Análise qualitativa quanto à diferença da imunoexpressão do gene c-Kit em melanomas melânicos e amelânicos da cavidade oral em cães

Melanomas são as mais frequentes neoplasias malignas da cavidade bucal de cães. Sabe-se que a proliferação de células e expressão de conexina diferem em melanomas melanóticos e amelanóticos da cavidade bucal de cães. Este estudo analisou a expressão da proteína c-Kit em melanomas melanóticos e amelanóticos da cavidade bucal canina. Um total de 34 melanomas bucais caninos (19 melanóticos e 15 amelanóticos) foram coletados. Os melanomas amelanóticos apresentaram evolução mais rápida e maior incidência de metástase. Foi constatado um número significativamente maior de células positivas para c-Kit em neoplasias amelanóticas. Além disso, a intensidade de imunomarcação de c-Kit foi predominantemente mais forte em melanomas amelanóticos. Estes resultados confirmam um papel potencial para c-Kit em melanomas orais caninos, com diferenças claras em padrões de expressão entre os dois tipos histológicos de tumor, melanóticos e amelanóticos. Este trabalho destaca a importância de um estudo detalhado das mutações c-Kit em melanomas orais caninos para ser possível a melhor compreensão dos mecanismos moleculares envolvidos no desenvolvimento da doença.Melanomas are the most common oral malignancy in dogs. Cell proliferation and connexin expression has been shown to differ in canine melanotic and amelanotic oral melanomas. This study aimed to analyze the c-Kit protein expression in melanotic and amelanotic melanomas from canine buccal cavity. A total of 34 canine buccal melanomas (19 melanotic and 15 amelanotic).were collected. The amelanotic melanomas presented faster evolution and higher incidence of metastasis than melanotic tumors. A significantly higher number of c-Kit positive cells were observed in amelanotic neoplasms. In addition, the intensity of c-Kit immunolabeling was predominantly stronger in amelanotic melanomas. These results confirm a potential role for c-Kit in canine oral melanomas with clear differences in expression patterns between the two histological types of tumor, melanotic and amelanotic. This study highlights the importance of a detailed study of c-Kit mutations in canine oral melanomas to better understand the molecular mechanisms implicated in the development of this disease

Canine mammary tumors in Santos, Brazil: clinicopathological and survival profile

Os tumores das glândulas mamárias são as neoplasias mais comuns em cadelas em nosso país; no entanto, são poucos os trabalhos brasileiros dedicados ao estudo clinicopatológico e de sobrevida nesta doença. O presente trabalho teve por objetivo o estudo clínico e patológico dos tumores mamários caninos na Região Metropolitana de Santos, uma área no estado de São Paulo com uma população canina estimada em 120 mil animais. Dados de 14.298 cães foram coletados retrospectivamente dos prontuários médicos do Hospital Veterinário da Universidade Metropolitana de Santos – São Paulo – Brasil. Durante o período do estudo, foram atendidas 317 fêmeas com diagnóstico histopatológico de neoplasia, dos quais, 170 se referiam a lesões mamárias epiteliais distribuídas em 13 tumores benignos, 152 malignos (89,4% dos diagnósticos) e 5 lesões epiteliais não-neoplásicas (hiperplasia ductal). O tumor mais frequente foi o carcinoma tubular (38,2% dos tumores malignos) e tumores de grau I, respondendo por 73,0% do total diagnosticado. Estudos de sobrevida apontaram para o estadiamento clínico das neoplasias mamárias caninas como importante fator prognóstico, e na análise multivariada, diâmetro do tumor, grau histológico, quimioterapia adjuvante e recorrência apresentaram-se como covariáveis com valor preditivo de sobrevida. Levando-se em conta a elevada prevalência de carcinoma tubular simples na população canina de Santos, pode-se considerá-la como promissor modelo translacional para o estudo da doença.Tumors of the mammary glands are the most common neoplasms in dogs in our country; however, there are few Brazilian reports dedicated to clinicopathological and survival studies about this disease. This report aims the clinical and pathological study of canine mammary tumors in the Santos Metropolitan Region, an area in Sao Paulo state with an estimated canine population of 120,000 animals. Data of 14,298 dogs were collected retrospectively from the medical records of the Veterinary Medical Teaching Hospital of the Metropolitan University of Santos – São Paulo – Brazil. During the study period, from records of 317 females with histopathological diagnosis of neoplasia, 170 were mammary epithelial lesions distributed in 13 benign tumors, 152 malignant (89.4% of diagnosis) and 5 non-neoplasic epithelial lesions (ductal hyperplasia). The highest prevalent malignant tumor was tubular carcinoma (38.2% of diagnosis) and Grade I tumors, corresponding to 73.0% of all diagnosis. The results have shown clinical staging of canine mammary neoplasms as an important prognostic survival factor and, in a multivariate analysis, tumor diameter, tumor grade, adjuvant chemotherapy and recurrence as covariates with predictive value for survival. Moreover, the high prevalence of tubular carcinoma qualifies the canine population ofSantosas a promising model for the translational study of this disease

Modulação por PGE Ind.3 no perfil de subpopulações celulares e citocinas na evolução do Tumor Ascítico de Ehrlich (TAE)

O presente trabalho objetivou avaliar o envolvimento das prostaglandinas no crescimento tumoral, influxo inflamatório e secreação de citocinas durante a evolução do Tumor Ascítico de Ehrlich (TAE). Para tanto, camundongos foram inoculados com 1 x 103 células tumorais (ip) e tratados com indometacina (1mg/Kg,1x/dia,ip) ou com diluente (0,1 ml,1x/dia,ip). Decorridos 1, 3, 6, 10 e 13 dias os animais foram sacrificados e avaliados quanto ao influxo inflamatório diferencial, secreção de TNF-a, IL1-a, IL-2, IL- 4, IL-6, IL-10 e IL-13 e níveis de PGE2 no lavado peritoneal.. Dois grupos controle adicionais foram constituídos de animais não portadores de TAE tratados com indometacina ou diluente, seguindo o mesmo protocolo. Os resultados obtidos demonstraram que o implante do TAE induz produção de PGE2 durante toda sua evolução; aumento do número de células neoplásicas a partir do 10o dia e diminuição do influxo de células mesoteliais no 10º dia e de basófilos no 10º e 13º dia pós implante neoplásico. Em relação as citocinas o TAE induziu produção de IL-6 no 10º e 13º dia e de IL 2 no 13º dia, não alterando de modo significativo o perfil das outras citocinas estudadas. O tratamento de animais portadores de TAE com indometacina, foi eficaz em inibir o crescimento tumoral e a síntese de PGE2 a partir do 10o dia de crescimento neopásico, e promoveu aumento significativo no influxo de neutrófilos segmentados e de células nucleadas, apenas em tempos iniciais da evolução tumoral. Ainda, o tratamento com indometacina promoveu síntese de IL-13 e inibição significativa de IL-6 no 13o dia de crescimento tumoral, não alterando as outras citocinas analisadas. No grupo não portador de tumor tratado com indometacina observamos aumento no influxo de neutrófilos segmentados no 1º dia... .The aim of the present study was investigate the prostaglandin involvement during the growth of Ehrlich Ascites Tumor (EAT), using as parameters: tumoral growth, inflammatory influx and cytokine profile. Mice were inoculated with 1 x 103 tumor cells (ip) and treated with indomehacin (1mg/Kg,1x/day,ip) or diluent (0,1ml,1x/day,ip). After 1, 3, 6, 10 and 13 days the animals were sacrificed and evaluated in relation to inflammatory influx, secretion of TNF -a , IL1-a, IL-2, IL-4, IL-6, IL-10 and IL-13 and PGE2 level, in peritoneal cavity. Two groups no bearing EAT were treated with indomethacina or diluent as control ,following the same protocol. The results demonstrated that EAT implant induces PGE2 production during all evolution; increases tumoral cells number from the 10th day and decreases the mesotelial cells on 10th day and basophils cells on 10th and 13rd day.The cytokine profile showed EAT induces production of IL 6 from 10th day and of IL 2 on 13rd day, the other studied cytokines were not affected in a significant way. The indomethacin treatment of EAT bearing mice inhibited the tumoral growth and PGE2 synthesis from the 10th day and promoted significant increase on the neutrophils influx and total inflammatory cells, just in initial times of the tumoral evolution. Indomethacin treatment also promoted IL-13 synthesis and significant inhibition of IL-6 on 13rd day of EAT growth, but did not altered the others cytokines. The indomethacin treatment of animals do not bearing EAT increases neutrophils influx on the 1st day, lymphocytes on the 3rd day, eosinophils on 10th day; and no detected alteration was detected on cytokine profile Taken together, the results suggest that EAT growth is modulate by PGE2 and the inhibition of the tumoral growth could be partly related with suppression of IL-6 and liberation of IL-13.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Modulation of Cytokines Production by Indomethacin Acute Dose during the Evolution of Ehrlich Ascites Tumor in Mice

The aim of the present study was to investigate the influence of a nonselective COX1/COX2 inhibitor (indomethacin) on tumor growth of Ehrlich Ascites Tumor (EAT) in mice, using as parameters the tumor growth and cytokine profile. Mice were inoculated with EAT cells and treated with indomethacin. After 1, 3, 6, 10, and 13 days the animals were evaluated for the secretion of TNFα, IL-1α, IL-2, IL-4, IL-6, IL-10, and IL-13 and PGE2 level in peritoneal cavity. The results have shown that EAT induces PGE2 production and increases tumor cells number from the 10th day. The cytokine profile showed EAT induces production of IL-6 from 10th day and of IL-2 on 13th day; the other studied cytokines were not affected in a significant way. The indomethacin treatment of EAT-bearing mice inhibited the tumor growth and PGE2 synthesis from the 10th day. In addition, the treatment of EAT-bearing mice with indomethacin has stimulated the IL-13 production and has significantly inhibited IL-6 in the 13th day of tumor growth. Taken together, the results have demonstrated that EAT growth is modulated by PGE2 and the inhibition of the tumor growth could be partly related to suppression of IL-6 and induction of IL-13

Pelger-Huёt Anomaly in a Bitch Basenji

Background: Pelger-Huët anomaly (PHA) is characterized by morphological changes in all granulocytes, being more evident in neutrophils. Granulocytic function in these animals remains unchanged. Hereditary form of PHA should be differentiated from pseudo-PHA,a condition acquired from infections and/or inflammation conditions. Recognition of PHA is important to avoid misleading leukogram interpretation, since hyposegmentation of neutrophils can be confused with left shift, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The objective of this case report was to demonstrate the importance of laboratory diagnosis in PHA. Case: A 11-month-old bitch Basenji, was presented to perform preoperative evaluation for elective ovariosalpingohisterectomy at Veterinary Hospital (HUVET) of the Universidade Federal Fluminense (UFF). Tutor reported that animal was healthy, vaccination status was current, had deworming protocol applied and had not made use of medications recently. Animal presented normophagy, normodipsia, normuria and normochezia. Upon physical examination, animal was alert consciousness level, with adequate hydration, hyperemic oral mucosa, a less than 2 s capillary perfusion time, normal lymph nodes (submandibular, pre-scapular, inguinal and popliteal), rectal temperature of 39.2°C, heart rate of 160 beats per minute and respiratory rate of 60 movements per minute, possibly due to the animal’s agitated state. Abdominal palpation showed no changes. Physical examination presented no alterations. Preoperative exams included complete blood count (CBC) and biochemistry profile (ALT, AP, glucose, creatinine, urea, total proteins and fractions). Samples were sent to Hospital's Veterinary Clinical Pathology Laboratory (LABHUVET/UFF) for analysis. CBC was performed using automatic method. Blood smears were stained with hematological stain and then a cytomorphological evaluation was performed. The first CBC revealed 23% of neutrophils with nuclear hyposegmentation and 44% of neutrophils were bandsA follow up was performed after 9 months, and a Complete Blood Count was performed again in which 12% of neutrophils showed nuclear hyposegmentation with mature chromatin pattern, 40% of neutrophils were bands, 1% of meta-myelotcytes neutrophils, 1% of myelocytes neutrophils and, also, eosinophils with nuclear hyposegmentation. Animal was healthy, and had no alterations on physical examination suggesting a diagnosis of PHA. Discussion: Recognition of PHA is important to avoid misleading leukogram interpretation, since neutrophils hyposegmentation can be confused with left shift, which is considered severe with poor prognosis, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The diagnosis of PHA was considered by the shape of the leukocytes nuclei, without evidence of inflammatory disease, during the patient follow up. Therefore, this anomaly should be considered as a differential diagnosis of left shift, thus avoiding unnecessary clinical and therapeutic procedures. Guidance in face of this hereditary hematological syndrome is important. The responsible guardian of the animal must not allow it to act as a breeder in order to interrupt possible transmission of this anomaly to offspring, because there is a fatal form when it comes to homozygotes. Keywords: canine, dog, hereditary anomaly, WBC, nuclear hyposegmentation, neutrophils, left shift

Pelger-Huёt Anomaly in a Bitch Basenji

Background: Pelger-Huët anomaly (PHA) is characterized by morphological changes in all granulocytes, being more evident in neutrophils. Granulocytic function in these animals remains unchanged. Hereditary form of PHA should be differentiated from pseudo-PHA,a condition acquired from infections and/or inflammation conditions. Recognition of PHA is important to avoid misleading leukogram interpretation, since hyposegmentation of neutrophils can be confused with left shift, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The objective of this case report was to demonstrate the importance of laboratory diagnosis in PHA. Case: A 11-month-old bitch Basenji, was presented to perform preoperative evaluation for elective ovariosalpingohisterectomy at Veterinary Hospital (HUVET) of the Universidade Federal Fluminense (UFF). Tutor reported that animal was healthy, vaccination status was current, had deworming protocol applied and had not made use of medications recently. Animal presented normophagy, normodipsia, normuria and normochezia. Upon physical examination, animal was alert consciousness level, with adequate hydration, hyperemic oral mucosa, a less than 2 s capillary perfusion time, normal lymph nodes (submandibular, pre-scapular, inguinal and popliteal), rectal temperature of 39.2°C, heart rate of 160 beats per minute and respiratory rate of 60 movements per minute, possibly due to the animal’s agitated state. Abdominal palpation showed no changes. Physical examination presented no alterations. Preoperative exams included complete blood count (CBC) and biochemistry profile (ALT, AP, glucose, creatinine, urea, total proteins and fractions). Samples were sent to Hospital's Veterinary Clinical Pathology Laboratory (LABHUVET/UFF) for analysis. CBC was performed using automatic method. Blood smears were stained with hematological stain and then a cytomorphological evaluation was performed. The first CBC revealed 23% of neutrophils with nuclear hyposegmentation and 44% of neutrophils were bandsA follow up was performed after 9 months, and a Complete Blood Count was performed again in which 12% of neutrophils showed nuclear hyposegmentation with mature chromatin pattern, 40% of neutrophils were bands, 1% of meta-myelotcytes neutrophils, 1% of myelocytes neutrophils and, also, eosinophils with nuclear hyposegmentation. Animal was healthy, and had no alterations on physical examination suggesting a diagnosis of PHA. Discussion: Recognition of PHA is important to avoid misleading leukogram interpretation, since neutrophils hyposegmentation can be confused with left shift, which is considered severe with poor prognosis, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The diagnosis of PHA was considered by the shape of the leukocytes nuclei, without evidence of inflammatory disease, during the patient follow up. Therefore, this anomaly should be considered as a differential diagnosis of left shift, thus avoiding unnecessary clinical and therapeutic procedures. Guidance in face of this hereditary hematological syndrome is important. The responsible guardian of the animal must not allow it to act as a breeder in order to interrupt possible transmission of this anomaly to offspring, because there is a fatal form when it comes to homozygotes. Keywords: canine, dog, hereditary anomaly, WBC, nuclear hyposegmentation, neutrophils, left shift

Metabolic Symbiosis and Immunomodulation: How Tumor Cell-Derived Lactate May Disturb Innate and Adaptive Immune Responses

The tumor microenvironment (TME) is composed by cellular and non-cellular components. Examples include the following: (i) bone marrow-derived inflammatory cells, (ii) fibroblasts, (iii) blood vessels, (iv) immune cells, and (v) extracellular matrix components. In most cases, this combination of components may result in an inhospitable environment, in which a significant retrenchment in nutrients and oxygen considerably disturbs cell metabolism. Cancer cells are characterized by an enhanced uptake and utilization of glucose, a phenomenon described by Otto Warburg over 90 years ago. One of the main products of this reprogrammed cell metabolism is lactate. “Lactagenic” or lactate-producing cancer cells are characterized by their immunomodulatory properties, since lactate, the end product of the aerobic glycolysis, besides acting as an inducer of cellular signaling phenomena to influence cellular fate, might also play a role as an immunosuppressive metabolite. Over the last 10 years, it has been well accepted that in the TME, the lactate secreted by transformed cells is able to compromise the function and/or assembly of an effective immune response against tumors. Herein, we will discuss recent advances regarding the deleterious effect of high concentrations of lactate on the tumor-infiltrating immune cells, which might characterize an innovative way of understanding the tumor-immune privilege