Immunostaining and gene expression of epidermal growth factor receptors (HER1/HER2) in canine cutaneous squamous cell carcinoma.

Cutaneous squamous cell carcinoma (cSCC) is a neoplasm type often diagnosed in dogs. However, studies focused on further investigating its molecular biology, mainly biomarkers to help implementing new therapies, remain scare in the literature. Thus, immunostaining and the gene expression of epidermal growth factor receptors (HER1 and HER2) in canine cSCC presenting different cell differentiation degrees were herein assessed. Thirty-two (32) canine cSCC were selected, classified based on to their cell differentiation degree and subjected to immunohistochemical study to assess HER1 and HER2 immunostaining intensity and distribution. In addition, HER1 and HER2 gene expression was investigated through real-time PCR. Membranous and cytoplasmic immunostaining were observed in both markers. HER2 prevailed in poorly differentiated cSCC; there was positive protein expression correlation between both markers. Mean HER1 gene expression was higher in moderately differentiated, whereas mean HER2 gene expression was higher in poorly differentiated cSCC. Moreover, there was gene expression correlation between markers, regardless of cell differentiation degree. Thus, HER2 protein immunostaining and gene expression were higher in poorly differentiated canine cSCC and it enabled understanding that increase observed in this epidermal growth factor receptor is proportional to this neoplasm's cell differentiation degree in canine species. Results in the current study helped better understanding canine cSCC's molecular biology; however, it is relevant studying other markers aiming to investigate signaling pathways.

Sensitivity of PCR in conjunctival swab samples for the diagnosis of canine visceral leishmaniasis: a systematic review and meta-analysis.

To compare the sensitivity of conjunctival swab (CS) and conventional samples (blood, spleen, liver, lymphoid and cutaneous tissue) in the diagnosis of canine visceral leishmaniasis (CVL) by polymerase chain reaction (PCR), a systematic review and meta-analysis was carried out using PubMed, Science Direct, Scopus, Web of Science, VHL/BVS (Virtual Health Library), CAPES, and Scielo databases. Articles published from 2002 to 2022 were considered and the review was updated in Jul 2023. From the total of 371 identified studies, 8 met all the eligibility criteria and were included in this review. Data from 658 CVL-positive dogs and 2541 PCR results were considered. Using a random effect model, data on the sensitivity of the test was compared between intervention (CS samples) and comparison (all the other samples) groups. Overall, the use of CS in the PCR diagnosis of CVL produced 12% higher sensitivity (p=0.013) in the test than all the other samples in combination. The animals' clinical condition did not influence (p>0.142) this overall result. However, when CS was individually compared to each of the conventional samples, the consistent result was observed (p=0.012) only in the CS versus bone marrow comparison. Given their rapid acquisition, minimal invasiveness, and lower cost relative to conventional samples, CS samples present a promising alternative for the molecular diagnosis of CVL.

Macroscopic and histological aspects of the pharynx and larynx of the giant anteater (Myrmecophaga tridactyla Linnaeus, 1758).

Giant anteater (Myrmecophaga tridactyla) is an endangered species that resides in much of Latin America, but it has been losing its habitat, especially in the Cerrado biome, where it constantly suffers traumas resulting from fires and roadkill. The anatomical knowledge of structures of the respiratory system is important for a better morphophysiological understanding of the species. Thus, this study aimed to perform the macroscopic and histomorphological description of the pharynx and larynx of the giant anteater. Twelve adult giant anteaters were used, three of them fixed in buffered formalin for further dissection and pharynx and larynx macroscopic analysis of structures. From the other animals, samples of the pharynx and larynx were collected and prepared for histological evaluation under optical microscope. Macroscopically, their pharynx and soft palate are extensive, and the anatomical location of these structures and the larynx differs greatly from that described in other species. The larynx, although more caudal, was similar to that of other animals. Histologically, the epithelium of these regions varied between the pseudostratified ciliated columnar and the non-keratinized stratified squamous epithelium. Laryngeal cartilages were composed of elastic (epiglotti) and hyaline cartilages (arytenoid, cricoid and thyroid cartilage), with an ossification process and glandular clusters around the hyaline cartilage. The distinct anatomical location of the pharynx and larynx of Myrmecophaga tridactyla is the main macroscopic finding of this study, besides the length of the pharynx and soft palate of these animals.

Comparative Pathobiology of Canine and Human Prostate Cancer: State of the Art and Future Directions.

First described in 1817, prostate cancer is considered a complex neoplastic entity, and one of the main causes of death in men in the western world. In dogs, prostatic carcinoma (PC) exhibits undifferentiated morphology with different phenotypes, is hormonally independent of aggressive character, and has high rates of metastasis to different organs. Although in humans, the risk factors for tumor development are known, in dogs, this scenario is still unclear, especially regarding castration. Therefore, with the advent of molecular biology, studies were and are carried out with the aim of identifying the main molecular mechanisms and signaling pathways involved in the carcinogenesis and progression of canine PC, aiming to identify potential biomarkers for diagnosis, prognosis, and targeted treatment. However, there are extensive gaps to be filled, especially when considering the dog as experimental model for the study of this neoplasm in humans. Thus, due to the complexity of the subject, the objective of this review is to present the main pathobiological aspects of canine PC from a comparative point of view to the same neoplasm in the human species, addressing the historical context and current understanding in the scientific field.

Fibroma cutâneo atípico em uma vaca / Atypical cutaneous fibroma in a cow

Background: Cutaneous fibroma is a benign neoplasm affecting the fibroblasts and collagen matrix that develops in the dermis or subcutaneous tissue. This neoplasm is uncommon in cattle, and few reports have described the treatment and resolution of this neoplasm. Despite its benign character, a veterinarian should consider cutaneous fibroma in the differential diagnosis of skin tumors. This report aims to describe a rare case of large fibroma in the scapular region in a cow, with emphasis on the clinical-surgical and anatomopathological aspects of the condition. Case: A 3-year-old Girolando 3/4 cow was attended to at a rural property in Lagamar-MG, Brazil. According to the owner, the animal presented with a small mass in the right scapular region that grew progressively over 1 year and 6 months. Clinical examination revealed an exuberant and painless increase in volume on palpation in the proximal region of the right thoracic limb, which, in its vertical axis, extended from the proximal end of the scapula to near the olecranon tuberosity, and, in its horizontal axis, extended from the 6th intercostal space to the scapulohumeral joint, reaching the dimensions 66 cm and 62 cm, respectively. It presented with multiple nodules that were firm in consistency with extensive areas of ulceration. Neoplasia was suspected, and surgical excision was decided upon. The cow was sedated and restrained in the left lateral decubitus position. Trichotomy and antisepsis of the operative field were performed followed by an infiltrative anesthetic block around the tumor. The tumor was excised maintaining a safety margin of 1 cm. Dermorrhaphy was not possible, and healing by secondary intention was awaited. In the postoperative period, antibiotic therapy with benzathine penicillin, analgesia with meloxicamand dipyrone and daily dressing of the wound were performed. There were no postoperative complications and complete healing occurred approximately 100 days after surgery. One year after the surgical procedure, the owner reported that the cow did not present with recurrence of the neoplasm. The resected tumor weighed 11.2 kg, and, when cut, presented with solid conformation and whitish coloration. Tumor fragments were harvested, fixed in 10% formalin, and sent for histopathological examination, which revealed neoproliferation of remarkable cellular density composed of dense, well vascularized fibrocollagenous connective tissue arranged in multidirectional bundles and undulating pattern. Mild cellular pleomorphism was identified, and no mitosis figures were observed. Alcian blue staining was negative for mucopolysaccharides, differing from Masson's trichrome staining, which widely stained the fibrocollagenous tissue blue. In view of these findings, the diagnosis of cutaneous fibroma was confirmed. Discussion: Cutaneous fibromas are benign neoplasms of fibrous tissue, and they are uncommon in cattle and may be associated with bovine papillomavirus and/or trauma. Although the origin of cutaneous fibroma is not clear, the present report stands out due to the large size of the tumor mass. The complete healing of the surgical wound, the absence of recurrence one year after surgery and the return of the animal to dairy production demonstrate that the surgical treatment was adequate. The macro- and microscopic characteristics of the cutaneous fibroma in this case corroborate with other cases reported in the literature. Large cutaneous fibroma is uncommon in bovines, and may hinder surgical excision and prolong healing time, as well as the complete recovery of the animal. Moreover, the differential diagnosis with other neoplasms of fibroblastic origin is relevant, especially for those with malignant biological behavior, such as fibrosarcoma and myxosarcoma.

Histological and immunohistochemical evaluations of the bone marrow from femur and sternal manubrium of dogs reactive for leishmaniasis by DPP® and ELISA tests / Avaliações histológica e imunoistoquímica da medula óssea do fêmur e do manúbrio esternal de cães reagentes para leishmaniose aos testes DPP® e ELISA

As the bone marrow is one of the most organs affected by canine visceral leishmaniasis (CVL), samples from this are frequently taken for parasitological tests, with occurrence of myelodysplastic changes, with consequent anemia, leukopenia, and thrombocytopenia. Thus, this study aimed to investigate the histological and immunohistochemical changes in the bone marrow of the femur and sternal manubrium of dogs reactive for leishmaniasis by DPP® and ELISA tests. For this, thirteen canines from the epidemiological routine for CVL carried out by the Directorate of Zoonosis Surveillance of Goiânia (DVZ), GO, Brazil, were subjected to anatomopathological examination. 46.2% of bone marrow samples from the femur showed a higher proportion of the red series, and 53.9% of bone marrow of the sternal manubrium evidenced a higher proportion of the red series. Also, there were varied macrophage hyperplasia, hemosiderosis, and megakaryocytic emperipolesis. Amastigote forms of Leishmania spp. in the bone marrow of the femur and sternal manubrium to histopathological and immunohistochemical evaluations were observed, with good agreement them, but without difference in the parasite intensity between the bone marrow of these anatomical sites. It was concluded that bone marrow of the femur and sternal manubrium of dogs reactive for leishmaniasis by DPP® and ELISA tests has histological changes resulting from the disease, regardless of the parasite presence or intensity, with macrophage hyperplasia, hemosiderosis, and emperipolesis being the main medullary changes in these animals. Also, the bone marrow of the femur and sternal manubrium are useful anatomical sites for the diagnosis of CVL by direct methods.

Como a medula óssea é um dos órgãos mais acometidos pela leishmaniose visceral canina (LVC), amostras desta são frequentemente colhidas para exames parasitológicos, sendo possível a ocorrência de alterações mielodisplásicas, com consequente anemia, leucopenia e trombocitopenia. Assim, este estudo teve como objetivo investigar alterações histológicas e imunoistoquímicas na medula óssea do fêmur e manúbrio esternal de cães reativos para leishmaniose aos testes DPP® e ELISA. Para isso, 13 caninos da rotina epidemiológica para LVC realizada pela Diretoria de Vigilância de Zoonoses de Goiânia (DVZ), GO, Brasil, foram submetidos ao exame anatomopatológico. 46,2% e 53,9% das amostras de medula óssea do fêmur e do manúbrio esternal apresentaram maior proporção da série vermelha, respectivamente. Além disso, havia variados graus de hiperplasia macrofágica, hemossiderose e emperipolese megacariocítica. Formas amastigotas de Leishmania spp. na medula óssea do fêmur e do manúbrio esternal às avaliações histopatológicas e imunoistoquímicas foram observadas, com boa concordância entre essas, mas sem diferença na intensidade parasitária entre a medula óssea desses sítios anatômicos. Conclui-se que a medula óssea do fêmur e do manúbrio esternal de cães reativos para leishmaniose aos testes DPP® e ELISA apresenta alterações histológicas decorrentes da doença, independente da presença ou intensidade do parasito, sendo hiperplasia de macrófagos, hemossiderose e emperipolese as principais alterações medulares nesses animais. Além disso, a medula óssea do fêmur e do manúbrio esternal compreendem sítios anatômicos úteis ao diagnóstico de LVC por métodos diretos.

Lipossarcoma pleomórfico esplênico primário em uma cadela / Primary splenic pleomorphic liposarcoma in a bitch

Background: Liposarcoma is a malignant neoplasm of lipoblasts with low incidence in dogs, representing 1.7% of tumorsdiagnosed in the spleen. In veterinary medicine, this neoplasm is classified morphologically into the myxoid, well-differentiated, undifferentiated and pleomorphic subtypes, the latter being one of the most aggressive forms, mainly in cavityorgans. This report refers to a primary splenic pleomorphic liposarcoma in a female dog, addressing anatomopathologicaland immunohistochemical aspects.Case: A 14-year-old, 35 kg bitch mongrel with history of absence of defecation, progressive weight loss, difficulty walking, sensitivity to abdominal palpation, prostration, pale mucous membranes, tachypnea and abdominal distention waspresented to diagnosis. The condition evolved to death and, on necroscopy, there was an increase in splenic volume withneoformation of whitish and reddish color, measuring 32 × 27 cm in its largest axes and weighing 8.9 kg. The neoformationexhibited areas of firm and soft consistency, and sectioning revealed focal areas of extensive necrosis and cavity collectionsof different diameters that allowed the flow of liquid serous contents with a brownish red color. Microscopy showed cellsof neoplastic morphology infiltrating the splenic parenchyma, mostly with slightly acidophilic cytoplasm and few intracytoplasmic lipid vacuoles, which varied in size and distribution. The nucleus of the cells was large, eccentric and irregular,with round to oval morphology, grossly lacy chromatin and single or multiple evident nucleoli. These cells exhibited markedanisocytosis, anisokaryosis and pleomorphism, with more than one mitotic figure per high magnification field visible. Mildinflammatory infiltrate, predominantly lymphocytic, permeated the neoplastic cells, and marked depletion of lymphoidfollicles and atrophy of the red pulp were found in the remaining splenic parenchyma. Immunohistochemical tests...(AU)

Morphological and Molecular Characterization of Proliferative Inflammatory Atrophy in Canine Prostatic Samples.

Proliferative inflammatory atrophy (PIA) is an atrophic lesion of the prostate gland that occurs in men and dogs and is associated with a chronic inflammatory infiltrate. In this study, we retrospectively reviewed canine prostatic samples from intact dogs, identifying 50 normal prostates, 140 cases of prostatic hyperplasia, 171 cases of PIA, 84 with prostate cancer (PC), 14 with prostatic intraepithelial neoplasia (PIN) and 10 with bacterial prostatitis. PIA samples were then selected and classified according to the human classification. The presence of PIA lesions surrounding neoplastic areas was then evaluated to establish a morphological transition from normal to preneoplastic and neoplastic tissue. In addition, the expression of PTEN, P53, MDM2 and nuclear androgen receptor (AR) were analyzed in 20 normal samples and 20 PIA lesions by immunohistochemistry and qPCR. All PIA lesions showed variable degrees of mononuclear cell infiltration around the glands and simple atrophy was the most common histopathological feature. PIA was identified between normal glands and PC in 51 (61%) out of the 84 PC samples. PIA lesions were diffusely positive for molecular weight cytokeratin (HMWC). Decreased PTEN and AR gene and protein expression was found in PIA compared to normal samples. Overall, our results strongly suggest that PIA is a frequent lesion associated with PC. Additionally, this finding corroborates the hypothesis that in dogs, as is the case in humans, PIA is a pre neoplastic lesion that has the potential to progress into PC, indicating an alternative mechanism of prostate cancer development in dogs.

Effects of the Latex of Synadenium grantii Hook F. (Euphorbiaceae) on a Preclinical Model of Canine Prostate Cancer.

Prostatic cancer (PC) stands out in terms of its occurrence, pathophysiology, and unfavorable prognostics in humans and dogs. Natural drugs bear an integrative potential for conventional antineoplastic treatments. In this context, the bioproducts of Synadenium grantii have been empirically used in different parts of Brazil for the integrative treatment of prostate cancer in humans. However, there is no availability of scientific evidence of the antitumor effects of S. grantii. Therefore, this study aimed to investigate the bioactive compounds in the latex of S. grantii using the high-resolution mass spectrophotometry (HRMS) and to evaluate its cytotoxic effects on primary canine PC cell cultures. Four fragments of phorbol ester were identified as potential bioactive compounds using the HRMS. With the help of an MTT ([3-(4,5-dimethyldiazol-2-yl)-2,5 diphenyltetrazolium bromide]) assay, two canine prostatic carcinoma cell lines (PC 1 and PC2) showed a decrease in the tumor cell count, with an Inhibitory concentration 50 (IC50)of 0.8469 and 0.6068 mg/ml, respectively, for PC1 and PC2. In conclusion, the latex of S. grantii contains phorbol esters in its composition, and its aqueous solution has a cytotoxic effect on canine metastatic PC cells in vitro.

Lipossarcoma pleomórfico esplênico primário em uma cadela / Primary splenic pleomorphic liposarcoma in a bitch

Background: Liposarcoma is a malignant neoplasm of lipoblasts with low incidence in dogs, representing 1.7% of tumorsdiagnosed in the spleen. In veterinary medicine, this neoplasm is classified morphologically into the myxoid, well-differentiated, undifferentiated and pleomorphic subtypes, the latter being one of the most aggressive forms, mainly in cavityorgans. This report refers to a primary splenic pleomorphic liposarcoma in a female dog, addressing anatomopathologicaland immunohistochemical aspects.Case: A 14-year-old, 35 kg bitch mongrel with history of absence of defecation, progressive weight loss, difficulty walking, sensitivity to abdominal palpation, prostration, pale mucous membranes, tachypnea and abdominal distention waspresented to diagnosis. The condition evolved to death and, on necroscopy, there was an increase in splenic volume withneoformation of whitish and reddish color, measuring 32 × 27 cm in its largest axes and weighing 8.9 kg. The neoformationexhibited areas of firm and soft consistency, and sectioning revealed focal areas of extensive necrosis and cavity collectionsof different diameters that allowed the flow of liquid serous contents with a brownish red color. Microscopy showed cellsof neoplastic morphology infiltrating the splenic parenchyma, mostly with slightly acidophilic cytoplasm and few intracytoplasmic lipid vacuoles, which varied in size and distribution. The nucleus of the cells was large, eccentric and irregular,with round to oval morphology, grossly lacy chromatin and single or multiple evident nucleoli. These cells exhibited markedanisocytosis, anisokaryosis and pleomorphism, with more than one mitotic figure per high magnification field visible. Mildinflammatory infiltrate, predominantly lymphocytic, permeated the neoplastic cells, and marked depletion of lymphoidfollicles and atrophy of the red pulp were found in the remaining splenic parenchyma. Immunohistochemical tests...

Análises anatomopatológica e imuno- histoquímica do baço e linfonodo de cães soropositivos para leishmaniose em testes sorológicos / Anatomomopathological and immunohistochemical analyses of the spleen and lymph node of dogs seropositives for leishmaniasis in serological tests

A leishmaniose canina (CanL) é uma zoonose causada pelo protozoário da espécie Leishmania infantum. O baço e os linfonodos sofrem alterações morfológicas durante o CanL. Esta pesquisa teve como objetivo realizar um estudo anatomopatológico e imuno-histoquímico desses órgãos em cães reativos para leishmaniose aos testes de Imunoensaio Cromatográfico “Dual Path Platform” (DPP®) e Ensaio de Imunoabsorção Enzimática (ELISA). Vinte e sete cães foram avaliados ao exame anatomopatológico, com 92,6% exibindo alterações à avaliação macroscópica, especialmente esplenomegalia e linfadenomegalia. Todos os cães apresentaram alterações no baço não relacionadas à carga parasitária, sendo a esplenite granulomatosa a alteração mais grave. Hiperplasia cortical e paracortical difusa e hiperplasia e hipertrofia dos cordões medulares foram observadas nos linfonodos. Formas amastigotas de Leishmania spp. foram encontradas no baço e linfonodo às avaliações histopatológica e imuno-histoquímica, com boa concordância entre os métodos (k = 0,55, p = 0,00124), mas não foi observada diferença na intensidade parasitária entre esses órgãos à imuno-histoquímica (p = 0,23). Conclui-se que baço e linfonodo de cães reativos para leishmaniose aos testes DPP® e ELISA apresentam alterações histomorfológicas decorrentes da doença, independente da carga parasitária, assim como esses órgãos apresentam carga parasitária semelhante ao método imuno-histoquímico.

Canine leishmaniasis (CanL) is a zoonosis caused by the protozoan of the species Leishmania infantum. The spleen and lymph nodes undergo morphological changes during CanL. This research aimed to perform an anatomopathological and immunohistochemical study of these organs in dogs reactive to leishmaniasis in the Dual-path Platform chromatographic immunoassay (DPP®) and Enzyme Immunoabsorption Assay (ELISA). Twenty-seven dogs were evaluated for anatomopathological examination with 92.6% showing changes at gross evaluation, specially splenomegaly and lymphadenomegaly. All dogs showed changes in the spleen unrelated to the parasitic load, with granulomatous splenitis being the most severe change. Diffuse cortical and paracortical hyperplasia, and hyperplasia and hypertrophy of the medullary cords were observed in the lymph node. Amastigote forms of Leishmania spp. were found in the spleen and lymph node at histopathological and immunohistochemical evaluations, with good agreement between these evaluations (k = 0.55, p = 0.00124), but no difference was observed in the parasitic intensity of these organs at immunohistochemistry (p = 0.23). It was concluded that spleen and lymph node from dogs reactive to leishmaniasis on the DPP® and ELISA tests show histomorphological changes resulting from the disease, independent to the parasitic load, as well as these organs show similar parasitic load at immunohistochemical test.

Elucidando a imunovigilância e imunoedição tumoral: uma revisão abrangente / Elucidating tumor immunosurveillance and immunoediting: a comprehensive review

A ação do sistema imunológico contra as enfermidades neoplásicas tem se tornado uma das principais fontes de pesquisa na atualidade. As vias biológicas desse sistema são conhecidas por contribuir na limitação da progressão e na eliminação do tumor, e são delineadas por conceitos e mecanismos de imunovigilância e imunoedição. A imunovigilância é considerada o processo pelo qual o sistema imunológico reconhece e inibe o processo neoplásico. O conceito de imunoedição tem origem no sentido de que o sistema imune é capaz de moldar o perfil antigênico do tumor devido à pressão seletiva, baseada nas etapas de eliminação, equilíbrio e evasão tumoral. A resposta imunológica ocorre contra antígenos tumorais e modificações do microambiente tumoral, envolvendo diferentes componentes do sistema imune inato, como células T, células natural Killer, linfócitos B e macrófagos. Nesse sentido, conhecer esses conceitos e compreender seus respectivos mecanismos torna-se essencial na investigação de novas estratégias de prevenção e combate ao câncer. Dessa forma, esta revisão apresenta aspectos históricos e definições de imunovigilância e imunoedição tumoral, com ênfase em sua importância e aplicabilidade, assim como aos diferentes métodos utilizados em imunoterapia.

The action of the immune system against neoplastic diseases has become one of the main sources of research. The biological pathways of this system are known to contribute in limiting the progression and elimination of the tumor, and are delineated by concepts and mechanisms of immunosurveillance and immunoediting. Immunosurveillance is considered the process by which the immune system recognizes and inhibits the neoplastic process. The concept of immunoediting arises in the sense that immune system is able to shape the antigenic profile of the tumor due to selective pressure, based on the stages of tumor elimination, balance and evasion. The immune response occurs against tumor antigens and changes in the tumor microenvironment, involving different components of the innate immune system, such as T cells, natural Killer cells, B lymphocytes and macrophages. In this sense, knowing these concepts and understanding their respective mechanisms becomes essential in the investigation of new strategies for cancer prevention and cure. Thus, this review presents historical aspects and definitions of immunosurveillance and tumor immunoediting, with emphasis on its importance and applicability, such as on the different methods used in immunotherapy.

Autologous platelet-rich plasma action on skin autografts in horses / Ação do plasma rico em plaquetas autólogo em autoenxertos de pele em equinos

ABSTRACT: This study evaluated the action of autologous platelet-rich plasma (PRP) on cutaneous wounds, containing skin autografts, in the gluteal region of horses. Seven healthy horses were used. Two 6 x 6cm cutaneous wounds were produced on each side of the gluteal region. Eight days after wound induction, grafts were performed with skin fragments harvested from the neck, as well as the application of PRP, prepared by double-centrifugation protocol. Wounds with autografts on the left side received PRP (group T), and those with autografts on the right side did not receive treatment (group C). Macroscopic and microscopic evaluations were performed, considering the integration of autografts and retraction of wound edges, as well as neovascularization, inflammatory infiltrate, young fibroblasts, collagenization, reepithelization and autografts integration. There was no difference between the groups (P > 0.05) in relation to most macroscopic and microscopic variables. However, neovascularization was significantly greater (p = 0.0191) in group T, on the 14th day after grafting. It is concluded that PRP favors the process of skin repair with autografts in horses, since it increases the neovascularization in the initial phase of wound healing. Furthermore, the PRP seems to positively influence the integration of the skin autografts and the retraction of the wound edges.

RESUMO: Este estudo avaliou a ação do plasma rico em plaquetas (PRP) autólogo em feridas cutâneas, contendo autoenxertos de pele em equinos. Foram utilizados sete equinos hígidos, nos quais foram produzidas duas feridas cutâneas de 6 x 6cm, em cada um dos lados da região glútea. Oito dias após a indução das feridas, foram realizados enxertos com fragmentos de pele colhidos do pescoço, assim como a aplicação do PRP, preparado através de protocolo de dupla centrifugação. As feridas com autoenxertos do lado esquerdo receberam PRP (grupo T), e as com autoenxertos do lado direito não receberam tratamento (grupo C). Foram realizadas avaliações macroscópica e microscópica, considerando as variáveis integração dos autoenxertos e retração das bordas da ferida, além de neovascularização, infiltrado inflamatório, fibroblastos jovens, colagenização, reepitelização e integração dos autoenxertos. Não houve diferença entre os grupos (p > 0,05) em relação à maioria das variáveis macroscópicas e microscópicas. Contudo, a neovascularização foi significativamente maior (P = 0,0191) no grupo T, no 14º dia após a realização da enxertia. Conclui-se que o PRP favorece o processo de reparo da pele com autoenxertos em equinos, já que aumenta a neovascularização na fase inicial da cicatrização da ferida. Ainda, o PRP parece influenciar positivamente a integração dos autoenxertos de pele e a retração das bordas da ferida.

Anatomomopathological and immunohistochemical analyses of the spleen and lymph node of dogs seropositives for leishmaniasis in serological tests / Análises anatomopatológica e imuno-histoquímica do baço e linfonodo de cães soropositivos para leishmaniose em testes sorológicos

Canine leishmaniasis (CanL) is a zoonosis caused by the protozoan of the species Leishmania infantum. The spleen and lymph nodes undergo morphological changes during CanL. This research aimed to perform an anatomopathological and immunohistochemical study of these organs in dogs reactive to leishmaniasis in the Dual-path Platform chromatographic immunoassay (DPP®) and Enzyme Immunoabsorption Assay (ELISA). Twenty-seven dogs were evaluated for anatomopathological examination with 92.6% showing changes at gross evaluation, specially splenomegaly and lymphadenomegaly. All dogs showed changes in the spleen unrelated to the parasitic load, with granulomatous splenitis being the most severe change. Diffuse cortical and paracortical hyperplasia, and hyperplasia and hypertrophy of the medullary cords were observed in the lymph node. Amastigote forms of Leishmania spp. were found in the spleen and lymph node at histopathological and immunohistochemical evaluations, with good agreement between these evaluations (k = 0.55, p = 0.00124), but no difference was observed in the parasitic intensity of these organs at immunohistochemistry (p = 0.23). It was concluded that spleen and lymph node from dogs reactive to leishmaniasis on the DPP® and ELISA tests show histomorphological changes resulting from the disease, independent to the parasitic load, as well as these organs show similar parasitic load at immunohistochemical test.

A leishmaniose canina (CanL) é uma zoonose causada pelo protozoário da espécie Leishmania infantum. O baço e os linfonodos sofrem alterações morfológicas durante o CanL. Esta pesquisa teve como objetivo realizar um estudo anatomopatológico e imuno-histoquímico desses órgãos em cães reativos para leishmaniose aos testes de Imunoensaio Cromatográfico "Dual Path Platform" (DPP®) e Ensaio de Imunoabsorção Enzimática (ELISA). Vinte e sete cães foram avaliados ao exame anatomopatológico, com 92,6% exibindo alterações à avaliação macroscópica, especialmente esplenomegalia e linfadenomegalia. Todos os cães apresentaram alterações no baço não relacionadas à carga parasitária, sendo a esplenite granulomatosa a alteração mais grave. Hiperplasia cortical e paracortical difusa e hiperplasia e hipertrofia dos cordões medulares foram observadas nos linfonodos. Formas amastigotas de Leishmania spp. foram encontradas no baço e linfonodo às avaliações histopatológica e imuno-histoquímica, com boa concordância entre os métodos (k = 0,55, p = 0,00124), mas não foi observada diferença na intensidade parasitária entre esses órgãos à imuno-histoquímica (p = 0,23). Conclui-se que baço e linfonodo de cães reativos para leishmaniose aos testes DPP® e ELISA apresentam alterações histomorfológicas decorrentes da doença, independente da carga parasitária, assim como esses órgãos apresentam carga parasitária semelhante ao método imuno-histoquímico.

Tumor de células granulares pulmonar em um equino / Pulmonar granular cell tumor in a horse

Background: The occurrence of neoplasms in horses is relatively low. Granular cell tumor is a seldom diagnosed neoplasm, usually benign, of mesenchymal origin. Controversies exist regarding its origin, which is possibly from Schwann cells or cells with neuroendocrine differentiation. Despite being one of the main primary neoplasms in the lungs of horses, the number of cases is low in comparison to that of secondary lung tumors. Thus, this study proposes to report the anatomopathological aspects in a horse with granular cell tumor of primary pulmonary origin. Case: An 11-year-old female Quarter Horse breed underwent exploratory right lateral thoracotomy after presenting with chronic respiratory changes. During the operation, tumor masses were found in the right and left caudal pulmonary lobes. Due to the severity of clinical respiratory signs and the extent of the lesions, the animal was subjected to euthanasia and anatomopathological examination. Upon necroscopic examination, a tumor mass was found in the middle third of the left caudal lobe, rounded to flattened, measuring 10.0 × 8.0 cm in height and length, white in color, of firm consistency, smooth and regular surface and rising to the lung surface. When sectioned, the mass showed to be composed of multiple firm and dense circular lobes, separated by a thin layer of connective tissue. The tumor invaded the lumen of nearby segmental and subsegmental bronchi, which were partially or totally obstructed by the mass. In the right lung, multiple similar nodules were observed, accompanied by peritumoral hemorrhage. Histopathological analysis of the new formation revealed a dense cluster of cells that expanded over the lung parenchyma. The neoplastic cells were pleomorphic, moderately cohesive, without defined borders, with abundant cytoplasm, densely eosinophilic and finely granular. Intracytoplasmic granules were well evidenced by periodic acid Schiff staining (PAS). The cell nucleus was rounded to oval, excentric, markedly basophilic and with dense chromatin. There was moderate anisocytosis and mild anisokaryosis, with rare mitotic figures. Immunohistochemical analysis revealed positive staining for anti-vimentin and anti-S100 antibodies, confirming the diagnosis of granular cell tumor. Discussion: Granular cell tumors have no predisposition as to breed, sex or age. However, most of the described cases are reported in female horses aged around 13 years. The advanced age of the diagnosed animals may be related to late definitive diagnosis, since the clinical signs are nonspecific and treated palliatively like other respiratory diseases. Macroscopically, this tumor is more common in the multinodular form and, as observed in this case, it has a greater capacity for infiltration. Histologically, the visualization of large, polygonal cells, with a wide cytoplasm containing eosinophilic granules leads to the diagnosis of granular cell tumor. However, PAS staining and immunohistochemical tests were essential for the diagnostic conclusion in this report, confirming the presence of cytoplasmic granules and the mesenchymal and neuroectodermal origin of this neoplasm, respectively. Thus, considering the low occurrence of pulmonary granular cell tumor, the description of this case contributes to the basis of the knowledge of medical-veterinary professionals about this tumor in its clinical and diagnostic aspects.

Influencing factors for preparation of platelet-rich plasma in horses / Fatores que influenciam o preparo do plasma rico em plaquetas na espécie equina

There is no consensus in the literature as to the best platelet-rich plasma (PRP) acquisition method or the ideal platelet concentration to stimulate tissue repair. Besides that, most studies do not provide a clear and replicable description of the methods used, which makes standardization and result comparison difficult. Thus, this study aimed to accurately describe the method and evaluate factors influencing PRP preparation in equines. In this regard, two protocols were used, P1 and P2, based on two centrifugation methods, which differed in blood volume, speed and time, and platelet pipetting and resuspension. In conclusion, factors such as centrifugation strength and time are essential to obtain PRP with adequate platelet count. However, other factors such as tube type and plasma pipetting and resuspension methods directly interfere with the replicability of the technique, and hence influencing PRP applicability.(AU)

Não há consenso na literatura quanto ao melhor método de obtenção do plasma rico em plaquetas (PRP) ou à concentração plaquetária ideal com o objetivo de estimular o reparo tecidual. Além disso, grande parte dos trabalhos não faz descrição clara e reprodutível dos métodos utilizados, o que dificulta a padronização e a comparação dos resultados. Dessa forma, este trabalho teve por objetivo descrever detalhadamente a metodologia e avaliar fatores que influenciam o preparo do PRP em equinos. Para isso, foram utilizados dois protocolos, P1 e P2, baseados em duas centrifugações, que diferiram quanto ao volume de sangue colhido, velocidade e tempo das centrifugações e método de pipetagem e ressuspensão das plaquetas. Conclui-se que fatores como força e tempo de centrifugação são essenciais à obtenção de um PRP com concentração plaquetária adequada. Contudo, fatores adicionais como tipo de tubo e métodos de pipetagem e ressuspensão do plasma interferem diretamente na repetibilidade da técnica, influenciando a aplicabilidade prática do PRP.(AU)

Immunoexpression of metalloproteinases 9 (MMP-9) and 2 (MMP-2) and their inhibitors (TIMP-1 and TIMP-2) in normal and neoplastic canine mammary tissue / Imunomarcação de metaloproteinases 9 (MMP-9) e 2 (MMP-2) e seus inibidores (TIMP-1) e (TIMP-2) no tecido mamário canino e neoplásico

The aim of this study was to perform the immunostaining of MMP-9 and MMP-2 and its inhibitors, TIMP-1 and TIMP-2, on normal and neoplastic canine mammary tissue in order to evaluate the behavior of these proteins in extracellular matrix (ECM) remodeling in different neoplastic mammary types. Thus, 48 samples of canine mammary tissue were analyzed, 14 of which complex carcinomas, 13 tubulopapillary carcinomas, six single adenomas and 15 normal mammary tissue. There were differences in MMP-9, TIMP-1 and TIMP-2 according to mammary histomorphology, and MMP-9 presented increased immunoexpression in epithelial and stromal cells in tubulopapillary and complex carcinomas. TIMP-1 exhibited reduced immunostaining in the stromal cells of the complex carcinomas and TIMP-2 enhanced immunostaining in the epithelial cells of tubulopapillary carcinomas. There was a positive correlation between MMP-9 and TIMP-1 in epithelial and stromal cells regarding immunostaining intensity and number of labeled cells in the normal breast. There was a positive correlation between MMP-9 and TIMP-2 in the epithelial cells of tubulopapillary carcinomas. It is concluded that balanced activity between MMP-9, MMP-2, TIMP-1 and TIMP-2 maintains normal canine mammary tissue homeostasis while increased immunoexpression of MMP-9 and TIMP-2 and reduced TIMP- 1 in carcinomas suggest a favorable condition for tumor evolution.(AU)

Este estudo teve como objetivo realizar a imunomarcação de MMP-9 e MMP-2 e seus inibidores, TIMP-1 e TIMP-2, no tecido mamário canino normal e neoplásico, a fim de avaliar o comportamento dessas proteínas no remodelamento da matriz extracelular (MEC) em diferentes tipos neoplásicos mamários. Foram analisadas 48 amostras de tecido mamário canino, sendo 14 carcinomas complexos, 13 carcinomas tubulopapilares, seis de adenomas simples e 15 mamas sem alterações. Houve diferença em MMP-9, TIMP-1 e TIMP-2 de acordo com a histomorfologia mamária, sendo que MMP-9 apresentou maior imunoexpressão em células epiteliais e estromais em carcinomas tubulopapilares e complexos. TIMP-1 exibiu menor imunomarcação nas células estromais dos carcinomas complexos e TIMP-2 maior imunomarcação nas células epiteliais dos carcinomas tubulopapilares. Houve correlação positiva entre MMP-9 e TIMP-1 nas células epiteliais e estromais quanto à intensidade de imunomarcação e número de células marcadas na mama normal. Houve correlação positiva entre MMP-9 e TIMP-2 nas células epiteliais dos carcinomas tubulopapilares. Conclui-se que a atividade equilibrada entre MMP-9, MMP-2, TIMP-1 e TIMP-2 mantém a homeostase do tecido mamário canino normal enquanto a imunoexpressão aumentada de MMP-9 e TIMP-2 e a imunoexpressão reduzida de TIMP-1 nos carcinomas sugere condição propícia à evolução tumoral.(AU)

Immunoexpression of metalloproteinases 9 (MMP-9) and 2 (MMP-2) and their inhibitors (TIMP-1 and TIMP-2) in normal and neoplastic canine mammary tissue / Imunomarcação de metaloproteinases 9 (MMP-9) e 2 (MMP-2) e seus inibidores (TIMP-1) e (TIMP-2) no tecido mamário canino e neoplásico

The aim of this study was to perform the immunostaining of MMP-9 and MMP-2 and its inhibitors, TIMP-1 and TIMP-2, on normal and neoplastic canine mammary tissue in order to evaluate the behavior of these proteins in extracellular matrix (ECM) remodeling in different neoplastic mammary types. Thus, 48 samples of canine mammary tissue were analyzed, 14 of which complex carcinomas, 13 tubulopapillary carcinomas, six single adenomas and 15 normal mammary tissue. There were differences in MMP-9, TIMP-1 and TIMP-2 according to mammary histomorphology, and MMP-9 presented increased immunoexpression in epithelial and stromal cells in tubulopapillary and complex carcinomas. TIMP-1 exhibited reduced immunostaining in the stromal cells of the complex carcinomas and TIMP-2 enhanced immunostaining in the epithelial cells of tubulopapillary carcinomas. There was a positive correlation between MMP-9 and TIMP-1 in epithelial and stromal cells regarding immunostaining intensity and number of labeled cells in the normal breast. There was a positive correlation between MMP-9 and TIMP-2 in the epithelial cells of tubulopapillary carcinomas. It is concluded that balanced activity between MMP-9, MMP-2, TIMP-1 and TIMP-2 maintains normal canine mammary tissue homeostasis while increased immunoexpression of MMP-9 and TIMP-2 and reduced TIMP- 1 in carcinomas suggest a favorable condition for tumor evolution.

Este estudo teve como objetivo realizar a imunomarcação de MMP-9 e MMP-2 e seus inibidores, TIMP-1 e TIMP-2, no tecido mamário canino normal e neoplásico, a fim de avaliar o comportamento dessas proteínas no remodelamento da matriz extracelular (MEC) em diferentes tipos neoplásicos mamários. Foram analisadas 48 amostras de tecido mamário canino, sendo 14 carcinomas complexos, 13 carcinomas tubulopapilares, seis de adenomas simples e 15 mamas sem alterações. Houve diferença em MMP-9, TIMP-1 e TIMP-2 de acordo com a histomorfologia mamária, sendo que MMP-9 apresentou maior imunoexpressão em células epiteliais e estromais em carcinomas tubulopapilares e complexos. TIMP-1 exibiu menor imunomarcação nas células estromais dos carcinomas complexos e TIMP-2 maior imunomarcação nas células epiteliais dos carcinomas tubulopapilares. Houve correlação positiva entre MMP-9 e TIMP-1 nas células epiteliais e estromais quanto à intensidade de imunomarcação e número de células marcadas na mama normal. Houve correlação positiva entre MMP-9 e TIMP-2 nas células epiteliais dos carcinomas tubulopapilares. Conclui-se que a atividade equilibrada entre MMP-9, MMP-2, TIMP-1 e TIMP-2 mantém a homeostase do tecido mamário canino normal enquanto a imunoexpressão aumentada de MMP-9 e TIMP-2 e a imunoexpressão reduzida de TIMP-1 nos carcinomas sugere condição propícia à evolução tumoral.

Outbreak of monensin poisoning in cattle due to supplementation error / Surto de intoxicação por monensina em bovinos devido a erro de suplementação

Iatrogenic intoxications occur in domestic animals. Mortality was observed in a group of calves, reaching 20 deaths until the day of the technical visit, with or without sudden clinical signs, on a farm in the city of Pires do Rio, State of Goiás, Brazil. Deaths after the visit were not accounted systematically. Clinical signs included muscle weakness, ataxia, recumbency, bilateral jugular distention, and death. Suspected diagnosis of poisoning by ionophore antibiotics was made based on clinical and laboratory findings from nine animals, which included high plasma CK and LDH levels, as well as on anatomopathological findings of a recently dead calf, which showed myocardial and skeletal muscle degeneration and necrosis. Monensin overdose (25.5 µg/kg in skeletal muscle and 209.4 µg/kg in the liver) was detected by liquid-chromatography and mass-spectrophotometry analysis, contributing to the confirmation of ionophore poisoning diagnosis.(AU)

Intoxicações iatrogênicas ocorrem em animais domésticos. Um lote de bezerros apresentou mortalidade, chegando a 20 mortes até o dia da visita técnica, após sinais clínicos súbitos ou ausência destes, em uma propriedade rural do município de Pires do Rio, estado de Goiás, Brasil. As mortes após a visita não foram sistematicamente contabilizadas. Os sinais incluíam fraqueza muscular, ataxia, decúbito, distensão bilateral da veia jugular e morte. Baseado nos achados clínicos e laboratoriais de nove bovinos, que incluíram níveis de CK e LDH elevados no plasma, assim como degeneração e necrose miocárdica e na musculatura esquelética à avaliação anatomopatológica em um dos bezerros mortos, que apresentou degeneração e necrose em miocárdio e músculo esquelético. Uma sobredosagem de monensina (25,5 µg/g no músculo esquelético e 209,4 µg/g no fígado) foi detectada com análise por cromatografia líquida e espectrofotometria de massas, contribuindo para a confirmação do diagnóstico de intoxicação por ionóforos.(AU)

Imunomarcação de TGF-β em próstatas caninas normais e com lesões proliferativas / Immunoexpression of TGF-β in normal canine prostates and with proliferative lesions

O fator de crescimento transformador-β (TGF-β), um mediador do crescimento prostático, induz a angiogênese e inibe a proliferação celular. Neste estudo, esse marcador foi utilizado com o objetivo de avaliar sua imunomarcação no tecido normal e com lesões proliferativas benignas, pré-neoplásicas e neoplásicas da próstata canina. Para isso, foram selecionadas 54 glândulas com histomorfologia normal, hiperplasia prostática benigna (HPB) epitelial, HPB estromal, atrofia inflamatória proliferativa (PIA), neoplasia intraepitelial prostática (PIN) e carcinoma, utilizadas para a confecção de um bloco de microarranjo tecidual (Tissue Microarray - TMA). As lâminas de TMA foram submetidas à técnica de imunoistoquímica com o anticorpo anti-TGF-β, sendo avaliada a intensidade de imunomarcação nas células epiteliais e estromais. Houve imunomarcação de TGF-β no tecido normal e naqueles com lesões proliferativas. Maior imunomarcação de TGF-β foi constatada nas células do tecido prostático normal e com HPB, enquanto as células prostáticas com PIA, PIN e carcinoma exibiram menor imunomarcação dessa citocina, o que sugere a ação do TGF-β na manutenção da homeostase do tecido normal e com lesão proliferativa benigna e na progressão das lesões proliferativas pré-malignas e malignas da próstata canina.(AU)

The transforming growth factor-β (TGF-β), a mediator of prostatic growth induces angiogenesis and inhibits cell proliferation. It was used in this study in order to evaluate its expression in normal prostatic tissue and those with benign proliferative lesions, pre-malignant and malignant prostatic diseases. A total of 54 glands with normal histomorphology, epithelial benign prostatic hyperplasia (BPH), stromal BPH, prostatic inflammatory atrophy (PIA), prostatic intraepithelial neoplasia (PIN), and adenocarcinoma were selected and used in the tissue microarray block (TMA). The TMA slides were subjected to the immunohistochemistry with anti-TGF-β and its staining intensity in epithelial and stromal cells was evaluated. There was TGF-β immunostaining by both normal tissues and those with proliferative lesions. Higher TGF-β immunostaining was observed in cells from normal prostatic tissues and with HPB, whereas prostatic cells with PIA, PIN and carcinoma exhibited lower immunostaining of this cytokine, suggesting the action of TGF-β in the maintenance of normal tissue homeostasis and with benign proliferative lesion and in the progression of pre-malignant and malignant proliferative lesions of the canine prostate.(AU)