Safety and efficacy of subcutaneous alpha-tocopherol in healthy adult horses.

Vitamin E is essential for neuromuscular function. The primary treatment, oral supplementation with natural ('RRR') α-tocopherol, is not effective in all horses. The objectives of this pilot study were to evaluate the safety and efficacy of a subcutaneously administered RRR-α-tocopherol preparation. Horses were randomly assigned in a cross-over design to initially receive RRR-α-tocopherol (5000 IU/450 kg of 600 IU/mL) subcutaneously (n = 3) or orally (n = 3) or were untreated sentinels (n = 2). Tissue reactions following injection in Phase I of the study necessitated adjustment of the preparation with reduction of the RRR-α-tocopherol concentration to 500 IU/mL in Phase 2. Following an 8-week washout period, horses received the reciprocal treatment route with the new preparation (5000 IU/450 kg of 500 IU/mL). Serum, CSF and muscle α-tocopherol concentrations were determined by high-performance liquid chromatography over a 14-day period during each phase. Serum and CSF α-tocopherol concentrations increased significantly postinjection only when the 500 IU/mL product was administered (P<0.0001). There was no significant difference in the muscle concentration of α-tocopherol following either treatment. All eight horses had marked tissue reaction to subcutaneous injection, regardless of product concentration. Whilst we have demonstrated that this route may be a useful alternative to oral supplementation, the marked tissue reaction makes use of such products limited at this time to only the most refractory of cases.

Effects of lidocaine on equine ejaculated sperm and epididymal sperm post-castration.

In equids, it is common to inject lidocaine into the testicles at the time of routine castration to provide analgesia. The effects of lidocaine on equine sperm have not been evaluated in vitro or on epididymal sperm collected following castration. The aims of this study were to determine effects of clinically relevant doses of lidocaine on equine spermatozoa in vitro using freshly collected semen and to compare the characteristics of epididymal spermatozoa after routine castration with or without intra-testicular lidocaine administration. We hypothesized that increasing concentrations of lidocaine would decrease total motility (TM), progressive motility (PM), velocity of the average path (VAP), velocity of the curved line (VCL), linearity (LIN), normal morphology (M) and membrane integrity (MI). We also hypothesized that injection of intra-testicular lidocaine would decrease TM, PM, VAP, VCL, LIN, M, and MI following routine castration, epididymal flushing and cryopreservation. In experiment 1, sperm was collected from four stallions and mixed with lidocaine at concentrations of 1 µg/ml, 10 µg/ml, 100 µg/ml, 1,000 µg/ml and 10,000 µg/ml. M and MI were compared to the control sample at 0 and 48 h. Motility parameters were analyzed at 0, 2, 4, 6, 24, and 48 h. In experiment 2, 12 stallions were castrated under general anesthesia. One testicle was removed without the use of intra-testicular lidocaine and the other testicle was removed 10 min after injection of 10 ml of 2% lidocaine. Results: In experiment 1, fresh sperm showed no significant difference (p < 0.05) compared to control at either 1 µg/ml or 10 µg/ml concentrations of lidocaine. There were significant decreases in PM, VAP, VCL, and LIN at concentrations of 100µg/ml-10,000 µg/ml and for TM at lidocaine concentrations of 1,000-10,000 µg/ml compared to control. Morphology did not change at any lidocaine concentration. Membrane integrity decreased significantly at 10,000 µg/ml lidocaine. In the second experiment 1.03 ±â€¯0.42 µg/ml lidocaine was detected in the epididymal flush of stallions treated with lidocaine. There were no significant differences in any measured parameters between the control and the lidocaine treated testicles. Intra-testicular lidocaine injection at the time of castration did not affect any measured parameters after epididymal flush. Lidocaine concentrations higher than 100 µg/ml in-vitro resulted in decreased motility parameters of the spermatozoa independent of exposure time.

18 F-sodium fluoride positron emission tomography of the racing Thoroughbred fetlock: Validation and comparison with other imaging modalities in nine horses.

BACKGROUND: Early and accurate detection of stress remodelling in racehorses is of utmost importance to prevent catastrophic injuries. Current imaging techniques have limitations in assessing early changes predisposing to catastrophic breakdowns. Positron emission tomography (PET) using 18 F-sodium fluoride (18 F-NaF) is a sensitive method for the detection of early bone turnover and may improve early recognition of subtle injuries. OBJECTIVES: To validate the clinical use of 18 F-NaF PET in Thoroughbred racehorses, to assess the value of PET in the detection of bone lesions and to compare PET results with findings of other advanced imaging modalities, clinical examination and pathology. STUDY DESIGN: Experimental exploratory study. METHODS: Twenty fetlocks from nine Thoroughbred racehorses were imaged using 18 F-NaF PET, computed tomography (CT) and scintigraphy. Five fetlocks were also imaged with magnetic resonance imaging and four fetlocks were also examined histologically. Imaging findings were independently reviewed by three board certified radiologists. Imaging, clinical and histopathological findings were correlated. RESULTS: PET imaging was well-tolerated by all horses. PET detected focal areas of 18 F-NaF uptake in instances where other imaging modalities did not identify abnormalities, in particular in the proximal sesamoid bones. Maximal standardised uptake values could be measured to quantify the activity of lesions. Areas of 18 F-NaF uptake corresponded to regions of increased vascularity and increased osteoblastic activity. MAIN LIMITATIONS: Limited number of cases. CONCLUSIONS: 18 F-NaF PET imaging of the Thoroughbred fetlock is feasible and compares favourably with other imaging modalities in detecting stress remodelling in Thoroughbred racehorses. PET appears to be a beneficial imaging modality when used for early detection of stress remodelling in an effort to prevent catastrophic musculoskeletal injuries in this population of horses.

Generation of an equine biobank to be used for Functional Annotation of Animal Genomes project.

The Functional Annotation of Animal Genomes (FAANG) project aims to identify genomic regulatory elements in both sexes across multiple stages of development in domesticated animals. This study represents the first stage of the FAANG project for the horse, Equus caballus. A biobank of 80 tissue samples, two cell lines and six body fluids was created from two adult Thoroughbred mares. Ante-mortem assessments included full physical examinations, lameness, ophthalmologic and neurologic evaluations. Complete blood counts and serum biochemistries were also performed. At necropsy, in addition to tissue samples, aliquots of serum, ethylenediaminetetraacetic acid (EDTA) plasma, heparinized plasma, cerebrospinal fluid, synovial fluid, urine and microbiome samples from all regions of the gastrointestinal and urogenital tracts were collected. Epidermal keratinocytes and dermal fibroblasts were cultured from skin samples. All tissues were grossly and histologically evaluated by a board-certified veterinary pathologist. The results of the clinical and pathological evaluations identified subclinical eosinophilic and lymphocytic infiltration throughout the length of the gastrointestinal tract as well as a mild clinical lameness in both animals. Each sample was cryo-preserved in multiple ways, and nuclei were extracted from selected tissues. These samples represent the first published systemically healthy equine-specific biobank with extensive clinical phenotyping ante- and post-mortem. The tissues in the biobank are intended for community-wide use in the functional annotation of the equine genome. The use of the biobank will improve the quality of the reference annotation and allow all equine researchers to elucidate unknown genomic and epigenomic causes of disease.

18 F-sodium fluoride positron emission tomography of the equine distal limb: Exploratory study in three horses.

BACKGROUND: Positron emission tomography (PET) is a cross-sectional, functional imaging modality that has recently become available to the horse. The use of 18 F-sodium fluoride (18 F-NaF), a PET bone tracer, has not previously been reported in this species. OBJECTIVES: To assess the feasibility of 18 F-NaF PET in the equine distal limb and explore possible applications in the horse in comparison with other imaging modalities. STUDY DESIGN: Exploratory descriptive study involving three research horses. METHODS: Horses were placed under general anaesthesia prior to intravenous (i.v.) administration of 1.5 MBq/kg of 18 F-NaF. Positron emission tomography imaging of both front feet and fetlocks was performed using a portable scanner. Computed tomography (CT) of the distal limb was performed under a separate anaesthetic episode. Bone scintigraphy and magnetic resonance imaging (MRI) were subsequently performed under standing sedation. Images obtained from PET and other imaging modalities were independently assessed and the results correlated. RESULTS: Positron emission tomography images were obtained without complication. The radiation exposure rate was similar to equine bone scintigraphy. Positron emission tomography detected focal 18 F-NaF uptake in areas where other imaging modalities did not identify any abnormalities. This included sites of ligamentous attachment, subchondral compact bone plate and the flexor cortex of the navicular bone. 18 F-NaF uptake was identified in some, but not all, osseous fragments and areas of osseous formation, suggesting a distinction between active and inactive lesions. MAIN LIMITATIONS: A small number of horses were included and histopathology was not available. CONCLUSIONS: 18 F-NaF PET imaging of the equine distal limb provides useful additional information when compared with CT, MRI and scintigraphy and has the potential for both research and clinical applications in the horse. The Summary is available in Chinese - see Supporting information.

Blood and Cerebrospinal Fluid α-Tocopherol and Selenium Concentrations in Neonatal Foals with Neuroaxonal Dystrophy.

BACKGROUND: Equine neuroaxonal dystrophy/equine degenerative myeloencephalopathy (NAD/EDM) is a neurodegenerative disorder affecting genetically predisposed foals maintained on α-tocopherol (α-TP)-deficient diet. OBJECTIVE: Intramuscular α-TP and selenium (Se) administration at 4 days of age would have no significant effect on serum or cerebrospinal fluid (CSF) α-TP in healthy foals. Serum and CSF α-TP, but not Se, would be significantly decreased in NAD/EDM-affected foals during first year of life. ANIMALS: Fourteen Quarter horse foals; 10 healthy foals supplemented with 0.02 mL/kg injectable α-TP and Se (n = 5) or saline (n = 5) at 4 days of age and 4 unsupplemented NAD/EDM-affected foals. METHODS: Complete neurologic examinations were performed, blood and CSF were collected before (4 days of age) and after supplementation at 10, 30, 60, 120, 180, 240, and 360 days of age. Additional blood collections occurred at 90, 150, 210, and 300 days. At 540 days, NAD/EDM-affected foals and 1 unsupplemented healthy foal were euthanized and necropsies performed. RESULTS: Significant decreases in blood, CSF α-TP and Se found in the first year of life in all foals, with most significant changes in serum α-TP from 4-150 days. Dam α-TP and Se significantly influenced blood concentrations in foals. Injection of α-TP and Se did not significantly increase CSF Se, blood or CSF α-TP in healthy foals. NAD/EDM-affected foals had significantly lower CSF α-TP through 120 days. CONCLUSIONS AND CLINICAL IMPORTANCE: Injection of α-TP and Se at 4 days of age does not significantly increase blood or CSF α-TP. Despite all 14 foals remaining deficient in α-TP, only the 4 genetically predisposed foals developed NAD/EDM.

Mitochondrial metabolism reveals a functional architecture in intact islets of Langerhans from normal and diabetic Psammomys obesus.

The cells within the intact islet of Langerhans function as a metabolic syncytium, secreting insulin in a coordinated and oscillatory manner in response to external fuel. With increased glucose, the oscillatory amplitude is enhanced, leading to the hypothesis that cells within the islet are secreting with greater synchronization. Consequently, non-insulin-dependent diabetes mellitus (NIDDM; type 2 diabetes)-induced irregularities in insulin secretion oscillations may be attributed to decreased intercellular coordination. The purpose of the present study was to determine whether the degree of metabolic coordination within the intact islet was enhanced by increased glucose and compromised by NIDDM. Experiments were performed with isolated islets from normal and diabetic Psammomys obesus. Using confocal microscopy and the mitochondrial potentiometric dye rhodamine 123, we measured mitochondrial membrane potential oscillations in individual cells within intact islets. When mitochondrial membrane potential was averaged from all the cells in a single islet, the resultant waveform demonstrated clear sinusoidal oscillations. Cells within islets were heterogeneous in terms of cellular synchronicity (similarity in phase and period), sinusoidal regularity, and frequency of oscillation. Cells within normal islets oscillated with greater synchronicity compared with cells within diabetic islets. The range of oscillatory frequencies was unchanged by glucose or diabetes. Cells within diabetic (but not normal) islets increased oscillatory regularity in response to glucose. These data support the hypothesis that glucose enhances metabolic coupling in normal islets and that the dampening of oscillatory insulin secretion in NIDDM may result from disrupted metabolic coupling.

Juvenile coho salmon locomotion and mosaic muscle are modified by 3',3',5'-tri-iodo-L-thyronine (T3).

Studies of maximum aerobic swimming performance in smolting juvenile salmonids indicate that these animals may be aerobically compromised during downstream migration. To test our hypothesis that hyperthyroid status contributes to decreased swimming performance through modification of muscle contractility in juvenile (112 mm mean total length) coho salmon (Oncorhynchus kisutch), we measured swimming performance and isolated muscle bundle contractility of fish implanted with 3',3',5'-tri-iodo-l-thyronine (T3) pellets, of fish implanted with sham pellets and of fish with no pellet implantation (control group). After 3 weeks (N=12-13), critical swimming speeds (maximum aerobic swimming speed or U(crit)) were measured. Muscle bundles (N=15-16) were dissected from the hypaxial musculature and stimulated to measure the force and velocity of an isometric twitch and tetani. T3-treated fish demonstrated visible morphological changes associated with smoltification. Mean values of U(crit) were significantly decreased and the prolonged contraction (tetani) and twitch rates of contraction, relaxation and maximum force were significantly increased by T3 treatment compared with both the sham and control fish. Hematocrit, body mass and body length were not significantly affected by T3 treatment. In conclusion, we suggest that the reported decrease in U(crit) during salmonid smoltification may be mediated by endogenous T3-induced contractile modification of mosaic muscle fibers.

The causes of failure of lumbar transpedicular spinal instrumentation and fusion: a prospective study.

A prospective study was made of 101 patients who underwent transpedicular spinal instrumentation and fusion of the lumbar spine. All were reviewed by an independent observer. The objectives were to determine the causes of clinical failure and to recommend preventive measures. Forty-two primary and 59 revision operations were performed. The average age was 46 years and follow up 4 years. The variable screw placement and Cotrel-Dubousset systems were used. Posterior lumbar interbody fusion was carried out in 24 patients. The outcome was satisfactory after primary operations in 67% and after revisions in 46%. Nerve root injuries due to screw placement occurred in 4% (2% permanent and 2% transient). Instrumentation-induced foraminal stenosis developed in 2%. Proper surgical technique can avoid these complications. Predictive factors for failure are: abnormal psychology, symptomatic epidural fibrosis, inadequate decompression of lateral stenosis, surgical complications and the use of allografts.

The importance of the blood supply in the healing of tibial fractures.

A clinical study of the effects of nonischemic arterial injuries to the leg on tibial fracture healing is presented. In addition, the literature published during the past 100 years describing the vascular supply to the tibia is reviewed, and the San Francisco General Hospital treatment protocol for difficult open tibial fractures is outlined. Based on the finding that the delayed and nonunion rate is three times higher when one of the three arteries of the leg is disrupted, a modification of the Gustilo fracture classification is proposed.

Delayed unions and nonunions of open tibial fractures. Correlation with arteriography results.

To investigate the importance of arterial integrity in tibial fracture healing, the authors retrospectively assessed prognosis as a function of arterial injury in a large series of tibial fracture patients. The records of 114 patients who were treated for an open fracture of the tibia at a San Francisco hospital between 1981 and 1991 were selected for review; in each case, the injury had been caused by a blunt trauma, and arteriography had been performed to evaluate a suspected arterial injury in the ipsilateral extremity. Sixty-two patients had had a normal arteriogram and 52 had demonstrated occlusion of one or two of the three arteries that supply the foot. The group of patients who had demonstrated arterial occlusion had a significantly greater incidence of delayed union or nonunion (24 of 52 compared with ten of 62) and notably more cases of osteomyelitis (nine of 52 compared with four of 62); when the patients with osteomyelitis are excluded, the difference between the groups is still significant. This study is the first that indicates that patients with open fractures of the tibia who demonstrate arterial occlusion in their ipsilateral extremity may be more prone to delayed union or nonunion. More vigilant follow-up evaluation may therefore be warranted in such patients to better anticipate the need for exchange rodding or bone grafting.