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EPITHELIAL-MESENCHYMAL TRANSITION IN CANINE MAMMARY TUMOURS: THE ROLE OF MYOEPITHELIAL CELLS G. Beha * , G. Sarli * , B. Brunetti * , F. Sassi y , D. Ferrara * , F. Morandi * and C. Benazzi * * University of Bologna and y Institute for Cancer Research and Treatment, Torino, Italy Introduction: In malignant epithelial tumours the epithelial-mesen- chymal transition (EMT) phenomenon is reported. In the mammary gland, two cell types are present: luminal epithelial and basal myoe- pithelial cells, the latter hypothesized as being precursors of mesen- chymal tissue. A panel of immunohistochemical markers of myoepithelial cells was applied to four different types of neoplasms with the aim of testing the role of myoepithelial cells in EMT in mam- mary tumours of the dog. Materials and Methods: Monoclonal and polyclonal antibodies specific for ER, p63, vimentin (VIM) and a-smooth muscle actin (a-SMA) were used on mammary gland neoplasms (five benign and three malignant myoepitheliomas, eight carcinomas in mixed tu- mours and nine complex carcinomas) and an algorithm was built to characterize the resting and the motile phenotype of myoepithelial cells. Results: ER labelled some basal (resting) and stellate (motile) myoe- pithelial cells. Myoepithelial markers, namely p63, VIM and a-SMA, labelled basal myoepithelial cells. Stellate myoepithelium lost the la- belling with p63, but maintained positivity for VIM and a-SMA, which are also typical of EMT. Conclusions: This affinity, together with the increased transition of myoepithelium from resting basal to motile stellate cells, may be in- dicative of a transition from polarized immotile myoepithelial cells into highly migratory fibroblast-like cells. INTRAOCULAR PRIMITIVE NEUROECTODERMAL TUMOURS IN TWO HORSES K. Brandes * and J.P. Teifke y * Animal Pathology Augsburg and y Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany Introduction: Intraocular tumours of neuroepithelial origin are rare in horses. In the present study morphological and immunohistochem- ical features of two primary intraocular neoplasms of neuroectoder- mal origin are presented. Materials and Methods: Case 1: the posterior eye segment of a 1.5-year-old male warmblood horse with a white mass (5 3 2 cm) lifting the retina and occupying 40% of the vitreous body. Case 2: a 4-year-old male Icelandic horse with a 3.5 2.5 2 cm white neoplasm expanding the ciliary body and growing by expan- sion into the vitreous and infiltrating the iris. Results: Microscopically, the tumour from case 1 consisted of densely arranged polygonal cells forming few rosette-like structures with subretinal and intravitreal spreading. The tumour from case 2 was composed of polygonal cells forming numerous rosette-like struc- tures and a spindle cell component embedded in a myxomatous ma- trix. Several immunohistochemical markers (vimentin, GFAP, S-100 and neurofilament) were applied. Conclusions: On the basis of the histopathological features a medul- loepithelioma was diagnosed in case 1 and a teratoid medulloepithe- lioma in case 2. EVALUATION OF MALIGNANCY OF CANINE MAST CELL TUMOUR USING COMPUTER-AIDED MORPHOMETRY S. Curtseit, E. C. Popa, G. Dinescu, A. Feghiu, L. Tudor and M. Militaru University of Agronomic Science and Veterinary Medicine, Bucharest, Romania Introduction: Mast cell tumours (MCTs) are potentially malignant neoplastic processes and their histopathological grading often proves to be subjective. Consequently, the morphometry of these tumours was analyzed by computer in order to grade and predict the behav- iour of MCTs. Materials and Methods: Eighteen canine cutaneous MCTs were analysed morphometrically with regard to mean cellular area and pe- rimeter, mean nuclear area and perimeter, nucleus/cytoplasm ratio and regularity factor using May Grunwald Giemsa-stained cytopa- thology smears. Lesions were graded as I, II or III according to the cellular morphology and degree of degranulation. The smears were analysed with an Olympus BX41 microscope coupled to a com- puter equipped with Cell B analysis system. Results: There were significant differences between the cellular area of grade I and II tumours and between grade I and III tumours. There were no significant differences in cellular perimeter between grades. Measurements of the nuclear perimeter increased with an in- crease in grade of the MCT. The nucleus/cytoplasm ratio had higher values in grade III MCTs than in grade II tumours. There were in- significant differences between the values of regularity factor. Conclusions: The study adds new data to cellular morphometry of MCT, correlating the values of selected parameters with the grade of the tumour. In combination with the rapid and cheap cytopathology technique, cellular morphometry becomes a useful tool for evaluation of tumour malignancy. TRACHEAL NEUROENDOCRINE CARCINOMA IN A CAT B. Osi nska and I. Dolka Warsaw University of Life Sciences-SGGW, Warsaw, Poland Introduction: Neuroendocrine carcinoma is rare in the larynx and extremely rare in the trachea. Human patients with laryngeal neuro- endocrine carcinoma are mainly elderly male smokers. Materials and Methods: A 7-year-old, male, Main Coon cat pre- sented with intensifying dyspnoea of 6-months duration. Tracheo- scopy showed a deformation of the tracheal mucous membrane. The segment of the trachea with the deformation was removed surgi- cally and the specimen was fixed in 10% neutral buffered formalde- hyde and embedded in paraffin wax, sectioned and stained with HE, periodic acideSchiff (PAS) and the Grimelius silver method. Immunohistochemistry was performed with monoclonal anti-human cytokeratin (clone MNF116), monoclonal anti-human vimentin (clone 3B4), polyclonal rabbit anti-human chromogranin A and poly- clonal rabbit anti-human calcitonin. Results: Microscopical investigation showed round and fusiform cells with hyperchromatic nuclei, arranged in nests, sheets or gland- like patterns. Immunohistochemical labelling for chromogranin A and cytokeratin was positive in some tumour cells and calcitonin ex- pression was not observed. Some vimentin-positive and argyrophilic cells were observed. Conclusions: The tumour was recognized as a neuroendocrine carcinoma. 80 ESVP/ECVP Proceedings 2011 146:1, 2012

Intraocular Primitive Neuroectodermal Tumours in Two Horses

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80 ESVP/ECVP Proceedings 2011 146:1, 2012

EPITHELIAL-MESENCHYMAL TRANSITION IN CANINEMAMMARY TUMOURS: THE ROLE OF MYOEPITHELIAL

CELLSG. Beha*, G. Sarli*, B. Brunetti*, F. Sassi y, D. Ferrara*,

F. Morandi* and C. Benazzi**University of Bologna and yInstitute for Cancer Research and Treatment,

Torino, ItalyIntroduction: In malignant epithelial tumours the epithelial-mesen-chymal transition (EMT) phenomenon is reported. In the mammarygland, two cell types are present: luminal epithelial and basal myoe-pithelial cells, the latter hypothesized as being precursors of mesen-chymal tissue. A panel of immunohistochemical markers ofmyoepithelial cells was applied to four different types of neoplasmswith the aim of testing the role of myoepithelial cells in EMT in mam-mary tumours of the dog.Materials and Methods: Monoclonal and polyclonal antibodiesspecific for ER, p63, vimentin (VIM) and a-smooth muscle actin(a-SMA) were used on mammary gland neoplasms (five benignand three malignant myoepitheliomas, eight carcinomas in mixed tu-mours and nine complex carcinomas) and an algorithm was built tocharacterize the resting and the motile phenotype of myoepithelialcells.Results: ER labelled some basal (resting) and stellate (motile) myoe-pithelial cells. Myoepithelial markers, namely p63, VIM and a-SMA,labelled basal myoepithelial cells. Stellate myoepithelium lost the la-belling with p63, but maintained positivity for VIM and a-SMA,which are also typical of EMT.Conclusions: This affinity, together with the increased transition ofmyoepithelium from resting basal to motile stellate cells, may be in-dicative of a transition from polarized immotile myoepithelial cellsinto highly migratory fibroblast-like cells.

INTRAOCULAR PRIMITIVE NEUROECTODERMALTUMOURS IN TWO HORSESK. Brandes* and J.P. Teifkey

*Animal Pathology Augsburg and yFederal Research Institute for Animal

Health, Greifswald-Insel Riems, GermanyIntroduction: Intraocular tumours of neuroepithelial origin are rarein horses. In the present study morphological and immunohistochem-ical features of two primary intraocular neoplasms of neuroectoder-mal origin are presented.Materials and Methods: Case 1: the posterior eye segment ofa 1.5-year-old male warmblood horse with a white mass (5� 3 �2 cm) lifting the retina and occupying 40% of the vitreous body.Case 2: a 4-year-old male Icelandic horse with a 3.5� 2.5� 2 cmwhite neoplasm expanding the ciliary body and growing by expan-sion into the vitreous and infiltrating the iris.Results: Microscopically, the tumour from case 1 consisted ofdensely arranged polygonal cells forming few rosette-like structureswith subretinal and intravitreal spreading. The tumour from case 2was composed of polygonal cells forming numerous rosette-like struc-tures and a spindle cell component embedded in a myxomatous ma-trix. Several immunohistochemical markers (vimentin, GFAP, S-100and neurofilament) were applied.Conclusions: On the basis of the histopathological features a medul-loepithelioma was diagnosed in case 1 and a teratoid medulloepithe-lioma in case 2.

EVALUATION OF MALIGNANCY OF CANINE MAST CELLTUMOUR USING COMPUTER-AIDED MORPHOMETRY

S. Curtseit, E. C. Popa, G. Dinescu, A. Feghiu,L. Tudor and M. Militaru

University of Agronomic Science and Veterinary Medicine, Bucharest, RomaniaIntroduction: Mast cell tumours (MCTs) are potentially malignantneoplastic processes and their histopathological grading often provesto be subjective. Consequently, the morphometry of these tumourswas analyzed by computer in order to grade and predict the behav-iour of MCTs.Materials and Methods: Eighteen canine cutaneous MCTs wereanalysed morphometrically with regard to mean cellular area and pe-rimeter, mean nuclear area and perimeter, nucleus/cytoplasm ratioand regularity factor using May Gr€unwald Giemsa-stained cytopa-thology smears. Lesions were graded as I, II or III according tothe cellular morphology and degree of degranulation. The smearswere analysed with an Olympus BX41 microscope coupled to a com-puter equipped with Cell B analysis system.Results: There were significant differences between the cellular areaof grade I and II tumours and between grade I and III tumours.There were no significant differences in cellular perimeter betweengrades. Measurements of the nuclear perimeter increased with an in-crease in grade of the MCT. The nucleus/cytoplasm ratio had highervalues in grade III MCTs than in grade II tumours. There were in-significant differences between the values of regularity factor.Conclusions: The study adds new data to cellular morphometry ofMCT, correlating the values of selected parameters with the grade ofthe tumour. In combination with the rapid and cheap cytopathologytechnique, cellular morphometry becomes a useful tool for evaluationof tumour malignancy.

TRACHEAL NEUROENDOCRINE CARCINOMA IN A CATB. Osi�nska and I. Dolka

Warsaw University of Life Sciences-SGGW, Warsaw, PolandIntroduction: Neuroendocrine carcinoma is rare in the larynx andextremely rare in the trachea. Human patients with laryngeal neuro-endocrine carcinoma are mainly elderly male smokers.Materials and Methods: A 7-year-old, male, Main Coon cat pre-sented with intensifying dyspnoea of 6-months duration. Tracheo-scopy showed a deformation of the tracheal mucous membrane.The segment of the trachea with the deformation was removed surgi-cally and the specimen was fixed in 10% neutral buffered formalde-hyde and embedded in paraffin wax, sectioned and stained withHE, periodic acideSchiff (PAS) and the Grimelius silver method.Immunohistochemistry was performed with monoclonal anti-humancytokeratin (clone MNF116), monoclonal anti-human vimentin(clone 3B4), polyclonal rabbit anti-human chromogranin A and poly-clonal rabbit anti-human calcitonin.Results: Microscopical investigation showed round and fusiformcells with hyperchromatic nuclei, arranged in nests, sheets or gland-like patterns. Immunohistochemical labelling for chromogranin Aand cytokeratin was positive in some tumour cells and calcitonin ex-pression was not observed. Some vimentin-positive and argyrophiliccells were observed.Conclusions: The tumour was recognized as a neuroendocrinecarcinoma.