Resumo
Hematological and biochemical alterations in animals with neoplasms may result from the direct effects of tumor growth or paraneoplastic syndromes. The objective of this study was to evaluate these hematological and biochemical alterations in female dogs with mammary tumors and with inflammatory carcinoma (IC). Blood samples were collected from 43 female dogs divided into three groups according to clinical staging: Group 1 (G1) - animals in initial stage (T1,2,3N0M0, N=17), Group 2 (G2) - animals in advanced stage (T1,2,3N1M0,1, N=15) and Group 3 (G3) - animals presenting IC (N=11). Hematological and biochemical parameters obtained were related to patients' clinical staging. Among alterations, the most common were anemia, neutrophilic leukocytosis, monocytosis, increased ALT, AST, and hypoalbuminemia, mainly in dogs in advanced clinical staging and with inflammatory carcinoma.
Aterações hematológicas e bioquímicas em animais portadores de neoplasias podem resultar dos efeitos diretos do crescimento tumoral ou de síndromes paraneoplásicas. O objetivo deste estudo foi avaliar as alterações hematológicas e bioquímicas em cadelas com neoplasias mamárias e carcinoma inflamatório (IC). Foram coletadas amostras de sangue de 43 cadelas divididas em três grupos de acordo com o estadiamento clínico: grupo 1 (G1) - cadelas em estádio inicial (T1,2,3N0M0, N=17), grupo 2 (G2) - cadelas em estádio avançado (T1,2,3N1M0,1, N=15) e grupo 3 (G3) - cadelas que apresentaram IC (N=11). Os parâmetros hematológicos e bioquímicos obtidos foram relacionados com o estadiamento clínico das pacientes. Entre as alterações, as mais comuns foram anemia, leucocitose neutrofílica, monocitose, aumento de ALT, AST e hipoalbuminemia, principalmente em cadelas em estadiamento clínico avançado e com carcinoma inflamatório.
Assuntos
Animais , Cães , Fenômenos Bioquímicos , Contagem de Células Sanguíneas , Neoplasias da Mama , Carcinoma , Doenças do Cão , AnemiaResumo
In breast cancer treatment, chemotherapy resistance is a major problem where many receptive tumors rebound and develop resistance. When provided in combination, cancer drugs are most successful, thus reducing the risk of developing resistant cancer cells. However, the evaluation of combination therapies has increased rapidly in recent years. Consequently, by repurposing old treatments, the discovery of additional medicines that may interact synergistically with chemotherapy is considered a current medical aim through discovering a new cancer medication or therapeutic strategy. The purpose of this research is to increase the anti-cancer activity of carboplatin (CP) by increasing the apoptotic effect of breast cancer cells (MCF-7) during in vitro experiments in combination with oxytetracycline. Our results showed a high synergistic effect between oxytetracycline and carboplatin, MCF-7 representative cell treated with carboplatin with/without different concentrations of oxytetracycline (5% and 10% of IC50). Oxytetracycline, which potentiated the action of carboplatin and/or had notable activity was reported as a single agent. This research demonstrated the synergistic relationship between oxytetracycline and carboplatin in viability assays. Surprisingly, our findings suggest that inhibiting treatment strategies can extend carboplatin's therapeutic window, potentially allowing for cancer therapy.(AU)
No tratamento do câncer de pulmão a resistência à quimioterapia é o maior problema no qual muitos tumores receptivos apresentam um rebote e desenvolvem a resistência. Quando oferecidas em combinações, as drogas anticancerígenas apresentam maior taxa de sucesso, reduzindo assim o risco de desenvolvimento de células cancerígenas resistentes. Contudo, a avaliação das terapias de combinação tem crescido muito rapidamente. Consequentemente, por reaproveitamento de tratamentos antigos, a descoberta de novos medicamentos adicionais que podem interagir sinergicamente com a quimioterapia que é considerada como auxílio médico na corrente busca à descoberta de novas medicações anticancerígenas ou estratégias terapêuticas. O propósito da presente pesquisa é aumentar a atividade anticancerígena da Carboplatina (CP) pelo incremento do efeito apoptótico de células de câncer pulmonar (MCF-7) em experimentos in vitro pela combinação com oxitetraciclina. Os resultados obtidos confirmaram elevado efeito sinérgico entre oxitetraciclina e Carboplatina em células MCF-7 representativas tratadas com Carboplatina, com e sem diferentes concentrações de oxitetraciclina (5% e 10% de IC50). A oxitetracilina que potencializou a ação da Carboplatina e/ou teve uma notável atividade relatada como um agente isolado. A pesquisa demonstrou a relação sinérgica entre oxitetraxiclina e Carboplatina nos ensaios de viabilidade. Surpreendentemente, os resultados obtidos sugeriram que as estratégias de tratamento inibidor podem aplicar um janela terapêutica da Carboplatina com potencial para a terapia do câncer.(AU)
Assuntos
Oxitetraciclina/farmacologia , Carboplatina/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Técnicas In Vitro/veterinária , Sinergismo Farmacológico , Células MCF-7Resumo
Although feline mammary carcinoma is not the most prevalent among the species, its aggressive behavior represents a low life expectancy, compared with most undifferentiated types of breast cancer. Tissue stiffness induced by the accumulation of collagen fibers is related to a risk factor for carcinogenesis in healthy women and aggressiveness in those with breast cancer, which can also occur in cats. The objective of this work is to identify the relationship between stromal collagen density and aggressiveness of mammary carcinoma in cats, according to the peripheral and central tissue distribution by the Picrossirius Red histochemical method. Image.J® and MatLab® software were used for digital image processing. The mean values of kurtosis and entropy attributes were grouped into a control group, and low and high-grade carcinoma groups, analyzed with one-way ANOVA and Bonferronis multiple comparison test (p <0.01). Interpretation of stromal dynamics is important to evaluate both central and peripheral locations. According to entropy, there was a significant increase in the peripheral density in the carcinoma groups in relation to the control group, which can be justified by blood support. The same can be said of the central region, with a significant gain in collagen fibers from the tumors, indicated by kurtosis. The results suggest the presence of increases in stromal density in mammary carcinomas of cats, regardless of their graduation, and occurring in both regions.
Assuntos
Animais , Gatos , Colágeno/análise , Gatos , Imuno-Histoquímica , Neoplasias Mamárias AnimaisResumo
Although feline mammary carcinoma is not the most prevalent among the species, its aggressive behavior represents a low life expectancy, compared with most undifferentiated types of breast cancer. Tissue stiffness induced by the accumulation of collagen fibers is related to a risk factor for carcinogenesis in healthy women and aggressiveness in those with breast cancer, which can also occur in cats. The objective of this work is to identify the relationship between stromal collagen density and aggressiveness of mammary carcinoma in cats, according to the peripheral and central tissue distribution by the Picrossirius Red histochemical method. Image.J® and MatLab® software were used for digital image processing. The mean values of kurtosis and entropy attributes were grouped into a control group, and low and high-grade carcinoma groups, analyzed with one-way ANOVA and Bonferronis multiple comparison test (p <0.01). Interpretation of stromal dynamics is important to evaluate both central and peripheral locations. According to entropy, there was a significant increase in the peripheral density in the carcinoma groups in relation to the control group, which can be justified by blood support. The same can be said of the central region, with a significant gain in collagen fibers from the tumors, indicated by kurtosis. The results suggest the presence of increases in stromal density in mammary carcinomas of cats, regardless of their graduation, and occurring in both regions.(AU)
Assuntos
Animais , Gatos , Gatos , Colágeno/análise , Neoplasias Mamárias Animais , Imuno-HistoquímicaResumo
The present work evaluated the immunomodulatory effect of thalidomide (Thal) at different doses on tumor-associated macrophages (TAMs) using a mouse model of human breast cancer. Mice were inoculated with 4T1 cells in the left flank and treated with Thal once a day at concentrations of 50, 100, and 150mg/kg body weight from the 5th day until the 28th day of tumor inoculation. The tumors were sized, proliferation index and TAMs count were evaluated in primary tumors and metastatic lungs. In addition, the metastasis rate was evaluated in the lungs. Thal at 150mg/kg significantly decreased tumor growth, proliferation index, and TAMs infiltration in primary tumors. Conversely, a higher number of TAMs and lower proliferation index were observed in metastatic lungs in mice treated with 150mg/kg of Thal. Furthermore, Thal at 150mg/kg significantly decreased the metastatic nodules in the lungs. Our findings demonstrated that Thal treatment considerably decreased the primary tumor and lung metastasis in mice associated with different TAM infiltration effects in these sites.(AU)
No presente trabalho, foi avaliado o efeito imunomodulador de diferentes doses de talidomida em macrófagos associados ao tumor (TAMs), em um modelo murino de câncer de mama. Camundongos foram inoculados com células 4T1, na região do flanco esquerdo, e tratados com talidomida, uma vez ao dia, nas doses de 50, 100 e 150mg/k, por massa corporal, do quinto dia ao 28º dia de inoculação tumoral. Os tumores foram medidos, o índice de proliferação celular e a contagem de TAMs foram avaliados nos tumores primários e nos pulmões com metástases. Além disso, a taxa de metástases pulmonares também foi avaliada. A talidomida na dose de 150mg/kg diminuiu significativamente o crescimento tumoral, o índice de proliferação celular e a infiltração de TAMs nos tumores primários. Por outro lado, maior número de TAMs e menor índice de proliferação celular foram observados nos pulmões metastáticos, em camundongos tratados com 150mg/kg de talidomida. Ademais, a talidomida na dose de 150mg/kg diminuiu significativamente os nódulos metastáticos nos pulmões. Os resultados demonstraram que o tratamento com talidomida diminuiu o crescimento tumoral e as metástases pulmonares em camundongos, associado com diferentes efeitos na infiltração de TAMs nesses locais.(AU)
Assuntos
Animais , Camundongos , Talidomida/análise , Neoplasias Mamárias Animais/tratamento farmacológico , Macrófagos/efeitos dos fármacos , Imunomodulação , Metástase NeoplásicaResumo
Background: Studies pointed out that the tumor-infiltrating lymphocytes (TILs) have considerable importance in caninemammary tumor (CMT). On the other hand, cancer cells sometimes find ways to use immune checkpoint proteins as ashield to avoid being identified and attacked by the immune system as programmed death 1 ligand 1 (PD-L1). In this study,it was investigated the relationship between PD-L1 expression, stromal tumor-infiltrating lymphocytes (TILs) in caninemammary tumor (CMT), and the association with clinical and pathological characteristics of the tumors.Materials, Methods & Results: PD-L1 expression and TILs were assessed in 23 female dogs with CMT. The tumors weregrouped into simple carcinoma (CA, n = 8) and complex carcinoma (CC, n = 15). Stromal TILs were assessed using twothresholds as TILs-Low representing < 50% of infiltrate within stromal area and TILs-High representing ≥ 50% of stromalarea. Clinicopathological data of CMT was characterized according to key parameters, as well as survival rates. TILs evaluation within tumor stroma revealed that 65.2% (n = 15) of tumors had TILs-Low. PD-L1 expression and stromal TILs weresignificantly associated (P = 0.009). PD-L1 expression was observed in 39% (n = 9) of all tumors of which 17.4% (n = 4)were from CA group and 21.7% (n = 5) were from CC group. PD-L1 expression within TILs was observed in 39% (n =9) of the tumors. PD-L1 in malignant epithelium was present in all lymph node metastasis (n = 5). PD-L1 was associatedwith involvement of regional lymph nodes (P = 0.034). Survival curves demonstrated TILs-Low had higher (P = 0.010)overall survival (OS) compared with TILs-High, and PD-L1+ and PD-L1 (P = 0.06) did not differed. The clinicopathological variables significantly correlated with OS by univariate analysis were the histological grade (P = 0.009), lymphnode involvement (P = 0.004), stromal...
Assuntos
Feminino , Animais , Cães , Cães , Linfócitos , Neoplasias da Mama/veterinária , Proteínas de Membrana , Mastectomia Segmentar/veterináriaResumo
Background: Studies pointed out that the tumor-infiltrating lymphocytes (TILs) have considerable importance in caninemammary tumor (CMT). On the other hand, cancer cells sometimes find ways to use immune checkpoint proteins as ashield to avoid being identified and attacked by the immune system as programmed death 1 ligand 1 (PD-L1). In this study,it was investigated the relationship between PD-L1 expression, stromal tumor-infiltrating lymphocytes (TILs) in caninemammary tumor (CMT), and the association with clinical and pathological characteristics of the tumors.Materials, Methods & Results: PD-L1 expression and TILs were assessed in 23 female dogs with CMT. The tumors weregrouped into simple carcinoma (CA, n = 8) and complex carcinoma (CC, n = 15). Stromal TILs were assessed using twothresholds as TILs-Low representing < 50% of infiltrate within stromal area and TILs-High representing ≥ 50% of stromalarea. Clinicopathological data of CMT was characterized according to key parameters, as well as survival rates. TILs evaluation within tumor stroma revealed that 65.2% (n = 15) of tumors had TILs-Low. PD-L1 expression and stromal TILs weresignificantly associated (P = 0.009). PD-L1 expression was observed in 39% (n = 9) of all tumors of which 17.4% (n = 4)were from CA group and 21.7% (n = 5) were from CC group. PD-L1 expression within TILs was observed in 39% (n =9) of the tumors. PD-L1 in malignant epithelium was present in all lymph node metastasis (n = 5). PD-L1 was associatedwith involvement of regional lymph nodes (P = 0.034). Survival curves demonstrated TILs-Low had higher (P = 0.010)overall survival (OS) compared with TILs-High, and PD-L1+ and PD-L1(P = 0.06) did not differed. The clinicopathological variables significantly correlated with OS by univariate analysis were the histological grade (P = 0.009), lymphnode involvement (P = 0.004), stromal...(AU)
Assuntos
Animais , Feminino , Cães , Cães , Neoplasias da Mama/veterinária , Linfócitos , Proteínas de Membrana , Mastectomia Segmentar/veterináriaResumo
Background: Cancer is the second most common fatal disease in the world, behind cardiovascular disorders in the first place. It accounts for around 0.3 million deaths per year in India due to the lack of proper diagnostic facilities, prevention and treatment. Current therapeutic methods do not provide adequate protection and affect normal cells along with cancerous ones. Thus, there is a need for some alternative therapeutic strategy, preferably from natural products, which have been traditionally used for treatment of various diseases in the country. Methods: In this study, we have conjugated purified NN-32 toxin from Naja naja venom with gold nanoparticles and its anticancer potential was evaluated against human breast cancer cell lines. UV-Vis spectroscopy, dynamic light scattering, transmission electron microscopy, atomic force microscopy and zeta potential analysis were the techniques used for characterization of GNP-NN-32. Results: GNP-NN-32 showed dose- and time-dependent cytotoxicity against breast cancer cell lines (MCF-7 and MDA-MB-231). NN-32 and GNP-NN-32 induced apoptosis in both breast cancer cell lines. The results of CFSE cell proliferation study revealed that NN-32 and GNP-NN-32 arrested cell division in both MCF-7 and MDA-MB-231 cell lines resulting in inhibition of proliferation of these cancer cells. Conclusion: GNP-NN-32 showed an anticancer potential against human breast cancer cell lines. Analysis of detailed chemical characterization along with its cytotoxic property might help to perceive a new dimension of the anti-cancer potential of GNP-NN-32 that will enhance its biomedical function in near future.(AU)
Assuntos
Animais , Venenos Elapídicos , Naja naja , AntineoplásicosResumo
Background:Cancer is the second most common fatal disease in the world, behind cardiovascular disorders in the first place. It accounts for around 0.3 million deaths per year in India due to the lack of proper diagnostic facilities, prevention and treatment. Current therapeutic methods do not provide adequate protection and affect normal cells along with cancerous ones. Thus, there is a need for some alternative therapeutic strategy, preferably from natural products, which have been traditionally used for treatment of various diseases in the country.Methods:In this study, we have conjugated purified NN-32 toxin from Naja naja venom with gold nanoparticles and its anticancer potential was evaluated against human breast cancer cell lines. UV-Vis spectroscopy, dynamic light scattering, transmission electron microscopy, atomic force microscopy and zeta potential analysis were the techniques used for characterization of GNP-NN-32.Results:GNP-NN-32 showed dose- and time-dependent cytotoxicity against breast cancer cell lines (MCF-7 and MDA-MB-231). NN-32 and GNP-NN-32 induced apoptosis in both breast cancer cell lines. The results of CFSE cell proliferation study revealed that NN-32 and GNP-NN-32 arrested cell division in both MCF-7 and MDA-MB-231 cell lines resulting in inhibition of proliferation of these cancer cells.Conclusion:GNP-NN-32 showed an anticancer potential against human breast cancer cell lines. Analysis of detailed chemical characterization along with its cytotoxic property might help to perceive a new dimension of the anti-cancer potential of GNP-NN-32 that will enhance its biomedical function in near future.(AU)
Assuntos
Animais , Naja naja , Venenos Elapídicos/uso terapêutico , Anticarcinógenos/análise , Nanopartículas , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/terapia , Citometria de FluxoResumo
Background: Mammary tumours represent about 50 to 70% of all neoplasms in female dogs and their occurrence is directlyrelated to the reproductive status and patient´s age. The purpose of this research was to apply the Brazilian consensus ondiagnosis, prognosis and treatment of canine mammary tumours and to define the regional epidemiological aspects ofcanine mammary gland tumours in Vitoria metropolitan region (ES, Brazil) between 2012 and 2016 and to correlate themacroscopic characteristics such as lesion size and location of the neoplasm with histopathological diagnosis, tumoursgrade and lymph node metastasis.Materials, Methods & Results: Data were collected from the archives of the Laboratory of Animal Pathology of UVV andmedical records of patients attended at the Veterinary Hospital Prof. Ricardo Alexandre Hippler in 5 years (2012 to 2016). Theanimals were separated into groups by age to facilitate classification in the group with the highest occurrence of neoplasms. Theevaluation of the macroscopic characteristics was performed through the histopathological record described in the pathologylaboratory, for each patient, at the time of the initial evaluation. After descriptive analysis, data was correlated using Spearmanntest, and frequency dispersion was evaluated using chi-square test, both in the software Graph Pad Prism v. 6.01. This studyincluded 255 bitches and diagnosis of 486 lesions, once 48.6% of the dogs had more than one lesion, classified according tothe Brazilian Consensus for Canine Mammary Tumours and graded according to Elston and Ellis system. 86.8% of lesionswere consistent with actual mammary neoplasms, of which 67% were malignant and 20% were benign. Non-neoplastic lesionscorresponded to 7.2% of cases and 5.8% were extra-mammary neoplasms, with an increased incidence of lipomas (39.3%) andmast cell tumours (32.1%). Cross-breed dogs represented 26.7% of cases. Poodles (25.5%), Pinschers (9.8%) and Dachshund(4.7%)...(AU)
Assuntos
Animais , Feminino , Cães , Neoplasias Mamárias Animais/epidemiologia , Estadiamento de Neoplasias/veterinária , Técnicas Histológicas/veterinária , Fatores EtáriosResumo
Background: Mammary tumours represent about 50 to 70% of all neoplasms in female dogs and their occurrence is directlyrelated to the reproductive status and patient´s age. The purpose of this research was to apply the Brazilian consensus ondiagnosis, prognosis and treatment of canine mammary tumours and to define the regional epidemiological aspects ofcanine mammary gland tumours in Vitoria metropolitan region (ES, Brazil) between 2012 and 2016 and to correlate themacroscopic characteristics such as lesion size and location of the neoplasm with histopathological diagnosis, tumoursgrade and lymph node metastasis.Materials, Methods & Results: Data were collected from the archives of the Laboratory of Animal Pathology of UVV andmedical records of patients attended at the Veterinary Hospital Prof. Ricardo Alexandre Hippler in 5 years (2012 to 2016). Theanimals were separated into groups by age to facilitate classification in the group with the highest occurrence of neoplasms. Theevaluation of the macroscopic characteristics was performed through the histopathological record described in the pathologylaboratory, for each patient, at the time of the initial evaluation. After descriptive analysis, data was correlated using Spearmanntest, and frequency dispersion was evaluated using chi-square test, both in the software Graph Pad Prism v. 6.01. This studyincluded 255 bitches and diagnosis of 486 lesions, once 48.6% of the dogs had more than one lesion, classified according tothe Brazilian Consensus for Canine Mammary Tumours and graded according to Elston and Ellis system. 86.8% of lesionswere consistent with actual mammary neoplasms, of which 67% were malignant and 20% were benign. Non-neoplastic lesionscorresponded to 7.2% of cases and 5.8% were extra-mammary neoplasms, with an increased incidence of lipomas (39.3%) andmast cell tumours (32.1%). Cross-breed dogs represented 26.7% of cases. Poodles (25.5%), Pinschers (9.8%) and Dachshund(4.7%)...
Assuntos
Feminino , Animais , Cães , Estadiamento de Neoplasias/veterinária , Neoplasias Mamárias Animais/epidemiologia , Fatores Etários , Técnicas Histológicas/veterináriaResumo
Breast cancer is the neoplasm with both the highest incidence and mortality rate among women worldwide. Given the known snake venom cytotoxicity towards several tumor types, we evaluated the effects of BthTX-I from Bothrops jararacussu on MCF7, SKBR3, and MDAMB231 breast cancer cell lines. Methods: BthTX-I cytotoxicity was determined via MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide assay. Cell death was measured by a hypotonic fluorescent solution method, annexin-V-FITC/propidium iodide staining and by apoptotic/autophagic protein expression. Cancer stem cells (CSCs) were quantified by flow cytometry using anti-CD24-FITC and anti-CD44-APC antibodies and propidium iodide. Results: BthTX-I at 102 µg/mL induced cell death in all cell lines. The toxin induced apoptosis in MCF7, SKBR3, and MDAMB231 in a dose-dependent manner, as confirmed by the increasing number of hypodiploid nuclei. Expression of pro-caspase 3, pro-caspase 8 and Beclin-1 proteins were increased, while the level of the antiapoptotic protein Bcl-2 was diminished in MCF7 cells. BthTX-I changed the staining pattern of CSCs in MDAMB231 cells by increasing expression of CD24 receptors, which mediated cell death. Conclusions: BthTX-I induces apoptosis and autophagy in all breast cancer cell lines tested and also reduces CSCs subpopulation, which makes it a promising therapeutic alternative for breast cancer.(AU)
Assuntos
Humanos , Células-Tronco Neoplásicas , Neoplasias da Mama , Apoptose , Bothrops , Venenos Elapídicos/síntese química , Citometria de FluxoResumo
Background:Breast cancer is the neoplasm with both the highest incidence and mortality rate among women worldwide. Given the known snake venom cytotoxicity towards several tumor types, we evaluated the effects of BthTX-I from Bothrops jararacussu on MCF7, SKBR3, and MDAMB231 breast cancer cell lines.Methods:BthTX-I cytotoxicity was determined via MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide assay. Cell death was measured by a hypotonic fluorescent solution method, annexin-V-FITC/propidium iodide staining and by apoptotic/autophagic protein expression. Cancer stem cells (CSCs) were quantified by flow cytometry using anti-CD24-FITC and anti-CD44-APC antibodies and propidium iodide.Results:BthTX-I at 102 µg/mL induced cell death in all cell lines. The toxin induced apoptosis in MCF7, SKBR3, and MDAMB231 in a dose-dependent manner, as confirmed by the increasing number of hypodiploid nuclei. Expression of pro-caspase 3, pro-caspase 8 and Beclin-1 proteins were increased, while the level of the antiapoptotic protein Bcl-2 was diminished in MCF7 cells. BthTX-I changed the staining pattern of CSCs in MDAMB231 cells by increasing expression of CD24 receptors, which mediated cell death.Conclusions:BthTX-I induces apoptosis and autophagy in all breast cancer cell lines tested and also reduces CSCs subpopulation, which makes it a promising therapeutic alternative for breast cancer.(AU)
Assuntos
Animais , Bothrops , Venenos de Crotalídeos/administração & dosagem , Venenos de Crotalídeos/química , Venenos de Crotalídeos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/terapia , Células-Tronco Neoplásicas/efeitos dos fármacos , ApoptoseResumo
In recent decades, snake venom disintegrins have received special attention due to their potential use in anticancer therapy. Disintegrins are small and cysteine-rich proteins present in snake venoms and can interact with specific integrins to inhibit their activities in cell-cell and cell-ECM interactions. These molecules, known to inhibit platelet aggregation, are also capable of interacting with certain cancer-related integrins, and may interfere in important processes involved in carcinogenesis. Therefore, disintegrin from Crotalus durissus collilineatus venom was isolated, structurally characterized and evaluated for its toxicity and ability to interfere with cell proliferation and migration in MDA-MB-231, a human breast cancer cell line. Methods: Based on previous studies, disintegrin was isolated by FPLC, through two chromatographic steps, both on reversed phase C-18 columns. The isolated disintegrin was structurally characterized by Tris-TricineSDS-PAGE, mass spectrometry and N-terminal sequencing. For the functional assays, MTT and wound-healing assays were performed in order to investigate cytotoxicity and effect on cell migration in vitro, respectively. Results: Disintegrin presented a molecular mass of 7287.4 Da and its amino acid sequence shared similarity with the disintegrin domain of P-II metalloproteases. Using functional assays, the disintegrin showed low cytotoxicity (15% and 17%, at 3 and 6 µg/mL, respectively) after 24 h of incubation and in the wound-healing assay, the disintegrin (3 µg/mL) was able to significantly inhibit cell migration (24%, p < 0.05), compared to negative control. Conclusion: Thus, our results demonstrate that non-RGD disintegrin from C. d. collilineatus induces low cytotoxicity and inhibits migration of human breast cancer cells. Therefore, it may be a very useful molecular tool for understanding ECM-cell interaction cancer-related mechanisms involved in an important integrin family that highlights molecular aspects of tumorigenesis. Also, non-RGD disintegrin has potential to serve as an agent in anticancer therapy or adjuvant component combined with other anticancer drugs.(AU)
Assuntos
Venenos de Serpentes , Crotalus , Desintegrinas , Neoplasias da MamaResumo
Background:In recent decades, snake venom disintegrins have received special attention due to their potential use in anticancer therapy. Disintegrins are small and cysteine-rich proteins present in snake venoms and can interact with specific integrins to inhibit their activities in cell-cell and cell-ECM interactions. These molecules, known to inhibit platelet aggregation, are also capable of interacting with certain cancer-related integrins, and may interfere in important processes involved in carcinogenesis. Therefore, disintegrin from Crotalus durissus collilineatus venom was isolated, structurally characterized and evaluated for its toxicity and ability to interfere with cell proliferation and migration in MDA-MB-231, a human breast cancer cell line.Methods:Based on previous studies, disintegrin was isolated by FPLC, through two chromatographic steps, both on reversed phase C-18 columns. The isolated disintegrin was structurally characterized by Tris-Tricine-SDS-PAGE, mass spectrometry and N-terminal sequencing. For the functional assays, MTT and wound-healing assays were performed in order to investigate cytotoxicity and effect on cell migration in vitro, respectively.Results:Disintegrin presented a molecular mass of 7287.4 Da and its amino acid sequence shared similarity with the disintegrin domain of P-II metalloproteases. Using functional assays, the disintegrin showed low cytotoxicity (15% and 17%, at 3 and 6 μg/mL, respectively) after 24 h of incubation and in the wound-healing assay, the disintegrin (3 μg/mL) was able to significantly inhibit cell migration (24%, p < 0.05), compared to negative control.Conclusion:Thus, our results demonstrate that non-RGD disintegrin from C. d. collilineatus induces low cytotoxicity and inhibits migration of human breast cancer cells. Therefore, it may be a very useful molecular tool for understanding ECM-cell interaction cancer-related mechanisms involved in...(AU)
Assuntos
Humanos , Animais , Venenos de Crotalídeos/análise , Desintegrinas/análise , Movimento Celular , Adesão Celular , Neoplasias da Mama/tratamento farmacológico , CrotalusResumo
O objetivo deste estudo foi avaliar a utilização de dreno de sucção fechado em feridas cirúrgicas de mastectomias unilaterais totais em cadelas. Foram utilizadas 18 cadelas com peso médio de 14,87±9,29kg, idade média 9,34±2,68 anos, provenientes da rotina hospitalar, as quais foram selecionadas por meio de avaliação clínica e oncológica. Os animais foram alocados em dois grupos: Convencional (GC, n=9) que foram submetidas à cirurgia de mastectomia unilateral total, sem colocação de dreno e grupo Dreno (GD, n=9) que foram submetidas à mastectomia unilateral total com colocação do dreno de sucção fechado (Biovácuo?), sendo o diâmetro do dreno determinado conforme peso do animal. Foram realizadas cinco avaliações durante o período pós-operatório: avaliação clínica diária dos animais através da frequência cardíaca (FC), frequência respiratória (f), temperatura retal (TR), coloração de mucosas, tempo de preenchimento capilar (TPC), grau de desidratação, hemograma, perfil bioquímico (ureia, creatinina, colesterol, triglicérides e glicose); avaliação qualitativa do dreno; avaliação qualitativa da ferida cirúrgica, sendo esta também realizada em 14 e 30 dias quanto à presença ou não de exsudato, edema, integridade da pele, deiscência, necrose e fibrose cicatricial; avaliação quantitativa do fluido drenado e avaliação qualitativa do fluido drenado, quanto à celularidade, tipo e morfologia celular. Houve aumento dos valores médios de densidade, neutrófilos, linfócitos, e aumento do valor de mediana de fibrinogênio do fluido drenado em 72 horas em relação às 24 horas no grupo GD. Ocorreu aumento do exsudato em 24, 48 e 72 horas no GD quando comparado ao GC. Houve aumento do volume de fluido drenado em 60 e 72 horas em relação às 12 horas no grupo GD. Houve redução do valor médio de eritrócitos em 72 horas em relação ao basal no grupo GD. Observou-se aumento nos valores médios de hemoglobina e hematócrito em 24 horas no grupo GC quando comparado ao GD. Houve aumento no valor médio de neutrófilos segmentados no basal no GD quando comparado ao GC. Conclui-se que a técnica de colocação de dreno de sucção fechado apresenta maior teor de exsudato. Recomenda-se que o vácuo do dreno seja refeito de 12 em 12 horas, sendo inicialmentrealizado 6 horas após o término da cirurgia. O dreno de sucção fechado não gera desconforto adicional ao paciente; recomenda-se que o mesmo seja removido 72 horas após o término da cirurgia, quando a produção de fluido diminui, porém mesmo assim alguns indivíduos podem apresentar formação de seroma no pós-operatório tardio.(AU)
The aim of this study was to evaluate the use of closed suction drain in surgical wounds of total unilateral mastectomy in dogs. Eighteen dogs were used with an average weight of 14.87±9.29kg, average age 9.34±2.68 years, from the hospital routine, which were selected through clinical and oncological evaluation. The animals were divided into two groups: Conventional (CG, n=9) which underwent total unilateral mastectomy surgery without drain and Drain group (DG, n=9) which underwent complete unilateral mastectomy with placement closed suction drain (Biovácuo?). The diameter of the drain was determined according to the animal weight. Five evaluations were performed during the postoperative period: clinical evaluation of the animals daily through the heart rate (HR), respiratory rate (RR), rectal temperature (RT), color of mucous membranes, capillary refill time (CRT), degree of dehydration, blood count, chemistry profile (urea, creatinine, cholesterol, triglycerides and glucose); qualitative assessment of the drain; qualitative assessment of the surgical wound, is also being held on 14 and 30 days for the presence or absence of exudate, edema, skin integrity, dehiscence, necrosis and scarring; quantitative evaluation of the drained fluid; qualitative assessment of fluid drained, as the cellularity type and cell morphology. Surgical time was recorded in both groups. There were an increase in average values of density, neutrophils, lymphocytes, and increased fibrinogen median value of drained fluid in 72 hours compared to 24 hours in DG group. There was an increase of exudate within 24, 48 and 72 hours in DG when compared to GC. There was an increase in the volume of drained fluid in 60 and 72 hours compared to 12 hours in DG. There was a reduction in the average value of red blood cells within 72 hours from baseline in DG group. There was an increase in average hemoglobin and hematocrit in 24 hours in the control group compared to the DG. There was an increase in the average value of neutrophils targeted at baseline in DG when compared to CG. It follows that, the closed suction drain placement technique presents more exudate. It is recommended that the drain vacuum is redone 12 in 12 hours, initially being performed 6 hours after surgery. The closed suction drain does not generate additional discomfort to the patient; it is recommended that it be removed 72 hours after the surgery, when the fluid production decreases, but even so some individuals may present seroma formation in the late postoperative.(AU)
Assuntos
Animais , Feminino , Cães , Drenagem/tendências , Canal de Drenagem do Solo , Cães/anormalidades , Cães/cirurgia , Mastectomia/estatística & dados numéricosResumo
O objetivo deste estudo foi avaliar a utilização de dreno de sucção fechado em feridas cirúrgicas de mastectomias unilaterais totais em cadelas. Foram utilizadas 18 cadelas com peso médio de 14,87±9,29kg, idade média 9,34±2,68 anos, provenientes da rotina hospitalar, as quais foram selecionadas por meio de avaliação clínica e oncológica. Os animais foram alocados em dois grupos: Convencional (GC, n=9) que foram submetidas à cirurgia de mastectomia unilateral total, sem colocação de dreno e grupo Dreno (GD, n=9) que foram submetidas à mastectomia unilateral total com colocação do dreno de sucção fechado (Biovácuo?), sendo o diâmetro do dreno determinado conforme peso do animal. Foram realizadas cinco avaliações durante o período pós-operatório: avaliação clínica diária dos animais através da frequência cardíaca (FC), frequência respiratória (f), temperatura retal (TR), coloração de mucosas, tempo de preenchimento capilar (TPC), grau de desidratação, hemograma, perfil bioquímico (ureia, creatinina, colesterol, triglicérides e glicose); avaliação qualitativa do dreno; avaliação qualitativa da ferida cirúrgica, sendo esta também realizada em 14 e 30 dias quanto à presença ou não de exsudato, edema, integridade da pele, deiscência, necrose e fibrose cicatricial; avaliação quantitativa do fluido drenado e avaliação qualitativa do fluido drenado, quanto à celularidade, tipo e morfologia celular. Houve aumento dos valores médios de densidade, neutrófilos, linfócitos, e aumento do valor de mediana de fibrinogênio do fluido drenado em 72 horas em relação às 24 horas no grupo GD. Ocorreu aumento do exsudato em 24, 48 e 72 horas no GD quando comparado ao GC. Houve aumento do volume de fluido drenado em 60 e 72 horas em relação às 12 horas no grupo GD. Houve redução do valor médio de eritrócitos em 72 horas em relação ao basal no grupo GD. Observou-se aumento nos valores médios de hemoglobina e hematócrito em 24 horas no grupo GC quando comparado ao GD... (AU)
The aim of this study was to evaluate the use of closed suction drain in surgical wounds of total unilateral mastectomy in dogs. Eighteen dogs were used with an average weight of 14.87±9.29kg, average age 9.34±2.68 years, from the hospital routine, which were selected through clinical and oncological evaluation. The animals were divided into two groups: Conventional (CG, n=9) which underwent total unilateral mastectomy surgery without drain and Drain group (DG, n=9) which underwent complete unilateral mastectomy with placement closed suction drain (Biovácuo?). The diameter of the drain was determined according to the animal weight. Five evaluations were performed during the postoperative period: clinical evaluation of the animals daily through the heart rate (HR), respiratory rate (RR), rectal temperature (RT), color of mucous membranes, capillary refill time (CRT), degree of dehydration, blood count, chemistry profile (urea, creatinine, cholesterol, triglycerides and glucose); qualitative assessment of the drain; qualitative assessment of the surgical wound, is also being held on 14 and 30 days for the presence or absence of exudate, edema, skin integrity, dehiscence, necrosis and scarring; quantitative evaluation of the drained fluid; qualitative assessment of fluid drained, as the cellularity type and cell morphology. Surgical time was recorded in both groups. There were an increase in average values of density, neutrophils, lymphocytes, and increased fibrinogen median value of drained fluid in 72 hours compared to 24 hours in DG group. There was an increase of exudate within 24, 48 and 72 hours in DG when compared to GC. There was an increase in the volume of drained fluid in 60 and 72 hours compared to 12 hours in DG. There was a reduction in the average value of red blood cells within 72 hours from baseline in DG group. There was an increase in average hemoglobin and hematocrit in...(AU)
Assuntos
Animais , Feminino , Cães , Drenagem/tendências , Canal de Drenagem do Solo , Cães/anormalidades , Cães/cirurgia , MastectomiaResumo
ABSTRACT: The aim of this study was to evaluate the use of closed suction drain in surgical wounds of total unilateral mastectomy in dogs. Eighteen dogs were used with an average weight of 14.87±9.29kg, average age 9.34±2.68 years, from the hospital routine, which were selected through clinical and oncological evaluation. The animals were divided into two groups: Conventional (CG, n=9) which underwent total unilateral mastectomy surgery without drain and Drain group (DG, n=9) which underwent complete unilateral mastectomy with placement closed suction drain (Biovácuo?). The diameter of the drain was determined according to the animal weight. Five evaluations were performed during the postoperative period: clinical evaluation of the animals daily through the heart rate (HR), respiratory rate (RR), rectal temperature (RT), color of mucous membranes, capillary refill time (CRT), degree of dehydration, blood count, chemistry profile (urea, creatinine, cholesterol, triglycerides and glucose); qualitative assessment of the drain; qualitative assessment of the surgical wound, is also being held on 14 and 30 days for the presence or absence of exudate, edema, skin integrity, dehiscence, necrosis and scarring; quantitative evaluation of the drained fluid; qualitative assessment of fluid drained, as the cellularity type and cell morphology. Surgical time was recorded in both groups. There were an increase in average values of density, neutrophils, lymphocytes, and increased fibrinogen median value of drained fluid in 72 hours compared to 24 hours in DG group. There was an increase of exudate within 24, 48 and 72 hours in DG when compared to GC. There was an increase in the volume of drained fluid in 60 and 72 hours compared to 12 hours in DG. There was a reduction in the average value of red blood cells within 72 hours from baseline in DG group. There was an increase in average hemoglobin and hematocrit in 24 hours in the control group compared to the DG. There was an increase in the average value of neutrophils targeted at baseline in DG when compared to CG. It follows that, the closed suction drain placement technique presents more exudate. It is recommended that the drain vacuum is redone 12 in 12 hours, initially being performed 6 hours after surgery. The closed suction drain does not generate additional discomfort to the patient; it is recommended that it be removed 72 hours after the surgery, when the fluid production decreases, but even so some individuals may present seroma formation in the late postoperative.
RESUMO: O objetivo deste estudo foi avaliar a utilização de dreno de sucção fechado em feridas cirúrgicas de mastectomias unilaterais totais em cadelas. Foram utilizadas 18 cadelas com peso médio de 14,87±9,29kg, idade média 9,34±2,68 anos, provenientes da rotina hospitalar, as quais foram selecionadas por meio de avaliação clínica e oncológica. Os animais foram alocados em dois grupos: Convencional (GC, n=9) que foram submetidas à cirurgia de mastectomia unilateral total, sem colocação de dreno e grupo Dreno (GD, n=9) que foram submetidas à mastectomia unilateral total com colocação do dreno de sucção fechado (Biovácuo?), sendo o diâmetro do dreno determinado conforme peso do animal. Foram realizadas cinco avaliações durante o período pós-operatório: avaliação clínica diária dos animais através da frequência cardíaca (FC), frequência respiratória (f), temperatura retal (TR), coloração de mucosas, tempo de preenchimento capilar (TPC), grau de desidratação, hemograma, perfil bioquímico (ureia, creatinina, colesterol, triglicérides e glicose); avaliação qualitativa do dreno; avaliação qualitativa da ferida cirúrgica, sendo esta também realizada em 14 e 30 dias quanto à presença ou não de exsudato, edema, integridade da pele, deiscência, necrose e fibrose cicatricial; avaliação quantitativa do fluido drenado e avaliação qualitativa do fluido drenado, quanto à celularidade, tipo e morfologia celular. Houve aumento dos valores médios de densidade, neutrófilos, linfócitos, e aumento do valor de mediana de fibrinogênio do fluido drenado em 72 horas em relação às 24 horas no grupo GD. Ocorreu aumento do exsudato em 24, 48 e 72 horas no GD quando comparado ao GC. Houve aumento do volume de fluido drenado em 60 e 72 horas em relação às 12 horas no grupo GD. Houve redução do valor médio de eritrócitos em 72 horas em relação ao basal no grupo GD. Observou-se aumento nos valores médios de hemoglobina e hematócrito em 24 horas no grupo GC quando comparado ao GD. Houve aumento no valor médio de neutrófilos segmentados no basal no GD quando comparado ao GC. Conclui-se que a técnica de colocação de dreno de sucção fechado apresenta maior teor de exsudato. Recomenda-se que o vácuo do dreno seja refeito de 12 em 12 horas, sendo inicialmentrealizado 6 horas após o término da cirurgia. O dreno de sucção fechado não gera desconforto adicional ao paciente; recomenda-se que o mesmo seja removido 72 horas após o término da cirurgia, quando a produção de fluido diminui, porém mesmo assim alguns indivíduos podem apresentar formação de seroma no pós-operatório tardio.
Resumo
O antígeno CA 15-3 é uma proteína presente no soro utilizado no acompanhamento de mulheres com câncer de mama, essencialmente na detecção de metástases. Os objetivos deste estudo foram avaliar a efetividade e a viabilidade da utilização do marcador tumoral CA 15-3 em cadelas, comparando-se os valores do marcador entre cadelas sem e com neoplasia mamária, avaliando-se alterações nos valores do marcador após a mastectomia, e suas correlações entre o tipo histológico. Foi realizada a quantificação sérica do marcador tumoral CA 15-3 (teste de eletroquimioluminescência), em vinte cadelas hígidas (grupo controle) e vinte cadelas com neoplasia mamária (grupo teste). Os animais com neoplasia tiveram a dosagem do marcador realizada antes e 10 dias após a mastectomia. Ainda, foi realiza a citologia vaginal no momento da mastectomia e foram estabelecidos três grupos de acordo com a fase estral de cada cadela, Diestro, Proestro e Anestro. As massas tumorais foram encaminhadas para exame histopatológico. A avaliação dos dados de citologia vaginal entre os grupos Diestro, Proestro e Anestro pelo teste de ANOVA não demonstrou diferença estatística significativa entre os valores encontrados. E na análise para a comparação dos valores do marcador tumoral com os tipos histológicos de neoplasias, divididas em dois grupos, benignas e malignas, utilizando o teste de Mann-Whitney Rank Sum Test, o teste não demonstrou diferença estatística significativa visto que p>0,05. Os valores encontrados do marcador no grupo controle foram uma média de 0,19+0,39 U/mL, no grupo pré-mastectomia 1,56+0,39 U/mL e pós-mastectomia 0,66+0,27 U/mL. Em análise estatística com a comparação de grupo pré e pós-mastectomia, e do grupo controle com o grupo pré e pós-mastectomia observou-se significância com p< 0,005. Assim, observou-se diferença nos valores do marcador antes e depois da remoção cirúrgica da neoplasia, sugerindo seu possível uso como controle de crescimento tumoral pós-mastectomia individual. Porém há muita variação dos resultados nos diferentes métodos existentes, e não há ainda um padrão dos valores de referência para cada método, sendo necessários mais estudos sobre o uso dos marcadores.(AU)
The CA 15-3 antigen is a protein present in the serum, used to monitor women with breast cancer, mainly in metastatic disease detection. The objective of this study was to evaluate the effectiveness and feasibility of the CA 15-3 tumor marker in dogs, comparing the marker values between dogs with or without breast cancer, to estimate changes in marker values after mastectomy, and their correlation between the histological types. Serum quantification of the tumor marker CA 15-3 (electrochemiluminescence test) was performed in twenty healthy bitches and twenty others with mammary neoplasia. Bitches with cancer had the content of the tracer performed before and 10 days after mastectomy. The vaginal cytology was performed at the moment of the mastectomy, dividing the animals into three different groups (diestrus, proestrus and anestrus). All the mammary tumors were examined histopathologically. The evaluation of the vaginal cytology data of the groups Diestro, Proestro and Anestro by the ANOVA test did not show a statistically significant difference between the values found. In the analysis histological types of tumor marker values of neoplasms, divided into two groups, benign and malignant, using the Mann-Whitney Rank Sum Test, there was no statistical significant difference at p>0.05. The values of the marker in the control group had an average of 0.19+0.39 U/mL, of the pre-mastectomy group 1.56+0.39 U/mL, and of the post-mastectomy group 0.66+0.27 U/mL. The statistic was performed comparing groups pre- and post-mastectomy, and the control group with group pre- and post-mastectomy with a statistical significance p< 0.005 in both tests. There was a difference of marker values before and after surgical removal of the neoplasia, suggesting its possible use in post-mastectomy tumor control. But exist variation of results with the different existing methods, and there will be still a standard reference value for each method.(AU)
Assuntos
Animais , Feminino , Cães , Neoplasias da Mama/veterinária , Biomarcadores Tumorais/análise , Cães/anormalidades , Mucina-1 , EletroquimioterapiaResumo
O antígeno CA 15-3 é uma proteína presente no soro utilizado no acompanhamento de mulheres com câncer de mama, essencialmente na detecção de metástases. Os objetivos deste estudo foram avaliar a efetividade e a viabilidade da utilização do marcador tumoral CA 15-3 em cadelas, comparando-se os valores do marcador entre cadelas sem e com neoplasia mamária, avaliando-se alterações nos valores do marcador após a mastectomia, e suas correlações entre o tipo histológico. Foi realizada a quantificação sérica do marcador tumoral CA 15-3 (teste de eletroquimioluminescência), em vinte cadelas hígidas (grupo controle) e vinte cadelas com neoplasia mamária (grupo teste). Os animais com neoplasia tiveram a dosagem do marcador realizada antes e 10 dias após a mastectomia. Ainda, foi realiza a citologia vaginal no momento da mastectomia e foram estabelecidos três grupos de acordo com a fase estral de cada cadela, Diestro, Proestro e Anestro. As massas tumorais foram encaminhadas para exame histopatológico. A avaliação dos dados de citologia vaginal entre os grupos Diestro, Proestro e Anestro pelo teste de ANOVA não demonstrou diferença estatística significativa entre os valores encontrados. E na análise para a comparação dos valores do marcador tumoral com os tipos histológicos de neoplasias, divididas em dois grupos, benignas e malignas, utilizando o teste de Mann-Whitney Rank Sum Test, o teste não demonstrou diferença estatística significativa visto que p>0,05. Os valores encontrados do marcador no grupo controle foram uma média de 0,19+0,39 U/mL, no grupo pré-mastectomia 1,56+0,39 U/mL e pós-mastectomia 0,66+0,27 U/mL. Em análise estatística com a comparação de grupo pré e pós-mastectomia, e do grupo controle com o grupo pré e pós-mastectomia observou-se significância com p< 0,005. Assim, observou-se diferença nos valores do marcador antes e depois da remoção cirúrgica da neoplasia, sugerindo seu possível uso como controle de crescimento tumoral pós-mastectomia individual. Porém há muita variação dos resultados nos diferentes métodos existentes, e não há ainda um padrão dos valores de referência para cada método, sendo necessários mais estudos sobre o uso dos marcadores.(AU)
The CA 15-3 antigen is a protein present in the serum, used to monitor women with breast cancer, mainly in metastatic disease detection. The objective of this study was to evaluate the effectiveness and feasibility of the CA 15-3 tumor marker in dogs, comparing the marker values between dogs with or without breast cancer, to estimate changes in marker values after mastectomy, and their correlation between the histological types. Serum quantification of the tumor marker CA 15-3 (electrochemiluminescence test) was performed in twenty healthy bitches and twenty others with mammary neoplasia. Bitches with cancer had the content of the tracer performed before and 10 days after mastectomy. The vaginal cytology was performed at the moment of the mastectomy, dividing the animals into three different groups (diestrus, proestrus and anestrus). All the mammary tumors were examined histopathologically. The evaluation of the vaginal cytology data of the groups Diestro, Proestro and Anestro by the ANOVA test did not show a statistically significant difference between the values found. In the analysis histological types of tumor marker values of neoplasms, divided into two groups, benign and malignant, using the Mann-Whitney Rank Sum Test, there was no statistical significant difference at p>0.05. The values of the marker in the control group had an average of 0.19+0.39 U/mL, of the pre-mastectomy group 1.56+0.39 U/mL, and of the post-mastectomy group 0.66+0.27 U/mL. The statistic was performed comparing groups pre- and post-mastectomy, and the control group with group pre- and post-mastectomy with a statistical significance p< 0.005 in both tests. There was a difference of marker values before and after surgical removal of the neoplasia, suggesting its possible use in post-mastectomy tumor control. But exist variation of results with the different existing methods, and there will be still a standard reference value for each method.(AU)