This is a case of medulloepithelioma in a 14-year-old mixed breed gelding horse, presenting a large abnormal mass of tissue involving the entire extension of the right eye. Ophthalmic examination showed deformation and swelling of the eye. The animal showed signs of pain on palpation of the organ, but the specific examination of the systems did not reveal any other changes. Due to the extension of the apparently neoplastic mass and the discomfort experienced by the animal, transpalpebral enucleation procedure was decided. The excised tissue was sent for histopathological analysis, wherein a neoplastic proliferation of neuroectodermal cells was noted. The neoplasm was poorly delimited, unencapsulated, infiltrative, sustained by moderate fibrovascular stroma, and formed cords and rosettes with cells arranged in palisades around the luminal structures (Flexner-Wintersteiner rosettes), suggestive of medulloepithelioma. The immunohistochemical profile was also performed, confirming the diagnosis. The neoplastic cells were immunolabeled to vimentin, S100 protein (S100), and specific neuro enolase (NSE), but not for pan cytokeratin (AE1AE3), glial fibrillary acidic protein (GFAP), and cytokeratin 8/18 (CK8/18). Five months postoperatively, the animal was healthy, without any relapse or evidence of metastasis.
RESUMOAvaliou-se o congelamento do plasma rico em plaquetas (PRP) de equinos, a -196ºC em nitrogênio líquido, utilizando-se como crioprotetor o DMSO em duas concentrações (3% e 6%), e, como ponto final, a avaliação da morfologia e da agregometria plaquetária. Foram utilizadas 12 amostras de PRP em duas repetições. Previamente ao congelamento, as amostras foram submetidas a um resfriamento lento (-0,07ºC/minuto) até a temperatura final de 4-5ºC. A criopreservação do PRP equino, incluindo um resfriamento lento a 4-5ºC, previamente ao congelamento a -197ºC em nitrogênio líquido, foi similar para as concentrações do crioprotetor DMSO a 3% ou 6%, quando avaliado o percentual de ativação e de agregação plaquetária. . Palavras-chave: equino, plasma rico em plaquetas (PRP), criopreservação, DMSO
ABSTRACT
Equine platelet-rich plasma (PRP) frozen at -196°C in liquid nitrogen
Platelet-rich plasma (PRP) has been proposed as an agent to accelerate the healing process and stimulate the regenerative capacity of tissues due to its abundance of growth factors. A large variety of kits and protocols are available to obtain PRP by different cell-separation systems. However, the lack of standardization may lead to inconsistent results. The aim of this study was to characterize cellular composition, platelet parameters using the ADVIA 120 flow cytometer, and TGF-β1 concentration from the PRP product obtained through a closed system, using simple centrifugation. Six clinically healthy horses were used in this study. The protocol in the closed system resulted in approximately 1.6-fold higher platelet and approximately 2.0-fold lower white blood cell concentrations in comparison with whole blood values. The evaluated system was efficient in concentrating platelets and in retrieving a small number of leukocytes, using a protocol of single centrifugation at low speed.
The aim of this study was to standardize a simple, manual platelet-rich plasma (PRP) protocol in Catalonian donkeys using single-spin tube centrifugation as a treatment for jenny endometritis. The objective was to obtain a blood product with a moderate concentration of platelets (2 or 3 times baseline physiologic values) and a low WBC (White Blood Cells) concentration. Blood was drawn from six Catalonian donkeys using acid citrate dextrose (ACD) as an anticoagulant, and then processed by single centrifugation at 133× g for two different centrifugation times (10 and 15 min). The PRP samples were evaluated by flow cytometry, and TGF-β1 (Transforming Growth Factor-Beta1) concentrations were determined by enzyme-linked immunosorbent assay (ELISA). The 10 min centrifugation protocol resulted in a slightly greater release of TGF-β1 (6044.79 ng/mL), a 2.06-fold increase in platelet concentration, and a 15-fold reduction in leukocyte concentration when compared to the initial values. The 15 min centrifugation time resulted in a 2.44-fold increase in baseline platelet concentration, a reduction in WBC count by a factor of 20, and slightly lower TGF levels (5206 ng/mL). We conclude that both protocols are adequate for the obtention of PRP, and both may have an acceptable therapeutic potential for use in this species, although this needs to be further validated.
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