Progress in ultrahigh energy resolution EELS.

Electron energy loss spectroscopy (EELS) in the electron microscope has progressed remarkably in the last five years. Advances in monochromator and spectrometer design have improved the energy resolution attainable in a scanning transmission electron microscope (STEM) to 4.2 meV, and new applications of ultrahigh energy resolution EELS have not lagged behind. They include vibrational spectroscopy in the electron microscope, a field that did not exist 5 years ago but has now grown very substantially. Notable examples include vibrational mapping with about 1 nm spatial resolution, analyzing the momentum dependence of vibrational states in very small volumes, determining the local temperature of the sample from the ratio of energy gains to energy losses, detecting hydrogen and analyzing its bonding, probing radiation-sensitive materials with minimized damage by aloof spectroscopy and leap-frog scanning, and identifying biological molecules with different isotopic substitutions. We review the instrumentation advances, provide a summary of key applications, and chart likely future directions.

Use of routine histopathology and factor VIII-related antigen/von Willebrand factor immunohistochemistry to differentiate primary hemangiosarcoma of bone from telangiectatic osteosarcoma in 54 dogs.

Hemangiosarcoma (HSA) of bone and telangiectatic osteosarcoma (tOSA) can appear similar histologically, but differ in histogenesis (malignant endothelial cells versus osteoblasts), and may warrant different treatments. Immunohistochemistry (IHC) for endothelial cell marker factor VIII-related antigen/von Willebrand factor (FVIII-RAg/vWF) is a well-documented ancillary test to confirm HSA diagnoses in soft tissues, but its use in osseous HSA is rarely described. Archived samples of 54 primary appendicular bone tumours previously diagnosed as HSA or tOSA were evaluated using combination routine histopathology (RHP) and IHC. Approximately 20% of tumours were reclassified on the basis of FVIII-RAg/vWF immunoreactivity, typically from an original diagnosis of tOSA to a reclassified diagnosis of HSA. No sample with tumour osteoid clearly identified on RHP was immunopositive for FVIII-RAg/vWF. RHP alone was specific but not sensitive for diagnosis of HSA, compared with combination RHP and IHC. Routine histopathological evaluation in combination with FVIII-RAg/vWF IHC can help differentiate canine primary appendicular HSA from tOSA.

In vitro effects of Yunnan Baiyao on canine hemangiosarcoma cell lines.

Yunnan Baiyao is a Chinese herbal medicine that has been utilized for its anti-inflammatory, haemostatic, wound healing and pain relieving properties in people. It has been utilized in the veterinary profession to control bleeding in dogs with hemangiosarcoma (HSA) and has been anecdotally reported to prolong survival times in dogs with this neoplasm. This study evaluated the in vitro activity of Yunnan Baiyao against three canine HSA cell lines after treatment with increasing concentrations of Yunnan Baiyao (50, 100, 200, 400, 600 and 800 µg mL(-1) ) at 24, 48 and 72 h. Mean half maximum inhibitory concentration (IC50 ) at 72 h for DEN, Fitz, SB was 369.9, 275.9 and 325.3 µg mL(-1) , respectively. Caspase-3/7 activity increased in correlation with the IC50 in each cell line which was confirmed by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL, APO-BRDU Kit; BD Biosciences, San Jose, CA, USA) assay. VEGF in cell supernatant was also quantified. Overall, the study found that Yunnan Baiyao causes dose and time dependent HSA cell death through initiation of caspase-mediated apoptosis, which supports future studies involving Yunnan Baiyao.

Traumatic tympanic bulla fracture.

A Pekingese dog was presented for evaluation of head trauma with ventral head and neck swelling, puncture wounds, palpable mandibular fractures, and loss of menace, severe miosis, and loss of palpebral reflex of the right eye. Computed tomography confirmed multiple mandibular and zygomatic fractures, a right ear canal avulsion, and a complete right tympanic bulla fracture with ventral displacement. The tympanic bulla fracture was managed conservatively. Topical lubrication and antibiotic ointment was prescribed for the right eye. A subtotal hemimandibulectomy was performed to address the mandibular fractures. A temporary oesophagostomy feeding tube was placed. No short-term complications developed as a result of the fractured bulla and avulsed ear canal being left in situ, and no complications were reported 18 months after the injury. To the authors' knowledge this is the first report of a traumatic tympanic bulla fracture in the dog.

Radiosensitivity and capacity for radiation-induced sublethal damage repair of canine transitional cell carcinoma (TCC) cell lines.

Understanding the inherent radiosensitivity and repair capacity of canine transitional cell carcinoma (TCC) can aid in optimizing radiation protocols to treat this disease. The objective of this study was to evaluate the parameters surviving fraction at 2 Gy (SF(2) ), α/ß ratio and capacity for sublethal damage repair (SLDR) in response to radiation. Dose-response and split-dose studies were performed using the clonogenic assay. The mean SF(2) for three established TCC cell lines was high at 0.61. All the three cell lines exhibited a low to moderate α/ß ratio, with the mean being 3.27. Two cell lines exhibited statistically increased survival at 4 and 24 h in the dose-response assay. Overall, our results indicate that the cell lines are moderately radioresistant, have a high repair capacity and behave similarly to a late-responding normal tissue. These findings indicate that the radiation protocols utilizing higher doses with less fractionation may be more effective for treating TCC.

Prognostic factors for cutaneous and subcutaneous soft tissue sarcomas in dogs.

Soft tissue sarcomas (STSs) develop from mesenchymal cells of soft tissues, and they commonly occur in the skin and subcutis of the dog. Although phenotypically diverse with frequently controversial histogenesis, STSs are considered as a group because they have similar features microscopically and clinically. Following resection, local recurrence rates are low in general but vary according to histologic grade and completeness of surgical margins. Complete margins predict nonrecurrence. Even most grade I STSs with "close" margins will not recur, but propensity for recurrence increases with grade. The frequency of metastasis has not been accurately estimated, but it is believed to be rare for grade I STSs and most likely to occur with grade III STSs. However, metastasis does not necessarily equate with poor survival. High mitotic index is prognostic for reduced survival time. Further research is needed to determine more precise estimates for recurrence rates and survival as related to completeness of surgical margins and to delineate potential differences in metastatic rate and median survival time between grades. Other potential indicators of prognosis that presently require further investigation include histologic type, tumor dimension, location, invasiveness, stage, markers of cellular proliferation, and cytogenetic profiles. Common issues limiting prognostic factor evaluation include biases from retrospective studies, small sample sizes, poor verification of metastasis, inconsistent STS classification and use of nomenclature, difficulties in differentiating STS phenotype, and diversity of the study population (stage of disease and treatment status).

Recommended guidelines for submission, trimming, margin evaluation, and reporting of tumor biopsy specimens in veterinary surgical pathology.

Neoplastic diseases are typically diagnosed by biopsy and histopathological evaluation. The pathology report is key in determining prognosis, therapeutic decisions, and overall case management and therefore requires diagnostic accuracy, completeness, and clarity. Successful management relies on collaboration between clinical veterinarians, oncologists, and pathologists. To date there has been no standardized approach or guideline for the submission, trimming, margin evaluation, or reporting of neoplastic biopsy specimens in veterinary medicine. To address this issue, a committee consisting of veterinary pathologists and oncologists was established under the auspices of the American College of Veterinary Pathologists Oncology Committee. These consensus guidelines were subsequently reviewed and endorsed by a large international group of veterinary pathologists. These recommended guidelines are not mandated but rather exist to help clinicians and veterinary pathologists optimally handle neoplastic biopsy samples. Many of these guidelines represent the collective experience of the committee members and consensus group when assessing neoplastic lesions from veterinary patients but have not met the rigors of definitive scientific study and investigation. These questions of technique, analysis, and evaluation should be put through formal scrutiny in rigorous clinical studies in the near future so that more definitive guidelines can be derived.

Recommended guidelines for the conduct and evaluation of prognostic studies in veterinary oncology.

There is an increasing need for more accurate prognostic and predictive markers in veterinary oncology because of an increasing number of treatment options, the increased financial costs associated with treatment, and the emotional stress experienced by owners in association with the disease and its treatment. Numerous studies have evaluated potential prognostic and predictive markers for veterinary neoplastic diseases, but there are no established guidelines or standards for the conduct and reporting of prognostic studies in veterinary medicine. This lack of standardization has made the evaluation and comparison of studies difficult. Most important, translating these results to clinical applications is problematic. To address this issue, the American College of Veterinary Pathologists' Oncology Committee organized an initiative to establish guidelines for the conduct and reporting of prognostic studies in veterinary oncology. The goal of this initiative is to increase the quality and standardization of veterinary prognostic studies to facilitate independent evaluation, validation, comparison, and implementation of study results. This article represents a consensus statement on the conduct and reporting of prognostic studies in veterinary oncology from veterinary pathologists and oncologists from around the world. These guidelines should be considered a recommendation based on the current state of knowledge in the field, and they will need to be continually reevaluated and revised as the field of veterinary oncology continues to progress. As mentioned, these guidelines were developed through an initiative of the American College of Veterinary Pathologists' Oncology Committee, and they have been reviewed and endorsed by the World Small Animal Veterinary Association.

Paraneoplastic T cell lymphocytosis associated with a thymoma in a dog.

A four-year-old male neutered Australian shepherd dog was diagnosed with a thymoma and concurrent mature T cell lymphocytosis. The lymphocytosis consisted of a mixed population of T cells expressing either CD4 or CD8 or neither marker, and the result of polymerase chain reaction for antigen receptor rearrangement was negative. The peripheral lymphocytosis resolved within 24 hours following thoracotomy and thymectomy. Similar cases have been reported in man, but the aetiology of the increased circulating lymphocytes remains unclear. Although peripheral lymphocytosis is an uncommon paraneoplastic syndrome associated with thymomas, thymoma should be considered as a differential when the increased lymphocytes consist of a mixed population of T cells.

Hypobaric intrathecal anaesthesia for partial hemipelvectomy in a dog.

OBJECTIVE: To report the intrathecal use of a hypobaric anaesthetic solution for partial hemipelvectomy in a nine-year-old, neutered female, Golden Retriever dog, weighing 34 kg. METHODS: Under inhalational anaesthesia, with the dog lying in lateral recumbency and the surgical side uppermost, 1.9 ml of a hypobaric solution containing 3.42 mg of bupivacaine and 0.66 mg of morphine were administered in the subarachnoid space at L5-6 level 30 minutes before surgery. Following the intrathecal injection the dog was maintained for five minutes in a 10 degrees head-down position, then for three minutes in a 10 degrees head-up position. RESULTS: Apart from a transient increase in heart and respiratory rates during resection of the sartorius muscle, which was treated with a plasma Target Controlled Infusion (TCI) of fentanyl, spinal anaesthesia provided cardiovascular stability and excellent relaxation of the surgical site. Neither motor blockade nor proprioceptive deficit were apparent in the contra-lateral hind limb at recovery, 200 minutes after injection. Postoperatively, rescue analgesia was not required in the 48 hours following surgery. CLINICAL SIGNIFICANCE: In dogs, the use of intrathecal hypobaric bupivacaine and morphine as a part of a balanced anaesthetic protocol should be considered during unilateral major orthopaedic surgeries of the pelvis and hind limb, as it allowed a reduction in the dose administered compared to isobaric solutions, providing selective spinal anaesthesia, excellent long-lasting analgesia, and rapid recovery of ambulation.

Oesophageal leiomyosarcoma in dogs: surgical management and clinical outcome of four cases.

Oesophageal leiomyosarcoma has yet to be reported in dogs. This retrospective case series describes the case management and clinical outcome of four dogs with oesophageal leiomyosarcoma treated by marginal excision alone. Histological features used to determine tumour grade included capsular invasion, percent necrosis, pleomorphism and mitotic rate. All tumours were designated grade 1 leiomyosarcoma. Excision of all grossly evident tumour tissue was achieved in two of the four cases; however, histopathologic evaluation showed tumour cells at the surgical margins in one of these two cases. Two dogs had grossly incomplete excision. Two dogs died from unrelated conditions, one 3 years and 5.5 months after surgery, the other at 65 days. One dog had persistent mega-oesophagus and was lost to follow-up 388 days after surgery and one dog is still alive (last follow-up 405 days after surgery). Despite large tumour size and incomplete excision, surgical removal of low-grade leiomyosarcomas can result in long-term resolution of clinical signs.

Panniculitis, polyarthritis and osteomyelitis associated with pancreatic neoplasia in two dogs.

A 12-year-old crossbred dog (case 1) and a 12-year-old Shetland sheepdog (case 2) were presented with a history of lameness and distal limb swelling. Physical examination revealed joint effusions and asymmetrical swellings of the extremities. In case 1, a diagnosis of arthritis and cellulitis was made on fine-needle aspiration biopsy of the synovium and subcutis. In case 2, bone biopsies and synovial aspirates diagnosed osteomyelitis and arthritis. A diagnosis of pancreatic disease was made on the findings of marked elevations of serum lipase concentrations and ultrasonographic identification of pancreatic masses in both cases. Both the cases were non-responsive to symptomatic management and were subsequently euthanased. Postmortem examination confirmed the diagnosis of panniculitis, arthritis and osteomyelitis in both cases. A pancreatic exocrine adenoma was identified in case 1 and a pancreatic adenocarcinoma with widespread metastases in case 2. To the authors' knowledge the association of panniculitis, polyarthritis and osteomyelitis with pancreatic disorders has not been reported previously in canine clinical cases.

Combined clotrimazole irrigation and depot therapy for canine nasal aspergillosis.

OBJECTIVES: To evaluate the effect of short duration 1 per cent clotrimazole flush when combined with 1 per cent clotrimazole cream instilled into the frontal sinuses for the treatment of nasal aspergillosis in 14 dogs. METHODS: Fourteen dogs with clinical, radiological, serological and rhinoscopic findings consistent with nasal aspergillosis were treated by frontal sinus trephination and a short, five-minute flushing of 1 per cent topical clotrimazole solution followed by a 1 per cent clotrimazole cream instilled as a depot agent. RESULTS: Twelve of the 14 dogs (86 per cent) responded well to treatment and either had no clinical signs after treatment or had signs consistent with mild rhinitis during a minimum follow-up period of six months. Only one dog required multiple treatments. Treatment was well tolerated by all patients, with minimal complications. CLINICAL SIGNIFICANCE: This treatment compares favourably to previously published data using one-hour topical clotrimazole or enilconazole flushing treatment protocols. The treatment technique significantly reduced treatment time under anaesthesia.

Acquired idiopathic laryngeal paralysis as a prominent feature of generalised neuromuscular disease in 39 dogs.

A retrospective evaluation of 17 dogs treated surgically for idiopathic acquired laryngeal paralysis demonstrated a marked variability in outcome, with many dogs continuing to exhibit weakness and exercise tolerance. In a subsequent prospective study, 22 consecutive affected dogs were tested for neurological deficits by neurological examination, electrodiagnostic evaluation, and blood analysis to measure thyroxine and thyroid-stimulating hormone and to detect any evidence of myasthenia gravis. Clinical neurological deficits and/or electrodiagnostic abnormalities were found in each case. There was limited evidence that specific neurological deficits were associated with a poor prognosis for full recovery of exercise tolerance. None of the dogs was positive for anti-acetylcholine receptor antibodies, and only three showed evidence of thyroid dysfunction.

Pancreatic mixed acinar-endocrine carcinoma in a dog.

A 10.5-year-old crossbreed dog was presented with a history of hypoglycaemic episodes and elevated serum insulin concentration. A pancreatic mass was removed at surgery along with an enlarged draining lymph node. An unresectable hepatic nodule was also present. Immunohistochemistry confirmed the pancreatic and lymph node masses as functional mixed acinar-endocrine carcinoma, previously unreported in domestic species. Persistent hypoglycaemia and hyperinsulinaemia post-operatively was highly suggestive of the hepatic mass being a functional metastasis. The dog was managed on prednisolone and remained asymptomatic 9 months post-operatively. This tumour type has only been rarely reported in human patients and may highlight the need for more rigorous immunohistochemical staining of pancreatic masses in veterinary species to identify the prevalence of this tumour type.

An immunohistochemical investigation of 18 cases of feline nasal lymphoma.

This report details clinical, histopathological and immunohistochemical findings in 18 cats with chronic nasal disease diagnosed as nasal lymphoma. Eight of the cats were female and 10 were male, with a median age of 10.5 years (range 7-14 years). Three of the cats were Siamese, one was Burmese, and the rest were non-pedigree. The duration of clinical signs before referral ranged from 30 to 540 days (median 88.5 days). The most common clinical signs were nasal discharge, stertor and sneezing. Nasal radiographs were abnormal in 14/16 cases examined. Abnormal masses were detected endoscopically in 13/18 cases. Nine cats received multi-agent chemotherapy or radiation therapy, or both, with survival times ranging from 14 to >541 days. Biopsy material from these 18 cats was examined by light microscopy, and serial sections were subjected to immunohistochemical labelling for the T lymphocyte marker CD3 and the B lymphocyte marker CD79a. In 13 tissues, expression of class II molecules of the major histocompatibility complex and the myelomonocytic antigen MAC387 was also determined. Twelve of the tumours were classified as diffuse large B-cell lymphomas, four as lymphoblastic B-cell lymphomas, and one as a follicular B-cell lymphoma. The tumour cells within these lesions all expressed CD79a, and (where tested) most also expressed MHC class II. One tumour was an anaplastic large cell neoplasm, in which the neoplastic cells expressed MHC class II alone in the absence of either lymphoid marker. There was a variable infiltration of reactive small T lymphocytes into these tumours, and zones of necrosis within the tumour tissue were sometimes heavily infiltrated by MAC387+ phagocytic cells.

Total ear canal ablation in the cat: indications, morbidity and long-term survival.

Fifty-two total ear canal ablation (TECA) procedures in 44 cats were reviewed. The indication for surgery was neoplasia in 41 per cent of the cats, 86 per cent of which had ceruminous gland adenocarcinoma. Chronic inflammatory or polypoid disease accounted for 50 per cent of surgical procedures. Postoperative complications included Horner's syndrome (42 per cent) and facial paralysis (56 per cent) and these were permanent in 14 per cent and 28 per cent of cases, respectively, with the rest resolving in the ensuing weeks or months. The higher incidence of Horner's syndrome and facial paralysis in the cat, compared to the dog, was attributed to greater fragility of the feline tympanic plexus and facial nerve. The median survival time of cats with ceruminous gland adenocarcinoma was 50-3 months, and did not differ significantly from that for inflammatory or polypoid disease. A potential prognostic indicator for this tumour was the mitotic index (MI): cases with MI < or = 2 survived significantly longer than those with MI > or = 3.

Extrahepatic biliary tract surgery in the cat: a case series and review.

Four cases of extrahepatic biliary tract surgery in the cat are described. The causes of the disease were inflammation of the gallbladder, distal common bile duct (CBD) or major duodenal papilla, and traumatic avulsion of the CBD. Bile peritonitis was present in two of the cats. Biliary enterostomy was performed in three cats, two of which were euthanased at five weeks and three months postsurgery; the third was alive at the time of writing, four months postsurgery. Cholecystectomy was curative in one cat. A literature review reveals high early mortality following biliary diversion, with only 50 per cent of cases surviving more than two weeks, and 23 per cent surviving more than six months. Surviving cats had repeated intermittent vomiting and anorexia that responded to antibiotics. No postoperative mortality was seen when biliary diversion was avoided. Whenever biliary enterostomy or temporary diversion methods are performed, a poorer prognosis should be offered due to the increased likelihood of postoperative complications and mortality.

Treatment of urethral sphincter mechanism incompetence in 11 bitches with a sustained-release formulation of phenylpropanolamine hydrochloride.

Between 1995 and 1999, urethral sphincter mechanism incompetence was diagnosed in 11 bitches. They had been treated with phenylpropanolamine hydrochloride at the recommended dose rate, but had shown no response or had become refractory to treatment. They were treated with phenylpropanolamine hydrochloride in a sustained-release formulation combined with diphenylpyraline hydrochloride. The urinary incontinence resolved fully in six of the bitches, two of which remained continent after the treatment was withdrawn; two showed a marked improvement on daily treatment, but the other three bitches failed to respond and underwent colposuspension.