An assessment of intra- and interobserver agreement of reporting orthopaedic findings on presale radiographs of Thoroughbred yearlings.

REASONS FOR PERFORMING STUDY: Radiography is commonly used in clinical practice but agreement of reporting of radiographically detected orthopaedic findings in horses has rarely been studied. OBJECTIVES: To assess agreement within and between observers for reporting of orthopaedic findings on presale radiographs of Thoroughbred yearlings. STUDY DESIGN: Retrospective analysis of archived radiographs. METHODS: Four veterinary radiology specialists each twice examined 167 sets of radiographs for orthopaedic findings in the fore feet, fore and hind fetlocks, carpi, tarsi and stifles. There were 27 findings analysed for agreement. Kappa statistic (κ), percentage of positive agreement (Ppos) and percentage of negative agreement are reported. RESULTS: An excellent percentage of negative agreement was observed for all findings, with the exception of regular vascular channels of the proximal sesamoid bones. Ppos and κ results were variable. The presence of extra carpal bones, osseous cyst-like lesions of the ulnar carpal bone, sagittal ridge defects of the third metacarpus, fracture of the fore and hind proximal sesamoid bones, regular vascular channels in the hind proximal sesamoids, osteochondrosis lesions of the distal intermediate ridge and/or medial malleoli of the tibia, and osseous cyst-like lesions in the medial femoral condyle was consistently observed with an intra- and interagreement κ≥0.5 and Ppos≥50%. Lucency within the proximal sesamoids consistently had an observed intra- and interagreement κ<0.4 and Ppos<40%. CONCLUSIONS: Observation of orthopaedic findings on yearling repository radiographs showed generally excellent agreement on the absence of findings, but variable agreement on the presence of findings. Agreement was good for larger and easy to categorise radiographic findings. More accurate definitions and training need to be developed to improve agreement within and between observers for orthopaedic findings with poor or fair to good agreement.

Biliary excretion of technetium-99m-sestamibi in wild-type dogs and in dogs with intrinsic (ABCB1-1Delta mutation) and extrinsic (ketoconazole treated) P-glycoprotein deficiency.

P-glycoprotein (P-gp), the product of ABCB1 gene, is thought to play a role in the biliary excretion of a variety of drugs, but specific studies in dogs have not been performed. Because a number of endogenous (ABCB1 polymorphisms) and exogenous (pharmacological P-gp inhibition) factors can interfere with normal P-gp function, a better understanding of P-gp's role in biliary drug excretion is crucial in preventing adverse drug reactions and drug-drug interactions in dogs. The objectives of this study were to compare biliary excretion of technetium-99m-sestamibi ((99m)Tc-MIBI), a radio-labelled P-gp substrate, in wild-type dogs (ABCB1 wild/wild), and dogs with intrinsic and extrinsic deficiencies in P-gp function. Dogs with intrinsic P-gp deficiency included ABCB1 mut/mut dogs, and dogs with presumed intermediate P-gp phenotype (ABCB1 mut/wild). Dogs with extrinsic P-gp deficiency were considered to be ABCB1 wild/wild dogs treated with the P-gp inhibitor ketoconazole (5 mg/kg PO q12h x 9 doses). Results from this study indicate that ABCB1 mut/mut dogs have significantly decreased biliary excretion of (99m)Tc-MIBI compared with ABCB1 wild/wild dogs. Treatment with ketoconazole significantly decreased biliary excretion of (99m)Tc-MIBI in ABCB1 wild/wild dogs. P-gp appears to play an important role in the biliary excretion of (99m)Tc-MIBI in dogs. It is likely that concurrent administration of a P-gp inhibitor such as ketoconazole will decrease P-gp-mediated biliary excretion of other substrate drugs as well.

Digital radiography.

Digital radiography has been used in human medical imaging since the 1980s with recent and rapid acceptance into the veterinary profession. Using advanced image capture and computer technology, radiographic images are viewed on a computer monitor. This is advantageous because radiographic images can be adjusted using dedicated computer software to maximize diagnostic image quality. Digital images can be accessed at computer workstations throughout the hospital, instantly retrieved from computer archives, and transmitted via the internet for consultation or case referral. Digital radiographic data can also be incorporated into a hospital information system, making record keeping an entirely paperless process. Digital image acquisition is faster when compared to conventional screen-film radiography, improving workflow and patient throughput. Digital radiography greatly reduces the need for 'retake' radiographs because of wide latitude in exposure factors. Also eliminated are costs associated with radiographic film and x-ray film development. Computed radiography, charged coupled devices, and flat panel detectors are types of digital radiography systems currently available.

Liver histopathology and liver and serum alanine aminotransferase and alkaline phosphatase activities in epileptic dogs receiving phenobarbital.

Phenobarbital (PB) therapy is frequently associated with elevated serum alanine aminotransferase (ALT) and alkaline phosphatase (AP) activities in dogs without clinical signs of liver disease. The goal of this study was to determine if increased serum ALT and AP activities in clinically healthy PB-treated epileptic dogs are due to hepatic enzyme induction or to subclinical liver injury. Liver biopsies were obtained from 12 PB-treated dogs without clinical signs of liver disease but with elevated serum ALT and/or AP activities or both. Liver biopsies were obtained from eight healthy control dogs not receiving PB. Biopsies were evaluated histopathologically (all dogs) and liver homogenates were assayed for ALT (all dogs) and AP (six treated dogs, all controls) activities. As a positive control, liver cytochrome P4502B, an enzyme known to be induced by PB, was measured by benzyloxyresorufin-O-dealkylase activity and immunoblotting (five treated dogs, all controls). Serum AP isoenzyme analyses were performed. Results showed that ALT and AP activities in liver homogenates were not increased in treated dogs compared with controls, whereas the positive control for induction, CYP2B, was dramatically increased in treated dogs. Histopathological examination of liver biopsies revealed more severe and frequent abnormalities in treated dogs compared to controls, but similar types of abnormalities were found in both groups. Serum AP isoenzyme analyses in treated dogs demonstrated increased corticosteroid-induced and liver isoenzyme activities compared to controls. Results do not support induction of ALT or AP in the liver as the cause of elevated serum activities of these enzymes due to PB.

Metabolic changes and induction of hepatic lipidosis during feed restriction in llamas.

OBJECTIVES: To determine whether feed restriction induces hepatic lipidosis (HL) in llamas and to evaluate the metabolic changes that develop during feed restriction. ANIMALS: 8 healthy adult female llamas. PROCEDURE: Llamas were fed grass hay at a rate of 0.25% of their body weight per day for 13 to 28 days. Llamas were monitored by use of clinical observation, serum biochemical analyses, and ultrasound-guided liver biopsies. RESULTS: All 8 llamas lost weight and mobilized fat. Five llamas developed HL, including 4 that were nursing crias. During the period of feed restriction, mean serum concentration of bile acids and activities of aspartate aminotransferase (AST), sorbitol dehydrogenase (SDH), and gamma-glutamyl transferase (GGT) were significantly higher in llamas that developed HL, compared with llamas that did not. Mean insulin-to-cortisol concentration ratios were lower in llamas with HL before and up to 7 days of feed restriction, compared with those that did not develop HL. CONCLUSIONS AND CLINICAL RELEVANCE: HL in llamas may be induced by severe feed restriction, particularly in the face of increased energy demand. Llamas with weight loss attributable to inadequate dietary intake may develop biochemical evidence of hepatopathy and HL. Increases in serum concentration of bile acids and activities of GGT, AST, and SDH may indicate the development of HL in llamas and identify affected animals for aggressive therapeutic intervention.

Computed tomography for the diagnosis of a lumbosacral nerve sheath tumour and management by hemipelvectomy.

Computed tomography was used to define the gross extent of a lumbosacral plexus nerve sheath tumour in a 10-year-old crossbred dog. The tumour was also visualised ultrasonographically, which enabled a percutaneous biopsy to be taken. Hemipelvectomy was performed to achieve a clean margin of surgical excision.

Thoracic radiographic appearance in the normal llama.

The goal of this study was to collect quantitative and qualitative radiographic information of the normal adult llama thorax. Standing right-left lateral radiographs of the thorax of 16 normal llamas were made. Normal ratios of cardiac height, width, and height plus width to thoracic vertebrae 3-5 and thoracic height were calculated. Normal values determined for tracheal angle of divergence from the thoracic spine, cardiophrenic and cardiosternal contact are additional potential indicators of cardiac enlargement. Ratios of normal pulmonary artery and vein, caudal vena cava and trachea to the height of the fourth thoracic vertebra should allow identification of pathology of these structures. Observations regarding pulmonary vessels and airways, thoracic spine, sternebrae and portions of the gastrointestinal tract observed on thoracic radiographs are also included. It is proposed that these normal values and observations can be used to better evaluate diseases of the cardiovascular and respiratory systems of adult llamas.

The Saccharomyces cerevisiae LEP1/SAC3 gene is associated with leucine transport.

Leucine uptake by Saccharomyces cerevisiae is mediated by three transport systems, the general amino acid transport system (GAP), encoded by GAP1, and two group-specific systems (S1 and S2), which also transport isoleucine and valine. A new mutant defective in both group-specific transport activities was isolated by employing a gap1 leu4 strain and selecting for trifluoroleucine-resistant mutants which also showed greatly reduced ability to utilize L-leucine as sole nitrogen source and very low levels of [14C]L-leucine uptake. A multicopy plasmid containing a DNA fragment which complemented the leucine transport defect was isolated by selecting for transformants that grew normally on minimal medium containing leucine as nitrogen source and subsequently assaying [14C]L-leucine uptake. Transformation of one such mutant, lep1, restored sensitivity to trifluoroleucine. The complementing gene, designated LEP1, was subcloned and sequenced. The LEP1 ORF encodes a large protein that lacks characteristics of a transporter or permease (i.e., lacks hydrophobic domains necessary for membrane association). Instead, Lep1p is a very basic protein (pI of 9.2) that contains a putative bipartite signal sequence for targeting to the nucleus, suggesting that it might be a DNA-binding protein. A database search revealed that LEP1 encodes a polypeptide that is identical to Sac3p except for an N-terminal truncation. The original identification of SAC3 was based on the isolation of a mutant allele, sac3-1, that suppresses the temperature-sensitive growth defect of an actin mutant containing the allele act1-1. Sac3p has been previously shown to be localized in the nucleus. When a lep1 mutant was crossed with a sac3 deletion mutant, no complementation was observed, indicating that the two mutations are functionally allelic.

Use of computed tomography in the diagnosis of a cerebral abscess in a goat.

A 7-month-old La Mancha doe was presented with a right head tilt and ventral strabismus while circling to the left. History and physical examination led to a tentative diagnosis of a cerebral abscess. Computed tomography revealed a large, complex mass with ring enhancement in the left cerebral hemisphere, supporting the diagnosis. Postmortem examination confirmed the cerebral abscess. Bacterial cultures yielded heavy growth of Actinomyces pyogenes.

Upper airway obstruction in a llama caused by aberrant nasopharyngeal bots (Cephenemyia sp.).

A 9 month old female llama was presented with inspiratory dyspnea. Radiographically, there was a large soft tissue mass nearly occluding the nasopharynx. During endoscopic examination three nasopharyngeal bots were identified embedded in the mass. The larvae were removed and the patient treated with ivermectin. The patient was discharged one week later free of clinical respiratory disease. In follow-up radiographs made 6 weeks later, only residual radiopacity in the area of the mass remained.

RAS2/PKA pathway activity is involved in the nitrogen regulation of L-leucine uptake in Saccharomyces cerevisiae.

The aim of the present work is to study the participation of RAS2/PKA signal pathway in the nitrogen regulation of L-leucine transport in yeast cells. The study was performed on Saccharomyces cerevisiae isogenic strains with the normal RAS2 gene, the RAS2val19 mutant and the disrupted ras2::LEU2. These strains bring about different activities of the RAS2/PKA signal pathway, L-(14C)-Amino acid uptake measurements were determined in cells grown in a rich YPD medium with a mixed nitrogen source or in minimal media containing NH4+ or L-proline as the sole nitrogen source. We report herein that in all strains used, even in those grown in a minimal proline medium, the activity of the general amino acid permease (GAP1) was not detected. L-Leucine uptake in these strains is mediated by two kinetically characterized transport systems. Their KT values are of the same order as those of S1 and S2 L-leucine permeases. Mutation in the RAS2 gene alters initial velocities and Jmax values in both high and low affinity L-leucine transport systems. Activation of the RAS2/PKA signalling pathway by the RAS2val19 mutation, blocks the response to a poor nitrogen source whereas inactivation of RAS2 by gene disruption, results in an increase of the same response.

Isolation of a trifluoroleucine-resistant mutant of Saccharomyces cerevisiae deficient in both high- and low-affinity L-leucine transport.

A yeast mutant defective in permeases S1 and S2 which transport L-leucine was isolated from a parental strain already deficient in the general amino acid permease, GAP1. The mutant was selected as a spontaneous, trifluoroleucine-resistant (TFLR) strain. Full resistance depended upon the presence of two unlinked mutant genes designated let1 and let2. The let1 mutation completely inactivates the high-affinity leucine transport system defined kinetically as S1. Although the let2 mutation caused a marked decrease in the Jmax of the low-affinity transport system, S2, residual leucine transport in the let1 let2 gap1 mutant had the same KT as in the LET1 LET2 gap1 parent. The mutant exhibited a marked decrease in growth on minimal medium containing leucine, isoleucine or valine as a sole nitrogen source. Moreover, assimilation of methionine, phenylalanine, serine and threonine was decreased, whereas basic and acidic amino acids supported normal growth. This indicates that at least one of the leucine permeases has a fairly broad, but still limited, specificity. Reversion of the gap1 gene restored leucine transport. The revertant was sensitive to TFL when grown on proline but resistant when NH4+ was the nitrogen source. The previously published mutations (shr3, aat1, lup1 or raa) would not be related to either LET1 or LET2.