Matrix metalloproteinase 9 expression in canine mammary carcinomas

Mammary neoplasms are among the most common canine tumors, with high risk of invasion and metastasis. Important steps for these events are the loss of cell adhesion to the main tumor mass and extracellular matrix degradation. Matrix metalloproteinases (MMPs) are proteolytic enzymes containing zinc, which are capable of degrading and remodeling the surrounding extracellular matrix, facilitating these events. Involvement of MMPs has been demonstrated in many pathological processes, as well as in human and canine tumors, which has been related to malignancy and prognosis. The aim of this study was to characterize the immunohistochemical expression of matrix metalloproteinase 9 (MMP-9) in canine mammary gland tumors, as well as to verify its relation with the different histologic patterns. Thirty-one of the 41 tumors (75.61%) were positive. Twenty-two samples (70.97%) had diffuse cytoplasmic immunostaining and 9 (29.03%) had a finely granular pattern. Immunostaining intensity was strong in 21 (67.74%) tumors and weak in 10 (32.26%). No statistically significant differences were found between anaplastic carcinomas, carcinomas in a mixed tumor and simple carcinomas regarding positivity (p=0.9707), intensity (p=0.5386) and staining pattern (0.6135); between solid carcinomas and simple papillary carcinomas for positivity (p=0,7333), intensity (p=0.7333) and staining pattern (p=0.3037); or betweensolid carcinomas and simple tubular and papillary carcinomas for positivity (p=0.9682), intensity (p=0.8450) and staining pattern (p=0.5068). MMP-9 was detected with variable intensity and morphological patterns of cytoplasmic staining. However significant statistic differences were not found between the histological types or histopathological grades.

Matrix metalloproteinase 9 expression in canine mammary carcinomas

Mammary neoplasms are among the most common canine tumors, with high risk of invasion and metastasis. Important steps for these events are the loss of cell adhesion to the main tumor mass and extracellular matrix degradation. Matrix metalloproteinases (MMPs) are proteolytic enzymes containing zinc, which are capable of degrading and remodeling the surrounding extracellular matrix, facilitating these events. Involvement of MMPs has been demonstrated in many pathological processes, as well as in human and canine tumors, which has been related to malignancy and prognosis. The aim of this study was to characterize the immunohistochemical expression of matrix metalloproteinase 9 (MMP-9) in canine mammary gland tumors, as well as to verify its relation with the different histologic patterns. Thirty-one of the 41 tumors (75.61%) were positive. Twenty-two samples (70.97%) had diffuse cytoplasmic immunostaining and 9 (29.03%) had a finely granular pattern. Immunostaining intensity was strong in 21 (67.74%) tumors and weak in 10 (32.26%). No statistically significant differences were found between anaplastic carcinomas, carcinomas in a mixed tumor and simple carcinomas regarding positivity (p=0.9707), intensity (p=0.5386) and staining pattern (0.6135); between solid carcinomas and simple papillary carcinomas for positivity (p=0,7333), intensity (p=0.7333) and staining pattern (p=0.3037); or betweensolid carcinomas and simple tubular and papillary carcinomas for positivity (p=0.9682), intensity (p=0.8450) and staining pattern (p=0.5068). MMP-9 was detected with variable intensity and morphological patterns of cytoplasmic staining. However significant statistic differences were not found between the histological types or histopathological grades.(AU)

Evaluation of matrix metalloproteinases in blood from buck naturally infected with Caprine Arthritis Encephalitis in Semiarid region of Brazil / Avaliação das metaloproteinases de matriz no sangue de reprodutores caprinos naturalmente infectados com Artrite Encefalite Caprina na Região Semiárida do Brasil

Background: Studies have attempted to understand the importance of metalloproteinase (MMPs) in the pathogenesis ofdiseases caused by lentiviruses, being the human immunodeficiency virus (HIV) the most investigated. Despite advancesin studies with MMPs in others diseases, research correlating the presence and activity of gelatinases in animals affectedby caprine arthritis encephalitis virus (CAEV), a lentiviruses, are incipient and there is a need for research aiming to understand the dynamic of these enzymes in animals infected and its relation to pathological condition. The aim of this studywas to evaluate the presence and activity of the MMPs in blood serum of chronically infected bucks by CAEV.Materials, Methods & Results: The experiment was constituted by two groups (n = 5 each group). The first one wascomposed of five naturally infected bucks (4-5 years) and second group constituted of five seronegative bucks (3-4 years)for CAE. Serology was performed using the Western Blotting (WB) and confirmed by Polymerase Chain Reaction (PCR).These bucks belong to the experimental flock at Embrapa Goats and Sheep and the seropositive bucks were confirmed forCAE in the first two years. Blood samples were collected by puncturing the jugular vein from animals and evaluated byzymography (SDS-PAGE) using gelatin as substratum. Clinical examination was performed by evaluating the temperature (T), cardiac frequency (FC), respiratory frequency (FR) and clinical articular index (IAC). The IAC was calculatedby measuring the circumference of carpal joint and metacarpal bone height (difference between greater extent carpal andmetacarpal lesser extent). In infected and control groups were found molecular mass bands of 66 kDa (MMP-2), 72 kDa(pro-MMP-2), 86 kDa (MMP-9) and 92 kDa (pro-MMP-9). The correlation...(AU)

Evaluation of matrix metalloproteinases in blood from buck naturally infected with Caprine Arthritis Encephalitis in Semiarid region of Brazil / Avaliação das metaloproteinases de matriz no sangue de reprodutores caprinos naturalmente infectados com Artrite Encefalite Caprina na Região Semiárida do Brasil

Background: Studies have attempted to understand the importance of metalloproteinase (MMPs) in the pathogenesis ofdiseases caused by lentiviruses, being the human immunodeficiency virus (HIV) the most investigated. Despite advancesin studies with MMPs in others diseases, research correlating the presence and activity of gelatinases in animals affectedby caprine arthritis encephalitis virus (CAEV), a lentiviruses, are incipient and there is a need for research aiming to understand the dynamic of these enzymes in animals infected and its relation to pathological condition. The aim of this studywas to evaluate the presence and activity of the MMPs in blood serum of chronically infected bucks by CAEV.Materials, Methods & Results: The experiment was constituted by two groups (n = 5 each group). The first one wascomposed of five naturally infected bucks (4-5 years) and second group constituted of five seronegative bucks (3-4 years)for CAE. Serology was performed using the Western Blotting (WB) and confirmed by Polymerase Chain Reaction (PCR).These bucks belong to the experimental flock at Embrapa Goats and Sheep and the seropositive bucks were confirmed forCAE in the first two years. Blood samples were collected by puncturing the jugular vein from animals and evaluated byzymography (SDS-PAGE) using gelatin as substratum. Clinical examination was performed by evaluating the temperature (T), cardiac frequency (FC), respiratory frequency (FR) and clinical articular index (IAC). The IAC was calculatedby measuring the circumference of carpal joint and metacarpal bone height (difference between greater extent carpal andmetacarpal lesser extent). In infected and control groups were found molecular mass bands of 66 kDa (MMP-2), 72 kDa(pro-MMP-2), 86 kDa (MMP-9) and 92 kDa (pro-MMP-9). The correlation...

AVALIAÇÃO IN VIVO DO EFEITO DA CICLOFOSFAMIDA NO DESENVOLVIMENTO DO MELANOMA MURINO

A ciclofosfamida (CPA) é um quimioterápico antineoplásico que possui efeitos secundários em componentes do sistema imunológico. Devido sua ação imunomoduladora, capaz de influenciar respostas imunidade humoral e imunidade mediada por células, este fármaco é também prescrito para tratamento de doenças autoimunes. Sua utilização por longos períodos de tempo no tratamento destas doenças já foi correlacionado com o aumento de risco para desenvolvimento de câncer de pele não melanoma, porém este aumento ainda não foi associado ao melanoma. A capacidade de modular a resposta imunológica pode também favorecer ou inibir o desenvolvimento de tumores. No melanoma, estudos demonstraram que a CPA pode favorecer uma resposta antitumoral, no entanto, não há relatos sobre a influência deste fármaco no desenvolvimento da doença quando utilizada anterior ao seu desenvolvimento. Considerando os efeitos controversos da CPA no desenvolvimento de tumores, o intuito deste trabalho foi avaliar os efeitos do pré-tratamento com CPA sob o crescimento e progressão do melanoma murino, abordando tanto os mecanismos celulares, como moleculares. Para isto, camundongos C57BL/6 foram pré-tratados com CPA e, posteriormente, inoculados com células B16F10 na região subcutânea do dorso. Resultados obtidos no presente estudo e apresentados em formato de artigo que será submetido à revista Journal of Immunology Research, sugerem que tratamento com CPA reduz população de neutrófilos e macrófagos no microambiente tumoral, o que pode ser um dos mecanismos pelos quais o tratamento com CPA reduz MMP-9 e, consequentemente, reduz o crescimento e progressão do melanoma, melhorando a sobrevida destes animais.

Cyclophosphamide (CPA) is an antineoplastic chemotherapeutic that has an effect on the immune system. Due to its immunomodulatory action, capable of influencing humoral immunity responses and cell mediated immunity, this drug is also prescribed for the treatment of autoimmune diseases. Its use for long periods of time in the treatment of these diseases has already been correlated with the increased chances for development of non-melanoma skin cancer, however this increase was not associated with melanoma. The ability to modulate the immune response may also favor or inhibit the development of tumors In melanoma, studies have shown that CPA may favor an antitumor response, however, there are no reports on the influence of this drug on the development of the disease when used prior to its development. Considering the controversial effects of CPA on tumor development, the aim of this study was to evaluate the effects of pre-treatment with CPA under the growth and progression of murine melanoma, addressing both cellular and molecular mechanisms. For this, C57BL/6 mice were pre-treated with the drug and subsequently inoculated with B16F10 cells in the subcutaneous region of the dorsum. Results obtained in the present study and presented in an article format that will be submitted to the Journal of Immunology Research, suggest that CPA treatment reduces the neutrophil and macrophage population in the tumor microenvironment which may be one of the mechanisms by which CPA treatment reduces MMP-9 and consequently reduces the growth and progression of melanoma, improving the survival of these animals.

Ascorbic acid improve the survival and in vitro growth of isolated caprine preantral follicles

The present study aims to investigate the influence of two concentrations of ascorbic acid on the survival, growth , antral formation and m RNA expression of the matrix metalloproteinases - 9 (MMP - 9) and their tissue inhibitor - 2 (TIMP - 2) on caprine preantral follicles during long - term in vitro culture. Isolated preantral follicles were individ ually cultured without or with ascorbic acid at 50 μ g/ml (AA50) or 100 μ g/ml (AA100) during 18 days. The parameters evaluated were follicular viability, growth, antrum formation and extruded oocytes. The genes MMP - 9 and TIMP - 2 were quantified by real - time polymer ase chain reaction (qPCR) after 18 days of culture in the control medium (MEM + ) or ascorbic acid (50 or 100 μ g/ml) and in fresh control (non cultured) . At the end of culture, AA50 significantly increased the percentage of viable follicles compared w ith other treatments . Moreover, mean daily increase in follicular diameter (μm/day) was significantly higher in the presence of both concentrations of ascorbic acid than in MEM + alone. Higher rates of antral formation and lower percentages of extruded oocy tes were observed in medium containing AA50 compared with control medium. Real Time RT - PCR assays showed that AA50 increase s MMP - 9 expression significantly compared with fresh control and MEM + alone. In conclusion, ascorbic acid at 50 μ g/ml was very import ant for the maintenance of caprine preantral follicle viability and development after in vitro culture and influences in vitro the enzymes involved with basement membrane remodeling.

Ascorbic acid improve the survival and in vitro growth of isolated caprine preantral follicles

The present study aims to investigate the influence of two concentrations of ascorbic acid on the survival, growth , antral formation and m RNA expression of the matrix metalloproteinases - 9 (MMP - 9) and their tissue inhibitor - 2 (TIMP - 2) on caprine preantral follicles during long - term in vitro culture. Isolated preantral follicles were individ ually cultured without or with ascorbic acid at 50 μ g/ml (AA50) or 100 μ g/ml (AA100) during 18 days. The parameters evaluated were follicular viability, growth, antrum formation and extruded oocytes. The genes MMP - 9 and TIMP - 2 were quantified by real - time polymer ase chain reaction (qPCR) after 18 days of culture in the control medium (MEM + ) or ascorbic acid (50 or 100 μ g/ml) and in fresh control (non cultured) . At the end of culture, AA50 significantly increased the percentage of viable follicles compared w ith other treatments . Moreover, mean daily increase in follicular diameter (μm/day) was significantly higher in the presence of both concentrations of ascorbic acid than in MEM + alone. Higher rates of antral formation and lower percentages of extruded oocy tes were observed in medium containing AA50 compared with control medium. Real Time RT - PCR assays showed that AA50 increase s MMP - 9 expression significantly compared with fresh control and MEM + alone. In conclusion, ascorbic acid at 50 μ g/ml was very import ant for the maintenance of caprine preantral follicle viability and development after in vitro culture and influences in vitro the enzymes involved with basement membrane remodeling.(AU)