PGC-1 beta é um co-ativador de transcrição gênica responsável pela regulação do metabolismo celular, principalmente na biogênese e função mitocondrial, disponibilidade de substrato e síntese de lipídios. Nos últimos anos, outras isoformas de PGC-1 têm sido descritos como participantes na gênese e manutenção de tumores. Portanto, nosso objetivo foi determinar se o PGC-1beta está relacionado ao aumento da proliferação celular de células de melanoma. Inicialmente, foi demonstrado que os níveis de RNAm e proteína de PGC-1beta são muito mais elevados em linhagens de células de melanoma (Tm1 e Tm5) do que na linhagem parental de melanócitos não tumorais (Melan-a) como detectado por PCR quantitativa e western blotting. A fim de descobrir uma relação causal entre a expressão de PGC-1? e crescimento celular da linhagem Tm5, células de tal linhagem foram transfectadas com um oligonucleotídeo antisense (ASO) contra PGC-1beta. As células tratados com ASO apresentaram níveis mais baixos de RNAm e proteína PGC-1beta, além de redução em sua atividade avaliada pela expressão de genes PGC-1beta dependentes. Além disso, as células transfectadas apresentaram uma taxa de proliferação inferior em comparação com células de controle Tm5. Este fenômeno também foi observado in vivo. Quando injetadas em camundongos, as células Tm5 desenvolvem-se em um tumor que atinge 1,34 ± 0,20 cm3 após nove dias. Tumores tratados com ASO após o mesmo tempo apresentaram volume tumoral de 0,75 ± 0,05 cm3. Este crescimento não estava relacionada à necrose tumoral, mas sim com a proliferação reduzida de células. Finalmente, verificamos se o mesmo fenômeno seria observado em humanos. A expressão PGC-1beta foi muito maior em amostras de melanoma do que em nevos, alterações não-malignas da pele com alto conteúdo de melanina. Por conseguinte, conclui-se que a expressão PGC-1? está aumentada no melanoma, tanto murino e humano, e que o bloqueio da sua atividade leva à diminuição da proliferação celular e crescimento tumoral / PGC-1beta is a co-activator of gene transcription primarily responsible for the regulation of cellular metabolism, mainly in mitochondrial biogenesis and function and also substrate and lipid synthesis. In recent years, other isoforms of PGC-1 have been described as participating in the genesis and maintenance of tumors. Therefore, our objective was to determine whether PGC-1beta is related to increased proliferation of melanoma cells. Initially, it was demonstrated that mRNA and protein levels of PGC-1beta are much higher in melanoma cell lines (Tm1 and TM5) than in the non-tumoral parental lineage melanocytes (melan-a) as detected by quantitative PCR and Western blotting. In order to find a causal relationship between the expression of PGC-1beta and cell growth, Tm5 lineage cells were transfected with an antisense oligonucleotide (ASO) against PGC-1beta. The cells treated with ASO had lower levels of PGC-1beta mRNA and protein, as well as reduction in its activity detected by quantitation of PGC-1beta dependent genes expression. Furthermore, transfected cells showed a lower rate of proliferation compared to Tm5control cells. This phenomenon was also observed in vivo. When injected into mice, Tm5 cells develop a tumor which reaches 1.34 ± 0.20 cm3 after nine days. Tumors treated with ASO, after the same time, presented tumor volume of 0.75 ± 0.05 cm 3. This growth was not related to tumor necrosis, but with reduced cell proliferation. Finally, we checked whether the same phenomenon would be observed in humans. The PGC-1beta expression was much higher in melanoma samples than in nevi, a non-malignant skin alteration filled with melanin. Therefore, we concluded that PGC-1beta expression in melanoma is increased, both in murine and human, and that blocking its activity leads to decreased cell proliferation and tumor growth

Made available in DSpace on 2015-04-22T22:06:15Z (GMT). No. of bitstreams: 1 tullio.pdf: 2981239 bytes, checksum: d6f78923df4523264147690fa8713589 (MD5) Previous issue date: 2013-08-29 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Insects from the subfamily Phlebotominae (Diptera: Psychodidae) are of medical interest because they are vectors of protozoa of the genus Leishmania transmitted when the insect feeds. Approximately 500 species are known in the Americas and 133 registered in the state of Amazonas. Caused by different species of Leishmania, cutaneous leishmaniasis (ACL) affects humans and other animals, affecting skin and mucosa. This work is part of the project " Condições de vida e saúde de populações indígenas na Amazônia", PRONEX, which has an interdisciplinary approach to study eco-epidemiological and anthropological aspects of leishmaniasis in the Middle Purus, funded FAPEAM / CNPq, and is in progress in the municipality of Lábrea, where there are records of cases and suspected underreporting. Therefore, it was thought appropriate to the study sand flies of the municipality as a contribution to the knowledge of the epidemiology of ACL. The aim of this study was to investigate the entomological aspects related to the epidemiology of ACL in an indigenous area of the municipality of Lábrea. The collection of sand flies was carried out in February 2012 in the Aldeia Castanheira, Indian Reserve Caititú, municipality of Lábrea using light traps. After identification of insects, we performed detection and identification of Leishmania present in the collected sand flies using a molecular approach. A total of 1,267 sandflies was collected, 819 (64.6%) females and 448 (35.4%) males, distributed in 10 genera and 32 species. The most abundant genera were Psychodopygus, Nyssomyia and Trichophoromyia, the species Trichophoromyia ubiquitalis was the most abundant with 235 (18.5%) specimens, followed by Psychodopygus davisi with 228 (18%) and Nyssomyia antunesi with 135 (10.7%) individuals. The presence of Leishmania was detected in six species: Sciopemyia servulolimai, Trichophoromyia ubiquitalis, Evandromyia apurinan, Nyssomyia umbratilis, Nyssomyia yuilli yuilli, Psychodopygus davisi, the estimated minimum infection rate was 0.84%.The Leishmania species were: Leishmania (L.) amazonensis, Leishmania (V.) braziliensis and a sample identified only to the level of subgenus Leishmania (Viannia) sp. The Leishmania infection detected in vector species / suspected vectors found in this work, suggests the involvement of these species in the transmission cycle of leishmaniasis in the municipality. Future investigations should be made to help elucidate the epidemiology of cutaneous leishmaniasis in the municipality of Lábrea as well as the state of Amazonas. / Os insetos da subfamília Phlebotominae (Diptera: Psychodidae) são de interesse médico por serem vetores de protozoários do gênero Leishmania transmitidos no momento da alimentação do inseto. Cerca de 500 espécies são conhecidas paras as Américas e 133 registradas no estado do Amazonas. Causada por diferentes espécies de Leishmania, a leishmaniose tegumentar americana (LTA) acomete o homem e outros animais, afetando pele e mucosa. Este trabalho está inserido no projeto Condições de vida e saúde de populações indígenas na Amazônia , PRONEX, que possui abordagem interdisciplinar para estudar aspectos eco-epidemiológicos e antropológicos da LTA no Médio Purús, financiado pela FAPEAM/CNPq, e em andamento no município de Lábrea, estado do Amazonas onde há registros de casos e suspeita de subnotificações. Por isso, achou-se oportuno o estudo de flebotomíneos do município como contribuição para o conhecimento da epidemiologia da LTA. O objetivo deste trabalho foi estudar os aspectos entomológicos relacionados a epidemiologia da LTA em uma área indígena do município de Lábrea. A coleta de flebotomíneos foi realizada em fevereiro de 2012, na Aldeia Castanheira, Terra Iindígena Caititú, município de Lábrea, utilizando armadilhas com atrativo luminoso. Após a identificação dos insetos, foi realizada a detecção e identificação de leishmânias presentes nos flebotomíneos coletados utilizando uma abordagem molecular. Um total de 1.267 flebotomíneos foi coletado, 819 (64,6%) fêmeas e 448 (35,4%) machos, distribuídos em 10 gêneros e 32 espécies. Os gêneros mais abundantes encontrados foram Psychodopygus, Nyssomyia e Trichophoromyia, já a espécie Trichophoromyia ubiquitalis foi a mais abundante com 235 (18,5%) espécimes coletados, seguida de Psychodopygus davisi com 228 (18%) e Nyssomyia antunesi com 135 (10,7%) indivíduos. Foi detectada a presença de DNA de Leishmania em seis espécies: Sciopemyia servulolimai, Trichophoromyia ubiquitalis, Evandromyia apurinan, Nyssomyia umbratilis, Nyssomyia yuilli yuilli, Psychodopygus davisi, sendo a taxa de infecção mínima estimada foi de 0,84%. As espécies de leishmânias detectadas foram: Leishmania (L.) amazonensis, Leishmania (V.) braziliensis e uma amostra identificada somente ao nível de subgênero Leishmania (Viannia) sp. A infecção por Leishmania detectada em espécies vetoras encontradas neste trabalho, é um indício da participação das mesmas no ciclo de transmissão de LTA no município. Futuras investigações devem ser realizadas para ajudar a elucidar a epidemiologia da LTA não só no município de Lábrea mas para o estado do Amazonas.

Introdução - A leishmaniose tegumentar americana (LTA) é uma antropozoonose com o envolvimento de várias espécies de Leishmania, mamíferos reservatórios e flebotomíneos (vetores). Apresenta ampla distribuição no Brasil, com registro de casos em todas as unidades da Federação. É de interesse em saúde pública devido à gravidade de algumas de suas formas e alta prevalência em algumas regiões. Em áreas de transmissão na Grande São Paulo, têm sido registradas várias espécies de flebotomíneos, merecendo destaque Pintomyia fischeri, por ser altamente antropofílica e apresentar ampla distribuição no estado de São Paulo. Objetivo - Identificar a capacidade vetora dos flebotomíneos Pintomyia fischeri para Leishmania (Viannia) braziliensis, um dos agentes da leishmaniose tegumentar americana (LTA), comparando-a com a de Nyssomyia intermedia, vetor desse parasita na região Sudeste do Brasil. Métodos - Os seguintes parâmetros foram identificados: em campo - densidade das espécies em relação a um hospedeiro (hamster), com a instalação da armadilha de Disney em duas áreas (município do Embu na Grande São Paulo e município de Iporanga, na região do Vale do Ribeira) e teste de uma nova armadilha (apenas em Iporanga); em laboratório, a partir de infecção experimental de flebotomíneos por meio da alimentação em hamsters infectados pelo parasita - probabilidade diária de sobrevivência dos flebotomíneos, duração do ciclo gonotrófico, período de incubação extrínseca dos parasitas, proporção de flebotomíneos que se alimentam em fonte infectante e que se tornaram infectivos. Com estes dados calculou-se a capacidade vetora (novas infecções que a população vetora levará adiante ao se alimentar em um hospedeiro infectado), pela expressão V = m.a.b.Einf, onde V = capacidade vetora, m = proporção de fêmeas atraídas ao hospedeiro, a = proporção de fêmeas alimentando-se no hospedeiro/duração do ciclo gonotrófico, b = proporção de fêmeas infectadas que desenvolvem a forma infectante e Einf = expectativa de vida infectiva. Resultados Em se tratando do parâmetro observado em campo (m), densidade de flebotomíneos/hamster/dia, na armadilha de Disney em Iporanga esse valor para Ny. intermedia foi de 0,027 e para Pi. fischeri (0,011), em Embu, para Pi. fischeri (0,103) e para nova armadilha, apenas em Iporanga, Ny. intermedia (7,9) e Pi. fischeri (0,11). Nos parâmetros obtidos em laboratório: sobrevida infectiva (Einf): Ny. intermedia (0,84 dias) e Pi. fischeri (0,89 dias); proporção de fêmeas alimentadas em hamster: Ny. intermedia (0,77) e Pi. fischeri (0,67) e mediana do ciclo gonotrófico, para as duas espécies de 5 dias; período de incubação extrínseca para as duas espécies de 5 dias; e proporção de fêmeas infectadas por promastigotas que chegaram à forma infectante (b) para Ny. intermedia (0,90) e Pi. fischeri (0,311). A capacidade vetora obtida com a armadilha de Disney para Ny. intermedia em Iporanga foi de 0,0027 e para Pi. fischeri (0,00089) e no Embu para Pi. fischeri de 0, 0083 e para a armadilha nova, em Iporanga apenas, Ny. intermedia (0,8) e Pi. fischeri (0,0089). Conclusão - A capacidade vetora avaliada pelas armadilhas nova e a de Disney apresentou disparidades. Para Ny. intermedia, em Iporanga o valor da primeira foi 296,3 vezes maior que o da Disney e para Pintomyia fischeri foi 10 vezes maior. Na comparação entre o mesmo tipo de armadilha, em Iporanga para Ny. intermedia o valor da capacidade vetora obtido com a Disney foi 3,03 vezes superior ao de Pi. fischeri e na nova, foi 89,9 vezes. Para Pintomyia fischeri, os valores obtidos para a capacidade vetora considerando a Disney (Embu) foram 9,3 vezes maiores que os de Iporanga. A capacidade vetorial de Pi. fischeri (Embu) foi 3,07 vezes maior do que o de Ny. intermedia (Iporanga). Possivelmente as duas espécies atuem na transmissão da LTA no estado de São Paulo. Em Iporanga Ny. intermedia atua de maneira bem mais incisiva do que a Pi. fischeri. Por outro lado, no Embu, onde Ny. intermedia não tem sido capturada, a população de Pi. fischeri apresentou capacidade vetora superior à de Ny. intermedia (Iporanga), quando avaliadas segundo o mesmo tipo de armadilha para obtenção da densidade vetora. / Introduction American cutaneous leishmaniasis (ACL) is an anthroponosis involving several species of Leishmania, mammalian reservoirs and phlebotomine vectors. It is widespread in Brazil, cases having been registered in all the states. It constitutes a serious public health problem due to the gravity of some of its forms and its high prevalence in some regions. Several species, including Pintomyia fischeri that is dominant, widespread and anthropophilic, have been registered in areas of transmission within the metropolitan region of Greater São Paulo, Objective To identify the vectorial capacity of Pintomyia fischeri for Leishmania (Viannia) braziliensis, one of the agents of ACL, comparing it with that of Nyssomyia intermedia, a vector of this parasite in Southeastern Brazil. Methods The following parameters were identified: in the field density of the species in relation to a host (hamster), with the installation of a Disney trap in two areas: Embu municipality (within Greater São Paulo) and Iporanga municipality (in the Ribeira Valley region), and a new trap (in Iporanga); in the laboratory on the basis of the experimental infection of sandflies which fed on hamsters infected with the parasite; daily probability of survival of the sandflies; duration of the gonotrophic cycle; extrinsic period of incubation of the parasite; proportion of phlebotomines which fed on the infective host and which themselves became infectious. With these data the vectorial capacity (new infections which the vector population will transmit when it feeds on an infected host) was calculated, using the expression V = m.a.b.Einf, where V = vectorial capacity, m = proportion of females attracted to the host , a = proportion of females feeding on the host/duration of gonotrophic cycle, b = proportion of infected females which develop the infective forms of the parasite and Einf = infective life expectancy. Results As regards the parameter observed in the field (m), density of insects/hamster/day, in the Disney trap in Iporanga this value for Ny. intermedia was 0.027 and for Pi. fischeri, 0.011, in Embu, for Pi. fischeri , 0.103 and for the new trap in Iporanga, for Ny. intermedia, 7.9 and for Pi. fischeri, 0.11. In the laboratory: infective life expectancy (Einf): Ny. intermedia (0.84 days) and Pi. fischeri (0.89 days); proportion of females fed on the hamster: Ny. intermedia (0.67) and Pi. fischeri (0.77) and median of the duration of the gonotrophic cycle for both species, of 5 days; and proportion of females infected by promastigotes which attained infective form (b) for Ny. intermedia (0.90) and Pi. fischeri (0.59). The vectorial capacity obtained with the Disney trap for Ny. intermedia in Iporanga was 0.0027 and for Pi. fischeri, 0.00089 and in Embu for Pi. fischeri, 0.0083 and for the new trap only in Iporanga, Ny. intermedia (0.8) and Pi. fischeri (0.0089). Conclusion The vectorial capacity measured by the new and the Disney traps presented considerable differences. For Ny. intermedia in Iporanga the value of the former was 296.3 times greater than that of the Disney and for Pintomyia fischeri it was 10 times greater. In the comparison between traps of the same kind, in Iporanga for Ny. intermedia the value of vectorial capacity obtained with the Disney trap was 3.03 times greater than that of Pi. fischeri and in the new trap it was 89.9 times greater. For Pintomyia fischeri, the vectorial capacity obtained with Disney (Embu) was 9.3 times greater than that for Iporanga. The vectorial capacity of Pi. fischeri (Embu) was 3.07 times greater than that of Ny. intermedia (Iporanga). Possibly both species are active in the transmission of ACL in the state of São Paulo. In Iporanga Ny. intermedia acts in a much more incisive way than Pi. fischeri. On the other hand, in Embu, where Ny. intermedia was not captured, the Pi. fischeri population presented a vectorial capacity greater than that of Ny. intermedia (Iporanga) when assessed by the same type of trap for the calculation of vector density.

Introdução: a vasculopatia livedoide é uma doença cutânea rara que acomete principalmente mulheres adultas, cursando inicialmente com lesões purpúricas e necróticas intensamente dolorosas, acometendo extremidades inferiores, que podem evoluir com úlceras de tamanhos variados. Estas lesões costumam evoluir com cicatrizes atróficas, estreladas e com telangiectasias, denominadas cicatrizes de atrofia branca. Apesar da etiologia não estar totalmente esclarecida, os distúrbios da coagulação parecem ser o mecanismo fisiopatológico primário da vasculopatia livedoide. Inúmeras trombofilias já foram associadas à vasculopatia livedoide, porém cerca de metade dos casos permanece sem etiologia definida. Recentemente foi documentado aumento de lipoproteína(a) na vasculopatia livedoide. Níveis plasmáticos elevados de lipoproteína(a) são considerados um fator de risco causal independente para o desenvolvimento de doenças cardiovasculares e trombóticas, devido à sua similaridade estrutural com o plasminogênio. A deposição tecidual da lipoproteína(a) em vasos ateroscleróticos é bem descrita, porém na vasculopatia livedoide a participação da lipoproteína(a) não é conhecida. Objetivos: analisar os níveis séricos da lipoproteína(a) e a sua expressão tecidual na pele dos pacientes com vasculopatia livedoide e comparar os resultados com um grupo controle. Métodos: amostras de pele obtidas através de biópsia de pele de 38 pacientes com vasculopatia livedoide (27 do sexo feminino e 11 do sexo masculino) e 9 indivíduos do grupo controle (5 do sexo feminino e 4 do sexo masculino) foram avaliados para a presença de lipoproteína(a) através de imuno-histoquímica utilizando anticorpo policlonal anti- lipoprotéina(a). Os níveis plasmáticos da lipoprotéeíina(a) foram analisados por imunoturbidimetria e comparados com setenta pacientes portadores de outras doenças dermatológicas. Resultados: utilizando anticorpo policlonal contra lipoproteína(a), observou-se que a pele perilesional em pacientes com vasculopatia livedoide apresentava dez vezes mais lipoprotéina (a) do que a pele controle. O coeficiente de correlação de Pearson (r = 0,02) mostrou que os níveis teciduais de lipoproteína(a) não se correlacionaram com os níveis plasmáticos nos pacientes com vasculopatia livedoide. Níveis plasmáticos elevados de lipoproteína(a) foram observados nos portadores de vasculopatia livedoide, no entanto não pode ser demonstrada diferença estatística em relação ao grupo controle. Conclusões: achados de aumento da expressão tecidual de lipoproteína(a) na pele lesionada e níveis plasmáticos elevados desta proteína em pacientes com vasculopatia livedoide podem corroborar com a hipótese de que a lipoproteína(a) possa contribuir para a patogênese da vasculopatia livedoide através dos seus efeitos antifibrinolíticos, pró- trombóticos e ação direta no endotélio vascular, induzindo a formação do trombo / Background: livedoid vasculopathy is a chronic disorder that usually presents as recurrent reticulated purpura on the lower limbs, with recurrent painful, purpuric and/or necrotic macules that may lead to ulcerative lesions. These lesions usually heal into atrophic white scars, with depigmentation and telangiectasias, known as \"atrophie blanche\". Although the etiology of the livedoid vasculopathy is not fully understood, coagulation disorders appear to be the primary pathophysiological mechanism. Hereditary thrombophilia has been associated with livedoid vasculopathy, but about half of the cases remain without defined etiology. Recently, high serum levels of lipoprotein(a) in livedoid vasculopathy patients have been documented. Elevated plasma levels of lipoprotein(a) are an independent risk factor for the development of cardiovascular and thrombotic diseases due to their structural similarity with plasminogen. Lipoprotein(a) deposition in atherosclerotic vessels is well described, but its role in livedoid vasculopathy is unknown. Objectives: To analyze the serum levels of lipoprotein(a) and tissue expression in the skin of patients with livedoid vasculopathy and to compare the results with a control group. Methods: Skin biopsy samples from 38 patients (27 women-11 men) with active lesions diagnosed with livedoid vasculopathy and 9 samples of normal skin (5 women-4 men) from control individuals without livedoid vasculopathy were evaluated for skin expression of lipoprotein(a) by immunohistochemistry using polyclonal anti-lipoprotein(a) antibody. Plasma levels of lipoprotein(a) were analyzed by immunoturbidimetry and compared with seventy patients with other dermatological diseases. Results: We found lesional skin in patients with livedoid vasculopathy expressed tenfold higher lipoprotein(a) immunostaining than controls. High plasma levels of lipoprotein(a) were observed in livedoid vasculopathy patients, but we cannot observed a positive correlation (p = 0.02) between skin expression of Lipoprotein(a) and plasma levels of Lipoprotein(a) in patients with livedoid vasculopathy. Conclusions: Increased of lipoprotein(a) tissue expression on lesioned skin and elevated plasma levels of this protein in patients with livedoid vasculopathy may corroborate the hypothesis that lipoprotein(a) may contribute to the pathogenesis of livedoid vasculopathy through its antifibrinolytic effects, prothrombotic and direct action on the vascular endothelium, inducing thrombosis

Os protozoários do gênero Leishmania são os agentes etiológicos da leishmaniose cutânea e visceral. A infecção é transmitida por insetos hematófagos, nos quais os parasitos se desenvolvem sob a forma promastigota. Nos hospedeiros mamíferos, incluindo a espécie humana, as formas encontradas são as amastigotas que proliferam nos macrófagos; estas células podem controlar a infecção produzindo radicais derivados do oxigênio e do nitrogênio. O óxido nítrico (NO) é um dos mais potentes radicais com atividade leishmanicida. Existem várias moléculas já descritas que liberam NO em meio aquoso ou quando adicionadas a suspensões celulares. São chamadas de doadoras de NO. Algumas destas vêm sendo estudadas pelo seu potencial terapêutico no tratamento das leishmanioses cutâneas. A molécula S-nitrosoglutationa (GSNO) pertence a um grupo de moléculas conhecido com S-nitroso-tióis e é um doador de NO relativamente estável. A GSNO havia sido previamente testada em culturas de formas promastigotas de L. amazonensis e causou a morte do parasito. O possível uso da GSNO na terapia das leishmanioses cutâneas demanda o conhecimento de suas ações sobre as formas amastigotas encontradas nos hospedeiros mamíferos. Terapias complementares aos antimoniais usados no tratamento da leishmaniose cutânea são necessárias porque estes são apenas injetáveis, têm efeitos colaterais e ocorrem altos índices de desistência do tratamento. O objetivo do trabalho foi analisar o efeito da molécula GSNO nas culturas de células THP-1 (linhagem de monócitos humanos leucêmicos) infectadas com L.major e o efeito da sua administração tópica diretamente na lesão ulcerada leishmaniótica de camundongos infectados. Os experimentos com células THP-1 infectadas com L. major e tratadas com GSNO, mostram marcada redução do parasitismo intracelular, dose dependente, em comparação às culturas sem tratamento. O tratamento com GSNO foi testado em camundongos Balb/c 8 infectados com L. major e em camundongos C57BL/6 desprovidos do gene de IFN<font face=\"symbol\">g- (Knockout de IFN ou IFN-<font face=\"symbol\">g KO) infectados com L. braziliensis. Grupos de animais foram infectados no dorso e após 60 dias iniciou-se o tratamento tópico com GSNO diluída em PBS, em GEL F127 ou com os respectivos veículos, como controles; ainda, outros grupos foram tratados sistemicamente (via endovenosa) com o antibiótico com ação leishmanicida, anfotericina B, ou com anfotericina B associada à aplicação tópica de GSNO. Foram feitas duas medições semanais das lesões. Decorridos aproximadamente 60 dias após o início do tratamento, os animais foram sacrificados e retirados o linfonodo drenante (inguinal superficial) e a lesão para fazer a quantificação parasitária. O tratamento tópico da infecção por L.major com a molécula GSNO reduziu o tamanho da lesão cutânea, chegando a se observar fechamento da úlcera em alguns animais; a carga parasitária no linfonodo drenante e/ou na lesão também sofreu redução. Nos camundongos IFN-<font face=\"symbol\">g KO infectados por L. braziliensis, também ocorreu inibição da progressão da lesão pelo tratamento local com GSNO em comparação ao grupo não tratado. Como conclusão, conseguimos demonstrar que GSNO apresenta efeito leishmanicida sobre formas amastigotas de L. major em cultura celular e reduz a lesão cutânea causada por L.major ou por L. braziliensis em comparação aos grupos controle. / Protozoa of the genus Leishmania are the etiological agents of cutaneous and visceral leishmaniasis. The infection is transmitted by hematophagous insects in which the parasite multiplies and differentiates as promastigotes. In the mammalian hosts, including man, the parasite proliferates as amastigotes inside macrophages; production of reactive oxygen or nitrogen intermediates (ROI and RNI) by these cells can control the infection. Nitric oxide (NO) is a very toxic RNI active against leishmania. The are many molecules that liberate NO in solution or when added to cellular suspensions. They are collectively called NO donors. A few of them have been studied because of their potential therapeutic use in cutaneous leishmaniasis. The molecule S-nitrosoglutathione (GSNO) belongs to the S-nitroso-thiols and is a relatively stable NO donor. GSNO has been previously tested and caused the death of L. amazonensis promastigotes in liquid culture. A putative application of GSNO in the treatment of cutaneous leishmaniasis requires the understanding of its activities on amastigote forms which are the parasite forms found in the mammalian host. Because the classical leishmaniasis treatment relies on injectable antimonial drugs which have many side effects, many patients abandon the treatment. Therefore, additional therapy to accelerate the cure is needed as well as new drugs are much needed. The aim of the present work is to analyze the effect of the molecule GSNO in cultures of THP-1 cells (a leukemia human monocyte cell line) infected with Leishmania major as well as the effect of administering it topically into the lesion of infected mice. The experiments done on L. major-infected THP-1 cells showed marked dose-dependent reduction of parasitism when compared to untreated infected cells. The topical treatment with GSNO was done in BALB/c mice infected with L. major and in C57BL/6 mice deprived of the IFN-<font face=\"symbol\">g gene (IFN- <font face=\"symbol\">g KO) infected with L. braziliensis. Groups of mice were infected in the shaven dorsal skin and 60 days later the topical treatment with GSNO dissolved in PBS, in GEL F127 was started. Still other groups were treated systemically (i.v.) with Amphotericin B which is leishmanicidal or with Amphotericin B associated to topically applied GSNO. The lesions were measured twice a week. After 60 days the mice were killed and the draining lymph node and the lesion were removed for quantifying the parasite numbers. The topical treatment of L. major-induced ulcer with GSNO reduced the size of the ulcer leading in some mice to complete healing; the parasitic loads in the draining lymph node or in the lesion were also reduced. In L. braziliensis-infected IFN-<font face=\"symbol\">g KO mice, inhibition of lesion growth also occurred in the mice topically treated with GSNO in comparison to the untreated control group. In conclusion we showed that GSNO is leishmanicidal to L. major intracellular amastigotes and that the topical treatment reduces the size of the lesion caused in mice by L. major or by L. braziliensis in comparison to the control groups.

Thesis (M. OPT.) -- University of Limpopo, 2001 / Refer to document

Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2008.

Abstract Cutaneous T cell lymphoma (CTCL) is a rare, incurable, chronic disease accounting for approximately 3% of non-Hodgkin's lymphoma diagnoses every year. Patients with CTCL have skin lesions that can vary in severity putting patients at risk for developing symptoms that may impair their quality of life (QOL). The disease burden can lead to increased depressive symptoms, fatigue distress, and anxiety that the disease may be worsening. Seventy-five participants agreed to take part in an exploratory, prospective study to evaluate depressive symptoms, anxiety, fatigue distress, and spirituality as predictors of QOL in CTCL patients. Demographic variables including stage of disease, ethnicity, age, gender, marital status, level of education, and time since diagnosis, were also included in the analyses to assess for relationships. Bivariate correlations, t-tests, and regression analyses were conducted to assess for relationships among the predictor variables and QOL. The analyses revealed that the proposed model explained 64% of the variance, and depressive symptoms (t= -2.4, p= 0.020) and stage of disease (t= -3.0, p= 0.004) significantly predicted the QOL of CTCL patients. Evaluating for predictors that influence the QOL helps us to better understand the needs of the patients afflicted with CTCL. The importance of studying the QOL of the CTCL patients lies in the fact that nurses can assist in helping patients alleviate some of the symptoms they experience, thereby improving their QOL. Further study is warranted in developing interventions to assist in the preservation of QOL.

Inanga (Galaxias maculatus) are a culturally and economically important fish species in New Zealand and abroad. However, very little is known about their ability to deal with trace element contamination. As a scaleless fish with the ability to survive in relatively extreme environments, they may not fit toxicity models (such as the biotic ligand model; BLM) based on other fish species. The aim of this study was to determine how this fish responds to elevated trace elements in both the laboratory and field in order to determine the applicability of these toxicity models. In order to determine the impacts of stress on ion transport and subsequent metal toxicity, inanga were exposed to handling stress and measures of ion uptake were collected. Handling stress was shown to result in increased ventilation rates, resulting in stimulated sodium (Na+) efflux. A compensatory increase in Na+ influx was also measured as a result of this stress. Inanga largely recovered from this ionoregulatory stress within 2 hours, with full recovery after 24 hours. This was indicative of a rapid homeostatic response for maintaining ion balance. Enhanced Na+ uptake in response to this stress resulted in increased copper (Cu) uptake in Cu-contaminated water, suggesting stressed fish will accumulate more Cu (and likely other Na+ mimics) than an unstressed fish. These results suggest a heightened vulnerability of inanga to this type of contaminant as a result of exercise stress during migrations. A combination of field and laboratory studies was used in order to measure trace element accumulation in inanga. In situ field studies showed changes to aluminum (Al) and iron (Fe) body burdens when inanga were placed in streams of varying trace element concentrations along the West Coast of the South Island. However, other trace elements measured did not alter over the period of exposure (9-10 days). Biochemical biomarker analysis showed no changes in the activity of Na+/K+-ATPase (NKA), but a marker of lipid peroxidation (thiobarbituric acid reactive substances; TBARS) was elevated in one stream. Analysis suggested that stream pH was the major driver of this effect, whether directly or via changes to metal bioavailability. Subsequent laboratory exposures (96 h) of inanga to 1.2, 2.7, 10.8, and 44 µg L-1 dissolved Fe and 5.6, 23.3, 60.7, and 128.7 µg L-1 dissolved zinc (Zn) showed no difference in whole body trace element accumulation, ammonia excretion, ion influx (Ca2+ and Na+), and TBARS. There were significant differences in oxygen consumption (MO2) after Fe exposures, with increases in the 2.7 and 44 µg L-1 dissolved Fe exposures. Laboratory exposure results suggest inanga are relatively insensitive to short-term Fe and Zn exposures. Both in vivo (whole body partitioning) and in vitro (Ussing chamber) techniques were used to determine the influence of cutaneous ion transport on preventing trace element accumulation. Results suggest inanga use their skin as an additional site of calcium (Ca2+) and Na+ uptake. This is the first study to confirm these ion transport capabilities in inanga, and revealed that up to 48% of Na+ uptake may occur across the skin. Pharmacological inhibition of Ca2+ uptake was achieved by known Ca2+ channel blockers (verapamil and lanthanum). Furthermore Fe and Zn impaired cutaneous Ca2+ transport, indicating that ion transport pathways in the skin modulate in response to these metals.

The reactive control of fingertip forces maintaining grasp stability was examined in man during a prehensile task. Blindfolded subjects used the precision grip between the tips of index finger and thumb to restrain an object that was subjected to unpredictable load forces. These were delivered tangential to the parallel grip surfaces of the object. Load forces, grip forces (perpendicular to the grip surfaces) and position of the object were recorded.Subjects automatically adjusted the grip forces to loads of various amplitudes and rates. Thereby they maintained a reliable safety margin against frictional slips without using excessive grip forces. A rapid rise in grip force lasting about 0.2 s was triggered after a short delay following the onset of a sustained ramp load increase. This 'catch-up' response caused a quick restoration of an adequate grip:load force ratio that prevented frictional slips. If the ramp load continued to increase after the catchup response, the grip force also increased in parallel with the load change in a 'tracking' manner. Consequently, during the hold phases of 'ramp-and-hold' loads, the employed grip forces were approximately proportional to the load amplitude. Sensory information about the rate of change of the load force parametrically scaled the 'catchup' and 'tracking' responses.Following anesthetic block of sensory input from the digits, the grip responses were both delayed and attenuated or even abolished. To compensate for these impairments, subjects had to voluntarily maintain exceedingly high grip forces to prevent the object from slipping. The grip control improved slightly during hand and forearm support conditions that allowed marked wrist movements to occur in response to the loading. This indicates that signals from receptors in muscles, joints or skin areas proximal to the digits can to some extent be used to adjust grip forces during impaired digital sensibility. In contrast, these signals had only minor influence on the control during normal digital sensibility.Grip responses to loads delivered in various directions revealed that the load direction, in relation to gravity and to the hand's geometry, represents intrinsic task variables in the automatic processes that maintain a stable grasp. The load direction influenced both the response latencies and the magnitudes of the grip responses. The response latencies were shortest for loads in directions that were the most critical with regard to the consequences of frictional slippage, i.e., loads directed away from the palm or in the direction of gravity. Recordings of signals in cutaneous afferents innervating the finger tips demonstrated that these effects on the response latencies depended on differences in the time needed by the central nervous system to implement the motor responses. The short latencies in the most ‘criticar load directions may reflect the preparation of a default response, while additional central processing would be needed to execute the response to loads in other directions. Adjustments to local frictional anisotropies at the digit-object interface largely explained the magnitude effects.In conclusion, grip responses are automatically adjusted to the current loading condition during unpredictable loading of a hand held object. Subjects call up a previously acquired sensorimotor transform that supports grasp stability by preventing both object slippage and excessive grip forces. Cutaneous sensory information about tangential forces and frictional conditions at the digit-object interface is used to initiate and scale the grip responses to the current loading conditions, largely in a predictive manner. / <p>Diss. (sammanfattning) Umeå : Umeå universitet, 1995, Härtill 5 uppsatser</p> / digitalisering@umu