Linfonodos de 146 bovinos com lesões macroscópicas identificadas como sugestivas de tuberculose pelos Serviços de Inspeção Federal e Estadual de abatedouros do Estado de São Paulo, Brasil, no período de Maio de 2002 à Janeiro de 2004, conservadas em solução de borato de sódio em temperatura ambiente por no máximo 60 dias, negativos nos cultivos de micobactérias nos meios de Lowëstein-Jensen e Stonebrink com descontaminação por Petroff e HPC e conservadas por congelamento, foram submetidos a novos cultivos em Agar Sangue, Agar MacConkey e Agar Saboraud bem como ao exame histopatológico pelas colorações de Hematoxilina-Eosina e Ziëlh-Neelsen. Nos cultivos houve isolamentos de E. faecium, E. faecalis, E. solitarius, S. arlettae e S. epidermidis. Não houve correlação entre as bactérias isoladas e a localização dos respectivos linfonodos. Os exames histopatológicos classificaram 49 animais isentos de qualquer lesão microscópica, 4 positivos para micobactérias sp, 18 fortemente sugestivos para micobactérias sp e 45 como processos atribuídos a outros agentes que não micobactérias. Não houve correlação entre o aspecto histológico dos linfonodos e o tipo de microrganismo isolado. Os microrganismos isolados foram considerados como contaminantes que tiveram acesso aos materiais durante os procedimentos de abate. / Lymph nodes of 146 bovines presenting lesions identified macroscopically as suggestive of tuberculosis by the Federal and State Inspection Services of abbatoirs at the State of São Paulo, Brazil, from May, 2002 to January, 2004, that were preserved in sodium borate solution at room temperature for no longer than 60 days, negative to mycobacteria culturing on Lowenstein-Jensen and Stonebrink media, decontaminated by Petroff and HPC methods and kept frozen, were subjected to new cultivations on blood, MacConkey and Sabouraud agars as well as to histopathological examination by Hematoxilin-Eosin and Ziëlh-Neelsen coloration techniques. E. faecium, E. faecalis, E. solitarius, S. arlettaeand S. epidermidis were isolated by culturing. No correlation between the isolated bacteria and the respective lymph nodes localization was found. The histopathological classification of the animals was: 49 presented microscopically no lesion, 4 were Mycobacterium sp. positive, 18 were strongly suggestive of Mycobacterium sp. and 45 were attributed to agents other than mycobacteria. There was no correlation between the histological aspect of the lymphnodes and the microorganisms isolated. The isolated microorganisms were considered as contaminants that had access to these materials during the slaughter procedures.

A leprose dos citros é uma doença de grande importância econômica aos citricultores, devido aos prejuízos por ela causados. A doença provoca lesões localizadas nos ramos, frutos e folhas e pode causar grandes perdas aos pomares não tratados. A leprose dos citros possui etiologia viral sendo o vírus denominado Citrus leprosis virus (CiLV). Atualmente, atribuí-se a leprose dois tipos de vírus que causam sintomas morfológicos e citopatológicos distintos: o tipo nuclear (CiLV-N) e o tipo citoplasmático (CiLV-C). O presente trabalho teve como objetivo investigar as alterações anatômicas e comparar o perfil protéico entre as lesões causadas pelos dois tipos de vírus em laranjeira-doce [Citrus sinensis (L.) Osbeck]. Para as análises anatômicas foram coletadas amostras de tecidos sadios e de tecidos lesionados. As amostras foram fotografadas e em seguida fixadas em solução de Karnovsky, desidratadas em série etílica, infiltradas em resina hidroxi-etil-metacrilato (Leica Historesin), seccionadas em micrótomo rotativo a 5-7 µm de espessura, coradas com azul de Toluidina para as análises histológicas usuais. Também foram realizados testes histoquímicos para fenóis, compostos pécticos, proteína e lipídios. A captura de imagens digitais dos materiais preparados em lâminas foi realizada em microscópio equipado com câmera de vídeo. Para a análise do perfil protéico as amostras coletadas foram imediatamente criofixadas em nitrogênio líquido e armazenadas em refrigerador a - 80ºC para posterior extração protéica. As proteínas extraídas foram quantificadas e depois separadas por eletroforese em gel de policrilamida. As lesões foliares se iniciaram como pontuações necróticas, envolvidas por halos cloróticos que limitam o crescimento da lesão. Nos ramos, ocorreram dois tipos de lesões com fendas. As lesões de ramos e de folhas provenientes do campo com sintomas de CiLV-C apresentaram similaridades com as amostras inoculadas. No caso das lesões foliares causadas pelo CiLV-N observa-se a presença de três regiões distintas: o centro necrótico, o halo intermediário e o halo clorótico. As lesões nos frutos de ambos os tipos de leprose eram distintas. Ductos traumáticos gomosos foram observados nos ramos infectados pelo CiLV-C e nos feixes vasculares de folhas e frutos infectados pelo CiLV-N. Quanto ao perfil protéico, verificou-se um aumento na concentração protéica nos tecidos infectados, em especial, nas lesões causadas pelo CiLV-N. Foi possível observar que havia um padrão anatômico e histoquímico nas lesões causadas pela leprose em laranjeira doce independente do tipo de vírus, pois em todas as lesões foi verificada hipertrofia e hiperplasia celular, plasmólise, a redução de grãos de amido, o acúmulo de compostos lipídicos nas células da área necrosada e a presença de um halo, ao redor dessa área, constituído de células que acumulam proteínas. / The citrus leprosis of citrus has been considered a disease of great economic importance to citrus producers due to the damage that it causes. The disease is responsible for lesions found in the twigs, leaves and fruits, and it may cause great losses to the untreated orchards. The citrus leprosis has viral etiology being the virus called Citrus leprosis virus (CiLV). Currently, there are two types of virus attributed to leprosis that cause different morphological and cythopatological symptoms: the nuclear (CiLV-N) and cytoplasmic type (CiLV-C). This study aimed to investigate the anatomical changes and to compare the proteic profile between the injuries caused by two types of virus in sweet orange [Citrus sinensis (L.) Osbeck]. For anatomical analyses healthy tissue and different lesions were photographed and then fixed in Karnovsky solution, dehydrated in a graded ethylic series, embedded in hidroxy-ethyl methacrylate resin (Leica Historesin), sectioned (5 ?m thick), stained and mounted in synthetic resin. The digital images were capture in a microscope with video camera. For the analysis of protein profiles the samples were collected and immediately cryofixed in liquid nitrogen and stored in the refrigerated - 80 ° C. The proteins were extracted, quantified and then separated by gel electrophoresis of polyacrylamide gel. Foliar lesions began as necrotic spots surrounded by a yellow halo that inhibited the lesion expansion. The twigs presented two different lesions: depressed ones and pustules with ruptures. The collected-field lesions of leaves and twigs showed that symptoms of CiLV-C had a certain similarity with the inoculated samples. In the case of foliar lesions caused by CiLV-N there was the presence of three different regions: the necrotic center, the intermediate halo and chlorotic halo. The fruit lesions caused by both types of leprosis virus were different. Ductos traumáticos gomosos foram observados nos ramos infectados pelo CiLV-C e nos feixes vasculares de folhas e frutos infectados pelo CiLV-N. Gummous traumatic ducts in twigs infected by CiLV-C and in the vascular bundles of fruit and leaves infected with CiLV-N were observed. In the proteic profile, there was an increase in protein concentration in the infected tissues especially for injuries caused by CiLV-N. It was possible to observe that there was an anatomical and histochemical pattern in the lesions caused by leprosis in sweet orange independently of the type of virus, therefore in all the injuries was verified hyperplasia, hypertrophy, cell plasmolisis, the reduction of starch grains, the accumulation of lipids compounds in the cells of the necrotic areas and the presence of a halo, surrounding that area, constituted by cells that accumulate proteins.

Como a imunoproteção na Paracoccidioidomicose (PCM) é principalmente mediada por células T, investigamos o impacto da deficiência de CD28, molécula co-estimulatória de linfócitos T, na gravidade da infecção primária e secundária pelo Paracoccidioides brasiliensis. Quando comparados a camundongos C57Bl/6 normais (WT), camundongos CD28-deficientes (CD28KO) apresentaram infecção mais grave associada à produção diminuída de anticorpos e citocinas. Além disso, a pré-imunização de animais deficientes e normais resultou em imunoproteção equivalente. Inesperadamente, a sobrevida de animais CD28KO foi significativamente maior que a dos WT, apesar da sua elevada carga fúngica tecidual. Em conclusão, nosso trabalho mostrou que a deficiência de CD28 resulta em PCM mais grave, porém não letal, associada a resposta imune deficiente. Além disso, verificamos que carga fúngica elevada, na ausência da imunidade adaptativa efetora, não ocasiona diminuição de sobrevida, revelando que a resposta imune na PCM pode tanto proteger como ser deletéria aos hospedeiros. / As immunoprotection in Paracoccidioidomycosis (PCM) is mainly mediated by T cells, and CD28 is a costimulatory molecule for T lymphocytes, we investigated the influence of CD28 deficiency in primary and secondary PCM. Compared with normal C57BL/6 mice, CD28-deficient (CD28KO) mice developed a more severe infection associated with impaired antibody and cytokine production. In addition, CD28KO and WT mice previously immunized by the s.c. route developed equivalent immunoprotection when challenged by the pulmonary route. Interestingly, CD28KO mice presented increased mean survival time despite their elevated fungal loads in the lungs. In conclusion, our work showed, for the first time, that CD28-deficiency results in more severe, but not overwhelming, PCM. Furthermore, in the absence of effector adaptative immunity, elevated fungal loads do not cause lethal infections, revealing the protective and deleterious effects of immune responses to Paracoccidioides brasiliensis infected hosts.

O objetivo do trabalho foi avaliar a técnica de biópsia retal em equinos e as alterações histopatológicas encontradas na mucosa retal em animais clinicamente saudáveis e em animais com alterações do trato gastrintestinal. Foram utilizados 32 equinos divididos em três grupos: 10 animais clinicamente saudáveis (grupo 1), sete animais com alterações do trato gastrintestinal com diagnóstico estabelecido (grupo 2) e 15 animais com manifestações clínicas inespecíficas do trato gastrintestinal (grupo 3). Os equinos do grupo 1 foram biopsiados duas vezes, a segunda biópsia sendo realizada 48 horas após a primeira. As biópsias retais foram realizadas com os animais em estação, utilizando-se pinça de biópsia tipo jacaré. As amostras de mucosa retal foram coradas por hematoxilina-eosina (HE), azul de alciano com ácido periódico de Schiff a 2,5% e azul de toluidina. No grupo 1, não foram observadas diferenças significativas entre a primeira e a segunda biópsia, mas três equinos apresentaram pequenas alterações que podem ser decorrentes da palpação retal. Na primeira avaliação, sete animais desse grupo apresentaram padrão semelhante, permitindo estabelecer um padrão de normalidade histológica para a mucosa retal: epitélio superficial preservado; criptas uniformes e alinhadas perpendicularmente à superfície com grande número de células caliciformes com mucinas ácidas em maior quantidade; linfócitos intra-epiteliais na superfície do epitélio em número de até 15 linfócitos para cada 100 colonócitos; pequeno número de linfócitos, plasmócitos e eosinófilos infiltrando a lâmina própria e a submucosa. Os outros três equinos deste grupo apresentaram um aumento na intensidade das células inflamatórias (discreto a moderado na intensidade de linfócitos e plasmócitos na lâmina própria e submucosa, e moderado na intensidade dos eosinófilos na submucosa). Os equinos do grupo 2, todos com síndrome cólica, apresentaram as seguintes alterações histopatológicas: proctite linfoplasmocítica simples (4/7) e proctite eosinofílica (1/7). No grupo 3 (12 equinos com diarreia crônica e três com emagrecimento progressivo), apenas um animal apresentou mucosa retal com padrão de normalidade. Nos outros animais foram observadas: proctite linfoplasmocítica simples (6/15), proctite erosiva (3/15), proctite linfoplasmocítica com aumento de neutrófilos (2/15), proctite eosinofílica (2/15), proctite linfoplasmocítica com aumento de neutrófilos e eosinófilos (1/15) e infiltração por linfócitos atípicos/linfoma (1/15). Concluiu-se que: a técnica de biópsia retal avaliada é de fácil execução e segura; embora algumas alterações histológicas possam ser observadas na mucosa retal de equinos clinicamente saudáveis, causadas ou não por palpação retal, há um padrão de normalidade que deve ser usado para a avaliação de equinos com alterações gastrintestinais; apenas em alguns casos as alterações histológicas encontradas na mucosa retal em equinos permitem um diagnóstico definitivo, mas elas podem direcionar o raciocínio diagnóstico em outros, sendo este um importante exame complementar a ser indicado em casos de equinos com alterações gastrintestinais inespecíficas. / The purpose of this study was to asses the rectal biopsy techique in horses and the histopathological changes found in the rectal mucosa in clinically healthy animals and in animals with clinical signs of intestinal disorders. 32 horses were divided into three groups: 10 animals clinically healthy (group 1), seven animals with gastrointestinal disorders with an established diagnosis (group 2) and 15 animals with nonspecific clinical manifestations of the gastrointestinal tract (group 3). The horses in group 1 had two biopsies, the second biopsy was performed 48 hours after the first. Rectal biopsies were performed on the animals station, using alligator biopsy forceps. The samples of rectal mucosa were stained with hematoxylin end eosin (HE), alcian blue PAS pH 2.5% and toluidine blue. In group 1, no significant differences between the first and second biopsy were observed, but three horses showed small changes that may result from rectal palpation. In the first evaluation, seven animals of this group showed a similar pattern, establishing a standard of normality for histological rectal mucosa: Surface epithelial preserved; crypts of uniform length, diameter and perpendicular arrangement to the surface with acid mucins strongly dominated over neutral mucins in the goblet cell population; intraepithelial lymphocytes in the surface of the epithelium with sparse population of approximately 15 cells per stretch of 100 epithelial cells; sparse individual lymphocytes, plasma cells and eosinophils infiltrating the lamina propria and submucosa. The other three horses in this group had an increase in the intensity of inflammatory cells (mild to moderate in intensity of lymphocytes and plasma cells in the lamina propria and submucosa, and moderate intensity of eosinophils in the submucosa). The horses in group 2, all with colic syndrome, showed the following pathological changes: simple lymphocyticplasmacytic proctitis (4/7) and eosinophilic proctitis (1/7). In group 3 (12 horses with chronic diarrhea and three with progressive weight loss), only one animal showed rectal mucosa with normal histology. It was observed in the other animals: simple lymphocytic-plasmacytic proctitis (6/15), erosive proctitis (3/15), lymphocyticplasmacytic proctitis with increased neutrophils (2/15), eosinophilic proctitis (2/15), lymphocytic-plasmacytic proctitis with increased neutrophils and eosinophils (1/15) and atypical lymphocytes infiltration/lymphoma (1/15). It was concluded that: the rectal biopsy technique evaluated is effective and safe; although some histological changes can be observed in the rectal mucosa of clinically healthy horses, whether or not caused by rectal palpation, there is a normal range that should be used to the evaluation of horses with gastrointestinal disorders; only in some cases the histological changes found in the rectal mucosa in horses allow a definitive diagnosis, but they can lead to a diagnostic logic in others, which is an important complementary test to be indicated in cases of horses with nonspecific gastrointestinal disturbances.

O uso do frio pela medicina é muito antigo, utilizado inicialmente de forma empírica, teve grandes avanços nos séculos XIV e XX, evoluindo juntamente com a Física. O surgimento de novas técnicas e equipamentos, possibilitou à crioterapia tornar-se uma importante opção no tratamento de neoplasias, benignas e malignas, de diferentes órgãos (tais como fígado, próstata, pâncreas e mamas) e sistemas (ósseo, neurológico e tegumentar). O grande desenvolvimento tecnológico permitiu além do maior número de indicações, melhores resultados, mormente com o uso de técnicas de imagem para o acompanhamento do procedimento. Porém, poucos são os trabalhos que enfocam a monitorização e mensuração do congelamento, pela tomografia computadorizada, bem como avaliação da necrose obtida. No presente trabalho propôs-se avaliar as relações entre as medidas obtidas pela tomografia computadorizada, em 20 cães da raça Beagle, durante o congelamento por aplicação do nitrogênio pela sonda fechada (75 segundos), spray (75 e 135 segundos) e derramamento (75 segundos), comparando estas medidas com aquelas mensuradas pela histopatologia da necrose provocada, para posteriormente avaliar a efetividade dos métodos e criar protocolos de tratamento. Os valores de diâmetro, raio, profundidade e volume do congelamento foram comparados entre si e, com os respectivos valores da necrose após 96 horas do procedimento. Os valores foram submetidos a análise de variância de medidas repetidas e teste de Bonferroni (p<0,05). Constatou-se que: a técnica de sonda fechada oferece congelamento e subseqüente necrose mais superficial que os outros métodos, sendo a técnica menos efetiva; em todas as técnicas o volume, diâmetro e profundidade são inferiores no tecido necrosado quando comparados aos seus similares naquele congelado; em nenhuma das quatro técnicas a profundidade de congelamento foi equivalente ao seu raio. Finalmente, houve a possibilidade de desenvolvimento de protocolos alternativos de congelamento a partir da profundidade de necrose pretendida (hf), obtendo-se como resultado (sonda - Di = hf / 0,29; spray I; Di = hf / 0,31; spray II - Di = hf / 0,35; derramamento -Di = hf / 0,38) o diâmetro do congelamento (Di) necessário para o êxito do tratamento / The use of cold by Medicine is a very old practice, initially used empirically, which had significant advances in the XIV and XX centuries, developing along with Physics. The appearance of new techniques and equipment turned cryotherapy into an important option in the treatment of benign and malignant neoplasms in different organs (such as the liver, the prostate, the pancreas and breasts) and systems (bone, neurological and tegumentary). The large technological development resulted in a higher number of indications as well as better results, especially with the concomitant use of imaging techniques to follow the procedure. However, few studies focus on the monitoring and mensuration of freezing through computed tomography, as well as the assessment of the resulting necrosis. Our study aims at evaluating the association between the measurements obtained through computed tomography in 20 Beagle dogs during freezing by liquid nitrogen use through a probe (75 sec), spray (75 and 135 sec) and overflow (75 sec), comparing these measurements to those obtained through histopathological analysis of the resulting necrosis, for later evaluation of the effectiveness of the methods and design of better treatment protocols. The values of diameter, radius, depth and volume of freezing were compared among them and also with the respective necrosis values 96 hours after the procedure. The values were submitted to analysis of variance of repeated measurements and Bonferroni test (p<0.05). We observed that the closed probe technique causes a more superficial freezing and resulting necrosis when compared to the other methods, being the less effective technique. In all techniques, the volume, diameter and depth are lower in the necrotic tissue when compared to those in the frozen tissue; none of the four techniques showed that freezing depth was equivalent to its radius. Finally, alternative freezing protocols could be developed from the depth of the intended necrosis (hf), resulting in the freezing diameter (Di) (probe - Di = hf/0.29; spray I; Di = hf/0.31; spray II Di = hf/0.35; overflow Di = hf/0.38), which is necessary for therapy success

Orientadores: Ricardo Della Coletta, Ana Lucia Carrinho Ayroza Rangel / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-26T02:30:25Z (GMT). No. of bitstreams: 1 Sawazaki-Calone_Iris_D.pdf: 3631303 bytes, checksum: c364dea5f91c7e7903cb384af31b128a (MD5) Previous issue date: 2014 / Resumo: O carcinoma espinocelular (CEC) representa cerca de 95% de todas as neoplasias malignas que acometem a cavidade oral. Rotineiramente o tratamento e prognóstico desta doença são baseados na localização do tumor e no sistema TNM de classificação dos tumores malignos, no entanto há uma grande variação no comportamento biológico entre tumores no mesmo sítio e dentro do mesmo estadio clínico. Diante deste problema, vários sistemas de gradação histopatológica foram propostos para determinar o prognóstico e o plano do tratamento de pacientes com carcinoma espinocelular (CEC) oral. Este estudo avaliou quatro sistemas de gradação histopatológica ¿ (1) sistema OMS (Organização Mundial de Saúde), (2) sistema MG (gradação de malignidade de margens invasivas profundas), (3) modelo HR (modelo de risco histológico) e (4) escore de risco BD (ninho de células tumorais e profundidade de invasão) ¿ e comparou com dados clínico-patológicos e sobrevida em uma amostra de 113 pacientes com CEC oral primário, excluindo lábios. Os critérios de inclusão foram: pacientes diagnosticados e tratados entre 1998 e 2008, dados clínicos e demográficos completos, tratamento baseado em cirurgia radical com ou sem radioterapia e/ou quimioterapia pós-operatória e disponibilidade de todos os blocos de parafina. Associações significativas com sobrevida na análise univariada foram observadas com todos os sistemas de gradação histopatológica, excetuando o sistema MG. No entanto, aplicando a análise multivariada de COX, apenas o escore de risco BD foi significativamente associado com sobrevida livre de doença como um marcador prognóstico independente. A idade (>56 anos), o tamanho do tumor (estágio T3/T4) e a presença de metástase regional (estágio N+) foram também apontados como marcadores independentes da sobrevida dos pacientes. Nenhuma correlação clara entre os quatro sistemas de gradação foi observada aplicando o teste de correlação de Spearman. Os resultados do presente estudo revelaram uma associação significativa entre o escore de risco BD e a evolução clínica dos pacientes com CEC oral, reforçando a importância deste novo sistema de gradação histopatológica como possível ferramenta prognóstica no pós-operatório / Abstract: Squamous cell carcinoma (SCC) represents almost 95% of all malignant tumors that affect the oral cavity. Routinely the treatment and prognosis of this disease are based on its location and in the TNM classification of malignant tumors, however there is a great variation in the biological behavior among tumors at same location and clinical stage. In the view of those difficulties, several histopathological grading systems were proposed in order to determine the prognostic and the treatment plan of patients with oral squamous cell carcinoma (OSCC). This study evaluated four histopathological grading systems ¿ (1) WHO (World Health Organization) system, (2) MG (malignancy grading of the deep invasive margins) system, (3) HR (histological risk ) model and (4) BD (tumor budding and depth of tumor invasion) risk score ¿ and compared with clinicopathological data and survival in a sample of 113 patients with primary OSCC, excluding lips. The inclusion criteria included who were diagnosed and treated from 1998 to 2008, complete demographic and clinical data, treatment based on radical surgery with or without postoperative radiotherapy and/or chemotherapy, and availability of all paraffin-embedded blocks. Significant associations with survival were observed for all histopathological grading systems, with exception of the MG system. However, when multivariate regression analysis was applied, only BD risk score was significantly associated with disease-free survival as an independent prognostic marker. Age (>56 years), tumor size (T3/T4 stage) and presence of regional metastasis (N+ stage) were also independent markers of reduced survival. No clear correlation between the four grading systems was observed applying the Spearman¿s rank test. The results of the present study revealed a significant association between BD risk score and outcome of OSCC patients, reinforcing the importance of this new histopathological grading system as a possible postoperative prognostic tool / Doutorado / Patologia / Doutora em Estomatopatologia

Orientador: Elemir Macedo de Souza / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-18T22:06:21Z (GMT). No. of bitstreams: 1 Franca_AndreaFernandesEloydaCosta_D.pdf: 3442848 bytes, checksum: 1ed0dba42432e3b03b1b7c6c386bdf41 (MD5) Previous issue date: 2011 / Resumo: O lúpus eritematoso (LE) é uma doença auto-imune com espectro clínico variado. O lúpus eritematoso cutâneo (LEC) inclui o lúpus eritematoso agudo (LECA), o subagudo (LECSA) e o crônico (LECC). Lesões acrômicas podem ocorrer durante a evolução do LE, embora nunca tenham sido estudadas histologicamente. O objetivo deste trabalho é descrever os achados clínicos, laboratoriais e histológicos das lesões acrômicas no LE. Foram selecionados 12 pacientes com LE de um grupo de 220 atendidos no período de 2005 a 2008, sendo sete com LECC e cinco com LECSA. Doze pacientes com vitiligo e 10 controles de pele sã foram usados para comparação. As alterações histológicas encontradas foram: infiltrado inflamatório (75%); hiperceratose e espessamento da zona da membrana basal (ZMB) (66,7%); retificação da epiderme (58,3%); ceratinócitos apoptóticos epidérmicos, elastose e telangectasias (50%); fibrose (41,7%); degeneração vacuolar da ZMB (33,3%); rolhas córneas (16,7%). O diagnóstico histológico de LE foi possível em quatro casos. Melanina pela coloração de Fontana Masson (FM) foi vista em cinco casos e incontinência pigmentar em quatro. Melanócitos foram evidenciados em amostras de cinco doentes através da reação imunohistoquímica pelo HMB45 e Melan-A, com diferença estatística em relação aos controles. Quando comparado ao vitiligo, a diferença foi estatisticamente significante para os achados histológicos: ceratinócitos apoptóticos epidérmicos (p=0,014), espessamento da ZMB (p=0,009) e fibrose (p=0,037). Em relação à quantificação dos melanócitos, não houve diferença estatística entre o grupo LE e vitiligo usando os anticorpos Melan-A e HMB45. Concluímos que as lesões acrômicas no LE correspondem a lesões residuais, decorrentes de processo inflamatório liquenóide prévio que destrói os melanócitos. Não é possível diferenciar as lesões vitiligóides das duas dermatoses pela presença ou ausência de melanócitos, embora a repigmentação seja possível em ambas as doenças devido a presença de melanogênese ativa comprovada pela positividade pelo HMB45 / Abstract: Lupus Erythematosus (LE) is an autoimmune disorder with multiple clinical manifestations. Skin damage is a hallmark of the disease. Cutaneous LE (CLE) includes acute LE (ACLE), subacute LE (SCLE) and chronic LE (CCLE). Although achromic lesions are often found in patients with LEC, there are no detailed data about the histological features of such lesions. Therefore, we designed this study to determine clinical, laboratorial and histological profile of patients with LEC presenting achromic lesions. Between 2005 and 2008, we identified 12 individuals with LEC and acromic lesions from a larger group of 220 patients with LEC that were followed at the Dermatology outpatient clinic. There were seven patients with LECC and five with LECSA. Twelve patients with chronic stable vitiligo and 10 controls of unaffected skin were used for comparison. The most frequent histological abnormalities found in LEC-related achromic lesions were inflammatory infiltrates (75%); hyperkeratosis and thickening of the basement membrane (BM) (66.7%); epidermal flattening (58.3%); apoptotic epidermal keratinocytes, elastosis and vasodilation (50%); fibrosis (41.7%); hydropic degeneration of the basal cells (33.3%); follicular plugging (16.7%). These achromic lesions retained histological features that enabled the diagnosis of CLE to be established in four patients. Fontana Masson (FM) staining was positive for melanin in five cases and revealed pigmentary incontinence in four. Immunohistochemistry for HMB45 and Melan-A identified melanocytes in five CLE-related achromic lesions. Melanocyte counts were significantly smaller in achromic lesions when compared to unaffected skin samples. When compared to vitiligo, CLE-related achromic lesions showed more frequently apoptotic epidermal keratinocytes (p=0,014), thickening of the BM (p=0,009) and fibrosis (p=0,037). Melanocyte counts according to immunohistochemistry were similar in CLE and vitiligo groups. Our results indicate that CLE-related achromic lesions represent residual scars due to chronic lichenoid inflammation that leads to melanocyte destruction. Melanocyte count does not help to distinguish CLE-related achromic lesions and true vitiligo lesions. Despite this, HMB45 staining was sometimes positive in both conditions, which indicates active melanogenesis and suggests that repigmentation may be possible at least for some individuals / Doutorado / Clinica Medica / Doutor em Clínica Médica

M.Sc. / South Africa can be classified as a semi-arid country and the management of its precious water resources is essential. Environmental pollution, caused by the development of industry, technology and informal settlements, threaten the health status of many freshwater ecosystems. The health of all living organisms living in aquatic ecosystems is therefore subsequently affected by this decrease in water quality. The health of an ecosystem is thus often reflected by the health of its fauna. Fish are relatively sensitive to changes in their surrounding environment, including an increase in pollution. Fish health may as a result reflect, and give a good indication of the health status of the aquatic ecosystem in which the fish occurs. The initial toxic effects of the pollution may, however, only be evident on cellular or tissue level before significant changes can be identified in fish behaviour or external appearance. Histological analysis appears to be a very sensitive parameter and is crucial in determining cellular changes that may occur in target organs, including the liver. A histological investigation may therefore prove to be a cost-effective tool to determine the health of fish populations, hence reflecting the health of an entire aquatic ecosystem within a bio-monitoring process. Within the South African context, limited research has been conducted regarding the histology and histopathology of endemic fish species, and its value and effectiveness in aquatic monitoring. The aim of this study was to investigate and validate whether fish liver histology can be used as a monitoring tool, to indicate the health status of an aquatic ecosystem. In order to accomplish this, a preliminary histological study was done on the liver of Clarias gariepinus, a freshwater fish species endemic to the southern African region. Fish were collected in the Rietvlei Dam, Marais Dam and the Palala River, chosen as the three aquatic ecosystems for this study, due to their dissimilar pollution levels. Fish liver samples were fixed in 10% neutrally buffered formalin and prepared for light microscopy analysis using standard techniques for Haematoxylin and Eosin (H & E) staining. From the results obtained during this study, it was concluded that the liver histology of C. gariepinus did appear to be a sensitive monitoring tool of aquatic health. The histological investigation of the fish livers obtained in the Palala River, chosen as an unpolluted site, appeared to be in an overall healthier condition than the liver histology of fish obtained from the two relatively polluted sites, the Rietvlei Dam and the Marais Dam. This conclusion was obtained by means of an objective, quantitative analysis. The histological results in this study were quantified in terms of a histological index. An index value representing the specific histological characteristics of the liver was assigned to each individual specimen indicating either a healthy histological structure (index value of 0-2) or a possible pathological condition (index value of 3-6). According to the quantitative assessment, an average histological index value of 2.5 (indicating a relatively healthy histological condition) was calculated for the unpolluted site while a higher average index value of 3.2 (indicating a pathological condition) were calculated for both the relatively polluted sites. The fish liver histology did therefore reflect the health status of the aquatic ecosystem in which the fish specimen occurred. It was therefore concluded that the liver histology of C. gariepinus seemed to be a sensitive biomarker of environmental pollution, and hence an effective monitoring tool for estimating the health status of an aquatic ecosystem. / Dr. G.M. Pieterse

M.Sc. / The tigerfish, Hydrocynus vittatus member of the Alestidae, are ferocious and fierce predators capable of consuming prey up to 40% of their size. In South Africa H. vittatus can be found in the major east-flowing rivers from the Limpopo River to the Phongolo River flowing into the Pongolapoort Dam making up the southernmost population. The Pongolapoort Dam also known as Lake Jozini is located in the high-rainfall subtropical region of northern KwaZulu-Natal. The Pongolapoort Dam wall was completed in 1973 to make it the fifth largest dam in South Africa with a total surface area of 2 445.9 x 106 m2. It was primarily built for the irrigation of sugar-cane in the surrounding regions. The Fish Health Assessment Index is used as a quantitative index that identifies morphological alterations and assigns them a score for statistical comparison. The main purpose of the macroscopic Fish Health Assessment Index is to detect gross changes in the health of fish populations through necropsy early enough for remedial actions to be put into place. Histopathology is used as an important diagnostic tool for detecting infectious and non-infectious diseases and can therefore be used to assess the health of fish populations by analysing selected target organs. Catch and release (C&R) angling has become a very widely promoted conservation tool as sport angling continues to increase. Anglers release their fish under the assumption that the fish will survive; however, the end results have been shown to vary greatly, from death of the fish to sub-lethal end points such as exhaustive exercise, injury and/or air exposure which will affect the fitness of the fish. The first aim of the study was to determine the health status of H. vittatus and C. gariepinus from the Pongolapoort Dam by means of a histology-based fish health assessment protocol applied to a total of 45 H. vittatus specimens collected in February 2009 (n = 30) and July 2009 (n = 15) and a total of 19 C. gariepinus specimens collected in October 2009 (n = 10) and April 2010 (n = 9). In the H. vittatus specimens DDT was found in the axial muscle; the concentrations of these in February 2009 (5 403.9 ng∙g-1 lipid) and July 2009 (5 537.4 ng∙g-1 lipid). The Mean Index values showed that the Kidney Index (IK), Gill Index (IG) and Fish Index (IFISH) were higher in fish from the February survey while the Liver Index (IL) was higher in those collected during July. Liver alterations identified included intercellular oedema, granular degeneration, vacuolation, nuclear pleomorphism and lymphocyte infiltration. Kidney alterations included dilation of the glomerulus capillaries, vacuolation and hyaline droplet degeneration. Gill alterations identified included telangiectasia and hyperplasia of the secondary lamella, congestion, and rupture of pillar cells.

Rickettsiales-like organisms (RLOs) are thought to be related to bacteria in the order Rickettsiales. They have been reported to occur in the staghorn coral (Acropora cervicornis), and this study investigated trends of infection over time, and in relation to the health of infected corals. This study focuses on tissue samples taken mostly from visibly healthy A. cervicornis thickets in Broward County, Florida, and processed for histological examination. Samples were originally collected and analyzed to document reproduction during years 2001 through 2012, and tissue loss diseases (white-band disease [WBD] types I and II, and rapid tissue loss). The presence of suspect RLOs, the presence of ovoid bacterial aggregates in the basal body wall, and the condition of the coral tissue were examined in hematoxylin and eosin (H&E) and Giemsa-stained sections. Determination was made as to whether suspect RLO infection severity, location, or the presence of bacterial aggregates are correlated with changes in tissue condition associated with WBD. To better understand progression, trends, and periodicity in bacterial presence and coral tissue health, these data were then further analyzed for potential correlation with the month, year, and average monthly nighttime sea surface temperatures (categorized into ranges above, within, or below 24–29°C) when samples were collected. The severity of suspect RLO infections and the presence of bacterial aggregates in A. cervicornis varied over time, with no correlation with the location of suspect RLOs within the polyp. High suspect RLO prevalence was correlated with normal tissue conditions, while low suspect RLO prevalence was correlated with abnormal tissue conditions. However, high prevalence of bacterial aggregates was correlated to abnormal tissue conditions. Epidermal RLO and overall suspect RLO prevalence severity scores were significantly higher among samples collected when monthly average nighttime sea surface temperatures were below 24°C in contrast to samples collected when temperatures were between 24–29°C, suggesting direct or indirect effects of sea surface temperatures on infection severity. The areas of suspect RLO intracellular bodies within infected mucocytes were measured using digital image analysis software and found to be positively correlated with worsening coral tissue condition. Semiquantitative variable scores for histoslides stained with H&E were significantly different from those stained with Giemsa, indicating that these stains cannot be used interchangeably to study the presence of bacteria and the condition of coral tissue. Overall, the results of this study indicate that infection severity of suspect RLOs and the presence of bacterial aggregates are variable and correlated with the incidence of WBD-I in A. cervicornis. However, the exact nature of this relationship remains unclear. Further studies are necessary to interpret trends detected during this analysis to develop a better understanding of what contributes to the severe tissue-loss outbreaks and mortalities of A. cervicornis.