RESUMO
Morphological and immunohistochemical studies of solid arrangement canine mammary carcinomas have shown that the different histological types may be characterized by proliferation of epithelial and/or myoepithelial cells. However, little is known about immunophenotypes and the importance of inflammation as prognostic factors in these neoplasms. The objective of the present study was to characterize the immunophenotype and degree of inflammation in the solid type of canine mammary neoplasm and to investigate their association with metastasis, Ki-67 index, tumour size, necrosis and survival. Sixty-five carcinomas with solid pattern, basaloid carcinomas, solid papillary carcinomas, malignant adenomyoepitheliomas (MAMEs) or malignant myoepitheliomas (MMEs) were investigated. Luminal A, luminal B HER2 negative and HER2 positive, HER2 overexpressed and triple negative immunophenotypes were immunolabelled as were Ki-67 protein and cyclooxygenase-2 (Cox-2). Histological peritumoural and intratumoural inflammatory infiltrates were graded (distribution × intensity) and the presence of necrosis identified. We found a statistical difference between histological types and immunophenotypes, with MME and MAME having a higher occurrence of luminal A, whereas most neoplasms had the luminal B HER-negative immunophenotype. There was no correlation between immunophenotype and degree of peri- and intratumoural inflammation, nodal metastasis, necrosis or tumour size. An increased degree of peri- and intratumoural inflammation was significantly associated with lymph node metastasis, and more severe intratumoural inflammation was associated with the presence of tumour necrosis. Tumour size, Ki-67 index and Cox-2 score were not associated with inflammation in either peri- or intratumoural regions. No difference was observed in survival in relation to immunophenotype or degree of inflammation, but the Cox regression model revealed that nodal metastasis influenced the risk of death.
Assuntos
Doenças do Cão , Imunofenotipagem , Inflamação , Neoplasias Mamárias Animais , Animais , Cães , Feminino , Neoplasias Mamárias Animais/patologia , Doenças do Cão/patologia , Doenças do Cão/imunologia , Biomarcadores Tumorais/análiseRESUMO
We studied some fibrotic aspects of chronic interstitial pneumonitis in the lungs of dogs infected with Leishmania infantum. The lungs of eleven naturally infected dogs, twelve experimentally infected with two distinct strains of L. infantum (BH401 and BH46), and six uninfected (controls) dogs, were analyzed by histological, parasitological, and immunohistochemical studies. Conventional histology (HE), collagen deposition (Gomori's silver staining for reticulin collagen fibers), and immunohistochemistry for myofibroblast characterization were carried out based on the cellular expression of alpha-smooth muscle actin, vimentin, cytokeratin, E-cadherin, snail antigen homologue 1 (SNAI1) (Snail), and the cytokine expression of transforming growth factor-beta (TGF-ß). Parasitological screening was carried out using conventional polymerase chain reaction (PCR) and the immunohistochemical reaction of streptavidin-peroxidase for visualizing Leishmania amastigotes. Dogs naturally infected with L. infantum and experimentally infected with L. infantum BH401 strains showed intense interstitial pneumonitis characterized by thickening of the alveolar septa as a consequence of an intense diffuse and focal (plaques) chronic exudate of mononuclear cells associated with fibrogenesis. The expression of alpha-actin, vimentin, and TGF-ß was higher in the lung interstitium of all infected dogs than in the other two groups (BH46 strain and controls). Moreover, in both the naturally and experimentally infected dog (BH401 strain) groups, the expression of Snail was moderate to intense in contrast to the other groups. Based on these immunohistochemical results, we concluded that mesenchymal cells are active in promoting changes in the extracellular matrix in the lungs of dogs naturally and experimentally infected with L. infantum, but it depends on the virulence of the parasite.
RESUMO
Fibromatosis, or desmoid tumour, is characterized by excessive and infiltrative proliferation of connective tissue originating from aponeurotic muscle structures. Mammary fibromatosis is rare in humans and animals and its precise aetiology is unknown. A 10-year-old mixed-breed female dog developed a mass in the right cranial thoracic mammary gland (M1) and underwent lumpectomy. The mass was firm, with an irregular surface and distinct limits. Microscopically, it was a neoplastic proliferation of fusiform cells with low atypia, interspersed with abundant dense collagenous tissue, confirmed by histochemical staining with Gomori's trichrome and Masson's trichrome and immunopositivity for vimentin and smooth muscle actin, confirming mammary fibromatosis. Mammary fibromatosis in dogs needs further studies to elucidate its clinical, epidemiological and aetiopathogenic aspects.
Assuntos
Doenças do Cão , Fibroma , Fibromatose Agressiva , Humanos , Feminino , Cães , Animais , Fibromatose Agressiva/patologia , Fibromatose Agressiva/veterinária , Fibroma/veterinária , Músculos/patologiaRESUMO
Although it is well established that platelet-activated receptor (PAF) and protease-activated receptor 2 (PAR2) play a pivotal role in the pathophysiology of lung and airway inflammatory diseases, a role for a PAR2-PAFR cooperation in lung inflammation has not been investigated. Here, we investigated the role of PAR2 in PAF-induced lung inflammation and neutrophil recruitment in lungs of BALB/c mice. Mice were pretreated with the PAR2 antagonist ENMD1068, PAF receptor (PAFR) antagonist WEB2086, or aprotinin prior to intranasal instillation of carbamyl-PAF (C-PAF) or the PAR2 agonist peptide SLIGRL-NH2 (PAR2-AP). Leukocyte infiltration in bronchoalveolar lavage fluid (BALF), C-X-C motif ligand 1 (CXCL)1 and CXCL2 chemokines, myeloperoxidase (MPO), and N-acetyl-glycosaminidase (NAG) levels in BALF, or lung inflammation were evaluated. Intracellular calcium signaling, PAFR/PAR2 physical interaction, and the expression of PAR2 and nuclear factor-kappa B (NF-ÐB, p65) transcription factor were investigated in RAW 264.7 cells stimulated with C-PAF in the presence or absence of ENMD1068. C-PAF- or PAR2-AP-induced neutrophil recruitment into lungs was inhibited in mice pretreated with ENMD1068 and aprotinin or WEB2086, respectively. PAR2 blockade impaired C-PAF-induced neutrophil rolling and adhesion, lung inflammation, and production of MPO, NAG, CXCL1, and CXCL2 production in lungs of mice. PAFR activation reduced PAR2 expression and physical interaction of PAR2 and PAFR; co-activation is required for PAFR/PAR2 physical interaction. PAR2 blockade impaired C-PAF-induced calcium signal and NF-κB p65 translocation in RAW 264.7 murine macrophages. This study provides the first evidence for a cooperation between PAFR and PAR2 mediating neutrophil recruitment, lung inflammation, and macrophage activation.
Assuntos
NF-kappa B , Pneumonia , Camundongos , Animais , NF-kappa B/metabolismo , Fator de Ativação de Plaquetas/metabolismo , Receptor PAR-2/genética , Receptor PAR-2/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Aprotinina/metabolismo , Infiltração de Neutrófilos , Ativação Transcricional , Pneumonia/induzido quimicamenteRESUMO
This study investigated the effect of vegetable and fish oils with different n-3 / n-6 PUFAS ratios on the lipoprotein profile and on the development of murine breast cancer 4T1. Female Balb/c mice (6-7 weeks) received diets containing 4.0% fat during seven weeks. On the fourth week, animals were inoculated into the posterior left flank with 2.5 × 106 4T1 cells. Body weight and food intake were registered and the profile serum lipoproteins was determined. Tumor volume, histopathological and immunohistochemical studies, myeloperoxidase and N-acetylglucosaminidase activities, TNF-α, hemoglobin and VEGF levels were analysed. The highest n-3 / n-6 ratio was found in fish oil (15.8:1), followed by linseed (2.4:1), canola (1:2.1) and soybean (1:9.4) oils. Body weight, food and caloric intake, lipoprotein profile, tumor weight, tumor evolution and histopathological analysis were not different. Canola oil increased cell proliferation when compared to soybean oil, and fish oil changed the inflammatory response and increased VEGF in tumors compared to other groups. The type of fatty acid and the high ratio of n-3 / n-6 PUFAs in the diet influenced cell proliferation and inflammation in the tumor differentially, highlighting the increase of neutrophils and VEGF levels in animals fed on fish oil.
Assuntos
Ácidos Graxos Ômega-3 , Fator A de Crescimento do Endotélio Vascular , Animais , Feminino , Camundongos , Óleos de Plantas , Gorduras na Dieta , Ácidos Graxos Ômega-3/análise , Óleos de Peixe/metabolismo , Ácidos Graxos/análise , Lipoproteínas , Peso CorporalRESUMO
Emetic tartar (ET), was used in the treatment of leishmaniasis but its use was discontinued due to its low therapeutic index. Liposomes have been shown to be a promising strategy for delivery of bioactive substances in the region of interest, in order to reduce and/or eliminate undesirable effects. In the present study, liposomes containing ET were prepared and characterized to evaluate acute toxicity as well as their leishmanicidal action using BALB/c mice with an inoculum of Leishmania (Leishmania) infantum. Liposomes were composed of egg phosphatidylcholine and 3ß-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol, with an average diameter of 200 nm, zeta potential of +18 mV, and ET encapsulated into liposomes at a concentration near 2 g/L. Healthy mice were treated with ET or liposome containing ET (Lip-ET) in a single dose of 16 mg/kg of Sb3+ intravenously and observed for 14 days. The death of two animals in the ET-treated group and no deaths in the Lip-ET-treated group was observed. Higher hepatic and cardiac toxicity were observed in animals treated with ET when compared to animals treated with Lip-ET, blank liposomes (Blank-Lip) and PBS. The study of antileishmanial efficacy was conducted by intraperitoneal administration of Lip-ET, for ten consecutive days. It was observed by limiting dilution that treatments with liposomal formulations containing ET, as well as Glucantime®, led to a significant reduction in parasitic load in spleen and liver (p < 0.05) when compared to the untreated control group.
RESUMO
BACKGROUND: Cellular entry of SARS-CoV-2 has been shown to rely on angiotensin-converting enzyme 2 (ACE2) receptors, whose expression in the testis is among the highest in the body. Additionally, the risk of mortality seems higher among male COVID-19 patients, and though much has been published since the first cases of COVID-19, there remain unanswered questions regarding SARS-CoV-2 impact on testes and potential consequences for reproductive health. We investigated testicular alterations in non-vaccinated deceased COVID-19-patients, the precise location of the virus, its replicative activity, and the immune, vascular, and molecular fluctuations involved in the pathogenesis. RESULTS: We found that SARS-CoV-2 testicular tropism is higher than previously thought and that reliable viral detection in the testis requires sensitive nanosensors or RT-qPCR using a specific methodology. Through an in vitro experiment exposing VERO cells to testicular macerates, we observed viral content in all samples, and the subgenomic RNA's presence reinforced the replicative activity of SARS-CoV-2 in testes of the severe COVID-19 patients. The cellular structures and viral particles, observed by transmission electron microscopy, indicated that macrophages and spermatogonial cells are the main SARS-CoV-2 lodging sites, where new virions form inside the endoplasmic reticulum Golgi intermediate complex. Moreover, we showed infiltrative infected monocytes migrating into the testicular parenchyma. SARS-CoV-2 maintains its replicative and infective abilities long after the patient's infection. Further, we demonstrated high levels of angiotensin II and activated immune cells in the testes of deceased patients. The infected testes show thickening of the tunica propria, germ cell apoptosis, Sertoli cell barrier loss, evident hemorrhage, angiogenesis, Leydig cell inhibition, inflammation, and fibrosis. CONCLUSIONS: Our findings indicate that high angiotensin II levels and activation of mast cells and macrophages may be critical for testicular pathogenesis. Importantly, our findings suggest that patients who become critically ill may exhibit severe alterations and harbor the active virus in the testes.
Assuntos
COVID-19 , Testículo , Tropismo Viral , Animais , Humanos , Masculino , Angiotensina II/metabolismo , Chlorocebus aethiops , COVID-19/patologia , SARS-CoV-2 , Testículo/imunologia , Testículo/virologia , Células VeroRESUMO
Food allergy is a pathological condition that can lead to hives, swelling, gastrointestinal distress, cardiovascular and respiratory compromise, and even anaphylaxis. The lack of treatment resources emphasizes the necessity for new therapeutic strategies, and in this way, probiotics has been pointed out as an alternative, especially because of its immunomodulatory properties. The goal of this study was to evaluate the probiotic effect of Bifidobacterium longum subsp. longum 51A (BL51A) in a murine model of ovalbumin (OVA) food allergy, as well as to investigate the effect of the dose and viability of the bacteria on the proposed model. For this purpose, the probiotic effect was assessed by clinical, immunological, and histological parameters in mice treated or not with the BL51A and sensitized or not with OVA. Oral administration of BL51A prevented weight loss and reduced serum levels of IgE anti-OVA and of sIgA in the intestinal fluid. Also, it reduced the intestinal permeability, proximal jejunum damage, recruitment of eosinophils and neutrophils, and levels of eotaxin-1, CXCL1/KC, IL4, IL5, IL6, IL13, and TNF. Furthermore, the treatment was able to increase the levels of IL10. Investigating different doses administered, the level of 108 CFU showed the best results in terms of protective effect. In addition, the administration of the inactivated bacteria did not present any beneficial effect. Results demonstrate that BL51A promotes a systemic immunomodulatory protective effect in a murine model of food allergy that depends on the dose and viability of the bacteria, suggesting its use as probiotic in such disease.
Assuntos
Hipersensibilidade Alimentar , Probióticos , Animais , Camundongos , Modelos Animais de Doenças , Hipersensibilidade Alimentar/tratamento farmacológico , Hipersensibilidade Alimentar/prevenção & controle , Bifidobacterium , Inflamação/tratamento farmacológicoRESUMO
For the first time, compounds developed from the 1,2,3-triazole scaffold were evaluated as novel drugs to treat triple-negative breast cancer (TNBC). Four organic salts were idealized as nonclassical bioisosteres of miltefosine, which is used in the topical treatment for skin metastasizing breast carcinoma. Among them, derivative dhmtAc displayed better solubility and higher cytotoxicity against the human breast adenocarcinoma cell line and mouse 4T1 cell lines, which are representatives of TNBC. In vitro assays revealed that dhmtAc interferes with cell integrity, confirmed by lactate dehydogenase leakage. Due to its human peripheral blood mononuclear cell (PBMC) toxicity, dhmtAc in vivo studies were carried out with the drug incorporated in a long-circulating and pH-sensitive liposome (SpHL-dhmtAc), and the acute toxicity in BALB/c mice was determined. Free dhmtAc displayed cardiac and pulmonary toxicity after the systemic administration of 5 mg/kg doses. On the other hand, SpHL-dhmtAc displayed no toxicity at 20 mg/kg. The in vivo antitumor effect of SpHL-dhmtAc was investigated using the 4T1 heterotopic murine model. Intravenous administration of SpHL-dhmtAc reduced the tumor volume and weight, without interfering with the body weight, compared with the control group and the dhmtAc free form. The incorporation of the triazole compound in the liposome allowed the demonstration of its anticancer potential. These findings evidenced 1,3,4-trisubstituted-1,2,3-triazole as a promising scaffold for the development of novel drugs with applicability for the treatment of patients with TNBC.
Assuntos
Lipossomos , Neoplasias de Mama Triplo Negativas , Animais , Linhagem Celular Tumoral , Humanos , Leucócitos Mononucleares , Camundongos , Camundongos Endogâmicos BALB C , Relação Estrutura-Atividade , Triazóis/farmacologia , Neoplasias de Mama Triplo Negativas/tratamento farmacológicoRESUMO
Mammary neoplasms are the most frequently diagnosed tumours in female dogs and are classified into various histological types, including solid carcinomas. We proposed a subclassification of solid carcinomas based on morphological and immunohistochemical characteristics, and correlated the subtypes with prognostic factors. A total of 135 cases of solid mammary carcinoma were selected from 3,400 canine mammary neoplasms in the archives of the Laboratory of Comparative Pathology, Institute of Biological Sciences, Federal University of Minas Gerais, Brazil. Epidemiological and survival data were obtained, and immunolabelling for chromogranin A, pancytokeratin, cytokeratin 14, Ki67 and p63 was performed. Solid carcinomas were classified into six subgroups: malignant adenomyoepithelioma (68/135), carcinoma with solid pattern (22/135), malignant myoepithelioma (16/135), basaloid carcinoma (14/135), neuroendocrine carcinoma (10/135) and solid papillary carcinoma (5/135). Shorter survival time was associated with the presence of lymphatic invasion (P = 0.009) in the initial clinical staging (I-III). When considering all clinical stages (I-V), vascular invasion (P <0.001) and the presence of regional metastasis (P = 0.004) were important prognostic factors. Basaloid carcinoma and solid papillary carcinoma did not reach the median survival time for early-stage cases, and malignant myoepithelioma had the highest median survival in advanced stages. Carcinoma with a solid pattern was associated with a higher number of regional metastases. Distinguishing the various histological and immunophenotypic subtypes that exhibit a solid arrangement, using histological and immunohistochemical criteria, is essential for understanding the behaviour of these neoplasms and for the selection of more appropriate and specific therapies.
Assuntos
Carcinoma , Doenças do Cão , Neoplasias Mamárias Animais , Mioepitelioma , Animais , Brasil , Carcinoma/veterinária , Cães , Feminino , Imuno-Histoquímica , Mioepitelioma/veterináriaRESUMO
Cholangiocarcinoma (CCA) is the second most malignant neoplasm in the liver that arises from the biliary tree. CCA is associated with a poor prognosis, and the key players involved in its pathogenesis are still not well understood. Receptor tyrosine kinases (RTKs), such as epidermal growth factor receptor (EGFR), can mediate intracellular calcium (Ca2+) signaling pathways via inositol 1,4,5-trisphosphate (InsP3), activating inositol 1,4,5-trisphosphate receptors (ITPRs) and regulating tumor growth. ITPR isoform 3 (ITPR3) is the main intracellular Ca2+ release channel in cholangiocytes. The effects of intracellular Ca2+ are mediated by calcium-binding proteins such as Calmodulin and S100 calcium-binding protein A4 (S100A4). However, the clinicopathological and biological significance of EGFR, ITPR3 and S100A4 in CCA remains unclear. Thus, the present work investigates the immunoexpression of these three proteins in 59 CCAs from patients who underwent curative surgical treatment and correlates the data with clinicopathological features and survival. High ITPR3 expression was correlated with CA 19-9 levels, TNM stage and lymph node metastasis (N). Furthermore, ITPR3 expression was increased in distal CCA compared to control bile ducts and intrahepatic and perihilar CCAs. These observations were confirmed by proteomic analysis. ITPR3 and S100A4 clinical scores were significantly correlated. Furthermore, it was demonstrated that EGF induces calcium signaling in a cholangiocarcinoma cell line and ITPR3 colocalizes with nonmuscle myosin IIA (NMIIA). In summary, ITPR3 overexpression could contribute to CCA progression and it may represent a potential therapeutic target.
Assuntos
Neoplasias dos Ductos Biliares/patologia , Colangiocarcinoma/patologia , Receptores de Inositol 1,4,5-Trifosfato/genética , Proteína A4 de Ligação a Cálcio da Família S100/metabolismo , Neoplasias dos Ductos Biliares/genética , Sinalização do Cálcio , Colangiocarcinoma/genética , Progressão da Doença , Receptores ErbB/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Proteômica , Taxa de SobrevidaRESUMO
In this retrospective and prospective study, histopathological and immunohistochemical analyses of 62 cases of lymphomas in cats were performed to classify the anatomic forms and subtypes, according to the WHO guidelines, and correlate it to FeLV proviral DNA detected using PCR. The most common anatomical form was gastrointestinal (40.3%, 25/62), followed by multicentric (29%, 18/62), mediastinal (17.7%, 11/62) and extranodal (12,9%, 8/62). Among the lymphoma subtypes, diffuse large B-cell lymphoma (DLBCL) (30.6%, 19/62) was the most commonly diagnosed followed by peripheral T-cell lymphoma (PTCL) (29%, 18/62) and enteropathy associated T-cell lymphoma type 2 (14.5%, 9/62). DNA extraction from paraffin-embedded neoplastic tissue was obtained in 28 cases and FeLV proviral DNA was detected by PCR, in 23 of these. Of the cases presenting with FeLV proviral DNA, nine (32%) were of the multicentric form, five (22%) of the mediastinal and extranodal forms and four (17%) of the gastrointestinal form. The most frequent subtypes with FeLV proviral DNA, independent of the anatomical form, were DLBCL (39.1%, 9/23) and PTCL (34.7%, 8/23). The presence of the FeLV proviral DNA in 23 cats of this study, probably had association with the multicentric form of lymphoma and higher occurrence in the DLBCL and PTCL subtypes.
Neste estudo retrospectivo e prospectivo, análises histopatológicas e imuno-histoquímicas de 62 casos de linfomas em gatos foram realizadas para classificar as formas anatômicas o e subtipos do linfoma, de acordo com as diretrizes da OMS. Além disso, foi realizada a extração de DNA dos tumores incluídos na parafina para obtenção de DNA pró-viral do FeLV por PCR, e relacionada com os exames anteriores. A forma anatômica mais comum foi a gastrointestinal (40.3%, 25/62), seguida pela multicêntrica (29%, 18/62), mediastinal (17,7%, 11/62) e extranodal (12,9%, 8/62). Entre os subtipos de linfoma, o linfoma difuso de grandes células B (DLBCL) (30.6%, 19/62) foi o mais comumente diagnosticado, seguido por linfoma de células T periférico (PTCL) (29%, 18/62) e o linfoma de células T associado a enteropatia tipo 2 (14.5%, 9/62). A extração de DNA de tecido neoplásico emblocado em parafina foi obtida em 28 casos e o DNA pró-viral de FeLV foi detectado por PCR, em 23 deles. Dos casos com DNA pró-viral do FeLV, nove (32%) eram da forma multicêntrica, cinco (22%) das formas mediastinal e extranodal e quatro (17%) da forma gastrointestinal. Os subtipos mais frequentes com DNA pró-viral do FeLV, independente da forma anatômica, foram DLBCL (39.1%, 9/23) e PTCL (34.7%, 8/23). A presença do DNA pró-viral do FeLV em 23 gatos deste estudo, provavelmente teve associação com a forma multicêntrica do linfoma e maior ocorrência nos subtipos DLBCL e PTCL.
Assuntos
Animais , Gatos , Provírus , Leucemia Felina , Vírus da Leucemia Felina/isolamento & purificação , Linfoma/patologia , Doenças do Gato , Reação em Cadeia da Polimerase , Linfoma/veterináriaRESUMO
Diagnosis and prognosis of breast cancer is based on disease staging identified through histopathological and molecular biology techniques. Animal models are used to gain mechanistic insights into the development of breast cancer. C(3)1-TAg is a genetically engineered mouse model that develops mammary cancer. However, carcinogenesis caused by this transgene was characterized in the Friend Virus B (FVB) background. As most genetic studies are done in mice with C57BL/6 J background, we aimed to define the histological alterations in C3(1)-TAg C57BL/6 J animals. Our results showed that C3(1)-TAg animals with C57BL/6 J background develop solid-basaloid adenoid cystic carcinomas with increased fibrosis, decreased area of adipocytes, and a high proliferative index, which are triple-negative for progesterone, estrogen, and human epidermal growth factor receptor 2 (HER2) receptors. Our results also revealed that tumor development is slower in the C57BL/6 J background when compared with the FVB strain, providing a better model to study the different stages in breast cancer progression.
Assuntos
Antígenos Virais de Tumores/genética , Neoplasias da Mama/genética , Carcinoma Adenoide Cístico/genética , Modelos Genéticos , Animais , Antígenos Virais de Tumores/imunologia , Neoplasias da Mama/imunologia , Neoplasias da Mama/patologia , Carcinoma Adenoide Cístico/imunologia , Carcinoma Adenoide Cístico/patologia , Feminino , Vírus da Leucemia Murina de Friend/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos TransgênicosRESUMO
BACKGROUND: Invasive micropapillary carcinoma (IMPC) is a rare malignant breast tumor and a variant form of invasive ductal carcinoma that is an aggressive neoplasm of the human breast and canine mammary gland. The importance of the tumor microenvironment in cancer development has gradually been recognized, but little is known about the cell types outlining the cystic space of canine IMPC. This study aimed to characterize the neoplastic cells outlining the cystic space of IMPC. RESULTS: Immunohistochemistry (IHC), immunofluorescence (IF), superresolution and transmission electron microscopy (TEM) were used to assess the cell types in the cystic areas of IMPCs. Cells expressing the mesenchymal markers alpha-smooth muscle actin (αSMA), Vimentin, and S100A4 outlined the cystic space of IMPC. Furthermore, loss of epithelial cell polarity in IMPC was shown by the localization of MUC1 at the stroma-facing surface. This protein modulates lumen formation and inhibits the cell-stroma interaction. Immunohistochemical and IF staining for the myoepithelial cell marker p63 were negative in IMPC samples. Furthermore, associated with peculiar morphology, such as thin cytoplasmic extensions outlining cystic spaces, was observed under TEM. These observations suggested cells with characteristics of myoepithelial-like cells. CONCLUSIONS: The cells outlining the cystic space of IMPC in the canine mammary gland were characterized using IHC, IF and TEM. The presence of cells expressing αSMA, Vimentin, and S100A4 in the IMPC stroma suggested a role for tumor-associated fibroblasts in the IMPC microenvironment. The reversal of cell polarity revealed by the limited basal localization of MUC1 may be an important factor contributing to the invasiveness of IMPC. For the first time, the cystic space of canine mammary gland IMPC was shown to be delimited by myoepithelial-like cells that had lost p63 expression. These findings may enhance our understanding of the cellular microenvironment of invasive tumors to improve cancer diagnosis and treatment.
Assuntos
Carcinoma Papilar/veterinária , Doenças do Cão/patologia , Neoplasias Mamárias Animais/patologia , Animais , Biomarcadores Tumorais/metabolismo , Carcinoma Papilar/metabolismo , Carcinoma Papilar/patologia , Doenças do Cão/metabolismo , Cães , Feminino , Imunofluorescência/veterinária , Imuno-Histoquímica/veterinária , Glândulas Mamárias Animais/patologia , Neoplasias Mamárias Animais/metabolismo , Microscopia Eletrônica de Transmissão/veterinária , FenótipoRESUMO
Precise diagnosis and prognosis are key in prevention and reduction of morbidity and mortality in all types of cancers. Here we show that changes in the collagen fibres in the main histological subtypes of canine mammary gland carcinomas are directly associated with the tumour behaviour and the animal survival time and could become a useful tool in helping with diagnosis. Imaging by second harmonic generation and multiphoton excited fluorescence microscopy were performed to evaluate the collagen and cellular segment parameters in cancer biopsies. We present a retrospective study of 45 cases of canine mammary cancer analysing 836 biopsies regions including normal mammary gland tissue, benign mixed tumours, carcinoma in mixed tumour, carcinosarcoma, micropapillary carcinoma and solid carcinoma. The image analyses and the comparison between the tumour types allowed to assess the collagen fibre changes during tumour progression. We demonstrate that the collagen parameters correlate with the clinical and pathological data, the results show that in neoplastic tissues, the collagen fibres are more aligned and shorter as compared to the normal tissues. There is a clear association of the mean fibre length with the dogs survival times, the carcinomas presenting shorter collagen fibres indicate a worse survival rate.
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Colágeno/metabolismo , Neoplasias Mamárias Animais/patologia , Animais , Progressão da Doença , Cães , Feminino , Imageamento Tridimensional , Modelos Lineares , Neoplasias Mamárias Animais/diagnóstico por imagem , Prognóstico , Estatística como Assunto , Taxa de SobrevidaRESUMO
Brucella abortus is a Gram-negative bacterium responsible for a worldwide zoonotic infection-Brucellosis, which has been associated with high morbidity rate in humans and severe economic losses in infected livestock. The natural route of infection is through oral and nasal mucosa but the invasion process through host gut mucosa is yet to be understood. Studies have examined the role of NLRP6 (NOD-like receptor family pyrin domain-containing-6 protein) in gut homeostasis and defense against pathogens. Here, we investigated the impact of gut microbiota and NLRP6 in a murine model of Ba oral infection. Nlrp6-/- and wild-type (WT) mice were infected by oral gavage with Ba and tissues samples were collected at different time points. Our results suggest that Ba oral infection leads to significant alterations in gut microbiota. Moreover, Nlrp6-/- mice were more resistant to infection, with decreased CFU in the liver and reduction in gut permeability when compared to the control group. Fecal microbiota transplantation from WT and Nlrp6-/- into germ-free mice reflected the gut permeability phenotype from the donors. Additionally, depletion of gut microbiota by broad-spectrum-antibiotic treatment prevented Ba replication in WT while favoring bacterial growth in Nlrp6-/-. Finally, we observed higher eosinophils in the gut and leukocytes in the blood of infected Nlrp6-/- compared to WT-infected mice, which might be associated to the Nlrp6-/- resistance phenotype. Altogether, these results indicated that gut microbiota composition is the major factor involved in the initial stages of pathogen host replication and partially also by the resistance phenotype observed in Nlrp6 -/- mice regulating host inflammation against Ba infection.
Assuntos
Brucelose/fisiopatologia , Microbioma Gastrointestinal/fisiologia , Intestinos/microbiologia , Intestinos/fisiopatologia , Administração Oral , Animais , Antibacterianos/administração & dosagem , Brucella abortus , Brucelose/microbiologia , Transplante de Microbiota Fecal , Interações Hospedeiro-Patógeno , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Permeabilidade , Receptores de Superfície Celular/genética , Organismos Livres de Patógenos EspecíficosRESUMO
Resolution failure of exacerbated inflammation triggered by Influenza A virus (IAV) prevents return of pulmonary homeostasis and survival, especially when associated with secondary pneumococcal infection. Therapeutic strategies based on pro-resolving molecules have great potential against acute inflammatory diseases. Angiotensin-(1-7) [Ang-(1-7)] is a pro-resolving mediator that acts on its Mas receptor (MasR) to promote resolution of inflammation. We investigated the effects of Ang-(1-7) and the role of MasR in the context of primary IAV infection and secondary pneumococcal infection and evaluated pulmonary inflammation, virus titers and bacteria counts, and pulmonary damage. Therapeutic treatment with Ang-(1-7) decreased neutrophil recruitment, lung injury, viral load and morbidity after a primary IAV infection. Ang-(1-7) induced apoptosis of neutrophils and efferocytosis of these cells by alveolar macrophages, but had no direct effect on IAV replication in vitro. MasR-deficient (MasR-/-) mice were highly susceptible to IAV infection, displaying uncontrolled inflammation, increased viral load and greater lethality rate, as compared to WT animals. Ang-(1-7) was not protective in MasR-/- mice. Interestingly, Ang-(1-7) given during a sublethal dose of IAV infection greatly reduced morbidity associated with a subsequent S. pneumoniae infection, as seen by decrease in the magnitude of neutrophil influx, number of bacteria in the blood leading to a lower lethality. Altogether, these results show that Ang-(1-7) is highly protective against severe primary IAV infection and protects against secondary bacterial infection of the lung. These effects are MasR-dependent. Mediators of resolution of inflammation, such as Ang-(1-7), should be considered for the treatment of pulmonary viral infections.
Assuntos
Angiotensina I/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Fragmentos de Peptídeos/uso terapêutico , Infecções Pneumocócicas/tratamento farmacológico , Pneumonia Viral/tratamento farmacológico , Proteínas Proto-Oncogênicas/imunologia , Receptores Acoplados a Proteínas G/imunologia , Células A549 , Angiotensina I/farmacologia , Animais , Anti-Inflamatórios/farmacologia , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/imunologia , Cães , Humanos , Vírus da Influenza A , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Pulmão/patologia , Células Madin Darby de Rim Canino , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Fragmentos de Peptídeos/farmacologia , Peroxidase/imunologia , Fagocitose/efeitos dos fármacos , Infecções Pneumocócicas/imunologia , Infecções Pneumocócicas/patologia , Pneumonia Viral/imunologia , Pneumonia Viral/patologia , Proto-Oncogene Mas , Proteínas Proto-Oncogênicas/genética , Receptores Acoplados a Proteínas G/genética , Streptococcus pneumoniaeRESUMO
Feline injection site sarcomas (FISS) are aggressive, with high recurrence and rarely metastasising. The objective of this study was to evaluate, by immunohistochemistry, the expression of oestrogen (ER) and progesterone (PR) receptors in FISS and correlate them with clinical and histopathological aspects. This was a retrospective study with 51 cases of FISS. Immunohistochemistry was performed to detect vimentin, ER, PR and Ki67 expression. Clinical, histopathological and immunohistochemical characteristics were predictor variables and the expression of ER and PR were the dependent ones. Twenty-eight (55%) of the 51 FISS cases were female and 23 (45%) male with 10.7 ± 4.2 years and median tumour size of 3 cm (2.0-5.4). The trunk was the most affected site, with 38 cases (84%). Histological grade III was observed in 57% of the cases, considering differentiation score, necrosis and mitotic index. ER expression, positive in 64% of cases, was associated with the mitotic index (P = .05) and degree of pleomorphism (P = .04). PR was not associated with the variables and 63% of cases were negative for this receptor. Thus, ER expression can affect tumour growth. The knowledge on the FISS hormonal expression is important to clarify the pathophysiological mechanisms. Further studies are needed to predict the value of ER expression in the prognosis of FISS.
Assuntos
Doenças do Gato , Injeções/efeitos adversos , Receptores de Estrogênio/metabolismo , Sarcoma , Neoplasias de Tecidos Moles , Animais , Doenças do Gato/patologia , Gatos , Feminino , Antígeno Ki-67/metabolismo , Masculino , Receptores de Progesterona/metabolismo , Estudos Retrospectivos , Sarcoma/patologia , Sarcoma/veterinária , Neoplasias de Tecidos Moles/patologia , Neoplasias de Tecidos Moles/veterináriaRESUMO
A expressão de caderinas tem sido correlacionada ao desenvolvimento e agressividade de neoplasias epiteliais, entretanto, em tumores mamários caninos, seu prognóstico de importância é incerto. Devido a esse fato, a expressão das moléculas de adesão intracelular E e P-caderina e a correlação com a sobrevida global foram analisadas em 25 tumores de glândula mamária canina. A redução da expressão da caderina-E foi correlacionada com o tipo histológico, alto grau histológico e taxa de sobrevida global. A coloração de P-caderina foi maior em tumores malignos, sem relação com outras características clínico-patológicas de agressividade. Os resultados deste estudo sugerem uma relação entre menor expressão de E e P-caderina e pior prognóstico em tumores mamários caninos, incluindo menor sobrevida global.(AU)
Assuntos
Animais , Cães , Cães/anatomia & histologia , Cães/imunologia , Imuno-Histoquímica/veterinária , Caderinas , Neoplasias Mamárias Animais/diagnósticoRESUMO
A expressão de caderinas tem sido correlacionada ao desenvolvimento e agressividade de neoplasias epiteliais, entretanto, em tumores mamários caninos, seu prognóstico de importância é incerto. Devido a esse fato, a expressão das moléculas de adesão intracelular E e P-caderina e a correlação com a sobrevida global foram analisadas em 25 tumores de glândula mamária canina. A redução da expressão da caderina-E foi correlacionada com o tipo histológico, alto grau histológico e taxa de sobrevida global. A coloração de P-caderina foi maior em tumores malignos, sem relação com outras características clínico-patológicas de agressividade. Os resultados deste estudo sugerem uma relação entre menor expressão de E e P-caderina e pior prognóstico em tumores mamários caninos, incluindo menor sobrevida global.