peer reviewed | The purpose of this study was to assess the impact of freezing and thawing on MR images of equine feet examined ex vivo. Nine equine cadaver digits were first imaged at room temperature (T0). Among the 9 digits, 3 (group 1) were imaged in a 3 Tesla MR system after one and after 2 freezing-thawing cycles. Digits of group 1 were thawed in a cold room at 4°C for 36h. Three other digits (group 2) were imaged after one freezing-thawing cycle. Digits of group 2 were thawed in a cold room at 4°C and then rescanned after 24h at room temperature. The last 3 digits (group 3) were scanned after one freezing-thawing cycle. Digits of group 3 were thawed at room temperature for 24h. Sequences used were Spin Echo (SE) T1, Turbo Spin Echo (TSE) T2 and proton density (PD), Short Tau Inversion Recovery (STIR), Double Echo Steady State (DESS), 3D Gradient Echo (GE) T1 and 2D GE T2*. Images obtained on the fresh limbs at room temperature were subjectively compared side by side to images obtained at the different freezing-thawing cycles. A quantitative analysis to assess signal change between examinations was realized by measuring signal to noise ratio (SNR). Visibility and margination of the anatomical structures of the foot and overall image quality were subjectively considered unchanged except for the hoof where the lamina was considered less visible distally after freezing and thawing in the GE T2* and in TSE T2 and PD sequences. Quantitative analysis demonstrated SNR changes in the bone marrow only in the distal phalanx in the SE T1 sequence when the feet were thawed at room temperature. When the feet were thawed in a cold room at 4°C, bone marrow SNR changes were present in the SE T1, GE T1 and TSE PD sequences. Signal changes were significant in the synovial recess when the thawing process was made at 4°C and not when the thawing process was at ambient temperature. The soft tissue structures and the hoof capsule showed significant changes with an increase of SNR, except in STIR, after freezing and thawing at 4°C and at room temperature. SNR changes in the soft tissues were mainly present in GE sequences.

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Objective-To evaluate tendon injuries in horses over a 16-week period by use of ultrasonography and low-field magnetic resonance imaging (MRI). Sample-Tendons of 8 young adult horses. Procedures-The percentage of experimentally induced tendon injury was evaluated in cross section at the maximal area of injury by use of ultrasonography and MRI at 3, 4, 6, 8, and 16 weeks after collagenase injection. The MRI signal intensities and histologic characteristics of each tendon were determined at the same time points. Results-At 4 weeks after collagenase injection, the area of maximal injury assessed on cross section was similar between ultrasonography and MRI. In lesions of > 4 weeks' duration, ultrasonography underestimated the area of maximal cross-sectional injury by approximately 18%, compared with results for MRI. Signal intensity of lesions on T1-weighted images was the most hyperintense of all the sequences, lesions on short tau inversion recovery images were slightly less hyperintense, and T2-weighted images were the most hypointense. Signal intensity of tendon lesions was significantly higher than the signal intensity for the unaltered deep digital flexor tendon. Histologically, there was a decrease in proteoglycan content, an increase in collagen content, and minimal change in fiber alignment during the 16 weeks of the study. Conclusions and Clinical Relevance-Ultrasonography may underestimate the extent of tendon damage in tendons with long-term injury. Low-field MRI provided a more sensitive technique for evaluation of tendon injury and should be considered in horses with tendinitis of > 4 weeks' duration.

Objective—To compare values of urodynamic measurements of cats with idiopathic cystitis (IC) with previously published data for healthy female cats. Animals—11 female cats with IC. Procedures—2 sequential cystometrograms and 2 urethral pressure profiles were obtained for each cat. All tracings were evaluated for evidence of overactive urinary bladder (OAB). Maximum urethral pressure (MUP), maximum urethral closure pressure (MUCP), and functional profile length were recorded. Results—Only 3 cats had obvious micturition events. None of the 11 cats had evidence of OAB. Although not significant, threshold pressure was lower in cats with IC than in healthy cats (mean ± SD, 89.0 ± 12.0 cm H2O vs 75.7 ± 16.3 cm H2O, respectively); however, the total volume infused was significantly lower in cats with IC (4.8 ± 2.1 mL/kg vs 8.3 ± 3.2 mL/kg). The MUCP was significantly higher in cats with IC than in healthy cats (158.0 ± 47.7 cm H2O vs 88.9 ± 23.9 cm H2O, respectively). The MUP was also significantly higher in all portions of the urethra in cats with IC. Conclusions and Clinical Relevance—No evidence of OAB was identified in any cat evaluated; therefore, medications used to target this abnormality did not appear justified. The high MUCP in cats with IC suggested that α1-adrenoceptor antagonists or skeletal muscle relaxants may be useful in this disease, and if these data were applicable to male cats, then α1-adrenoceptor antagonism may help prevent recurrent obstructive IC. Further studies are indicated to determine the effects, if any, these drugs might have in cats with IC.

Objective—To evaluate the effects of pilot hole diameter and tapping on insertion torque and axial pullout strength of 4.0-mm cancellous bone screws in a synthetic canine cancellous bone substitute. Sample—75 synthetic cancellous bone blocks (15 blocks/group). Procedures—For groups 1 through 5, screw size-pilot hole diameter combinations were 3.5–2.5 mm (cortical screws), 4.0–2.5 mm, 4.0–2.5 mm, 4.0–2.0 mm, and 4.0–2.0 mm, respectively. Holes were tapped in groups 1, 2, and 4 only (tap diameter, 3.5, 4.0, and 4.0 mm, respectively). One 70-mm-long screw was inserted into each block; in a servohydraulic materials testing machine, the screw was extracted (rate, 5 mm/min) until failure. Mean group values of maximum insertion torque, axial pullout strength, yield strength, and stiffness were determined. Results—Mean maximum insertion torque differed significantly among the 5 groups; the group 5 value was greatest, followed by group 3, 4, 2, and 1 values. Group 3, 4, and 5 axial pullout strengths were similar and significantly greater than the group 2 value; all values were significantly greater than that for group 1. Group 5 and 4 yield strengths were similar and significantly greater than the group 3, 2, and 1 values. Stiffness in group 3 was similar to group 4 and 2 values but significantly greater than the group 5 value; all values were significantly greater than that for group 1. Conclusions and Clinical Relevance—These synthetic cancellous bone model findings suggested that tapping a 2.0-mm-diameter pilot hole when placing a 4.0-mm screw is the optimal insertion technique.

Objective-To develop and evaluate a sandwich ELISA incorporating rabbit antiserum specific for canine surfactant protein A (SP-A) for use in measuring concentrations of SP-A in serum of dogs Sample-Serum samples obtained from 6 healthy dogs and 3 dogs with pulmonary disease. Procedures-Rabbit antiserum was prepared against purified canine SP-A. The IgG fraction was isolated via protein G affinity chromatography and was then biotinylated. The sandwich ELISA was performed by use of anti-SP-A antibody (IgG) preabsorbed with sera from healthy dogs. Validity of the ELISA was confirmed by determination of the detection limit, precision, reproducibility, and accuracy. Serum SP-A concentrations were measured in 6 healthy dogs and 3 dogs with pulmonary disease. Results-Detection limit of the ELISA was 2.0 ng/mL. Within- and between-assay coefficients of variation ranged from 3.8% to 14.1% and from 15.5% to 35.6%, respectively. The observed-to-expected recovery ratio ranged from 77.1% to 89.9%. Serum SP-A concentrations measured by use of the ELISA were ≤ 2.3 ng/mL in the 6 healthy dogs, 25.6 ng/mL in a dog with severe cardiac pulmonary edema, 8.3 ng/mL in a dog with pneumonia, and 10.1 ng/mL in a dog with lung lobe torsion. Conclusions and Clinical Relevance-The sandwich ELISA was found to be useful for measuring purified canine SP-A concentrations and canine SP-A concentrations in serum samples. The ELISA was precise, reproducible, and accurate. The ELISA may be beneficial in assessing serum concentrations of canine SP-A as a potential biomarker of pulmonary diseases in dogs.

Objective—To evaluate the radiographic, computed tomographic (CT), and cadaveric anatomy of the head of boa constrictors. Animals—4 Boa constrictor imperator cadavers. Procedures—Cadavers weighed 3.4 to 5.6 kg and had a body length ranging from 189 to 221 cm. Radiographic and CT images were obtained with a high-detail screen-film combination, and conventional CT was performed with a slice thickness of 1.5 mm. Radiographic images were obtained in ventrodorsal, dorsoventral, and left and right laterolateral recumbency; CT images were obtained with the animals positioned in ventral recumbency directly laying on a plastic support. At the end of the radiographic and CT imaging session, 2 heads were sectioned following a stratigraphic approach; the other 2, carefully maintained in the same position on the plastic support, were moved into a freezer (-20°C) until completely frozen and then sectioned into 3-mm slices, respecting the imaging protocol. The frozen sections were cleaned and then photographed on each side. Anatomic structures were identified and labeled on gross anatomic images and on the corresponding CT or radiographic image with the aid of available literature. Results—Radiographic and CT images provided high detail for visualization of bony structures; soft tissues were not easily identified on radiographic and CT images. Conclusions and Clinical Relevance—Results provide an atlas of stratigraphic and cross-sectional gross anatomy and radiographic and CT anatomy of the heads of boa constrictors that might be useful in the interpretation of any imaging modality in this species.

Objective—To investigate the effect of opsonization of Rhodococcus equi with R equi-specific antibodies in plasma on bacterial viability and phagocyte activation in a cell culture model of infection. Sample—Neutrophils and monocyte-derived macrophages from 6 healthy 1-week-old foals and 1 adult horse. Procedures—Foal and adult horse phagocytes were incubated with either opsonized or nonopsonized bacteria. Opsonization was achieved by use of plasma containing high or low concentrations of R equi-specific antibodies. Phagocyte oxidative burst activity was measured by use of flow cytometry, and macrophage tumor necrosis factor (TNF)-α production was measured via an ELISA. Extracellular and intracellular bacterial viability was measured with a novel R equi-luciferase construct that used a luminometer. Results—Opsonized bacteria increased oxidative burst activity in adult horse phagocytes, and neutrophil activity was dependent on the concentration of specific antibody. Secretion of TNF-α was higher in macrophages infected with opsonized bacteria. Opsonization had no significant effect on bacterial viability in macrophages; however, extracellular bacterial viability was decreased in broth containing plasma with R equi-specific antibodies, compared with viability in broth alone. Conclusions and Clinical Relevance—The use of plasma enriched with specific antibodies for the opsonization of R equi increased the activation of phagocytes and decreased bacterial viability in the extracellular space. Although opsonized R equi increased TNF-α secretion and oxidative burst in macrophages, additional factors may be necessary for effective intracellular bacterial killing. These data have suggested a possible role of plasma antibody in protection of foals from R equi pneumonia.

Objective—To identify and characterize cytochrome P450 enzymes (CYPs) responsible for the metabolism of racemic ketamine in 3 mammalian species in vitro by use of chemical inhibitors and antibodies. Sample—Human, canine, and equine liver microsomes and human single CYP3A4 and CYP2C9 and their canine orthologs. Procedures—Chemical inhibitors selective for human CYP enzymes and anti-CYP antibodies were incubated with racemic ketamine and liver microsomes or specific CYPs. Ketamine N-demethylation to norketamine was determined via enantioselective capillary electrophoresis. Results—The general CYP inhibitor 1-aminobenzotriazole almost completely blocked ketamine metabolism in human and canine liver microsomes but not in equine microsomes. Chemical inhibition of norketamine formation was dependent on inhibitor concentration in most circumstances. For all 3 species, inhibitors of CYP3A4, CYP2A6, CYP2C19, CYP2B6, and CYP2C9 diminished N-demethylation of ketamine. Anti-CYP3A4, anti-CYP2C9, and anti-CYP2B6 antibodies also inhibited ketamine N-demethylation. Chemical inhibition was strongest with inhibitors of CYP2A6 and CYP2C19 in canine and equine microsomes and with the CYP3A4 inhibitor in human microsomes. No significant contribution of CYP2D6 to ketamine biotransformation was observed. Although the human CYP2C9 inhibitor blocked ketamine N-demethylation completely in the canine ortholog CYP2C21, a strong inhibition was also obtained by the chemical inhibitors of CYP2C19 and CYP2B6. Ketamine N-demethylation was stereoselective in single human CYP3A4 and canine CYP2C21 enzymes. Conclusions and Clinical Relevance—Human-specific inhibitors of CYP2A6, CYP2C19, CYP3A4, CYP2B6, and CYP2C9 diminished ketamine N-demethylation in dogs and horses. To address drug-drug interactions in these animal species, investigations with single CYPs are needed.