EFAR - Escola de Farmácia
URI permanente desta comunidadehttp://www.hml.repositorio.ufop.br/handle/123456789/451
Notícias
O curso de Farmácia em Ouro Preto foi criado em 1839, sendo a mais antiga Escola de Farmácia da América Latina.
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29 resultados
Resultados da Pesquisa
Item Risk profile for Leishmania infection in dogs coming from an area of visceral leishmaniasis reemergence.(2018) Leal, Gleisiane Gomes de Almeida; Carneiro, Mariângela; Pinheiro, Aimara da Costa; Marques, Luana Araújo; Ker, Henrique Gama; Reis, Alexandre Barbosa; Vital, Wendel CouraUntil the 1980s, visceral leishmaniasis was concentrated in poor rural areas of Brazil. The Vale do Rio Doce, located in the Southeastern Brazilian state of Minas Gerais, was an endemic area with high numbers of human and canine cases. Prophylactic measures adopted since the 1960s reduced the number of cases and the region became a ‘controlled endemic’ area. In the early 1990s, however, the program was interrupted, and the human disease reemerged in 2008. This cross-sectional study evaluated the prevalence and the risk profile of infection of dogs with Leishmania spp in this reemergence area of visceral leishmaniasis. Among a population of approximately 280,000 people, a total of 3835 dog owners were interviewed about socioeconomic conditions, housing, peridomicile features, and their dogs’ characteristics and behavior. Blood samples were collected from 5822 dogs of an estimated canine population of 20,000 and anti-Leishmaniasis antibodies were identified using Dual-Path Platform and ELISA. We observed that 1282 of the 5822 dogs were seropositive for the protozoan indicating a seroprevalence of 22%. The risk factors associated with Leishmania infection in dogs were: non-paved backyard (OR 1.4; 95%CI 1.2–1.7); the presence of dry leaves and decaying fruit in the backyard (OR 1.3; 95%CI 1.1–1.5); medium-sized (OR 1.3; 95% 1.1–1.5) or big-sized dogs (OR 1.8; 95%CI 1.5–2.3); short-haired dogs (OR 1.8; 95%CI 1.5–2.1); dogs that slept in the backyard (OR 2.6; 95%CI 1.8–3.6) or in the balcony (OR 1.6; 95%CI 1.1–2.3); and history of canine visceral leishmaniasis in the household (OR 1.3; 95%CI 1.1–1.5). Our results suggest a strong reemergence of canine visceral leishmaniasis after the discontinuation of the control programs. Also, the observed risk factors reinforce the role of health education and environmental management measures to the effective control of the disease.Item LBSapSal-vaccinated dogs exhibit increased circulating T-lymphocyte subsets (CD4+ and CD8+) as well as a reduction of parasitism after challenge with Leishmania infantum plus salivary gland of Lutzomyia longipalpis.(2014) Soares, Rodrigo Dian de Oliveira Aguiar; Roatt, Bruno Mendes; Ker, Henrique Gama; Moreira, Nádia das Dores; Mathias, Fernando Augusto Siqueira; Cardoso, Jamille Mirelle de Oliveira; Gontijo, Nelder de Figueiredo; Romero, Oscar Bruna; Carvalho, Andréa Teixeira de; Martins Filho, Olindo Assis; Oliveira, Rodrigo Corrêa de; Giunchetti, Rodolfo Cordeiro; Reis, Alexandre BarbosaBackground: The development of a protective vaccine against canine visceral leishmaniasis (CVL) is an alternative approach for interrupting the domestic cycle of Leishmania infantum. Given the importance of sand fly salivary proteins as potent immunogens obligatorily co-deposited during transmission of Leishmania parasites, their inclusion in an anti-Leishmania vaccine has been investigated in the last few decades. In this context, we previously immunized dogs with a vaccine composed of L. braziliensis antigens plus saponin as the adjuvant and sand fly salivary gland extract (LBSapSal vaccine). This vaccine elicited an increase in both anti-saliva and anti-Leishmania IgG isotypes, higher counts of specific circulating CD8+ T cells, and high NO production. Methods: We investigated the immunogenicity and protective effect of LBSapSal vaccination after intradermal challenge with 1 × 107 late-log-phase L. infantum promastigotes in the presence of sand fly saliva of Lutzomyia longipalpis. The dogs were followed for up to 885 days after challenge. Results: The LBSapSal vaccine presents extensive antigenic diversity with persistent humoral and cellular immune responses, indicating resistance against CVL is triggered by high levels of total IgG and its subtypes (IgG1 and IgG2); expansion of circulating CD5+, CD4+, and CD8+ T lymphocytes and is Leishmania-specific; and reduction of splenic parasite load. Conclusions: These results encourage further study of vaccine strategies addressing Leishmania antigens in combination with proteins present in the saliva of the vector.Item Immunological profile of resistance and susceptibility innaturally infected dogs by Leishmania infantum.(2014) Leal, Gleisiane Gomes de Almeida; Roatt, Bruno Mendes; Soares, Rodrigo Dian de Oliveira Aguiar; Carneiro, Cláudia Martins; Giunchetti, Rodolfo Cordeiro; Carvalho, Andréa Teixeira de; Martins Filho, Olindo Assis; Francisco, Amanda Fortes; Cardoso, Jamille Mirelle de Oliveira; Mathias, Fernando Augusto Siqueira; Oliveira, Rodrigo Corrêa de; Carneiro, Mariângela; Vital, Wendel Coura; Reis, Alexandre BarbosaVisceral leishmaniasis has a great impact on public health, and dogs are considered the maindomestic reservoir of Leishmania infantum, the causal parasite. In this study, 159 animalsnaturally infected by L. infantum from an endemic area of Brazil were evaluated through ananalysis of cellular responses, using flow cytometry, and of the hematological parameters.The results confirmed that disease progression is associated with anemia and reductionsin eosinophils, monocytes and lymphocytes. The investigation of the immune response,based on the immunophenotypic profile of peripheral blood, showed declines in the abso-lute numbers of T lymphocytes CD5+and their subsets (CD4+and CD8+) and a drop of Blymphocytes in asymptomatic seropositive (AD-II) and symptomatic seropositive (SD) dogs.Neutrophils, when stimulated with soluble antigen of L. infantum, showed higher synthesisof interferon (IFN)- _+in AD-II and SD groups, with decreased production of interleukinItem Molecular diagnosis of canine visceral leishmaniasis : a comparative study of three methods using skin and spleen from dogs with natural Leishmania infantum infection.(2013) Reis, Levi Eduardo Soares; Vital, Wendel Coura; Roatt, Bruno Mendes; Bouillet, Leoneide Érica Maduro; Ker, Henrique Gama; Brito, Rory Cristiane Fortes de; Resende, Daniela de Melo; Carneiro, Cláudia Martins; Reis, Alexandre BarbosaPolymerase chain reaction (PCR) and its variations represent highly sensitive and specificmethods for Leishmania DNA detection and subsequent canine visceral leishmaniasis (CVL)diagnosis. The aim of this work was to compare three different molecular diagnosis tech-niques (conventional PCR [cPCR], seminested PCR [snPCR], and quantitative PCR [qPCR])in samples of skin and spleen from 60 seropositive dogs by immunofluorescence antibodytest and enzyme-linked immunosorbent assay. Parasitological analysis was conducted byculture of bone marrow aspirate and optical microscopic assessment of ear skin and spleensamples stained with Giemsa, the standard tests for CVL diagnosis. The primers L150/L152and LINR4/LIN17/LIN19 were used to amplify the conserved region of the Leishmania kDNAminicircle in the cPCR, and snPCR and qPCR were performed using the DNA polymerasegene (DNA pol _) primers from Leishmania infantum. The parasitological analysis revealedparasites in 61.7% of the samples. Sensitivities were 89.2%, 86.5%, and 97.3% in the skin and81.1%, 94.6%, and 100.0% in spleen samples used for cPCR, snPCR, and qPCR, respectively.We demonstrated that the qPCR method was the best technique to detect L. infantum inboth skin and spleen samples. However, we recommend the use of skin due to the highsensitivity and sampling being less invasive.Item Cytokine and nitric oxide patterns in dogs immunized with LBSap vaccine, before and after experimental challenge with Leishmania chagasi plus saliva of Lutzomyia longipalpis.(2013) Resende, Lucilene Aparecida; Roatt, Bruno Mendes; Soares, Rodrigo Dian de Oliveira Aguiar; Viana, Kelvinson Fernandes; Mendonça, Ludmila Zanandreis de; Lanna, Mariana Ferreira; Lemos, Denise da Silveira; Oliveira, Rodrigo Corrêa de; Martins Filho, Olindo Assis; Fujiwara, Ricardo Toshio; Carneiro, Cláudia Martins; Reis, Alexandre Barbosa; Giunchetti, Rodolfo CordeiroIn the studies presented here, dogs were vaccinated against Leishmania (Leishmania) cha-gasi challenge infection using a preparation of Leishmania braziliensis promastigote proteinsand saponin as adjuvant (LBSap). Vaccination with LBSap induced a prominent type 1immune response that was characterized by increased levels of interleukin (IL-) 12 andinterferon gamma (IFN- _) production by peripheral blood mononuclear cells (PBMC) uponstimulation with soluble vaccine antigen. Importantly, results showed that this type ofresponsiveness was sustained after challenge infection; at day 90 and 885 after L. chagasichallenge infection, PBMCs from LBSap vaccinated dogs produced more IL-12, IFN- _ andconcomitant nitric oxide (NO) when stimulated with Leishmania antigens as comparedto PBMCs from respective control groups (saponin, LB- treated, or non-treated controldogs). Moreover, transforming growth factor (TGF)- _ decreased in the supernatant of SLcA-stimulated PBMCs in the LBSap group at 90 days. Bone marrow parasitological analysisrevealed decreased frequency of parasitism in the presence of vaccine antigen. It is con-cluded that vaccination of dogs with LBSap vaccine induced a long-lasting type 1 immuneresponse against L. chagasi challenge infection.Item Kinetics of cell migration to the dermis and hypodermis in dogs vaccinated with antigenic compounds of Leishmania braziliensis plus saponin.(2008) Souza, Juliana Vitoriano de; Reis, Alexandre Barbosa; Moreira, Nádia das Dores; Giunchetti, Rodolfo Cordeiro; Oliveira, Rodrigo Corrêa de; Carneiro, Cláudia MartinsThe search for new immunobiologicals against canine visceral leishmaniasis (CVL) has intensified in the last decade. However, it still remains to be elucidated that mechanisms of the innate immune response in situ after immunization (a.i.). The aim of this study was to investigate the kinetics of cell migration in the skin dogs with distinct antigenic compounds of the LBSap vaccine. Our major findings indicated that saponin adjuvant alone or combined with Leishmania braziliensis antigen induced strong local acute inflammatory reaction. However, these reactions not progressed to ulcerated lesions. Overall, the cell profile found in Sap and LBSap was composed of neutrophils, lymphocytes and eosinophils. There was also increased production of iNOS in Sap and LBSap groups. Thus, we can conclude that dogs immunized by LBSap and the saponin adjuvant elicited a potential innate-immune activations status compatible with effective control of the resistance to infection by Leishmania and contributing to a better understanding of the innate-immunity events induced by the LBSap vaccine.Item Dogs immunized with LBSap vaccine displayed high levels of IL-12and IL-10 cytokines and CCL4, CCL5 and CXCL8 chemokinesin the dermis.(2013) Souza, Juliana Vitoriano de; Moreira, Nádia das Dores; Souza, Daniel Menezes; Roatt, Bruno Mendes; Soares, Rodrigo Dian de Oliveira Aguiar; Mathias, Fernando Augusto Siqueira; Cardoso, Jamille Mirelle de Oliveira; Giunchetti, Rodolfo Cordeiro; Cota, Renata Guerra de Sá; Oliveira, Rodrigo Corrêa de; Carneiro, Cláudia Martins; Reis, Alexandre BarbosaThe complex interplay between cytokines and chemokines regulates innate and adaptive immuneresponses against pathogens; specifically, cytokine and chemokine expression drives activation ofimmune effector cells and their recruitment to tissue infection sites. Herein, we inoculated dogs withLeishmania braziliensis antigens plus saponin (the LBSap vaccine), as well as with the vaccine components,and then used real-time PCR to evaluate the kinetics of dermal expression of mRNAs of cytokines (IL-12,IFN- _, TNF- _, IL-4, IL-13, TGF- _ and IL-10) and chemokines (CCL2, CCL4, CCL5, CCL21 and CXCL8) 1, 12, 24and 48 h after inoculation. We also evaluated the correlation between cytokine and chemokine expres-sion and dermal cellularity. The LBSap vaccine induced high levels of IL-12 and IL-10 expression at 12 and24 h, respectively. Furthermore, we observed positive correlations between IL-12 and IL-13 expression,IFN- _ and IL-13 expression, and IL-13 and TGF- _ expression, suggesting that a mixed cytokine microen-vironment developed after immunization with the vaccine. Inoculation with the saponin adjuvant aloneinduced a chemokine and cytokine expression profile similar to that observed in the LBSap group. CCL4and CXCL8 chemokine expression was up regulated by the LBSap vaccine. CCL5 expression was initiallyhighest in the LBSap group, but at 48 h, expression was highest in the LB group. Information about thekinetics of the immune response to this vaccine gained using this dog model will help to elucidate themechanisms of and factors involved in a protective response against Leishmania infection and will aid inestablishing rational approaches for the development of vaccines against canine visceral leishmaniasis.Item Clinical forms of canine visceral Leishmaniasis in naturally Leishmania infantum – infected dogs and related myelogram and hemogram changes.(2013) Nicolato, Roney Luiz de Carvalho; Abreu, Raquel Trópia de; Roatt, Bruno Mendes; Soares, Rodrigo Dian de Oliveira Aguiar; Reis, Levi Eduardo Soares; Carvalho, Maria das Graças; Carneiro, Cláudia Martins; Giunchetti, Rodolfo Cordeiro; Bouillet, Leoneide Érica Maduro; Lemos, Denise da Silveira; Vital, Wendel Coura; Reis, Alexandre BarbosaHematological analysis has limited applications for disease diagnosis in Leishmania infantum–infected dogs, but it can be very important in evaluating the clinical forms of the disease and in understanding the evolution of canine visceral leishmaniasis (CVL) pathogenesis. Recently, we demonstrated that alterations in leucopoiesis and erythropoiesis are related to clinical status and bone marrow parasite density in dogs naturally infected by L. infantum. To further characterize these alterations, we evaluated the association between the hematological parameters in bone marrow and peripheral blood alterations in groups of L. infantum–infected dogs: asymptomatic I (AD-I: serum negative/PCR+), asymptomatic II (AD-II: serum positive), oligosymptomatic (OD), and symptomatic (SD). Results were compared with those from noninfected dogs (NID). The SD group was found to present a decrease in erythropoietic lineage with concomitant reductions in erythrocytes, hemoglobin, and hematocrit parameters, resulting in anemia. The SD group also had increased neutrophils and precursors and decreased band eosinophils and eosinophils, leading to peripheral blood leucopenia. In the AD-II group, lymphocytosis occurred in both the peripheral blood and the bone marrow compartments. The SD group exhibited lymphocytosis in the bone marrow, with lymphopenia in the peripheral blood. In contrast, the AD-I group, showed no significant changes suggestive of CVL, presenting normal counts in bone marrow and peripheral blood. Our results showed for the first time that important changes in hematopoiesis and hematological parameters occur during ongoing CVL in naturally infected dogs, mainly in symptomatic disease. Taken together, our results based on myelogram and hemogram parameters enable better understanding of the pathogenesis of the anemia, lymphocytosis, and lymphopenia, as well as the leucopenia (eosinopenia and monocytopenia), that contribute to CVL prognosis.Item Evaluation of change in canine diagnosis Protocol adopted by the visceral Leishmaniasis control program in Brazil and a new proposal for diagnosis.(2014) Vital, Wendel Coura; Ker, Henrique Gama; Roatt, Bruno Mendes; Soares, Rodrigo Dian de Oliveira Aguiar; Leal, Gleisiane Gomes de Almeida; Moreira, Nádia das Dores; Oliveira, Laser Antônio Machado de; Machado, Evandro Marques de Menezes; Morais, Maria Helena Franco; Oliveira, Rodrigo Corrêa de; Carneiro, Mariângela; Reis, Alexandre BarbosaThe techniques used for diagnosis of canine visceral leishmaniasis (CVL) in Brazil ELISA and IFAT have been extensively questioned because of the accuracy of these tests. A recent change in the diagnosis protocol excluded IFAT and included the Dual-Path Platform (DPP). We evaluated the prevalence and incidence rates of Leishmania spp. before and after the change in the protocol. In addition, based on our results, we propose a new alternative that is less expensive for the screening and confirmation of CVL. Plasma samples were obtained from a serobank from dogs evaluated in a cross-sectional study (1,226 dogs) and in a cohort study of susceptible animals (n = 447), followed for 26 months. Serology testing wasperformed using ELISA, IFAT, and DPP. The incidence and prevalence of CVL were determined by using the protocol of the Visceral Leishmaniasis Control and Surveillance Program until 2012 (ELISA and IFAT using filter paper) and the protocol used after 2012 (DPP and ELISA using plasma). The prevalence was 6.2% and the incidence was 2.8 per 1,000 dog-months for the protocol used until 2012. For the new diagnosis protocol for CVL resulted in an incidence of 5.4 per 1,000 dog-months and a prevalence of 8.1%. Our results showed that the prevalence and incidence of infection were far greater than suggested by the previously used protocol and that the magnitude of infection in endemic areas has been underestimated. As tests are performed sequentially and euthanasia of dogs is carried out when the serological results are positive in both tests, the sequence does not affect the number of animals to be eliminated by the Control Program. Then we suggest to municipalities with a large demand of exams to use ELISA for screening and DPP for confirmation, since this allows easier performance and reduced cost.Item Systemic and compartmentalized immune response in canine visceral leishmaniasis.(2009) Reis, Alexandre Barbosa; Martins Filho, Olindo Assis; Carvalho, Andréa Teixeira de; Giunchetti, Rodolfo Cordeiro; Carneiro, Cláudia Martins; Mayrink, Wilson; Tafuri, Washington Luiz; Oliveira, Rodrigo Corrêa deHuman visceral leishmaniasis (VL) and canine visceral leishmaniasis (CVL) are the most important emerging diseases with high prevalence in Latin American countries and are mainly caused by Leishmania (L.) chagasi (Syn = L. infantum). CVL has a great impact on Brazilian public health because domestic dogs are the most important VL peri-domicile reservoirs in both urban and peri-urban areas. Our findings highlight the complexity of cellular immunological events related to the natural infection from dogs by L. chagasi, additionally correlating major peripheral blood phenotypic markers with clinical status and tissues parasite density. Our main results demonstrated that lower frequency ofcirculating B cells and monocytes are important markers of severe CVL, whereas increased levels of CD8+ lymphocytes appear to be the major phenotypic feature of asymptomatic disease. Determination of the isotypes patterns during CVL demonstrated thatasymptomatic dogs and those with low parasitism are associated with an increase of IgG1, while the symptomatic dogs and those with high parasitism are associated with an increase of IgG, IgG2, IgM, IgA and IgE immunoglobulins. Pioneer findings obtained by our group showed a correlation between clinical status of CVL with degree of tissue parasite density. This data demonstrated that asymptomatic dogs presented low parasitism while symptomatic dogs are associated with high parasite load in various tissues such as skin, bone marrow and spleen. We have also investigated the association between tissue parasitism and CVL clinical forms. Regardless of clinical status, skin and spleen are the major sites of high parasite density during ongoing CVL. Furthermore, we demonstrated that bone marrow and spleen parasite density are the most reliable parasitological markers to decode the clinical status of CVL. In this article, we have reviewed some aspectsof the histopathological and immunological events occurring in natural and experimentalL. chagasi/L. infantum infection, pointing out the main L. chagasi-parasitized tissue. Wehave discussed the importance of the association between parasite density, immunological/ histopathological aspects and clinical status of the CVL, their current applications, challenges for the future and potential opportunities in CVL research.
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