A collection of twenty three selectively mono-protected di- and triamines, masked with the Boc, Fmoc or Ddiv protecting groups, were synthesised via continuous flow synthesis in a self-assembled meso-scale PTFE flow reactor. The continuous flow strategy offered direct access to the mono-protected compounds in good yields, especially in the case of the Fmoc carbamates which circumvented the use of another sacrificial protecting group. Two of the mono-Boc-protected carbamates were used as starting materials to generate N-alkylglycine monomers; synthesised via tandem mono-alkylation and Fmoc carbamation, linked by an in-line scavenging protocol using a silica-based trisamine scavenger resin. The final step of the monomer synthesis employed catalytic transfer hydrogenolysis using 20% Pd(OH)2/C and 1,4- cyclohexadiene. The three-step flow procedure gave access to two monomers, with one of them being a novel N-alkylglycine unit bearing a triethylene glycol bridge. The monomers were used as building blocks to assemble new oligo-N-alkylglycines (peptoids) via microwave-assisted solid phase synthesis. Three different types of peptoids were synthesised: (i) oligo-N-(6-aminohexyl)glycines (“standard” peptoids), (ii) oligo-N-{2-[2-(2-aminoethoxy)ethoxy]ethyl}glycines (“triethylene glycol” [TEG] peptoids) and (iii) hetero-oligomers of alternating “standard” and “TEG” monomers (“hybrid” peptoids). The peptoids were evaluated for their cellular permeability and cytotoxicity with HeLa, HEK-293 and CHO cells. All the peptoids were shown to be non-cytotoxic at 10 μM based on cell proliferation assays. In general, it was found that the cellular uptake of the hybrid peptoids outperformed their standard and TEG analogues. Flow cytometry and confocal microscopy results revealed that the hybrid nonamer had the highest cellular uptake efficiency of all the peptoids synthesised. At a concentration of 1 μM, it outperformed the second best molecular transporter (standard nonamer) by a factor of seven.

Calamagrostis phragmitoides belongs to taxonomically complex C. purpurea aggregate with circumpolar distribution area. The relations between morphologically similar taxa of this aggregate are not clear. Populations of C. purpurea agg. from Central Europe, Scandinavia and Siberia (including one plant from the North America) were genetically studied (cpDNA, ITS, ploidy level by FCM). Allozyme and morfological variability of populations C. phragmitoides and morphologically similar C. canescens was also studied in the Czech Republic. This study was supported by grant SGA PrF JU in 2014.

Orientador: Alexandra Ivo de Medeiros / Coorientador: Naiara Naiana Dejani / Banca: Alexandra Ivo de Medeiros / Banca: Cleslei Fernando Zanelli / Banca: Fabiani Gai Frantz / Resumo: A resolução de reações inflamatórias é caracterizada pela fagocitose de células mortas, processo denominado eferocitose. Como consequência deste processo, há a produção dos mediadores anti-inflamatórios, fator de transformação de crescimento (TGF-β), prostaglandina E2 (PGE2) e a interleucina-10 (IL-10). No entanto, atualmente sabe-se que a fagocitose de células apoptóticas infectadas direciona a produção de mediadores inflamatórios interleucina-6 (IL-6), interleucina-23 (IL-23) e TGF-β, Resultados obtidos recentemente por nosso grupo demonstram que a fagocitose de células apoptóticas infectadas (iAC) com Escherichia coli por células dendríticas promove, além da produção de TGF-β, interleucina-1β (IL-1β) e IL-6, a síntese de altos níveis de PGE2. Além disso, observamos que a PGE2, via receptor de prostaglandina E2 4 (EP4), inibe a diferenciação de linfócitos Th17 por meio da modulação da expressão do receptor de IL-1 (IL-1R) em linfócitos Th. Desta forma, o objetivo deste trabalho é elucidar o mecanismo pelo qual PGE2, oriunda da eferocitose de células apoptóticas infectadas, inibe a expressão de IL-1R e a diferenciação de linfócitos Th17. Os resultados obtidos demonstram que PGE2, via receptor EP4, induz a ativação de adenilato ciclase/PKA/EPAC, e que esta via de sinalização promove além da inibição de IL-1R, a inibição de um importante fator de transcrição envolvido na diferenciação de linfócitos Th17, STAT3. Sabe-se que a PGE2 pode exercer suas funções supressoras na difere... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The expression of inflammatory reactions is characterized by phagocytosis of dead cells, a process called eferocytosis. As a consequence of this process, there is the production of anti-inflammatory mediators, transforming growth factor (TGF-ȕ), prostaglandin E2 (PGE2) and an interleukin-10 (IL-10). However, it is now known that phagocytosis of infected apoptotic cells is one of the responsible for the production of interleukin-6 (IL-6), interleukin-23 (IL-23) and TGF-ȕ lung mediators. The results obtained by our group demonstrate that the phagocytosis of infected apoptotic cells (iCA) with Escherichia coli by dendritic cells promotes, in addition to the production of TGFȕ, interleukin-1ȕ (IL-1ȕ) and IL-6, high levels of PGE2. In addition, PGE2, via the prostaglandin E2 receptor (EP4), inhibits differentiation of Th17 lymphocytes by modulating IL-1 receptor (IL-1R) expression in Th lymphocytes. Thus, the objective of this work is to elucidate the mechanism by which PGE2, derived from the ectocytosis of infected apoptotic cells, inhibits the expression of IL-1R and differentiation of Th17 lymphocytes. The results obtained demonstrate that PGE2, via the EP4 receptor, induces the activation of adenylate cyclase / PKA / EPAC, and that this signaling pathway promotes in addition to the inhibition of IL-1R, the inhibition of an important transcription factor involved in the differentiation of Th17 lymphocytes, the STAT3. PGE2 is known to exert its suppressor functions in the differentiation of Th17 cells by the activation of SOCS, as well as to promote the activation of PI3K. However, despite the presence of PGE2 enhancing socs1 expression, inhibition of STAT3... (Complete abstract click electronic access below) / Mestre

Made available in DSpace on 2014-06-11T19:31:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-05-14Bitstream added on 2014-06-13T19:01:28Z : No. of bitstreams: 1 godoy_av_dr_jabo.pdf: 494650 bytes, checksum: d27d9898dce60f32a0eece2e8cdaf032 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O presente estudo teve como objetivos avaliar seqüencial e temporalmente o quadro leucocitário de dez cães portadores de linfoma em remissão, submetidos à alta dose de ciclofosfamida, durante o uso do estimulador de colônia de granulócitos (GCSF, filgrastine) seguido ou não do transplante autólogo de medula óssea (TMO), bem como quantificar células CD34+ no sangue periférico dos referidos cães. Para tal avaliação foram utilizados três grupos experimentais, sendo o grupo 1 (G1) formado por cinco animais em remissão de linfoma que passaram por alta dose de ciclofosfamida e TMO seguido do uso de G-CSF; o grupo 2 (G2) formado por cinco animais em remissão de linfoma que sofreram alta dose de ciclofosfamida seguida do uso de G-CSF e o grupo 3 (G3) formado por dez animais saudáveis que receberam apenas o G-CSF. O transplante consistiu na colheita de medula óssea, preparo e congelamento da bolsa que continha células medulares em suspensão, condicionamento do paciente com 500mg/m2 de ciclofosfamida, infusão das células hematopoéticas e aplicação do G-CSF. Para avaliar a recuperação hematopoética pós-transplante foi realizado leucograma e análise citométrica do sangue nos dias 7, 8, 9, 10 e 11 após condicionamento. O nadir médio dos neutrófilos segmentados (NS) no grupo com transplante de medula óssea (G1) foi 425 NS/mL, e no grupo sem TMO (G2) foi 637,4 NS/μL e ocorreu sete dias após alta-dose de quimioterápico em ambos os grupos. A duração média da neutropenia foi de três dias. Nenhum animal apresentou febre ou sepse após a alta dose de ciclofosfamida. A dose de 5μg/kg/dia durante quatro dias de filgrastine foi insuficiente na mobilização adequada de células CD34+ nos três grupos estudados, sendo necessários novos estudos para este propósito. Desta maneira, pode-se comprovar o fato de que o uso do G-CSF leva a reduções significativas na incidência... / The aims of this research were to provide an analysis of several counts of leucocytes values in dogs with lymphoma in remission phase, undergone to high-dose chemotherapy with cyclophosphamide, during treatment with granulocyte colonystimulating factor (G-CSF, filgrastine), followed or not by autologous bone marrow transplantation (BMT), as well to quantify CD34+ cells of peripheral blood from that dogs. For this purpose, three experimental groups were considered. Five dogs in clinical remission undergone to high-dose chemotherapy with cyclophosphamide and BMT, followed by G-CSF use were included in group 1 (G1), while group 2 (G2) was consisted by five dogs undergone to high-dose chemotherapy with cyclophosphamide, followed by G-CSF. Group 3 (G3) was composed of ten healthy dogs undergone to GCSF only. Transplantation consisted of bone marrow harvest, managing and freezing bags containing lifted marrow cells, cyclophosphamide conditioning (500mg/m2), hematopoietic cells infusion and treatment with G-CSF. After transplantation, hematopoietic recovery was evaluated by means of leukograms and flow cytofluorimetrical analysis on days 7, 8, 9, 10 and 11 after conditioning. Mean nadir neutrophil (NS) counts in group undergone transplantation (G1) was 425 NS/mL versus 637,4 NS/μL in group without BMT (G2). Nadir was observed on the seventh day after high-dose chemotherapy in both groups and neutropenia mean time was three days. Fever or sepsis was not observed in any dog after high-dose cyclophosphamide. Four days of filgrastine treatment in dose of 5μg/Kg, daily, was not enough to mobilize CD34+ cells appropriately in three groups analyzed, requiring new studies for this purpose. Considering these results, it is possible to conclude that G-CSF significantly reduces the incidence, severity and duration of neutropenia

Made available in DSpace on 2014-06-11T19:23:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-03-24Bitstream added on 2014-06-13T18:09:19Z : No. of bitstreams: 1 andreo_r_me_araiq.pdf: 430930 bytes, checksum: a9d88b788282568b9fdcb9f6baa590b6 (MD5) / A tripanosomíase americana ou doença de Chagas é uma doença parasitária causada pelo protozoário Trypanosoma cruzi. Atualmente a doença é considerada negligenciada devido à falta de investimentos para obtenção de fármacos capazes de tratar e curar a doença com menos efeitos colaterais do que o benzonidazol, utilizado até o momento. Desta forma, a busca por novos agentes terapêuticos contra a doença de Chagas é uma estratégia importante para eliminar ou, pelo menos, diminuir a incidência desta doença debilitante e que afeta aproximadamente 10 milhões de pessoas na América Latina. Neste trabalho foi avaliada a atividade tripanocida do ácido gálico e seus ésteres semi-sintéticos contra formas epimastigotas de Trypanosoma cruzi, assim como a citotoxicidade sobre células de mamíferos (HepG2). Informações adicionais foram obtidas sobre o perfil de produção de citocinas por macrófagos murinos não infectados e tratados com os galatos; também realizou-se o estudo do efeito apoptótico destas substâncias em formas epimastigotas de T. cruzi. Os resultados obtidos foram bastante interessantes e animadores, uma vez que, alguns ésteres de ácido gálico apresentaram atividade tripanocida maior do que aquela observada com benzonidazol e baixa citotoxicidade em células de mamíferos. Além disso, a avaliação do perfil de produção de citocinas, induzido pelas substâncias mostrou resposta favorável à resistência do hospedeiro contra a doença de Chagas. Nos experimentos realizados para avaliar a atividade apoptótica das substâncias em T. cruzi, aquelas que tiveram maior atividade tripanocida apresentaram baixa taxa de apoptose, o que sugere um mecanismo de ação mais relacionado com a característica surfactante destas substâncias; por outro lado, as que apresentaram menor atividade tripanocida, e provocaram apoptose, podem estar atuando como inibidoras da biossíntese de ergosterol / The American trypanosomiasis or Chagas disease is a parasitic disease caused by Trypanosoma cruzi protozoan. Currently, it is a neglected disease due to the lack of investments in order to obtain drugs able to treat and cure the disease with fewer side effects than benzonidazole, used until now. Thus, the search for new therapeutic agents against Chagas disease is an important strategy to eliminate or at least reduce the incidence of this debilitating disease that affects approximately 10 million people in Latin America. In this study were evaluated the trypanocidal activity of gallic acid and its semi synthetic esters against epimastigotes forms of Trypanosoma cruzi, as well as, the citotoxicity on mammalian cells (HepG2). Additional information was obtained about the profile of cytokine production by murine macrophages uninfected treated with gallates; also it was studied the apoptotic effect of these substances in epimastigotes forms of T. cruzi. The results obtained were very interesting and encouraging since some esters of gallic acid showed trypanocidal activity greater than that observed with benzonidazole and low citotoxicity in mammalian cells. Moreover, the evaluation of the profile of cytokine production induced by the substances showed a favorable response to the host resistance against Chagas disease. In the experiments performed to evaluate apoptosis activity of the substances in T. cruzi, those who had greater trypanocidal activity, presented a low apoptosis rate, which suggests a mechanism of action based on the surfactant feature of these substances; by the other hand, the substances that showed low trypanocidal activity and caused apoptosis, can act as inhibitors of the ergosterol biosynthesis

Made available in DSpace on 2014-06-11T19:23:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-18Bitstream added on 2014-06-13T20:50:57Z : No. of bitstreams: 1 anai_la_me_jabo.pdf: 941558 bytes, checksum: 3e8d781b74dbdeaa063ccb760a761d87 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O linfoma é o tumor de tecido hematopoético mais comum nos cães e um dos tumores malignos de maior ocorrência nesta espécie. É um ótimo modelo experimental para estudo devido a sua semelhança com o linfoma não-Hodgkin em humanos. Considerando a importância das alterações decorrentes da evolução desta neoplasia e aquelas ocorridas com o emprego da poliquimioterapia, avaliou-se a fórmula leucocitária absoluta e a contagem de células CD45+, no sangue de 25 cães linfomatosos, contrastando-as a partir do diagnóstico, uma vez por semana, durante as primeiras oito sessões quimioterápicas do protocolo de Madison-Wisconsin, e também foi comparada as contagens totais dos granulócitos, linfócitos e monócitos, obtidas em contador automático convencional e por intermédio da citometria de fluxo. Observou-se um maior número de animais com linfoma multicêntrico em estadio Vb. No leucograma, as principais alterações foram observadas no momento do diagnóstico e quando analisado o tratamento poliquimioterápico, ao longo do tempo, a maior mielosupressão foi imposta pela vincristina e suas associações. Cães linfomatosos podem ser avaliados tanto por intermédio de contadores eletrônicos como por meio da citometria de fluxo / Lymphoma is the most common hematopoietic tumor in dogs and one of the malignant tumors with highest occurrence in that species. It is a great experimental model to study since it has a great resemblance with the non-Hodgkin lymphoma in humans. Considering the importance of the alterations due to the evolution of this malignancy and the ones due to the use of polichemothearpy this work aimed to evaluate the absolute leucocyte count and the counts of CD45+ type cells in the blood of 25 lymphomatic dogs counterpointing them since diagnose, once a week, during the first eight chemotherapic sessions with the Madison-Wisconsin protocol. Furthermore the total counts of granulocytes, lymphocytes and monocytes obtained by conventional automatic counter and by flow cytometry were compared. It was observed a greater number of animals with multicentric lymphoma in stage Vb. The main alterations in the leukon were the ones observed at the moment of diagnose. When analyzing the polichemotherapy treatment the highest myelosupression was caused by vincristine and its associations. It is concluded that dogs with lymphoma can be evaluated by electronic counters as well as by flow cytometry

Made available in DSpace on 2014-06-11T19:23:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-18Bitstream added on 2014-06-13T20:51:05Z : No. of bitstreams: 1 gomes_mos_me_jabo.pdf: 622478 bytes, checksum: be8b00fa47ea963d66d4d14490453d61 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Considerando a importância da eubiota intestinal na saúde dos animais, o papel da dieta em sua composição e metabolismo tem sido estudado. Neste aspecto os prebióticos têm merecido destaque, incluindo-se a parede celular de leveduras (PCL), uma fonte natural de mananoligossacarídeos. Para o estudo de seu potencial prebiótico para cães, foram empregadas quatro dietas isonutrientes com inclusão de 0% (T0), 0,15% (T15), 0,30% (T30) e 0,45% (T45) de PCL. Foram utilizados oito Beagles adultos, divididos em dois quadrados latinos 4 X 4. Em cada período uma fase de adaptação de 15 dias precedeu 5 dias de coleta total de fezes para o ensaio de digestibilidade e 1 dia de coleta de fezes frescas para enumeração bacteriana, mensuração de pH e determinação das concentrações de ácidos graxos de cadeia curta e aminas bioativas. No dia 21 de cada período, amostra de sangue foi colhida para quantificação imunofenotípica por citometria de fluxo das subpopulações linfocitárias CD5+CD4+, CD5+, CD5+CD8+, CD45+ e CD45+CD21+. Os dados foram avaliados pelo Proc GLM do SAS, sendo as médias comparadas pelo teste de Tukey e por contraste polinomial e ortogonal (p<0,1). Os coeficientes de digestibilidade dos nutrientes não variaram com a inclusão de PCL (p>0,1). A inclusão de PCL não resultou em diferenças quanto às contagens de anaeróbios totais, aeróbios totais, bifidobactéria, lactobacilos, clostridios nas fezes dos cães (p>0.1). Verificou-se aumento linear na concentração fecal de butirato (p=0,055), redução linear das concentrações fecais de tiramina (P=0,1) e histamina (P=0,07) e redução quadrática de feniletilamina e triptamina (P=0,07). Os cães apresentaram, também, aumento linear na concentração de linfócitos CD5+ (P=0,10) e maior número de linfócitos CD45+CD21+ (P=0,053) mediante ingestão de PCL. Concluiu-se que a PCL apresenta efeito prebiótico... / Considering the importance of the intestinal microbiota to the animal health, the role of diet upon microbiota composition and metabolism have been studied. Prebiotics have been emphasized on this regard, including the yeasts cell walls (YCW), a natural source of mannanoligosaccharides. To study the prebiotic potential of this compound for dogs, four isonutrient diets were used with inclusion of 0% (T0), 0,15% (T15), 0,30% (T30) and 0,45% (T45) of YCW. Eight Beagles adults were used, arranged in two 4 X 4 Latin squares. In each replicate an adaptation phase of 15 days preceded a 5 days of total feces collection for digestibility trial and 1 day collection of fresh fecal samples for bacterial enumeration, pH measurement and determination of short chain fatty-acids and bioactive amines concentration. On day 21, a blood samples were collected for immunophenotypic quantification of lymphocyte subsets CD5+CD4+, CD5+, CD5+CD8+, CD45+ e CD45+CD21+ through flow cytometry. The data were evaluated using Proc GLM of SAS, means were compared by Tukey test and polinomial and orthogonal contrasts (p <0,1). The inclusion of YCW did not change the coefficient of total tract apparent digestibility of nutrients (p>0.1) nor the fecal bacterial enumeration (total aerobes and anaerobes, Bifidobacterium, Lactobacillus, Clostridium and Escherichia coli) in dog’s feces (p>0.1). Fecal butirate concentration increased linearly with YCW addition (p=0,055), fecal concentrations of tyramine (P=0,1) and histamine (P=0,07) decreased linearly, and phenylethylamine (P=0,07) and tryptamine (P=0,07) concentrations showed a quadratic reduction with YCW consumption. Dogs also presented a linear increase in lymphocytes CD5+ concentration (P=0,10) and a greater CD45+CD21+ lymphocytes number (P=0,053) with YCW addition. In conclusion YCW presents a prebiotic (changing bacterial end product formation) and immunostimulating effect for dogs.

Thesis (MTech (Medical Technology))--Cape Technikon, Cape Town, 2002 / Erythropoietic response is the first indication of bone marrow recovery following bone marrow or peripheral blood stem cell transplantation. Manual reticulocyte counting has not only proven to be outdated but an extremely crude method of analysis, particularly if accurate and reliable means of assessing erythroid response is required to assess bone marrow recovery. Automated methods allow for the quantification of maturation within each reticulocyte, by measuring the amount of RNA present. The method of choice for our reticulocyte analysis was the Reticulocyte Maturation Index (RMI). The RMI was obtained by dividing the number of immature reticulocytes counted by the total number of reticulocytes counted producing a reportable value of International Units (IU). A normal Reticulocyte Maturation Index is 0.20 to 0.50 IU. The aim of the study was multifold. We wanted to prove that the Reticulocyte Maturation Index (RMI) is indeed the fastest means to assess bone marrow recovery in various types of transplants, including Bone Marrow Transplant (BMT) and Peripheral Blood Stem Cell Transplant (PBSCT). We also wanted to draw comparisons between allogeneic and autologous transplants, as well as further assessing different disease types. This was done by measuring the Reticulocyte Maturation Index (RMI), Absolute Neutrophil Count (ANe) and the Platelet Count (PLT) within the various groups. We further wanted to assess the effect of preconditioning treatment, Mononuclear Counts (MNC) and Colony Forming Unit - Granulocyte and Monocyte Counts (CFU-GM) on the early RMI response. These comparisons resulted in a need to establish a working range to determine patients response therein, and final outcome of the transplants. Finally we wanted to establish whether the "day 14" marrow biopsy is necessary, particularly if the three peripheral blood parameters, RMI, ANC and PLT were used as routine procedure following transplantation. The Reticulocyte Maturation Index (RMI) was measured on the Coulter EPICS ProfIle II flow cytometer; the ANC and PLT were measured on the Technicon H2 Haematology System. All other results such as the Mononuclear Counts (MNC), Colony Forming Unit - Granulocyte and Monocyte counts (CFU-GM), "day 14" and "day 28" bone marrow biopsies were retrieved from laboratory records. Forty nine transplant patients were evaluated for RMI over a period of six months, at the Department of Haematology, Groote Schuur Hospital, Cape Town. Four patients failed to engraft; and were not used in the calculations; but were evaluated as an aspect of the study in the final analysis. Forty five patients were analysed to establish the values used in the study, these patients were divided into eleven groups.

Orientador: Valéria Marçal Félix de Lima / Banca: Rosemeri de Oliveira Vasconcelos / Banca: Cáris Maroni Nunes / Resumo: Os cães são o principal reservatório doméstico de Leishmania. (L.) chagasi. No hospedeiro vertebrado, incluindo o homem, o parasita pode causar leishmaniose visceral, transmitida aos humanos. As células T regulatórias (Treg) estão envolvidas na indução de supressão de mecanismos efetores relacionados à imunidade celular. Para investigar a possível implicação das células Treg durante a infecção por Leishmania spp., a presença de Treg no baço e no sangue periférico foi avaliada em cães naturalmente infectados e com sinais clínicos compatíveis com a doença. Quinze cães adultos provenientes do Centro de Controle de Zoonoses da Prefeitura Municipal de Araçatuba - SP, Brasil, com reação sorológica positiva para antígenos de Leishmania (L.) chagasi determinada por ELISA indireto e teste rápido rK39 (Kalazar Detect TM Rapid Test, InBios Inc-Seattle, WA-USA). Cinco cães saudáveis, provenientes de área não endêmica foram incluídos no estudo. As células T regulatórias foram quantificadas em amostras de baço e em células mononucleares do sangue periférico utilizando-se anticorpos monoclonais e citometria de fluxo. No baço, menor número de células Treg foi observado em cães infectados do que nos controles (p<0,05), enquanto que nenhuma diferença foi observada nas células mononucleares de sangue periférico. Esses resultados sugerem que as células Treg estão envolvidas na infecção por Leishmania spp. e que podem ter um papel na persistência do parasita e o estabelecimento da infecção crônica / Abstract: Dogs are the main domestic reservoirs of L. (L.) chagasi. Once in the vertebrate host, the parasite may cause visceral leishmaniasis, which can also be transmitted to humans. Regulatory T cells (Treg) have been shown to be involved in the direct induction of immunosupression of effector cellular immune response. To investigate the possible involvement of T reg cells during Leishmania infection, the presence of Treg cells from the spleen and peripheral blood mononuclear cells (PBMC) of dogs naturally infected with L. (L.) chagasi, and showing clinical signs, was quantified. A total of 15 adult dogs from the Center for Zoonosis Control of Araçatuba - SP, Brazil, positive for Leishmania (L.) chagasi by indirect ELISA and TM rapid test rK39 (Kalazar Detect TM Rapid Test, InBios Inc-Seattleadult healthy dogs from non endemic area were included in the sspleen and PBMC were used for quantification of Treg by flow cytometmonoclonal antibodies. In spleen, Treg levels in infected dogs control group (p<0.05); in PBMC, no differences were observed betgroups. These results suggest that Treg population is involved in infection and may play a role in promoting parasite persistence achronic infection / Mestre

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Candida tropicalis à uma levedura diplÃide causadora de infecÃÃes superficiais e/ou sistÃmicas, as quais podem ser adquiridas de maneira endÃgena ou exÃgena e podem acometer diversos ÃrgÃos. No Brasil, dentre as espÃcies de Candida spp., a C.tropicalis à a segunda espÃcie mais comumente isolada e no Cearà à pouco estudada. Nos dias atuais, temos vivenciado um aumento significativo das infecÃÃes fÃngicas invasivas. PorÃm as drogas antifÃngicas disponÃveis no mercado sÃo restritas a um pequeno nÃmero quando comparadas as antibacterianas. Logo, este fato unido ao aumento da frequÃncia de resistÃncia cruzada faz necessÃria a busca por novas estratÃgias terapÃuticas. A amiodarona (AMD) à usada classicamente para tratar pacientes com arritmia. Trabalhos recentes tÃm demonstrado uma ampla atividade antifÃngica desta droga quando associado ao fluconazol (FLC). No presente estudo induzimos resistÃncia em sete cepas de Candida tropicalis e avaliamos um eventual sinergismo entre FLC e AMD. A avaliaÃÃo da interaÃÃo das drogas foi determinada atravÃs do cÃlculo da Fractionary Inhibitory Concentration (FICI) e por meio da tÃcnica de citometria de fluxo, onde tambÃm avaliamos o provÃvel mecanismo de aÃÃo desse sinergismo. Os isolados utilizados no estudo pertencem ao LaboratÃrio de BioprospecÃÃo Experimental em Leveduras (LABEL) da UFC. Para o cumprimento da metodologia, as cepas foram recuperadas do estoque e identificadas por biologia molecular. A induÃÃo foi realizada adaptando-se o protocolo de Pinto e Silva (2009). Os testes de sensibilidade foram realizados utilizando-se o teste de microdiluiÃÃo em caldo padronizado pelo CLSI, segundo o documento M27-A3. Para avaliar o sinergismo utilizou-se a tÃcnica do checkboard. Por fim, para verificar o possÃvel mecanismo de aÃÃo do sinergismo, determinamos a integridade de membrana, potencial transmembrana mitocondrial, formaÃÃo de espÃcies reativas de oxigÃnio (ROS), teste do cometa, anÃlise da oxidaÃÃo de bases purinas e caspases. As cepas de C.tropicalis apÃs ~100 dias atingiram um CIM > 64Âg/mL. Os testes de sensibilidade apresentaram CIM > 64Âg/mL tanto para o FLC como para AMD. Das cepas testadas, seis apresentaram sinergismo. O tratamento FLC+AMD alterou a integridade da membrana plasmÃtica e mitocondrial, aumentou os nÃveis de ROS intracelularmente, causou danos ao DNA e, consequentemente, conduziu morte celular por apoptose. / Candida tropicalis is a diploid yeast causing superficial infections and / or systemic, which can be acquired endogenously or exogenously and can affect several organs. In Brazil, among the species of Candida spp. The C.tropicalis is the second most common species isolated in Cearà and is rarely studied. Nowadays, we have experienced a significant increase in invasive fungal infections. But antifungal drugs on the market are restricted to a small number when compared to the antibacterial. Therefore, this fact coupled with the increased frequency of cross-resistance is necessary to search for new therapeutic strategies. Amiodarone (AMD) is traditionally used to treat patients with arrhythmia. Recent studies have shown a broad antifungal activity of this drug when combined with fluconazole (FLC). In the present study we induced resistance in seven strains of Candida tropicalis and evaluate a possible synergism between FLC and AMD. The evaluation of the interaction of the drugs was determined by calculating the Fractionary Inhibitory Concentration (FICI) and by flow cytometry, where we also evaluated the likely mechanism of action of this synergism. The isolates used in this study belong to the Laboratory of Experimental Bioprospecting in yeast (LABEL) the UFC. For the fulfillment of the methodology, the strains were recovered from stock and identified by molecular biology. The induction was carried out adapting the protocol of Pinto and Silva (2009). Sensitivity tests were performed using the broth microdilution test standardized by the CLSI, the document M27-A3. To evaluate the synergism used the technique of checkboard. Finally, to determine the possible mechanism of action of synergism, we determine the membrane integrity, mitochondrial transmembrane potential, formation of reactive oxygen species (ROS), comet assay, analysis of the oxidation of purine bases and caspases. Strains C.tropicalis after ~ 100 days reached an MIC> 64&#956;g/mL. The sensitivity tests were MIC> 64&#956;g/mL both the FLC as AMD. Of the strains tested, six showed synergism. The treatment changed the FLC + AMD plasma membrane integrity and mitochondrial increased levels of intracellular ROS, cause DNA damage and therefore led cell death by apoptosis.