Imaging techniques in veterinary medicine. Part II: Computed tomography, magnetic resonance imaging, nuclear medicine.

Radiography and ultrasonography are the most used techniques in veterinary clinical practice, due to organizational, managerial and, mostly, economic reasons. However, in the last decades, Computed tomography (CT), Magnetic Resonance Imaging (MRI) and, to a lesser extent, Nuclear Medicine (MN) are increasingly used. As we said in the previous article, all the Diagnostic Imaging techniques are actually "indispensable" in Veterinary Medicine, where many patients do not show any symptoms.This second part describes Computed Tomography (CT), Magnetic Resonance (MRI) and Nuclear Medicine techniques in Veterinary Medicine are described.

Computed tomographic appearance of transcaval ureter in two dogs and three cats: A novel CVC congenital malformation.

Transcaval ureter is a rarely reported human congenital malformation of the prerenal segment of the inferior vena cava (IVC) not yet reported in veterinary medicine. The objective of this multicenter retrospective case series study was to describe the computed tomography (CT) features of transcaval ureters in dogs and cats. Patients referring to pre- and post-contrast CT exams of the abdomen and presenting this abnormality were retrospectively included. Multiple qualitative features were described for each ureteral abnormality detected. Three cats and two dogs with transcaval ureter were identified consisting of a segmental duplication of the CVC at the prerenal level creating a vascular ring through which the ureter extended, identified as a double-barrel gun sign. The malformation was divided into two types according to the symmetry of the caval branches and location in relation to the aorta, namely, type I symmetrical branches and right-lateral to the aorta, and type II asymmetrically branches and right-dorsal to the aorta. In one case, the malformation was associated with hydroureter and mild pyelectasis. In three cases, the anomaly was incidental and, in the remaining two cases, the clinical significance was uncertain. This is the first study describing the presence of transcaval ureter in dogs and cats. CT was a suitable method for the diagnosis of transcaval and a focal double-barrel gun sign of the CVC is proposed as the hallmark feature of this anomaly. The clinical relevance of this congenital vascular malformation is unclear and needs to be further investigated.

Use of dexmedetomidine repeated subcutaneous administration for balanced anaesthesia in horses.

BACKGROUND: A balanced anaesthetic protocol is a common concept in modern veterinary anaesthesia and aims to maintain good intraoperative cardiopulmonary function. In horses, alpha-2-agonists produce sedation and analgesia and have been shown to reduce inhalational anaesthetic requirements when administered intravenously. Furthermore, these drugs can improve recovery quality. Preliminary investigations of subcutaneous dexmedetomidine administration in humans demonstrated a reduced haemodynamic impact if compared with the intravenous route suggesting that dexmedetomidine is adequately absorbed with both administration routes. The aim of the study was to compare two different dexmedetomidine (DEX) administration routes: intravenous constant rate infusion (CRI) versus repeated subcutaneous (SC) injections on cardiopulmonary function and recovery in anaesthetized horses. RESULTS: No significant differences between groups in heart rate and systolic arterial pressure were detected. A significantly higher mean and diastolic arterial pressure were detected in the SC group at T25 (p = 0.04; p = 0.02), T75 (p = 0.02; p = 0.009), and T85 (p = 0.001; p = 0.005). In SC group there was a significantly lower dobutamine infusion rate (p = 0.03) and a significantly higher urinary output (p = 0.02). Moreover, recovery quality was higher (p = 0.01). CONCLUSIONS: Cardiopulmonary effects in both groups were comparable and within clinical ranges with less dobutamine requirement in the subcutaneous group. Recovery was of better quality with fewer attempts in horses receiving subcutaneous dexmedetomidine. The present study suggests that intravenous constant rate infusion and subcutaneous repeated administration of dexmedetomidine at indicated dosage can be useful in balanced anaesthesia without any systemic or local adverse effects; moreover, in healthy horses undergoing general anaesthesia, repeated subcutaneous dexmedetomidine administration may be a suitable alternative if constant rate infusion is not feasible.

Fall time may be a reliable discriminator between neoplastic and non-neoplastic urinary bladder lesions in dogs undergoing contrast-enhanced ultrasound: a pilot study.

Contrast-enhanced ultrasound (CEUS) can provide quantitative information on enhancement patterns and perfusion of lesions, based on time-intensity curves (TICs). No published studies have compared CEUS parameters in neoplastic and non-neoplastic urinary bladder lesions in dogs. The aim of the current prospective, pilot study was to quantitatively characterize the CEUS pattern of neoplastic and non-neoplastic urinary bladder lesions in dogs, assessing the influence of contrast arrival time (CAT) on the final appearance of the curves. Fourteen dogs with cyto-histopathological diagnoses were included (seven malignant and seven inflammatory lesions). B-mode ultrasound was performed followed by CEUS examination after an intravenous bolus injection of 0.04 mL/kg of contrast medium, and TICs were elaborated by dedicated software. Receiver operating characteristic curves (ROC) for each TIC parameter were obtained. Neoplastic lesions had subjectively shorter rise time (RT), time to peak (TTP) and fall time (FT) than inflammatory lesions. Based on ROC curve analyses, fall time ≥ 10.49 s was the most reliable parameter for diagnosing non-neoplastic disease in this small sample of dogs (area under the curve [AUC] 0.75, sensitivity 83.33%, specificity 66.67%). No difference was found between ROCs calculated for each parameter of TICs by adding or removing CAT. Results of the current study provide background for future, larger scale studies evaluating use of a CEUS FT threshold of 10.49 s as a possible discriminator for urinary bladder neoplastic lesions in dogs.

Imaging techniques in Veterinary Medicine. Part I: Radiography and Ultrasonography.

In Veterinary Medicine all the Imaging techniques are used and described but, due to organizational, managerial and, mostly, economical reasons, Radiography and Ultrasonography are the most used. Veterinary Radiology teaching has a relatively small number of educational credits in the degree courses but, nowday, educational opportunities are remarkably increased thanks to a number of post-degree courses and masters, organized both by the universities and private veterinary associations. The relevance of Diagnostic Imaging is particularly true in Veterinary Medicine, in which Radiology could be considered "indispensable" for diagnosis, prognosis and follow up. Furthermore, it should not be forgotten that the diagnostic image represents a "document" and, has a relevant role in legal medical debats. In this first part, Radiography and Ultrasonography are described.

A computed tomography-based method for the assessment of canine retrobulbar cone volume for ophthalmic anaesthesia.

OBJECTIVE: To develop a comprehensive formula for calculating the volume of local anaesthetic solution used for retrobulbar anaesthesia in dogs with different skull morphologies. STUDY DESIGN: Retrospective cohort imaging study. ANIMALS: Skull computed tomography (CT) images of 188 dogs of different breeds collected between January 2009 and December 2017. METHODS: Anatomical integrity of the orbit and adjacent structures, presenting complaint, clinical signs and CT findings were verified to exclude ocular abnormalities. The volume of the retrobulbar cone of 376 eyes was calculated using CT scans of the dogs' skulls. Additional data recorded included morphology of the skull, body weight, sex and size of the dogs, all of which were matched for possible association to the retrobulbar cone volume through univariable and multivariable linear regression models. Results of linear regression models were expressed as estimated beta coefficients with the corresponding 95% confidence intervals (95% CIs). RESULTS: Using univariate analysis, the retrobulbar cone volume was positively associated with weight and male sex. In addition, brachycephalic and dolichocephalic dogs showed a larger retrobulbar cone volume than mesocephalic dogs, while sex was no longer significantly associated with the retrobulbar cone volume. In multivariate analysis, when considering all variables in the model, weight emerged as the strongest predictor (beta coefficient: 0.062 mL kg-1, 95% CI: 0.056-0.067 mL kg-1, p < 0.001). CONCLUSIONS: and clinical relevance In the veterinary literature, there is no agreement on the precise volume of local anaesthetic solution that should be used to achieve intraconal retrobulbar anaesthesia in dogs. Here we suggest a formula to calculate the retrobulbar cone volume and, accordingly, the injection volume of local anaesthetic solution for effective retrobulbar anaesthesia.

Preoperative planar lymphoscintigraphy allows for sentinel lymph node detection in 51 dogs improving staging accuracy: Feasibility and pitfalls.

Sentinel lymph node (SLN) mapping is the current gold standard for the oncological staging of solid malignancies in humans. This prospective observational study describes the feasibility and the limits of preoperative lymphoscintigraphy for SLN detection in dogs with spontaneous malignancies and the improvements in staging accuracy. Client-owned dogs with confirmed malignant neoplasia and absence of distant metastasis were prospectively enrolled. Lymphoscintigraphy was performed after the peritumoral injection of Technetium-99m labeled nanocolloids. Regional dynamic and static images were acquired, with and without masking of the injection site with a lead shield. The dogs were then subjected to surgery for tumor excision and SLN extirpation. Intraoperative SLN detection was performed by combining methylene blue dye and a dedicated gamma probe. Overall, 51 dogs with a total of 60 solid malignant tumors were enrolled. Lymphoscintigraphy identified at least one SLN in 57 of 60 cases (95%). The SLN did not always correspond to the regional lymph node (35/57, 61.4%). The use of a lead shield, masking the injection site, markedly improved the SLN visibility. The median time of SLN appearance was 11.4 ± 9.3 min. No side effects were observed. Preoperative lymphoscintigraphy allows for SLN detection in dogs and can improve staging accuracy by either identifying the SLN in a different lymphosome than clinically expected or discriminating the draining node in uncertain cases. The combined use of preoperative and intraoperative techniques is recommended to increase the SLN detection rate.

To map or not to map the cN0 neck: Impact of sentinel lymph node biopsy in canine head and neck tumours.

Tumour stage is a prognostic indicator for canine malignant head and neck tumours (MHNT). However, consensus is lacking on nodal staging in the absence of clinically apparent nodal disease (cN0 neck). This prospective observational study aims to determine the diagnostic accuracy of radiopharmaceutical and blue dye for sentinel lymph node biopsy (SLNB), to assess the correspondence between sentinel lymph node (SLN) and clinically expected regional lymph node (RLN) and the impact on staging of the procedure in dogs with MHNT and cN0 neck. Twenty-three dogs with MHNT and cN0 neck underwent tumour excision and SLNB guided by preoperative lymphoscintigraphy and intraoperative gamma-probe and blue dye. Diagnostic performances and detection rate were calculated. Correspondence between SLN and RLN, number of nodes excised, histopathological status of the SLN and complications related to the procedure were recorded. The mapping technique identified at least one SLN in 19/23 dogs, with a detection rate of 83%. The SLN did not correspond to the RLN in 52% of dogs. Multiple nodes were removed in 61% of dogs. At histopathology, eight (42%) dogs had SLN+, of which four differed from the RLN. Only minor self-limiting complications occurred in five (22%) dogs. Radiopharmaceutical and blue dye guidance is accurate (sensitivity 88.9%; specificity 100%) for SLNB in dogs with MHNT and cN0 and allowed the extirpation of unpredictable and/or multiple SLN with minimal morbidity. Incorporation of SLNB in the management of MHNT is desirable to correctly stage the cN0 neck, owing the unpredictability of the lymphatic drainage.

Hind limb ossification centre hypoplasia and deformities induced by quadriceps contracture: Radiographic and Computed Tomographic study in 13 Doberman Pinscher littermates.

Quadriceps contracture (QC) is reported in dogs mainly as a complication of trauma or parasitic infection. QC causes progressive hind limb deviation, muscular hypotrophy and degenerative joint disease and, in puppies, bone hypoplasia. The aim of this clinical case series is to describe the radiographic and computed tomographic (CT) changes in hind limb ossification centres in thirteen 55 to 57-days-old Doberman Pinscher related littermates induced by QC after repeated intramuscular injections. The presence, size, and shape of ossification centres of affected and unaffected hind limbs were compared. Affected limbs were hyperextended and externally rotated, with genu recurvatum and proximo-medial patellar luxation. QC had no influence on the time of appearance of ossification centres however, it was associated with femoral head flattening, hip subluxation, flattening of the femoral distal epiphysis. The tibial plateau was tilted caudoproximally-craniodistally and wedged into the growth plate. Thirty-two out of fifty-five ossification centres (including diaphyseal and epiphyseal centres, such as femoral head and tibial plateau) were significantly smaller in affected limbs (p < 0.05). Lack of weight-bearing could account for the smaller size of ossification centres in affected tarsi and metatarsi. Progressive limb hyperextension and external rotation might have induced gradual loading withdrawal on the medial aspect of the foot justifying the reduced size observed only for the medial ossification centres of the digits of affected limbs. To the best of our knowledge, this is the first study describing CT findings of hind limb ossification centre changes in puppies with QC contracture.

Meniscus Matrix Remodeling in Response to Compressive Forces in Dogs.

Joint motion and postnatal stress of weight bearing are the principal factors that determine the phenotypical and architectural changes that characterize the maturation process of the meniscus. In this study, the effect of compressive forces on the meniscus will be evaluated in a litter of 12 Dobermann Pinschers, of approximately 2 months of age, euthanized as affected by the quadriceps contracture muscle syndrome of a single limb focusing on extracellular matrix remodeling and cell-extracellular matrix interaction (i.e., meniscal cells maturation, collagen fibers typology and arrangement). The affected limbs were considered as models of continuous compression while the physiologic loaded limbs were considered as controls. The results of this study suggest that a compressive continuous force, applied to the native meniscal cells, triggers an early maturation of the cellular phenotype, at the expense of the proper organization of collagen fibers. Nevertheless, an application of a compressive force could be useful in the engineering process of meniscal tissue in order to induce a faster achievement of the mature cellular phenotype and, consequently, the earlier production of the fundamental extracellular matrix (ECM), in order to improve cellular viability and adhesion of the cells within a hypothetical synthetic scaffold.

In vivo Diffusion Tensor Magnetic Resonance Tractography of the Sheep Brain: An Atlas of the Ovine White Matter Fiber Bundles.

Diffusion Tensor Magnetic Resonance Imaging (DTI) allows to decode the mobility of water molecules in cerebral tissue, which is highly directional along myelinated fibers. By integrating the direction of highest water diffusion through the tissue, DTI Tractography enables a non-invasive dissection of brain fiber bundles. As such, this technique is a unique probe for in vivo characterization of white matter architecture. Unraveling the principal brain texture features of preclinical models that are advantageously exploited in experimental neuroscience is crucial to correctly evaluate investigational findings and to correlate them with real clinical scenarios. Although structurally similar to the human brain, the gyrencephalic ovine model has not yet been characterized by a systematic DTI study. Here we present the first in vivo sheep (ovis aries) tractography atlas, where the course of the main white matter fiber bundles of the ovine brain has been reconstructed. In the context of the EU's Horizon EDEN2020 project, in vivo brain MRI protocol for ovine animal models was optimized on a 1.5T scanner. High resolution conventional MRI scans and DTI sequences (b-value = 1,000 s/mm2, 15 directions) were acquired on ten anesthetized sheep o. aries, in order to define the diffusion features of normal adult ovine brain tissue. Topography of the ovine cortex was studied and DTI maps were derived, to perform DTI tractography reconstruction of the corticospinal tract, corpus callosum, fornix, visual pathway, and occipitofrontal fascicle, bilaterally for all the animals. Binary masks of the tracts were then coregistered and reported in the space of a standard stereotaxic ovine reference system, to demonstrate the consistency of the fiber bundles and the minimal inter-subject variability in a unique tractography atlas. Our results determine the feasibility of a protocol to perform in vivo DTI tractography of the sheep, providing a reliable reconstruction and 3D rendering of major ovine fiber tracts underlying different neurological functions. Estimation of fiber directions and interactions would lead to a more comprehensive understanding of the sheep's brain anatomy, potentially exploitable in preclinical experiments, thus representing a precious tool for veterinaries and researchers.

Honeybee pupal length assessed by CT-scan technique: effects of Varroa infestation, developmental stage and spatial position within the brood comb.

Honeybee pupae morphology can be affected by a number of stressor, but in vivo investigation is difficult. A computed tomography (CT) technique was applied to visualize a comb's inner structure without damaging the brood. The CT scan was performed on a brood comb containing pupae developed from eggs laid by the queen during a time window of 48 hours. From the CT images, the position of each pupa was determined by recording coordinates to a common reference point. Afterwards, every brood cell was inspected in order to assess the developmental stage of the pupa, the presence of Varroa destructor, the number and progeny of foundress mites. Using data on 651 pupae, the relationships between varroa infestation status, developmental stage and spatial position of the pupa within the brood comb, and its length were investigated. Pupae at 8 post-capping days were shorter than pupae at 7 post-capping days. Pupae in infected cells were significantly shorter than those in varroa-free cells and this effect was linked both to mite number and stage and to the position in the comb. Overall, the results suggest that the CT-scan may represent a suitable non-invasive tool to investigate the morphology and developing status of honeybee brood.

Swine Meniscus: Are Femoral-Tibial Surfaces Properly Tuned to Bear the Forces Exerted on the Tissue?

IMPACT STATEMENT: The importance of the present study is linked to how the contact forces act on the knee meniscus in particular, considering the femoral condyles and tibial plateau: this can be useful as a base for the ultimate creation of tissue-engineered biphasic scaffolds, which can mimic the native tissue complex, for meniscal repair or regeneration.

Novel contributions in canine craniometry: Anatomic and radiographic measurements in newborn puppies.

The largest differences in intraspecific head shape among the Carnivora order are to be found in dogs. Based on their skull morphotypes, dog breeds are currently classified as dolichocephalic, mesaticephalic and brachycephalic. Due to the fact that some breeds have not been yet defined, this classification is incomplete; moreover, multi-breed studies on the skull morphology of puppies have never been performed. The aim of this work was to verify (i) whether differences in the skull conformation of purebred puppies are already present within the first week of age; (ii) whether radiographic and anatomic measures could be considered interchangeable, and (iii) to possibly classify puppies from non-categorized breeds thanks to their radiographic cranial measurements using neural nets. One hundred and thirty-seven dead puppies aged 0-7 days were examined considering their anatomic and radiographic measures. All linear measures and anatomic indices significantly differed among brachycephalic and non-brachycephalic puppies. Radiographic indices, with the exception of CI, identified the three skull morphotypes (p<0.05, for all comparisons). Radiographic and anatomic measures proved to be non-interchangeable in newborn puppies. Finally, nineteen puppies belonging to 5 non-categorized breeds could be classified thanks to neural nets in the three skull morphotypes with different probability (P between 0,66 and 0,95).

Dynamic tomographic studies of interscapular feline injection-site sarcoma: essential or useless practice?

Objectives Feline injection-site sarcomas (FISSs) are soft tissue tumours typically characterised by an interscapular location and highly infiltrative behaviour. CT is considered the modality of choice for FISS staging and double positioning (dynamic approach) was reported to successfully detect the exact extent of infiltration into the muscles. The aim of the present study was to investigate the utility of the dynamic approach in feline patients referred for preoperative staging of interscapular FISS. Methods Tumour volume estimates were compared between the ellipsoid and the semi-automated segmentation methods. Two radiologists blinded to the patient coding used images from each position to assess the extent of muscular infiltration. The distance between the neoplasm and the adjacent skeletal structures (scapulae, spinous processes) was recorded in both positions by a single radiologist. Results Fifty-nine of 84 neoplasms invaded the adjacent muscular structures, with up to 15 muscles infiltrated. Between the extended and flexed position the average estimated numbers of muscles infiltrated were 1.9 (extended) and 1.84 (flexed) for observer A and 1.89 (extended) and 1.85 (flexed) for observer B. Good agreement between observers was established, with higher tumour volumes detected via the ellipsoid method. Moreover, tumours with smaller volumes showed slightly decreased muscular infiltration. Marked difference in the recorded distance between the skeletal structures and the neoplasm in the two different positions was established (mean ± SD difference spinous processes: 9.74 ± 9.57 mm; mean ± SD difference scapulae: 15.15 ± 11.76 mm). Conclusions and relevance A dynamic approach should be used for a complete evaluation of the invasiveness of FISS along with appropriate methodology for tumour volume measurement, which could potentially alter the tomographic estimation of the real dimension of the neoplasms.

Use of the bolus tracking technique for the tomographic evaluation of the uretero-vesicular junction in dogs and assessment of dose records.

BACKGROUND: The aim of the work is the application of a bolus tracking technique for tomographic evaluation of the uretero-vesicular junction in dogs. Ten adult dogs (8-14 years) with variable body weight (2,8-32 kg) were enrolled in the prospective study. The patients were placed in sternal recumbency with a 10° elevated pelvis and the visualization of the uretero-vesicular junction was obtained with the bolus tracking technique after intravenous administration of non-ionic contrast medium. In the post-contrast late phase a region of interest was placed within the lumen of the distal ureters and the density values were monitored before starting the helical scan. RESULTS: The uretero-vesicular junction was clearly visible in 100% of patients with the visualization of the endoluminal ureteral contrast enhancement and bladder washout. At the end of the tomographic study an evaluation of the dose records was performed and compared to human exposures reported in literature for the pelvic region. The effective dose estimated for each patient (37,5-138 mSv) proved to be elevated, when compared to those reported in human patients. CONCLUSION: The bolus tracking technique could be applied for the visualization of the uretero-vesicular junction in non-pathological patients, placing the region of interest in the distal ureters. The high effective doses recorded in our study support the need of specific thresholds for veterinary patients, pointing out the attention for paediatric patient's exposure also in veterinary imaging.

Advances in the anatomic study of the interscapular region of the cat.

BACKGROUND: New clinical oriented approaches are supported by the integration of advanced imaging techniques, e.g. computed tomography and magnetic resonance, with gross anatomy imaging. The interscapular region of the cat is a typical site of a highly invasive infiltrative pathology, i.e. Feline Injection-Site Sarcoma. Even if chemotherapy and radiotherapy have been considered as pre-surgical therapies, extensive surgery is still the recommended treatment. Evidence suggested that the relationships between muscles, infiltrative mass and adjacent musculoskeletal structures could change according to the forelimb positions: a fact to be duly considered while planning the surgical approach. Anatomic and tomographic atlases provide only images of the interscapular region from cats positioned with their forelimbs extended cranially, which means that, they do not record musculoskeletal modifications due to the forelimb movements. Aim of this study was to provide detailed images of the changes occurring in the musculoskeletal structures of the interscapular region of cats according to their forelimb position by comparing cross-sectional gross anatomy, computed tomography and magnetic resonance imaging. RESULTS: We provide an atlas of normal cross-sectional anatomy, computed tomography and magnetic resonance imaging of the interscapular region of the cat, from the fifth cervical vertebra to the fifth thoracic vertebra. We compare and couple the slices obtained both in flexed and extended forelimb positioning with the animal maintained in sternal recumbency. CONCLUSION: This study shows a new and dynamic way to investigate the interscapular region of the cat and provides anatomical references for in vivo computed tomography and magnetic resonance imaging, considering changes in the muscular form according to the forelimb positioning. We believe that an in-depth anatomical knowledge of the interscapular region may be helpful to approach the study of any pathology located there and, in particular, to set up an appropriate therapy for the feline injection-site sarcoma.

Use of thermographic imaging in clinical diagnosis of small animal: preliminary notes.

INTRODUCTION: The authors, after a description of the physics of infrared thermographic technique (IRT), analyze the reading of images and the main applications in the veterinary field, compared to the existing literature on the subject and to their experimental researches. IRT lends itself to countless applications in biology, thanks to its characteristics of versatility, lack of invasiveness and high sensitivity. Probably the major limitation to its application in the animal lies in the ease of use and in its extreme sensitivity. MATERIALS AND METHODS: From September 2009 to October 2010, the experimental investigation with the thermo camera took into consideration 110 animals (92 dogs and 18 cats), without any selection criteria. All patients were brought to the Faculty of Veterinary Medicine in Milan University by the owner, to be examined by a specialist, or to undergo one of the following diagnostic procedures: X-rays, computed tomography, or ultrasound examinations; finally some patients were brought in for surgical procedures. With the consent of the owner, 1 to 10 thermographic images were recorded from each clinical case. RESULTS: In this first experimental investigation, thermography has shown a high sensitivity (100%), but a low specificity (44%). This figure excludes the use of thermal imaging technology to replace other imaging techniques such as radiography, computed tomography and magnetic resonance imaging. Furthermore, it does not show any ability to recognize the etiology of the disease, but only the thermal alteration, and this is restricting its use. However, this experimental study has demonstrated that thermography can be used in veterinary medicine, and specifically in dogs and cats. It is hoped that in the field of targeted diseases this technique will become an important tool for diagnostic purposes by using working protocols validated and repeatable.

Factors influencing wound healing complications after wide excision of injection site sarcomas of the trunk of cats.

OBJECTIVE: Wide surgery is the mainstay of the multimodal treatment of injection site sarcomas (ISS) in cats. The aim of the study was to analyze potential factors influencing the development of wound healing complications (WHC) in cats undergoing wide excision of ISS. STUDY DESIGN: Retrospective case series. ANIMALS: Forty-nine cats with ISS located on the trunk underwent wide excision after contrast-enhanced computed tomography planning. METHODS: The prognostic effect of covariates (sex, age, weight, body condition score (BCS), site, clinical dimension (CD), computed tomographic dimension (CTD), histotype, duration of surgery, surgical margin status, local anesthesia) on total, major and minor WHC was evaluated by univariate and bivariate analysis. Cox model was used for total WHC and Fine and Gray model was used for major and minor WHC. The relationship between duration of surgery and clinical and imaging variables was evaluated. RESULTS: The main factor associated to the risk of total and major WHC was surgical time. Based on univariate analysis, pattern of reconstruction, CDT, CD, weight, and BCS were significant prognostic factors for major WHC, but this was not confirmed when adjusted for other clinical variables in bivariate analysis. The duration of surgery was influenced by excision pattern and tumor CTD width. CONCLUSIONS: An increased duration of surgery as the consequence of complex surgical procedures represented the best predictor for the development of WHC.

Canine aural cholesteatoma: a histological and immunohistochemical study.

Canine aural cholesteatoma is an epidermoid cyst that forms in the middle ear cavity as a rare complication of otitis media but the aetiopathogenesis remains controversial. In the present study, 13 cases of canine aural cholesteatoma were investigated histologically and immunohistochemically and compared with cases of chronic otitis. The immunohistochemical investigation was performed using the following monoclonal antibodies: anti-cytokeratins (CK) 14, 16, 8/18, and 19, and anti-Ki67. The proliferative indexes (PIs) of cholesteatomata and otitis epithelium were calculated as the percentage of Ki67 positive nuclei/total nuclei. Histologically, the cholesteatomata were composed of a hyperplastic, hyperkeratotic epithelium (matrix) resting on a fibrous perimatrix, infiltrated by inflammatory cells and devoid of cutaneous adnexa. Immunohistochemically, the cholesteatoma epithelium was CK14- and CK16-positive, and CK8/18- and CK19-negative. A similar pattern of CK expression was found in otitis externa. In otitis media, ciliated epithelium stained CK8/18- and CK19-positive in all layers, CK14-positive in the basal layers, and CK16-negative. The mean PIs in cholesteatomata and otitides were 18.8 and 17.8, respectively. The immunohistochemical pattern of CK expression in cholesteatomata, when compared with chronic otitis, was suggestive of hyperproliferative epithelium, but its origin could not be demonstrated. Comparable PI values were obtained in cholesteatoma and in chronic otitis, which confirmed that Ki67 is a valuable indicator of a hyperproliferative state, but not a predictor of aggressiveness.