RESUMO
OBJECTIVES: We compared the proteomic signatures of the hippocampal lesion induced in three different animal models of mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE+HS): the systemic pilocarpine model (PILO), the intracerebroventricular kainic acid model (KA), and the perforant pathway stimulation model (PPS). METHODS: We used shotgun proteomics to analyze the proteomes and find enriched biological pathways of the dorsal and ventral dentate gyrus (DG) isolated from the hippocampi of the three animal models. We also compared the proteomes obtained in the animal models to that from the DG of patients with pharmacoresistant MTLE+HS. RESULTS: We found that each animal model presents specific profiles of proteomic changes. The PILO model showed responses predominantly related to neuronal excitatory imbalance. The KA model revealed alterations mainly in synaptic activity. The PPS model displayed abnormalities in metabolism and oxidative stress. We also identified common biological pathways enriched in all three models, such as inflammation and immune response, which were also observed in tissue from patients. However, none of the models could recapitulate the profile of molecular changes observed in tissue from patients. SIGNIFICANCE: Our results indicate that each model has its own set of biological responses leading to epilepsy. Thus, it seems that only using a combination of the three models may one replicate more closely the mechanisms underlying MTLE+HS as seen in patients.
Assuntos
Epilepsia do Lobo Temporal , Epilepsia , Animais , Benchmarking , Modelos Animais de Doenças , Epilepsia/patologia , Epilepsia do Lobo Temporal/patologia , Humanos , Proteoma , Proteômica , EscleroseRESUMO
This study describes an outbreak of alopecia in laboratory rats caused by the saprophytic fungi Scopulariopsis brevicaulis in a Brazilian lab animal facility. Initially, lesions were identified only in females during the lactation period, which showed circumscribed areas with alopecia without pruritus extensively at the abdomen and the medial side of the thighs. Later during the course of the infection, skin lesions affected both genders reaching about 10% of the colony. Skin scrapings, hair, and environmental samples from the facility were sent for analysis. The cultures were carried out in Sabouraud agar and over seven days fast-growing colonies varying in color from white to brown were observed. Micromorphology showed mycelium and characteristic ringworms of the fungus S. brevicaulis. There is a similar report of an outbreak of this fungus in Turkey. To date, this is the first report in the literature of the fungus S. brevicaulis causing disease in laboratory rats in Brazil.
Assuntos
Animais , Ratos , Surtos de Doenças/veterinária , Alopecia/veterinária , Alopecia/epidemiologia , Scopulariopsis , Animais de Laboratório/microbiologia , Ratos Wistar , AbdomeRESUMO
PURPOSE: To evaluate the effect of fractional carbon dioxide (CO2) laser treatment using lower power associated with pulse stacking within collagen fibers, using second harmonic generation microscopy and computerized image analysis. METHODS: Twenty male Wistar rats aging eight weeks were used. Each treatment area received a single-pass CO2 fractional laser with different parameters. The 20 animals were divided into two groups and euthanized after 30 and 60 days. Second harmonic generation images were obtained and program ImageJ was utilized to evaluate the collagen organization within all areas. Collagen anisotropy, entropy and optical density were quantified. RESULTS: Increased anisotropy over time was observed in all four areas, but only reached statistical significance (p = 0.0305) when the mildest parameters were used (area four). Entropy decreased over time in all areas, but without significance(p = 0.1779) in area four. Density showed an overtime increase only in area four, but no statistical significance was reached (p = 0.6534). CONCLUSIONS: When combined, the results obtained in this study regarding anisotropy, entropy and density tend to demonstrate that it is possible to achieve collagen remodeling with the use of lower power levels associated with stacked pulses.
Assuntos
Lasers de Gás , Microscopia de Geração do Segundo Harmônico , Envelhecimento da Pele , Animais , Dióxido de Carbono , Masculino , Ratos , Ratos Wistar , PeleRESUMO
Murine hepatitis virus (MHV) strain 3, one of the most important inducers of viral hepatitis, has been extensively studied as an organism to gain a better understanding of coronavirus biology and pathogenesis. Only one sequence is currently available. Another representative isolate has now been sequenced and added to the arsenal of MHV-3 variants.
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Mesial temporal lobe epilepsy (MTLE) is a chronic neurological disorder characterized by the occurrence of seizures, and histopathological abnormalities in the mesial temporal lobe structures, mainly hippocampal sclerosis (HS). We used a multi-omics approach to determine the profile of transcript and protein expression in the dorsal and ventral hippocampal dentate gyrus (DG) and Cornu Ammonis 3 (CA3) in an animal model of MTLE induced by pilocarpine. We performed label-free proteomics and RNAseq from laser-microdissected tissue isolated from pilocarpine-induced Wistar rats. We divided the DG and CA3 into dorsal and ventral areas and analyzed them separately. We performed a data integration analysis and evaluated enriched signaling pathways, as well as the integrated networks generated based on the gene ontology processes. Our results indicate differences in the transcriptomic and proteomic profiles among the DG and the CA3 subfields of the hippocampus. Moreover, our data suggest that epileptogenesis is enhanced in the CA3 region when compared to the DG, with most abnormalities in transcript and protein levels occurring in the CA3. Furthermore, our results show that the epileptogenesis in the pilocarpine model involves predominantly abnormal regulation of excitatory neuronal mechanisms mediated by N-methyl D-aspartate (NMDA) receptors, changes in the serotonin signaling, and neuronal activity controlled by calcium/calmodulin-dependent protein kinase (CaMK) regulation and leucine-rich repeat kinase 2 (LRRK2)/WNT signaling pathways.
Assuntos
Epilepsia do Lobo Temporal , Animais , Epilepsia do Lobo Temporal/patologia , Hipocampo/metabolismo , Pilocarpina/toxicidade , Proteômica , Ratos , Ratos WistarRESUMO
Purpose: To evaluate the effect of fractional carbon dioxide (CO2 ) laser treatment using lower power associated with pulse stacking within collagen fibers, using second harmonic generation microscopy and computerized image analysis. Methods: Twenty male Wistar rats aging eight weeks were used. Each treatment area received a single-pass CO2 fractional laser with different parameters. The 20 animals were divided into two groups and euthanized after 30 and 60 days. Second harmonic generation images were obtained and program ImageJ was utilized to evaluate the collagen organization within all areas. Collagen anisotropy, entropy and optical density were quantified. Results: Increased anisotropy over time was observed in all four areas, but only reached statistical significance (p = 0.0305) when the mildest parameters were used (area four). Entropy decreased over time in all areas, but without significance (p = 0.1779) in area four. Density showed an overtime increase only in area four, but no statistical significance was reached (p = 0.6534). Conclusion: When combined, the results obtained in this study regarding anisotropy, entropy and density tend to demonstrate that it is possible to achieve collagen remodeling with the use of lower power levels associated with stacked pulses.(AU)
Assuntos
Animais , Ratos , Dióxido de Carbono/uso terapêutico , Lasers de Gás/uso terapêutico , Colágeno , Envelhecimento da PeleRESUMO
Mesial temporal lobe epilepsy (MTLE) is a chronic neurological disorder affecting almost 40% of adult patients with epilepsy. Hippocampal sclerosis (HS) is a common histopathological abnormality found in patients with MTLE. HS is characterised by extensive neuronal loss in different hippocampus sub-regions. In this study, we used laser microdissection-based microproteomics to determine the protein abundances in different regions and layers of the hippocampus dentate gyrus (DG) in an electric stimulation rodent model which displays classical HS damage similar to that found in patients with MTLE. Our results indicate that there are differences in the proteomic profiles of different layers (granule cell and molecular), as well as different regions, of the DG (ventral and dorsal). We have identified new signalling pathways and proteins present in specific layers and regions of the DG, such as PARK7, RACK1, and connexin 31/gap junction. We also found two major signalling pathways that are common to all layers and regions: inflammation and energy metabolism. Finally, our results highlight the utility of high-throughput microproteomics and spatial-limited isolation of tissues in the study of complex disorders to fully appreciate the large biological heterogeneity present in different cell populations within the central nervous system.
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Conexinas/metabolismo , Epilepsia do Lobo Temporal/metabolismo , Hipocampo/metabolismo , Proteína Desglicase DJ-1/metabolismo , Proteômica/métodos , Receptores de Quinase C Ativada/metabolismo , Animais , Modelos Animais de Doenças , Epilepsia do Lobo Temporal/etiologia , Regulação da Expressão Gênica , Humanos , Microdissecção e Captura a Laser , Especificidade de Órgãos , Mapas de Interação de Proteínas , Ratos , Transdução de SinaisRESUMO
Norovirus is a highly prevalent pathogen that can infect a wide range of host species. Thus far, there have only been two reports of norovirus infection in rats. Diagnostic assays for the detection of norovirus are well established, but a specific molecular assay for the diagnosis of norovirus infection in laboratory rats has not yet been reported. In this study, we describe the development of a sensitive, semi-nested RT-PCR assay for detection of norovirus in fecal samples from Rattus norvegicus, reared in animal facilities under different sanitary barrier conditions. Additionally, we describe the first report of the presence of norovirus in rat colonies from Brazilian animal facilities.
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Infecções por Caliciviridae/diagnóstico , Fezes/virologia , Norovirus/isolamento & purificação , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Doenças dos Roedores/diagnóstico , Animais , Brasil , Reação em Cadeia da Polimerase/métodos , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: In recent decades some outbreaks of food-borne acute Chagas disease (ACD) in humans were identified by clinical and epidemiological characterization after association through the ingestion of açaí pulp probably contaminated with Trypanosoma cruzi. Whereas Belém and Abaetetuba stood out as important risk regions for disease transmission, the importance of Rhodnius pictipes, and Philander opossum for the biological cycle of T. cruzi, and data from agribusiness market of açaí, to study T. cruzi from vector and reservoir of the Brazilian Amazon region is critical for this context. Thus, the purpose of this study was to verify the infective capacity and the virulence of T. cruzi in açaí pulp from vector and reservoir at Pará State experimentally. METHODS: 105T. cruzi I in in natura açaí pulp from Belém at Pará State, at room temperature, after forced sieving, by intraperitoneal, gavage or oral route of inoculation in B6.129S7Rag1-/-tmMom/J Unib allowed food-borne ACD analysis using common light microscopy. PRINCIPAL FINDINGS: T. cruzi in in natura açaí pulp from R. pictipes (Val-De-Cans Forest, Belém, and Ajuaí River, Abaetetuba, Pará), and P. opossum (Combu Island, Belém, Pará) caused ACD and death between 17 and 52 days after experimental infections in murine immunodeficient hosts. CONCLUSIONS: T. cruzi from different sources and locations at Pará State in in natura açaí pulp retained its infective capacity and virulence, and can cause new outbreaks of ACD by oral transmission. Additionally, quality basic education will facilitate efficient hygiene practices throughout the açaí productive chain can eradicate food-borne ACD in the coming decades.
Assuntos
Doença de Chagas/transmissão , Euterpe/parasitologia , Parasitologia de Alimentos , Doenças Transmitidas por Alimentos/parasitologia , Trypanosoma cruzi/patogenicidade , Doença Aguda , Animais , Brasil/epidemiologia , Doença de Chagas/epidemiologia , Doença de Chagas/parasitologia , Reservatórios de Doenças/parasitologia , Vetores de Doenças , Feminino , Doenças Transmitidas por Alimentos/epidemiologia , Insetos Vetores/parasitologia , Masculino , Camundongos , Camundongos Endogâmicos , Gambás/parasitologia , Parasitemia/epidemiologia , Parasitemia/mortalidade , Rhodnius/parasitologia , VirulênciaRESUMO
Bartonella spp. are bacteria of worldwide distribution that cause asymptomatic to fatal infections in animals and humans. The most common zoonotic species is Bartonella henselae, for which cats are the major natural reservoir host. To better understand Bartonella sp. diagnostic limitations, we determined the frequency of bloodstream infection in 112 cats by comparing and combining the results of multiple conventional and nested PCRs from blood and liquid culture samples. Using liquid culture conventional PCR, Bartonella sp. DNA was amplified from 27.7% of samples (31/112) compared to 90.2% of samples (101/112) by combining nested PCR from blood and liquid culture, indicating that PCR testing of more than one type of sample provides better sensitivity than a standalone PCR and that bloodstream infection is very frequent among cats in southeastern Brazil. This study reinforces the need for multistep testing for Bartonella sp. infection to prevent false-negative diagnostic results, even in reservoir hosts such as cats that typically maintain higher bacteremia levels.