Domestic dogs are the main reservoir of Leishmania infantum, a causative agent of visceral leishmaniasis (VL). The number of human disease cases is associated with the rate of canine infection. Currently available drugs are not efficient at treating canine leishmaniasis (CanL) and months after the treatment most dogs show disease relapse, therefore the development of new drugs or new therapeutic strategies should be sought. In CanL, dogs lack the ability to mount a specific cellular immune response suitable for combating the parasite and manipulation of cytokine signaling pathway has the potential to form part of effective immunotherapeutic methods. In this study, recombinant canine cytokines (rcaIL-12, rcaIL-2, rcaIL-15 and rcaIL-7) and soluble receptor IL-10R1 (rcasIL-10R1), with antagonistic activity, were evaluated for the first time in combination (rcaIL-12/rcaIL-2, rcaIL-12/rcaIL-15, rcaIL-12/rcasIL-10R1, rcaIL-15/rcaIL-7) or alone (rcasIL-10R1) to evaluate their immunomodulatory capacity in peripheral blood mononuclear cells (PBMCs) from dogs with leishmaniasis. All the combinations of recombinant proteins tested were shown to improve lymphoproliferative response. Further, the combinations rcaIL-12/rcaIL-2 and rcaIL-12/rcaIL-15 promoted a decrease in programmed cell death protein 1 (PD-1) expression in lymphocytes. These same combinations of cytokines and rcaIL-12/rcasIL-10R1 induced IFN-γ and TNF-α production in PBMCs. Furthermore, the combination IL-12/IL-15 led to an increased in T-bet expression in lymphocytes. These findings are encouraging and indicate the use of rcaIL-12 and rcaIL-15 in future in vivo studies aimed at achieving polarization of cellular immune responses in dogs with leishmaniasis, which may contribute to the development of an effective treatment against CanL.
A B S T R A C TCurrently aquatic ecosystems have been altered significantly making it necessary the use of biological monitoring for these environments. In this perspective, this study aimed to analyze the influence of the use and occupation in sub-basin water quality of the Melo Machado Stream, in Araçatuba/SP, using benthic macroinvertebrates as bioindicators. The methodology was developed by means of satellite images geoprocessing, visit in locus and laboratory analysis. The results obtained in this study showed a decrease in the diversity of individuals directly proportional to the degree of land use and occupation, highlighting its impact on water quality in each sampling station. It is noteworthy that the results obtained in this study may support actions and strategies for environmental management of aquatic environments, ensuring the recovery of these environments are important elements in environmental quality. Keywords: Aquatic ecosystems, bioindicators, macroinvertebrates benthic. R E S U M OAtualmente os ecossistemas aquáticos têm sido alterados de maneira significativa tornando-se necessário o uso de medidas biológicas para monitoramento desses ambientes. Nesta perspectiva, o presente estudo objetivou analisar a influência do uso e ocupaçãodo solo na qualidade da água da sub-bacia do Córrego Machado de Melo, no município de Araçatuba/SP, utilizandomacroinvertebrados bentônicos como bioindicadores. A metodologia foi desenvolvida por meio de geoprocessamento de imagens desatélite, visita in locus e análises laboratoriais. Os resultados obtidos no presenteestudo revelaram uma diminuição da diversidade de indivíduos diretamenteproporcional ao grau do uso e ocupação do solo, evidenciando o seu impacto naqualidade da água em cada estação amostral. Vale ressaltar que os resultados obtidos nesta pesquisapoderão subsidiar ações e estratégias para a gestão ambiental de ambientesaquáticos continentais, assegurando a recuperação desses ambientes queconstituem importantes elementos na qualidade ambiental. Palavras-Chave: Ecossistemas aquáticos, bioindicadores, macroinvertebrados bentônicos. IntroduçãoA qualidade das águas de uma bacia hidrográfica pode ser influenciada por diversos fatores, dentre os impactos ambientais que podem alterar o ecossistema aquáticos. Rebouças et al. (2002) relatam que o desmatamento, o uso do solo
Eosinophils are traditionally associated with the immune response against helminth parasites. However, several studies have demonstrated that these cells have a role regarding protective immunity in leishmaniasis. Here, we examined the relationship between the presence of eosinophils and parasite load in biopsy samples from dogs, obtained through fine needle puncture and aspiration of lymph nodes. Histological slides containing biopsy material from lymph nodes of dogs with canine visceral leishmaniasis and healthy dogs were used to obtain baseline eosinophil counts. Subsequently, scrapings were taken from slides for DNA extraction and determination of parasite load, using real-time PCR (qRT-PCR). Additionally, production of nitric oxide (NO) and reactive oxygen species (ROS) levels by eosinophils in the peripheral blood of dogs with canine visceral leishmaniasis and healthy dogs was measured. The eosinophil percentage were higher in lymph nodes of infected group, and the parasite load showed a significant negative correlation with the eosinophil count. The production of NO and ROS by eosinophils in the peripheral blood was higher in the dogs with canine visceral leishmaniasis. All the results together suggest that eosinophils may participate in antileishmanial immunity in canine visceral leishmaniasis.
A B S T R A C TIn aquatic ecosystems, the presence of macrophytes offers survival conditions for colonization of macroinvertebrates. This study aimed to characterize the fauna of macroinvertebrates associated with Pistia stratiotes L. (Araceae), in the Capivaras Lagoon, located inside the Baguaçu Ecological Park, in the municipality of Araçatuba, and to relate them to the physical and chemical parameters of the water. The sampling of P. stratiotes was in three points; the samples were sorted and later identified. It was observed a high density of Oligochaeta and Chironomidae larvae in the three points sampled, which may be related to the higher capacity of these organisms tolerate adverse environmental conditions, such as the low concentration of oxygen dissolved in the water obtained in the physical-chemical analyzes. The small lake has a history of pollution and currently suffers from low levels of oxygenation and infestation of P. stratiotes forming a long "rug" enclosed throughout its cover. Considering find solutions to recover the integrity of the environment and,, mainly, to recover the oxygenation of the water, it is suggested more investigations in the area, in several areas and the drainage of the inlet and outlet water channel of the lagoon, beyond the drainage of input and output of water of the lagoon, to improve the rapid circulation, avoiding the establishment of large banks of aquatic macrophytes. Keywords: Aquatic ecosystems, macroinvertebrates, Pistia stratiotes. R E S U M OEm ecossistemas aquáticos, a presença das macrófitas oferece condições de sobrevivência para colonização de macroinvertebrados. Este estudo objetivou caracterizar a fauna de macroinvertebrados associados à Pistia stratiotes L. (Araceae), na Lagoa das Capivaras, localizada no interior do Parque Ecológico Baguaçu, no município de Araçatuba e relacioná-las aos parâmetros físicos e químicos da água. A coleta de P. stratiotes foi realizada em três pontos, as amostras foram triadas e posteriormente identificadas. Foi observada uma alta densidade de Oligochaeta e larvas de Chironomidae nos três pontos amostrados, a qual pode estar relacionada a maior capacidade desses organismos tolerarem condições adversas do ambiente, tais como a baixa concentração de oxigênio dissolvido na água determinada nas análises físico-químicas. O pequeno lago tem um histórico de poluição e, atualmente, sofre com baixos níveis de oxigenação e a infestação de P. stratiotes formando um longo "tapete" fechado em toda a sua cobertura. Visando encontrar soluções para recompor a integridade do ambiente e, principalmente, recuperar a oxigenação da água, sugerem-se mais estudos na área, além da drenagem do canal de entrada e de saída da água da lagoa, para que a ela circule com maior rapidez, impedindo a fixação de grandes bancos de macrófitas aquáticas. Palavras-Chave: Ecossistemas aquáticos, macroinvertebrados, Pistia stratiotes.
PGE 2 is a product derived from arachidonic acid, produced by the combination of cyclooxygenases (COX-1 and COX-2). 1 COX-1, expressed constitutively in most cells, is the dominant source of prostanoids that subserve housekeeping functions, such as gastric epithelial cytoprotection and homeostasis. 2 COX-2, induced by inflammatory stimuli, hormones and growth factors, is the most important source of prostanoid formation in inflammation and proliferative diseases, such as cancer. 2 PGE 2 is produced by various cell types. 3 PGE 2 is one of the most abundant PGs produced in the body, is most widely characterized in animal species and exhibits versatile biological activities.Under physiological conditions, PGE 2 is an important mediator of many biological functions, such as regulation of immune responses, blood pressure, gastrointestinal integrity and fertility. AbstractCanine leishmaniasis (CanL) is caused by the intracellular parasite Leishmania infantum. Prostaglandin E2 (PGE 2 ) exerts potent regulatory effects on the immune system in experimental model Leishmania infection, but this influence has not yet been studied in CanL. In this study, PGE 2 and PGE 2 receptor levels and the regulatory effect of PGE 2 on arginase activity, NO 2 , IL-10, IL-17, IFN-γ, TNF-α and parasite load were evaluated in cultures of splenic leucocytes obtained from dogs with CanL in the presence of agonists and inhibitors. Our results showed that splenic leucocytes from dogs with CanL had lower EP2 receptor levels than those of splenic leucocytes from healthy animals. We observed that NO 2 levels decreased when the cells were treated with a PGE 2 receptor agonist (EP1/EP2/EP3) or COX-2 inhibitor (NS-398) and that TNF-α, IL-17 and IFN-γ cytokine levels decreased when the cells were treated with a PGE 2 receptor agonist (EP2) or PGE 2 itself. The parasite load in splenic leucocyte cell cultures from dogs with CanL decreased after stimulation of the cells with PGE 2 . We conclude that Leishmania infection of dogs modulates PGE 2 receptors and speculate that the binding of PGE 2 to its receptors may activate the microbicidal capacity of cells. K E Y W O R D Sdogs, Leishmania spp., prostaglandin E2, receptors
In this study, we evaluated the performance of a new enzyme‐linked immunosorbent assay (ELISA) variant known as indirect “plasmonic ELISA” (pELISA) for the detection of Leishmania spp. infection. Serum samples from 170 dogs from an area where canine leishmaniosis (CanL) is endemic and from 26 healthy dogs from a nonendemic area were tested by indirect pELISA, and the results were compared to those of an indirect ELISA (both with recombinant antigen rK28) and those of an immunochromatographic test (dual‐path platform, TR‐DPP®) using real‐time PCR on blood samples or conjunctival swabs as the gold standard. The pELISA, indirect rK28 ELISA and the TR‐DPP® immunochromatographic test presented sensitivities of 94.7%, 89.5% and 79.0% and specificities of 100%, 92.7% and 91.5%, respectively. The analysis of the results revealed that the specificity of the indirect pELISA was greater than that of the method recommended by the Ministry of Health in Brazil and may increase the feasibility of diagnosis in resource‐constrained countries because it does not require sophisticated instruments to read. Thus, this method can be used as an additional tool for the detection of Leishmania spp. infection in these areas.
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