RESUMO
Glioblastoma, which is highly invasive and has a poor patient prognosis, is the most common type of brain tumor. Flavonoids have known antiproliferative and antineoplastic effects, such as apoptosis induction and tumor growth inhibition. We investigated the effects of treatment with three flavonoids (BAS-1, BAS-4, and BAS-6) isolated from the Amazon plant Brosimum acutifolium on the proliferation and migration of the C6 glioma cell line. Cytotoxicity was evaluated by MTT assay, and morphological changes were evaluated by phase-contrast microscopy and by transmission electron microscopy. Apoptosis was determined using Annexin V-FITC-propidium iodide (PI) staining. A hemolysis assay was used to evaluate plasma membrane injury. Antiproliferative effects were assessed by wound migration and colony formation assays. Mitochondrial transmembrane potential (ΔΨm) was determined using JC-1 dye and flow cytometry. To identify the flavonoid targets, western blotting was performed. BAS-1 and BAS-4 reduced C6 cell proliferation in a dose-dependent manner. BAS-6 showed no effect. Due to its high toxicity toward primary glial cells and its high hemolytic index, BAS-1 was not used in the remaining experiments. BAS-4 treatment did not induce cytotoxicity in primary glial cells; however, in glioma cells, it suppressed migration and invasion and led to apoptosis through mitochondrial damage, ΔΨm loss, cell cycle arrest, and reduced AKT phosphorylation, which is a component of the main cell survival pathway. We conclude that BAS-4 showed potential activity against glioma by inducing apoptosis mediated by ΔΨm loss and AKT pathway disruption, and future studies should further evaluate BAS-4 as a promising antineoplastic agent against glioblastoma.
Assuntos
Neoplasias Encefálicas/tratamento farmacológico , Flavonoides/farmacologia , Glioma/tratamento farmacológico , Moraceae/química , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Encefálicas/patologia , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Flavonoides/administração & dosagem , Flavonoides/isolamento & purificação , Citometria de Fluxo , Glioblastoma/tratamento farmacológico , Glioblastoma/patologia , Glioma/patologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos WistarRESUMO
Introducción: Las picaduras de serpientes constituyen un grave problema de Salud Pública. Objetivo: Describir los casos de mordeduras de serpientes notificados al Programa Nacional de Control de Zoonosis y Centro Antirrábico Nacional, Paraguay durante el año 2015. Método: Estudio observacional, descriptivo, retrospectivo y de corte transversal. Los criterios de inclusión fueron toda persona que haya sido mordida por alguna serpiente y que haya sido notificado al Programa Nacional de Control de Zoonosis y Centro Antirrábico Nacional durante el año 2015. Tipo de muestreo no probabilístico. Resultados: Fueron notificados 169 casos de mordeduras de serpientes. De acuerdo con las características sociodemográficas, la mayoría de los afectados fueron varones, con una edad promedio de 26,39 ±1.36. Los lugares más frecuentes donde ocurrieron los accidentes ofídicos fueron las chacras, y en el Departamento de San Pedro. En la mayoría de las mordeduras no se pudo identificar a la serpiente agresora, y cuando fueron identificadas, fueron las del genero Bothrops, las más frecuentes. En cuanto a la localización de las picaduras, fueron más frecuentes en los miembros inferiores, siendo necesario la hospitalización para el tratamiento. No se reportaron fallecidos. Conclusión: En el año 2015 fueron notificados 169 casos, los departamentos con mayor prevalência fueron San Pedro, Itapúa y Caazapá. El perfil del afectado es un varón con promedio de edad de 26 años, trabajador rural. Las serpientes del género Bothrops causaron el 32,5% de las mordeduras. Palabras clave: Mordeduras de serpiente; Envenenamiento; Epidemiología.
Introduction: Snake bites are a serious public health problem. Objective: To describe the cases of snake bites reported to the National Zoonosis Control Program and National Anti-Rabies Center, Paraguay during 2015. Method: Observational, descriptive, retrospective and cross-sectional study. The inclusion criteria were any person who has been bitten by a snake and it has been notified to the National Zoonosis Control Program and National Anti-Rabies Center during 2015. Type of non-probabilistic sampling. Results: 169 cases of snake bites were reported. According to the sociodemographic characteristics, the majority of those affected were males, with an average age of 26.39 ± 1.36. The most frequent places where theophidic accidents occurred were the farms, and in the Department of San Pedro. In most of the bites the aggressor snake could not be identified, and when they were identified, they were those of the genus Bothrops, the most frequent. As for the location of the bites, they were more frequent in the lower limbs, requiring hospitalization for treatment. No deaths were reported. Conclusion: In the year 2015, 169 cases were notified, the departments with the highest prevalence were San Pedro, Itapua and Caazapá. The profile of the affected is a male with an average age of 26 years, a rural worker. The snakes of the genus Bothrops caused 32.5% of the bites. Keywords: Snake bites; Poisoning; Epidemiology.
Assuntos
Humanos , Masculino , Feminino , Recém-Nascido , Lactente , Pré-Escolar , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Mordeduras de Serpentes/epidemiologia , Programas Nacionais de Saúde , Paraguai/epidemiologia , Prevalência , Estudos Transversais , Estudos RetrospectivosRESUMO
Os tumores da bainha perineural são pouco frequentes em animais domésticos. Relata-se o caso de um cão, sem raça definida, de cinco anos, fêmea, com histórico de aumento de volume em região retrobulbar do globo ocular direito (GOD) havia três semanas. Ao exame clínico, constatou-se a presença de uma neoformação retrobulbar direita com deslocamento do globo ocular cranialmente. Aos exames realizados durante o internamento, não foi possível localizar a origem e a extensão da neoformação. O animal veio a óbito por parada cardiorrespiratória e foi encaminhado para a necropsia. À avaliação macroscópica, observou-se neoformação esbranquiçada fixada à base do crânio em região selar, com possível origem no terceiro (III) par de nervos cranianos, a qual se infiltrava no encéfalo na altura do hipotálamo, estendia-se caudalmente em direção ao tronco encefálico e cranialmente à órbita direita, comprimindo, assim, o GOD. Microscopicamente consistia de feixes curtos entrelaçados ou enovelados de células fusiformes com pleomorfismo discreto a moderado, alternando-se a áreas de necrose multifocalmente, compatível com tumor da bainha perineural. Ao exame imuno-histoquímico, apresentou marcação fraca para S100 e GFAP e marcação positiva para vimentina, o que indica caráter maligno.(AU)
Tumors of the perineural sheath are uncommon in domestic animals. We report the case of a 5-year-old female dog with a history of increased volume in the retrobulbar region of the right eye (RE) three weeks ago. The clinical examination revealed a presence of a right retrobulbar neoformation with cranial ocular globe displacement. In the examinations carried out during hospitalization, it was not possible to locate a source and an extension of the neoformation. The animal died of cardiorespiratory arrest and was referred to an autopsy. The macroscopic evaluation revealed a whitish neoformation fixated to the base of the skull in a seal region, with a possible non-III origin of cranial nerves, infiltrating non-encephalon at the height of the hypothalamus, extending caudally towards the brainstem and cranially to the orbit right, compressing RE. Microscopically it consisted of short bundles intertwined or enovelados of spindle cells with discrete to moderate pleomorphism, alternating to areas of multifocal necrosis, compatible with tumor of the perineural sheath. Immunohistochemical examination showed weak marking for S100 and GFAP and positive marking for vimentin, indicating malignancy.(AU)
Assuntos
Animais , Cães , Bradicardia/veterinária , Cães/anormalidades , Síndrome de Horner/veterinária , Neoplasias/diagnósticoRESUMO
Os tumores da bainha perineural são pouco frequentes em animais domésticos. Relata-se o caso de um cão, sem raça definida, de cinco anos, fêmea, com histórico de aumento de volume em região retrobulbar do globo ocular direito (GOD) havia três semanas. Ao exame clínico, constatou-se a presença de uma neoformação retrobulbar direita com deslocamento do globo ocular cranialmente. Aos exames realizados durante o internamento, não foi possível localizar a origem e a extensão da neoformação. O animal veio a óbito por parada cardiorrespiratória e foi encaminhado para a necropsia. À avaliação macroscópica, observou-se neoformação esbranquiçada fixada à base do crânio em região selar, com possível origem no terceiro (III) par de nervos cranianos, a qual se infiltrava no encéfalo na altura do hipotálamo, estendia-se caudalmente em direção ao tronco encefálico e cranialmente à órbita direita, comprimindo, assim, o GOD. Microscopicamente consistia de feixes curtos entrelaçados ou enovelados de células fusiformes com pleomorfismo discreto a moderado, alternando-se a áreas de necrose multifocalmente, compatível com tumor da bainha perineural. Ao exame imuno-histoquímico, apresentou marcação fraca para S100 e GFAP e marcação positiva para vimentina, o que indica caráter maligno.(AU)
Tumors of the perineural sheath are uncommon in domestic animals. We report the case of a 5-year-old female dog with a history of increased volume in the retrobulbar region of the right eye (RE) three weeks ago. The clinical examination revealed a presence of a right retrobulbar neoformation with cranial ocular globe displacement. In the examinations carried out during hospitalization, it was not possible to locate a source and an extension of the neoformation. The animal died of cardiorespiratory arrest and was referred to an autopsy. The macroscopic evaluation revealed a whitish neoformation fixated to the base of the skull in a seal region, with a possible non-III origin of cranial nerves, infiltrating non-encephalon at the height of the hypothalamus, extending caudally towards the brainstem and cranially to the orbit right, compressing RE. Microscopically it consisted of short bundles intertwined or enovelados of spindle cells with discrete to moderate pleomorphism, alternating to areas of multifocal necrosis, compatible with tumor of the perineural sheath. Immunohistochemical examination showed weak marking for S100 and GFAP and positive marking for vimentin, indicating malignancy.(AU)
Assuntos
Animais , Cães , Bradicardia/veterinária , Cães/anormalidades , Síndrome de Horner/veterinária , Neoplasias/diagnósticoRESUMO
Peripheral nerve sheath tumours (PNSTs) are neoplastic growths derived from Schwann cells, perineural cells or both. Malignant PNSTs (MPNSTs) are uncommon in domestic animals. This report describes the concomitant occurrence of PNSTs in a 10-year-old female cocker spaniel with a clinical history of respiratory impairment. Grossly, there was a large infiltrative mass in the caudal lobe of the right lung; smaller nodules were observed in the other lobes of the right lung. Furthermore, a small encapsulated cutaneous nodule was observed on the left hindlimb. Histopathology of the pulmonary tumours revealed the proliferation of pleomorphic spindle-shaped cells with moderate mitotic index arranged in interwoven bundles and concentric Antoni A and Antoni B patterns; invasion of the adjacent pulmonary tissue was observed. The cutaneous nodule consisted of neoplastic mesenchymal cells in interwoven bundles with concentric whorls, but without the marked anisokaryosis, binucleation and infiltrative growth seen in the pulmonary tumour. Immunohistochemistry revealed that both tumours were immunoreactive for vimentin, glial fibrillary acidic protein and S100 protein, but were negative for factor VIII. These findings are indicative of a MPNST in the lung with a concomitant benign PNST in the limb. This case represents the first report of a primary MPNST in the lung of a dog. This neoplastic growth should be included in the differential diagnosis of primary malignant pulmonary tumours of dogs.
Assuntos
Doenças do Cão/patologia , Neoplasias Pulmonares/veterinária , Neoplasias Primárias Múltiplas/veterinária , Neurilemoma/veterinária , Neoplasias do Sistema Nervoso Periférico/veterinária , Neoplasias Cutâneas/veterinária , Animais , Cães , FemininoRESUMO
American tegumentary leishmaniasis is caused by different species of Leishmania. This protozoan employs several mechanisms to subvert the microbicidal activity of macrophages and, given the limited efficacy of current therapies, the development of alternative treatments is essential. Animal venoms are known to exhibit a variety of pharmacological activities, including antiparasitic effects. Crotoxin (CTX) is the main component of Crotalus durissus terrificus venom, and it has several biological effects. Nevertheless, there is no report of CTX activity during macrophage - Leishmania interactions. Thus, the main objective of this study was to evaluate whether CTX has a role in macrophage M1 polarization during Leishmania infection murine macrophages, Leishmania amazonensis promastigotes and L. amazonensis-infected macrophages were challenged with CTX. MTT [3-(4,5dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide] toxicity assays were performed on murine macrophages, and no damage was observed in these cells. Promastigotes, however, were affected by treatment with CTX (IC50 = 22·86 µg mL-1) as were intracellular amastigotes. Macrophages treated with CTX also demonstrated increased reactive oxygen species production. After they were infected with Leishmania, macrophages exhibited an increase in nitric oxide production that converged into an M1 activation profile, as suggested by their elevated production of the cytokines interleukin-6 and tumour necrosis factor-α and changes in their morphology. CTX was able to reverse the L. amazonensis-mediated inhibition of macrophage immune responses and is capable of polarizing macrophages to the M1 profile, which is associated with a better prognosis for cutaneous leishmaniasis treatment.
Assuntos
Crotoxina/farmacologia , Fatores Imunológicos/farmacologia , Leishmania/efeitos dos fármacos , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/parasitologia , Animais , Crotoxina/imunologia , Citocinas/efeitos dos fármacos , Citocinas/metabolismo , Concentração Inibidora 50 , Interleucina-6/biossíntese , Leishmania/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Óxido Nítrico/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Leishmaniasis are a neglected group of emerging diseases that have been found in 98 countries and are caused by protozoa of the genus Leishmania. The therapy for leishmaniasis causes several side effects and leads to drug-resistant strains. Natural products from plants have exhibited activities against Leishmania in various experimental models. Physalis angulata is a widely used plant in popular medicine, and in the literature it has well-documented leishmanicidal activity. However, its mechanism of action is still unknown. Thus, this study aims to evaluate the mechanism driving the leishmanicidal activity of an aqueous extract of P. angulata root (AEPa). AEPa was effective against both promastigotes and intracellular amastigote forms of Leishmania amazonensis. This effect was mediated by an increase of reactive oxygen species (ROS), but not of nitric oxide (NO). The increased production of ROS induces cell death by phenotypes seems by apoptosis cell death in Leishmania, but not autophagy or necrosis. In addition, morphological analysis of macrophages showed that AEPa induced a high number of cytoplasmic projections, increased the volume of cytoplasm and number of vacuoles, caused cytoskeleton alterations and resulted in high spreading ability. AEPa also promoted superoxide anion (O2(-)) production in both uninfected macrophages and those infected with Leishmania. Therefore, these results revealed that AEPa causes cell death by phenotypes seems by apoptosis cell death in L. amazonensis and modulates macrophage activation through morphofunctional alterations and O2(-) generation to induce Leishmania death.
Assuntos
Leishmania/fisiologia , Macrófagos Peritoneais/efeitos dos fármacos , Physalis/química , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Espécies Reativas de Oxigênio/metabolismo , Animais , Apoptose/efeitos dos fármacos , Autofagia , Leishmania/efeitos dos fármacos , Leishmania/imunologia , Estágios do Ciclo de Vida/efeitos dos fármacos , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos Peritoneais/imunologia , Macrófagos Peritoneais/parasitologia , Macrófagos Peritoneais/ultraestrutura , Camundongos , Necrose/parasitologia , FitoterapiaRESUMO
An 11-year-old female German shepherd dog was presented with a history of lameness and pain in the left forelimb. Clinical examination revealed ataxia of the hindlimbs and a subcutaneous mass in the left prescapular region. Radiography revealed metastatic foci in the left humerus, lung and abdomen. Gross necropsy examination revealed a firm, white mass in the left prescapular region. Multiple nodules with similar characteristics were observed in the lung, liver and spleen. Bone lysis was noted in the humerus and the fifth to seventh lumbar vertebrae. Microscopical examination revealed a proliferation of basal cells forming irregular islands of various sizes and surrounding extensive zones of keratinized 'ghost' cells. A definitive diagnosis of malignant pilomatricoma was made. This is a rare tumour in dogs with no previous report of metastasis to the spleen and liver.
Assuntos
Doenças do Cão/patologia , Doenças do Cabelo/veterinária , Pilomatrixoma/veterinária , Neoplasias Cutâneas/veterinária , Animais , Neoplasias Ósseas/secundário , Neoplasias Ósseas/veterinária , Carcinoma Basocelular/diagnóstico , Carcinoma Basocelular/veterinária , Diagnóstico Diferencial , Cães , Feminino , Doenças do Cabelo/diagnóstico , Doenças do Cabelo/patologia , Úmero/patologia , Vértebras Lombares/patologia , Pilomatrixoma/secundário , Neoplasias Cutâneas/patologiaRESUMO
A relação entre Helicobacter spp. e a presença de alterações histológicas na pars esophagea de suínos foi avaliada em 67 estômagos de animais em idade de abate. Para a identificação das helicobactérias, utilizou-se a técnica da PCR com primers específicos para o gênero Helicobacter. As alterações histológicas foram identificadas e classificadas como ulceração, erosão, degeneração epitelial, alongamento de papilas, hiperplasia, paraqueratose, intensidade do infiltrado inflamatório e aumento do número de folículos linfoides. As alterações mais frequentemente encontradas na pars esophagea foram a degeneração epitelial e o alongamento de papilas, observadas em 83,5 por cento (n=56) das amostras analisadas. Em 77,5 por cento (n=52) das amostras, observou-se paraqueratose e em 61,1 por cento (n=41) hiperplasia epitelial. Quarenta e sete (70,1 por cento) foram positivas na PCR para Helicobacter spp. Nessas amostras a erosão foi a lesão mais observada (40,2 por cento), seguida de ulceração da mucosa (11,9 por cento). Em 58,2 por cento das amostras positivas na PCR, não foram observadas ulcerações de mucosa. Observou-se associação significativa (P=0,003) entre a presença de Helicobacter spp. e a degeneração epitelial da pars esophagea de suínos em idade de abate.(AU)
The association between histological findings of gastric mucosa in pigs at slaughtering age and the presence of Helicobacter spp., identified by PCR, assay was investigated. Stomachs from 67 pigs were examined. Histological changes of pars esophagea were identified and classified as gastric ulcers, erosion, degeneration, distortion of papils, hyperplasia, paraqueratosis, and number of lymphoid follicles. Microscopic analysis revealed the most frequent alteration: 83.5 percent (n= 56) stomachs with epithelial degeneration and distortion of papils. Paraqueratosis of pars esophagea was observed in 77.5 percent (n=52) of the samples and epithelial hyperplasia in 61 percent (n=41). Forty-seven (70.1 percent) pigs were positive to Helycobacter spp. by PCR. Erosion of pars esophagea and ulceration were the most frequent findings in Helicobacter spp. PCR-positive pigs, occurring, respectively, in 40.2 percent and 11.9 percent. The frequency of animals without ulceration and Helicobacter spp. PCR-positive was 58.2 percent. It was observed a significant association (P=0.003) between Helicobacter spp. and epithelial degeneration of gastric mucosa in pigs at slaughtering age.(AU)
Assuntos
Animais , Helicobacter/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Estômago/anatomia & histologia , Estômago/patologia , Gastropatias/veterinária , Úlcera Gástrica/veterinária , SuínosRESUMO
A relação entre Helicobacter spp. e a presença de alterações histológicas na pars esophagea de suínos foi avaliada em 67 estômagos de animais em idade de abate. Para a identificação das helicobactérias, utilizou-se a técnica da PCR com primers específicos para o gênero Helicobacter. As alterações histológicas foram identificadas e classificadas como ulceração, erosão, degeneração epitelial, alongamento de papilas, hiperplasia, paraqueratose, intensidade do infiltrado inflamatório e aumento do número de folículos linfoides. As alterações mais frequentemente encontradas na pars esophagea foram a degeneração epitelial e o alongamento de papilas, observadas em 83,5 por cento (n=56) das amostras analisadas. Em 77,5 por cento (n=52) das amostras, observou-se paraqueratose e em 61,1 por cento (n=41) hiperplasia epitelial. Quarenta e sete (70,1 por cento) foram positivas na PCR para Helicobacter spp. Nessas amostras a erosão foi a lesão mais observada (40,2 por cento), seguida de ulceração da mucosa (11,9 por cento). Em 58,2 por cento das amostras positivas na PCR, não foram observadas ulcerações de mucosa. Observou-se associação significativa (P=0,003) entre a presença de Helicobacter spp. e a degeneração epitelial da pars esophagea de suínos em idade de abate.
The association between histological findings of gastric mucosa in pigs at slaughtering age and the presence of Helicobacter spp., identified by PCR, assay was investigated. Stomachs from 67 pigs were examined. Histological changes of pars esophagea were identified and classified as gastric ulcers, erosion, degeneration, distortion of papils, hyperplasia, paraqueratosis, and number of lymphoid follicles. Microscopic analysis revealed the most frequent alteration: 83.5 percent (n= 56) stomachs with epithelial degeneration and distortion of papils. Paraqueratosis of pars esophagea was observed in 77.5 percent (n=52) of the samples and epithelial hyperplasia in 61 percent (n=41). Forty-seven (70.1 percent) pigs were positive to Helycobacter spp. by PCR. Erosion of pars esophagea and ulceration were the most frequent findings in Helicobacter spp. PCR-positive pigs, occurring, respectively, in 40.2 percent and 11.9 percent. The frequency of animals without ulceration and Helicobacter spp. PCR-positive was 58.2 percent. It was observed a significant association (P=0.003) between Helicobacter spp. and epithelial degeneration of gastric mucosa in pigs at slaughtering age.