell response in 18 horses and ponies with insulin resistanceEQUINE VETERINARY JOURNAL, Issue 5 2008A. E. Durham Summary Reasons for performing study: Laminitis in equids is a very common debilitating disease, and insulin resistance (IR) and hyperinsulinaemia are increasingly recognised as important predisposing factors. Pharmacological modification of IR and hyperinsulinaemia might reduce the risk of laminitis. Hypothesis: Metformin, a drug commonly prescribed for treatment of human IR, may also decrease IR in equids. Methods: Eighteen horses and ponies with IR and recurrent laminitis were treated with 15 mg/kg bwt metformin per os q. 12 h. Each animal served as its own control by comparing pre- and post treatment proxies for IR, insulin sensitivity (IS) and pancreatic , cell function while controlling for possible dietary and managemental influences on IR. Results: Evidence of significantly improved IS and decreased pancreatic , cell secretion was found following metformin treatment. The magnitude of effect was greater at earlier resampling (6,14 days) than at later times (23,220 days). Apparent subjective clinical benefits were good but less favourable than effects on IR. Conclusions: Metformin is safe and appears to increase IS in equids. Potential relevance: Metformin may be indicated as a treatment for IR in equids. Further studies are required to define appropriate selection of subjects warranting therapy, dosing schedule and pharmacokinetics. [source] Racing performance following the laryngeal tie-forward procedure: A case-controlled studyEQUINE VETERINARY JOURNAL, Issue 5 2008J. Cheetham Summary Reasons for performing study: The laryngeal tie-forward procedure (LTFP) is becoming widely used for correction of dorsal displacement of the soft palate (DDSP) despite the absence of an evidence-based assessment of its efficacy. Hypotheses: The LTFP returns racing performance to preoperative baseline levels and to that of matched controls; and post operative laryngohyoid position is associated with post operative performance. Design and population: Case-controlled study of racehorses undergoing a LTFP for dorsal displacement of the soft palate at Cornell University between October 2002 and June 2007. Methods: The presence of at least one post operative start and race earnings ($) were used as outcome variables. Controls were matched by age, breed and sex from the third race prior to surgery. A novel radiographic reference system was used to determine laryngohyoid position pre- and post operatively. Data for definitively and presumptively diagnosed cases were analysed separately. Results: During the study interval, 263 racehorses presented, of which 106 were included in the study; 36 had a definitive diagnosis of DDSP and 70 a presumptive diagnosis. Treated horses were equally likely to race post operatively as controls in the equivalent race. Treated horses had significantly lower earnings in the race before surgery than matched controls. The procedure moved the basihyoid bone dorsally and caudally and the larynx dorsally and rostrally. A more dorsal post operative basihyoid position and more dorsal and less rostral laryngeal position were associated with an increased probability of racing post operatively. Conclusions: Horses undergoing a LTFP are as likely to race post operatively as matched controls. The procedure restores race earnings to preoperative baseline levels and to those of matched controls. Potential relevance: This study provides strong evidence supporting the use of the LTFP in racehorses. Further work is needed to determine the relationship between laryngohyoid conformation and nasopharyngeal stability in horses. [source] Bradykinin stimulates prostaglandin E2 production and cyclooxygenase activity in equine nonglandular and glandular gastric mucosa in vitroEQUINE VETERINARY JOURNAL, Issue 4 2008N. K. MORRISSEY Summary Reasons for performing study: There are few data available regarding regulation of prostaglandin (PG) generation by equine gastric mucosae and the role of the cyclooxygenase (COX) isoforms in their production. Objectives: To: 1) characterise and quantify PGE2 output in vitro; 2) examine the sensitivity of PGE2 production to exogenous bradykinin (BK) exposure; 3) determine the contribution of the COX-1 and COX-2 pathways to basal and BK-stimulated PGE2 production; and 4) measure if BK influences electrogenic ion transport in equine gastric mucosae in vitro. Methods: Full thickness gastric sheets were obtained from horses at post mortem, stripped of muscle layers and mounted in Ussing chambers. Tissues were exposed to bradykinin (BK, 0.1 ,mol/l) either alone, or following pretreatment with a selective COX-2 inhibitor (NS-398, 1 ,mol/l) or a nonselective COX inhibitor (piroxicam, 1 ,mol/l), or were untreated. Results: BK administration increased PGE2 output from the basolateral but not the apical faces of both tissue types. Piroxicam, but not NS-398, reduced basolateral PGE2 release below control levels in both tissue types. Both piroxicam and NS-398 pretreatment inhibited BK-stimulated PGE2 release. In separate experiments, BK was without effect upon electrophysiological parameters of tissues mounted in Ussing chambers. Conclusions: PGE2 is produced by the nonglandular and glandular equine gastric mucosae in vitro. Significantly more PGE2 is released basolaterally than apically. BK stimulated the production of PGE2 from the basolateral side of both tissue types. These findings suggest that COX-1 is a significant pathway for basal PGE2 production from the basolateral faces of both nonglandular and glandular equine gastric mucosae in vitro. Potential relevance: The identification of the cells responsible for basolateral PGE2 release, via both COX-1 and COX-2 pathways, under basal and BK-stimulated conditions requires further study. [source] Effect of paddock vs. stall housing on 24 hour gastric pH within the proximal and ventral equine stomachEQUINE VETERINARY JOURNAL, Issue 4 2008L. HUSTED Summary Reasons for performing study: Stall housing has been suggested as a risk factor for ulcer development in the equine stomach; however, the exact pathogenesis for this has not been established. Objectives: To investigate the effect of 3 environmental situations (grass paddock, stall alone or stall with adjacent companion) on pH in the proximal and the ventral stomach. Methods: Six horses with permanently implanted gastric cannulae were used in a randomised, cross-over, block design. Each horse rotated through each of three 24 h environmental situations. Horses remained on their normal diet (grass hay ad libitum and grain b.i.d.) throughout the study. Intragastric pH was measured continuously for 72 h just inside the lower oesophageal sphincter (proximal stomach) and via a pH probe in the gastric cannula (ventral stomach). Results: Neither proximal nor ventral 24 h gastric pH changed significantly between the 3 environmental situations. Mean hourly proximal gastric pH decreased significantly in the interval from 01.00,09.00 h compared to the interval from 13.00,20.00 h, regardless of environmental situation. Median hourly proximal pH only differed in the interval from 06.00,07.00 h compared to the interval 14.00,19.00 h. Neither mean nor median hourly ventral gastric pH varied significantly with the time of day. Conclusions: The change in housing status used in the current study did not affect acid exposure within either region of the equine stomach. The pH in the ventral stomach was uniformly stable throughout the study, while the proximal pH demonstrated a 24 h circadian pattern. Potential relevance: Since stall housing was not associated with prolonged acid exposure to the proximal stomach, this aspect alone does not explain the increased risk of squamous ulcer development. The circadian rhythm associated with proximal intragastric pH warrants further investigation. [source] The creation of an international audit and database of equine colic surgery: Survey of attitudes of surgeonsEQUINE VETERINARY JOURNAL, Issue 4 2008T. S. MAIR Summary Reasons for performing study: Currently, there is a lack of available evidence-based data concerning the optimum treatments for horses affected by different types of colic and this precludes the application of clinical audit in this area. In order to accumulate such data, a large-scale, multicentre database of the outcomes of colic surgery is proposed. The attitudes of surgeons is an important consideration in determining the feasibility of developing this database. Objectives: To assess attitudes and opinions of equine surgeons concerning clinical audit and to assess the perceived advantages and problems of setting up a large-scale international audit/database of colic surgery. Methods: Interviews were conducted with 30 equine surgeons (large animal/equine surgeons who are diplomates of either the American College of Veterinary Surgeons or the European College of Veterinary Surgeons). Questionnaires were sent by e-mail to 98 equine surgeons. Results: Face to face interviews were conducted (n = 30) and 43/98 completed questionnaires received (44%). The results of the 2 techniques were very similar. There was generally a high level of interest in the development of a large scale database of colic surgery, but perceived problems included time to collect and submit data, and confidentiality issues. A minority of surgeons reported that they were undertaking any form of specific monitoring of the results of colic surgery within their hospitals. Conclusions: There is a good level of interest among equine surgeons to develop a large scale database of colic surgery and most would be willing to contribute data from their own hospitals provided that data collection is quick and easy, and that confidentiality is maintained. Potential relevance: A large scale audit and database would provide relevant information to equine surgeons concerning the current success and complication rates of colic surgery. Such evidence-based data could be used in clinical audits within individual equine hospitals. The data would also be useful to identify trends within the discipline and could highlight areas that would benefit from active research. [source] Risk factors for epiploic foramen entrapment colic in a UK horse population: A prospective case-control studyEQUINE VETERINARY JOURNAL, Issue 4 2008D. C. ARCHER Summary Reasons for performing study: Epiploic foramen entrapment (EFE) is a common cause of small intestinal strangulation in the horse and its epidemiology requires further investigation. Objectives: To identify horse- and management-level risk factors for EFE and to explore reasons for the apparent seasonality of this condition. Hypothesis: Horses exhibiting certain behaviours and those exposed to particular management practices that vary seasonally are at increased risk of EFE. Methods: A prospective unmatched, multicentre case-control study was conducted over 24 months in the UK. Data on 77 cases and 216 control horses were obtained from 9 collaborating clinics and logistic regression was used to identify associations between horse and management variables and the likelihood of EFE. Results: In a final multivariable model crib-biting/ windsucking behaviour was associated with the largest increase in likelihood of EFE. A history of colic in the previous 12 months, increased stabling in the previous 28 days and height of the horse also increased the likelihood of EFE. Horses with access to a mineral/salt lick, those easily frightened and horses not fed at the same time as others were at reduced risk of EFE. Conclusions: Horses exhibiting certain behaviours, those with a previous history of colic and horses of greater height appear to be at inherently greater risk of EFE. The increase in likelihood of EFE with increased duration of stabling may explain the apparent seasonality of this condition. Potential relevance: These findings assist identification of horses at high-risk of EFE and provide information on management strategies that may reduce this risk. If the observed associations are causal, avoiding sudden increases in duration of stabling, not feeding horses in the same group at the same time and providing a mineral/salt lick may reduce the likelihood of EFE. The risk factors identified in this study provide important clues to the aetiology of EFE. [source] The pathology of bronchointerstitial pneumonia in young foals associated with the first outbreak of equine influenza in AustraliaEQUINE VETERINARY JOURNAL, Issue 3 2008J. C. PATTERSON-KANE Summary Objectives: The aim of this study was to describe post mortem lesions in EIV-infected foals. Methods: Post mortem examinations were conducted on 11 young foals (age 2,12 days) submitted to the Scone Veterinary Hospital, New South Wales, Australia over a 2-month period in 2007. The foals had presented with or developed fatal pneumonia, and were known or suspected to be EIV-positive. Equine influenza virus nucleic acid was detected in tissue specimens using an influenza A group reactive real-time reverse transcriptase PCR assay. Results: Grossly there was diffuse or extensive pulmonary consolidation. Histological changes included: bronchiolar and alveolar necrosis; neutrophilic infiltration; hyaline membrane formation; and hyperplasia and squamous metaplasia of airway epithelium. Tissues for 10 foals were EIV-positive, with a positive nasal swab from the remaining animal. Conclusions: This is the first detailed pathological description of bronchointerstitial pneumonia associated with EIV infection in young foals. It is also the first series of such cases in which a causative agent has consistently been detected. Potential relevance: Given the findings in this outbreak, and a previous outbreak in the UK in 1965 involving a similarly naive population, veterinary clinicians and pathologists should be aware that EIV can cause fatal bronchointerstitial pneumonia in young foals that do not have maternal immunity. The lesions did not differ from those previously reported in foals of various ages with bronchointerstitial pneumonia of other or undefined causes, indicating that this is most likely to be a stereotypical response to a variety of insults. Therefore, tissue specimens should be obtained from cases of pneumonia in young foals for virological and bacteriological testing. Reasons for performing study: The first outbreak of equine influenza virus (EIV) infection was confirmed in Australia in 2007. Some EIV-positive young foals died with broncho-interstitial pneumonia, a rare disease process in this age group that is often postulated to be caused by viral infection. [source] An epidemiological study of myopathies in Warmblood horsesEQUINE VETERINARY JOURNAL, Issue 2 2008L. M. HUNT Summary Reasons for performing study: There are few detailed reports describing muscular disorders in Warmblood horses. Objectives: To determine the types of muscular disorders that occur in Warmblood horses, along with presenting clinical signs, associated risk factors and response to diet and exercise recommendations, and to compare these characteristics between horses diagnosed with polysaccharide storage myopathy (PSSM), those diagnosed with a neuromuscular disorder other than PSSM (non-PSSM) and control horses. Methods: Subject details, muscle biopsy diagnosis and clinical history were compiled for Warmblood horses identified from records of biopsy submissions to the University of Minnesota Neuromuscular Diagnostic Laboratory. A standardised questionnaire was answered by owners at least 6 months after receiving the muscle biopsy report for an affected and a control horse. Results: Polysaccharide storage myopathy (72/132 horses) was the most common myopathy identified followed by recurrent exertional rhabdomyolysis (RER) (7/132), neurogenic or myogenic atrophy (7/132), and nonspecific myopathic changes (14/132). Thirty-two biopsies were normal. Gait abnormality, ,tying-up', Shivers, muscle fasciculations and atrophy were common presenting clinical signs. Forty-five owners completed questionnaires. There were no differences in sex, age, breed, history or management between control, PSSM and non-PSSM horses. Owners that provided the recommended low starch fat supplemented diet and regular daily exercise reported improvement in clinical signs in 68% (19/28) of horses with a biopsy submission and 71% of horses diagnosed with PSSM (15/21). Conclusions: Muscle biopsy evaluation was a valuable tool to identify a variety of myopathies in Warmblood breeds including PSSM and RER. These myopathies often presented as gait abnormalities or overt exertional rhabdomyolysis and both a low starch fat supplemented diet and regular exercise appeared to be important in their successful management. Potential relevance: Warmbloods are affected by a variety of muscle disorders, which, following muscle biopsy diagnosis can be improved through changes in diet and exercise regimes. [source] Liposome-mediated uptake of exogenous DNA by equine spermatozoa and applications in sperm-mediated gene transferEQUINE VETERINARY JOURNAL, Issue 1 2008B. A. BALL Summary Reasons for performing study: Sperm-mediated gene transfer has been reported as a method for production of transgenic animals in a variety of species, and this technique represents a possible method for production of transgenic equids. Objectives: To evaluate the uptake of exogenous DNA (enhanced green fluorescent protein; pEGFP) by equine spermatozoa and to assess the ability of transfected spermatozoa to introduce this transgene into early equine embryos. Methods: To evaluate incorporation of pEGFP into equine spermatozoa, washed spermatozoa were incubated with 32P-pEGFP, with or without lipofection. Spermatozoa were also transfected with fluorescently-labelled DNA (Alexa647 -pEGFP) and changes in sperm viability and DNA uptake were assessed. Mares were inseminated with pEGFP-transfected spermatozoa and embryos recovered. Expression of pEGFP was assessed by epifluorescence microscopy of embryos, and the presence of pEGFP DNA and mRNA was assessed by PCR and RT-PCR, respectively. Results: Liposome-mediated transfection increased the incorporation of 32P-pEGFP into spermatozoa compared to controls. Flow cytometric evaluation of spermatozoa after transfection with Alexa647 -pEGFP revealed a linear increase in the proportion of live, Alexa647+ spermatozoa with increasing DNA concentrations. After insemination with transfected spermatozoa, 8 embryos were recovered. There was no evidence of EGFP expression in the recovered embryos; however, PCR analysis revealed evidence of the pEGFP transgene in 2 of 5 embryos analysed. Conclusions: The incorporation of exogenous DNA by equine spermatozoa was enhanced by liposome-mediated transfection and this did not adversely affect sperm viability, acrosomal integrity or fertility. Although the EGFP transgene was detected in a proportion of Day 7,10 embryos, there was no evidence of expression of EGFP in these embryos. Potential relevance: Sperm-mediated gene transfer offers a potential technique for the generation of transgenic equids. [source] Superficial digital flexor tendon lesions in racehorses as a sequela to muscle fatigue: A preliminary studyEQUINE VETERINARY JOURNAL, Issue 6 2007M. T. Butcher Summary Reasons for performing study: Racing and training related lesions of the forelimb superficial digital flexor tendon are a common career ending injury to racehorses but aetiology and/or predisposing causes of the injury are not completely understood. Objectives: Although the injury takes place within the tendon, the lesion must be considered within the context of the function of the complete suspensory system of the distal limb, including the associated muscles. Methods: Both muscle and tendon function were investigated in vivo using implanted strain gauges in 3 Thoroughbred horses walking, trotting and cantering on a motorised treadmill. These data were combined with assessments of muscle architecture and fibre composition to arrive at an overview of the contribution of each muscle-tendon unit during locomotion. Results: The superficial digital flexor muscle has fatigue-resistant and high force production properties that allow its tendon to store and return elastic energy, predominantly at the trot. As running speed increases, deep digital flexor tendon force increases and it stabilises hyperextension of the fetlock, thus reinforcing the superficial digital flexor in limb load support. The deep digital flexor muscle has fast contracting properties that render it susceptible to fatigue. Conclusion: Based on these measurements and supporting evidence from the literature, it is proposed that overloading of the superficial digital flexor tendon results from fatigue of the synergistic, faster contracting deep digital flexor muscle. Potential relevance: Future research investigating distal limb system function as a whole should help refine clinical diagnostic procedures and exercise training approaches that will lead to more effective prevention and treatment of digital flexor tendon injuries in equine athletes. [source] |