Cross-species oncogenomics offers insight into human muscle-invasive bladder cancer.

BACKGROUND: In humans, muscle-invasive bladder cancer (MIBC) is highly aggressive and associated with a poor prognosis. With a high mutation load and large number of altered genes, strategies to delineate key driver events are necessary. Dogs and cats develop urothelial carcinoma (UC) with histological and clinical similarities to human MIBC. Cattle that graze on bracken fern also develop UC, associated with exposure to the carcinogen ptaquiloside. These species may represent relevant animal models of spontaneous and carcinogen-induced UC that can provide insight into human MIBC. RESULTS: Whole-exome sequencing of domestic canine (n = 87) and feline (n = 23) UC, and comparative analysis with human MIBC reveals a lower mutation rate in animal cases and the absence of APOBEC mutational signatures. A convergence of driver genes (ARID1A, KDM6A, TP53, FAT1, and NRAS) is discovered, along with common focally amplified and deleted genes involved in regulation of the cell cycle and chromatin remodelling. We identify mismatch repair deficiency in a subset of canine and feline UCs with biallelic inactivation of MSH2. Bovine UC (n = 8) is distinctly different; we identify novel mutational signatures which are recapitulated in vitro in human urinary bladder UC cells treated with bracken fern extracts or purified ptaquiloside. CONCLUSION: Canine and feline urinary bladder UC represent relevant models of MIBC in humans, and cross-species analysis can identify evolutionarily conserved driver genes. We characterize mutational signatures in bovine UC associated with bracken fern and ptaquiloside exposure, a human-linked cancer exposure. Our work demonstrates the relevance of cross-species comparative analysis in understanding both human and animal UC.

Tumour Incidence in Dogs in Germany: a Retrospective Analysis of 109,616 Histopathological Diagnoses (2014-2019).

Characterization of a tumour entity is based on the precise histopathological diagnosis taking into account the signalment of the diseased animal. The present study is a comprehensive, up-to-date statistical investigation on the type, frequency and breed distribution of neoplasia in dogs in Germany. The histopathological datasets of 109,616 German canine tissue samples (2014-2019) were processed and statistically examined in retrospect. Non-neoplastic diseases were found in 38,650 samples (35.3%) and 70,966 neoplasms (64.7%) were diagnosed. The most common neoplasms were mammary tumours (21.9%), benign epithelial skin tumours (15.4%), mast cell tumour (9.7%), histiocytoma (7.0%), soft tissue sarcoma (5.8%), lipoma (5.8%), melanocytic tumours (5.2%) and odontogenic tumours (4.7%). In general, Beagles, Magyar Vizslas, Boxers, Schnauzers, Spaniels, French Bulldogs and Golden Retrievers had an increased risk of neoplasia (odds ratio 1.17-1.46; all: P ≤0.001) compared with crossbreed dogs. In particular, Boxers, Golden Retrievers, Rottweilers and Schnauzers were often affected by malignant neoplasms, whereas some breeds (eg, West Highland White Terrier, Magyar Vizsla, Chihuahua, Dachshund and Yorkshire Terrier) were frequently affected by numerous benign tumour types. Despite the known risk of haemangiosarcoma in German Shepherd Dogs, other malignant tumours were rare in this breed. Depending on the type of tumour, some purebred dog breeds can have an increased, reduced or identical risk for certain neoplasms compared with crossbreeds. Discussion of breed predispositions to tumour diseases must therefore be conducted critically and with a view to clinical relevance.

Correlation of Histopathological Changes in the Left Atrium and Left Atrial Appendage with the Degree of Dilation in Cats.

Although atrial dilation is a common finding in feline cardiac disease, detailed investigations of atrial pathology are rare in cats. The aim of the study was to investigate the correlation between pathohistological findings, morphometric data and the degree of dilation of the left atrial appendage (LAA) in 53 cats. Based on the LAA volume, the samples were grouped into normal/control (group 0, ≤1 ml [n = 9]), mildly dilated (group 1, >1 to ≤2 ml [n = 16]), moderately dilated (group 2, >2 to ≤3 ml [n = 14]) and markedly dilated (group 3, >3 ml [n = 14]) groups, independent of the underlying disease. Samples from the LAA and the left atrium (LA) were histologically evaluated using haematoxylin and eosin- and Picrosirius red-stained sections, and morphometrically analysed using an image analysis system. The degree of endo-, myo- and epicardial fibrosis was directly correlated with increased LAA dilation. Due to cardiomyocyte hyperplasia and hypertrophy, the mean thickness of the atrial wall was significantly greater in groups 1 and 2 than in group 0. Conversely, group 3 had a lower mean atrial wall thickness than groups 1 and 2, which was attributed to increased transmural fibrosis and cardiomyocyte atrophy. These findings reflect intensive dynamic remodelling processes during LA and LAA dilation, indicating that reversibility appears to be limited in cases of severe left atrial dilation.

[Chronic bee paralysis virus - quo vadis? Incidence in Bavaria and consideration of therapeutic measures]. / Chronische-Bienenparalyse-Virus ­ quo vadis? Auftreten in Bayern und Betrachtung von Therapiemaßnahmen.

OBJECTIVE: Chronic bee paralysis virus (CBPV), a so far unclassified RNA virus that may cause neurological signs and hairless black syndrome in honey bees, has become increasingly prevalent in various European countries in recent years. The disease occasionally leads to immense losses of worker bees and a significant drop in performance, especially in strong colonies. The aim of this retrospective study was to analyse the spread of CBPV in Bavaria from 2018 to 2020 as well as to evaluate therapeutic measures. MATERIAL AND METHODS: In part 1 of the study, analysis results of the Tiergesundheitsdienst Bayern e. V. (Bavarian Animal Health Service) from 302 bee colonies were examined with regard to virus diagnostics and clinical signs with a focus on CBPV. In part 2, data collected with the help of a questionnaire regarding 105 CBPV-positive and symptomatic colonies were evaluated. RESULTS: In part 1, a significant (p = 0.004) increase in CBPV-positive samples from 2018 to 2020 was detected within Bavaria. In addition, the number of cases with clinical signs also increased during the last 2 years. Part 2 of the study showed a clustering of reports of initial onset of CBPV symptoms during springtime. The colonies mostly with a combination of symptoms recovered from the disease in 57 % of the cases. Therapeutic intervention was one decisive factor. A large number of different combinations of measures led to survival, and a lack of intervention mostly resulted in the death of the colony. In 62 % of the diseased and isolated colonies, isolation prevented further spread in the apiary. CONCLUSION UND CLINICAL RELEVANCE: Based on the results, CBPV occurred more frequently in connection with clinical signs in Bavaria in recent years. Due to the large number of applied combinations of measures, no evidence-based therapy recommendation may be derived based on the current evaluations. However, it has been shown that any therapeutic intervention is superior to withholding treatment. For precise therapy recommendations, trials with treatment under standard conditions are necessary.

Digital Lesions in Dogs: A Statistical Breed Analysis of 2912 Cases.

Breed predispositions to canine digital neoplasms are well known. However, there is currently no statistical analysis identifying the least affected breeds. To this end, 2912 canine amputated digits submitted from 2014-2019 to the Laboklin GmbH & Co. KG for routine diagnostics were statistically analyzed. The study population consisted of 155 different breeds (most common: 634 Mongrels, 411 Schnauzers, 197 Labrador Retrievers, 93 Golden Retrievers). Non-neoplastic processes were present in 1246 (43%), tumor-like lesions in 138 (5%), and neoplasms in 1528 cases (52%). Benign tumors (n = 335) were characterized by 217 subungual keratoacanthomas, 36 histiocytomas, 35 plasmacytomas, 16 papillomas, 12 melanocytomas, 9 sebaceous gland tumors, 6 lipomas, and 4 bone tumors. Malignant neoplasms (n = 1193) included 758 squamous cell carcinomas (SCC), 196 malignant melanomas (MM), 76 soft tissue sarcomas, 52 mast cell tumors, 37 non-specified sarcomas, 29 anaplastic neoplasms, 24 carcinomas, 20 bone tumors, and 1 histiocytic sarcoma. Predisposed breeds for SCC included the Schnauzer (log OR = 2.61), Briard (log OR = 1.78), Rottweiler (log OR = 1.54), Poodle (log OR = 1.40), and Dachshund (log OR = 1.30). Jack Russell Terriers (log OR = -2.95) were significantly less affected by SCC than Mongrels. Acral MM were significantly more frequent in Rottweilers (log OR = 1.88) and Labrador Retrievers (log OR = 1.09). In contrast, Dachshunds (log OR = -2.17), Jack Russell Terriers (log OR = -1.88), and Rhodesian Ridgebacks (log OR = -1.88) were rarely affected. This contrasted with the well-known predisposition of Dachshunds and Rhodesian Ridgebacks to oral and cutaneous melanocytic neoplasms. Further studies are needed to explain the underlying reasons for breed predisposition or "resistance" to the development of specific acral tumors and/or other sites.

Signalment and Clinical Data of Cats with Exocrine Pancreatic Insufficiency Diagnosed Using Feline Trypsin-like Immunoreactivity in Routine Diagnostics.

Serum feline trypsin-like immunoreactivity (fTLI) is commonly used to diagnose feline exocrine pancreatic insufficiency (EPI). This study aimed to describe signalment and clinical data of cats with EPI. Determination of TLI was performed using an in-house ELISA; the reference interval was defined using a Reference Limit Estimator. Groups were formed from 4813 cats (2019-2020), based on their fTLI concentration: 1 (<8 µg/L; decreased; n = 275), 2 (8-88 µg/L; reference interval; n = 4256), and 3 (>88 µg/L; increased; n = 282). Males and Domestic Shorthairs were most common in all groups. Group 3 had the highest (13 years), and group 1 had the lowest (9 years), median age. Clinical information was available for 200 cats (decreased fTLI: n = 87, lower reference interval (8-12 µg/L): n = 113). Treatment response was observed in 83% (decreased fTLI) and 66% (lower reference interval). EPI cats displayed weight loss (69%), diarrhoea (68%), vomiting (41%), anorexia (39%), poor hair coat (35%), lethargy (33%), and/or polyphagia (21%). The lower the serum fTLI concentration, the more often good treatment response was reported (p = 0.022) but there were no statistically significant clinical signs. In conclusion, fTLI is a helpful parameter to diagnose EPI but predicting treatment response based on signalment or clinical signs is not possible.

Histopathological findings and canine pancreatic lipase immunoreactivity in normal dogs and dogs with inflammatory and neoplastic diseases of the pancreas.

BACKGROUND: Diagnosis of pancreatic diseases in dogs is still challenging because of variable clinical signs, which do not always correspond with clinical pathology and histopathological findings. OBJECTIVES: To characterize inflammatory and neoplastic pancreatic diseases of dogs and to correlate these findings with clinical findings and canine pancreatic lipase immunoreactivity (cPLI) results. ANIMALS: Tissue specimens and corresponding blood samples from 72 dogs submitted for routine diagnostic testing. METHODS: Four groups were defined histologically: (1) normal pancreas (n = 40), (2) mild pancreatitis (n = 8), (3) moderate or severe pancreatitis (acute, n = 11; chronic, n = 1), and (4) pancreatic neoplasms (n = 12). An in-house cPLI ELISA (<180 µg/L, normal; >310 µg/L, pancreatitis) was performed. RESULTS: In dogs with normal pancreas, 92.5% of serum cPLI results were within the reference range and significantly lower than in dogs with mild acute pancreatitis, moderate or severe acute pancreatitis and pancreatic tumors. In dogs with moderate or severe acute pancreatitis, cPLI sensitivity was 90.9% (95% confidence interval [CI], 58.7%-99.8%). Most dogs (9/12) with pancreatic tumors (group 4) had additional pancreatic inflammation and cPLI results were increased in 10 dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: High cPLI indicates serious acute pancreatitis but underlying pancreatic neoplasms should also be taken into consideration. This study confirms the relevance of histopathology in the diagnostic evaluation of pancreatic diseases.

Evaluating the Histologic Grade of Digital Squamous Cell Carcinomas in Dogs with Dark and Light Haircoat-A Comparative Study of the Invasive Front and Tumor Cell Budding Systems.

Canine digital squamous cell carcinomas (CDSCC) are particularly aggressive when compared to their occurrence in other locations. Although these neoplasms are more frequently seen in dark-haired dogs, such as Giant Schnauzers, there are no data checking whether these tumors are histologically different between breeds. We histologically evaluated DSCC from 94 dogs. These were divided into two groups, namely, (1) dark-haired (N = 76) and (2) light-haired breeds (N = 18), further subdividing Group 1 into three subgroups, (1a) black breeds (n = 11), (1b) Schnauzers (n = 34) and (1c) black & tan breeds (n = 31). Adaptations from two different squamous cell carcinomas grading schemes from human and veterinary literature were used. Both systems showed significant differences when compared to Groups 1 and 2 in terms of final grade, invasive front keratinization, degree of invasion, nuclear pleomorphism, tumor cell budding, smallest tumor nest size and amount of tumor stroma. Group 2 was consistently better differentiated CDSCC than Group 1. However, there were no significant differences among the dark-haired breeds in any of the features evaluated. This study represents the first attempt to grade CDSCC while taking into account both phenotypical and presumptive genotypical haircoat color. In conclusion, CDSCC are not only more common in dark-haired dogs, they are also histologically more aggressive.

[Detection of BRAF mutation in canine prostatic diseases]. / Nachweis der BRAF-Mutation bei kaninen Prostataerkrankungen.

OBJECTIVE: In the literature, the BRAF mutation is reported to have been identified in 80 % of the examined canine prostate carcinomas (PCa). The objectives of this study were to test for the BRAF mutation in canine PCa in our cohort of canine patients, to determine the specificity and sensitivity of the test for this mutation, as well as to identify the association between the presence of the BRAF mutation and the histologic picture of PCa. Moreover, the method was to be established in cytology samples. MATERIAL AND METHODS: Biopsy samples (n = 70) and cytologic slides (n = 17) of 87 dogs with prostatic diseases were selected. Prostatic diseases were classified according to the literature as benign prostate hyperplasia (BPH, n = 22), prostatitis (n = 14), squamous cell metaplasia of the prostate (PM, n = 2), atrophy following castration (n = 3) und PCa (n = 46; histologic diagnosis n = 35, cytologic diagnosis n = 11). Additionally, the Gleason score was determined for each PCa. DNA isolation was performed using commercially available kits. Exon 15 was examined using the TaqMan® SNP assay. The specificity and sensitivity of the test were calculated. RESULTS: A Gleason score of 6 and 7 was shown in 1 PCa each, in 33 cases the score ranged between 8 and 10. Sufficient amount of good-quality DNA was isolated from all samples. 28/46 PCa were tested positive for the BRAF mutation (sensitivity 61 %). The BRAF mutation was not evident in any of the dogs with BPH, prostatitis, PM or atrophy (specificity 100 %). PCa positive for the BRAF mutation exhibited a significantly higher Gleason score (p = 0.002) in comparison to PCa without this mutation. CONCLUSION AND CLINICAL RELEVANCE: BRAF mutation analysis is a highly specific method and may aid in confirming the diagnosis of PCa in histologically and cytologically questionable cases. PCa positive for BRAF mutation exhibited more criteria of malignancy than PCa without this mutation. The clinical, therapeutic, and prognostic relevance of these findings needs to be evaluated by further studies.

Correlation of BRAF Variant V595E, Breed, Histological Grade and Cyclooxygenase-2 Expression in Canine Transitional Cell Carcinomas.

The presence of BRAF variant V595E, as well as an increased cyclooxygenase-2 (COX-2) expression in canine transitional cell carcinoma (TCC) are well-described in the literature. The aim of the present study was to investigate the correlation between breed (terrier versus non-terrier dogs), histological grade, COX-2 expression, and BRAF mutation in canine TCC. Therefore, transmural TCC biopsies from 65 dogs (15 terriers, 50 non-terriers) were graded histologically into low- and high-grade. Immunohistochemical evaluation of the intensity of COX-2 expression was performed using an immunoreactive score (IRS). Exon 15 of chromosome 16 was examined for the BRAF variant c.1799T>A by TaqMan® SNP assay. TCC was low-grade in 20 cases (one terrier, 19 non-terriers) and high-grade in 45 cases (14 terriers, 31 non-terriers). Contrary to humans, histological grade was not significantly correlated to the intensity of COX-2 expression. BRAF mutation was detected in 11/15 (73%) TCC of terriers and in 18/50 (36%) TCC of non-terriers. Histological grade and BRAF mutation were not correlated significantly (p = 0.2912). Terriers had a considerably higher prevalence of high-grade tumors (p < 0.0001), as well as of BRAF mutation (p ≤ 0.05) compared to non-terriers. In non-terriers, neoplasms with BRAF mutation showed a significantly higher intensity of COX-2 expression than those without BRAF mutation (p ≤ 0.05). In conclusion, in contrast to humans, testing for BRAF mutation in canine TCC is a sensitive diagnostic method especially in terriers (73%) and may be recommended as a screening test. However, evidence of BRAF mutation in canine TCC is not a predictor for the histological grade. Moreover, a positive correlation between histological grade and the intensity of COX-2 expression was not found. Further studies are necessary to clarify the clinical and prognostic relevance of the elevated intensity of COX-2 expression of TCC with BRAF mutation detected in non-terriers.

[Diagnostic value of the BRAF variant V595E in urine samples, smears and biopsies from canine transitional cell carcinoma]. / Diagnostische Aussagekraft der BRAF-Mutation V595E in Urinproben, Ausstrichen und Bioptaten beim kaninen Übergangszellkarzinom.

OBJECTIVE: Transitional cell carcinoma (TCC) is the most common malignant tumour of the canine urinary tract. Previously, the mutation of the BRAF gene V595E was identified in approximately 85 % of canine TCC cases by DNA sequencing of TCC tumour cells, both in frozen and paraffin-embedded tissue sections, as well as in urine. The objective of this study was to establish these methods in cytological smears and to investigate the prevalence of BRAF mutation V595E in canine TCC in our cohort of patients. MATERIAL AND METHODS: Biopsy samples (n = 43), urine (n = 48) and/or cytological smears (n = 31) from 66 dogs with TCC (n = 33), urinary bladder polyps (n = 7), cystitis (n = 23) or without bladder diseases (n = 3), submitted for routine diagnostics, were selected. DNA isolation from paraffin material, urine and cytological smears was performed using commercially available kits. Exon 15 was examined for the presence of the BRAF mutation c.1784T>A by Sanger sequencing. RESULTS: In 39/43 paraffin-embedded biopsies and 38/48 urine samples, a sufficient amount of good quality DNA was isolated. DNA isolation and sequencing were successful in 16/18 smears with a high cell count, but not in the 10/13 smears with low cellularity. In all cases from which different sample materials were available, the results of BRAF analysis were identical in paraffin-embedded tissue, cytological smears and/or urine. In 22/31 dogs (70.9 %) with TCC, the presence of the BRAF mutation was confirmed, whereas it could not be detected in animals without pathological findings or with cystitis or with a polyp. CONCLUSION AND CLINICAL RELEVANCE: BRAF mutation analysis is a new and good method to be able to mostly confirm a diagnosis of TCC in uncertain cases. Non-invasive diagnostic samples, including urine and urine sediment containing sufficient numbers of relevant cells as well as cytology aspirates and formalin-fixed biopsies can be used for analysis. However, it is important to note that only a positive identification of the mutation is diagnostic. Further research is necessary to investigate prognostic and therapeutic relevance of the variant and how this genetic analysis can be used as an early detection method for TCC.