A dissertation submitted to the Faculty of Health Sciences, University of Witwatersrand, in fulfilment of the requirements for the degree of Master of Science (Medicine) in the branch of Anatomical Pathology Johannesburg, 2012 / Objective To determine the effectiveness of the cell block technique for immunocytochemical diagnosis by comparing cytomorphologic preservation and immunocytochemistry (ICC) stains in paired cell block and conventional fine needle aspiration (FNA) samples. Study Design This was a prospective study. Material for both conventional smears and cell blocks were collected simultaneously during fine needle aspiration of 50 lesions comprising lymph node, lung and liver masses. Grading of cellularity, morphological preservation, architectural preservation, immunocytochemical staining intensity and presence of background staining were compared on paired FNA smears and cell block samples derived from the same case. Each arm of the paired analysis was performed blindly without knowledge of the grading outcome of the other. The Kappa statistic (Κ) was used to measure inter-rater agreement. Results The fifty samples evaluated included FNAs from the lung, 24/50 (48%); liver, 23/50 (46%) and lymph node, 3/50 (6%). The immunocytochemistry stains consisted of 44/50 (88%) CK7, 44/50 (88%) CK20, 18/50 (36%) TTF1, 10/50 (20%) synaptophysin, 10/50 (20%) Hepar-1 and 7/50 (14%) AE1/3. There was no overall agreement in preservation of cytomorphological detail and ICC staining between the two methods. The Papanicolaou stained conventional FNA smears fared better then cell block for the vi evaluation of nuclear and cytomorphologic characteristics; cells in the cell block were poorly preserved in many cases. The ICC stains worked better on the cell block samples due to lack of background and aberrant staining. Conclusion Conventional FNA smears and cell blocks complement each other. Our results indicate that it would be optimal to use both modalities in the diagnostic work-up of mass lesions amenable to FNA diagnosis; the former to assess morphology, and the latter for optimal immunocytochemistry results. In resource constrained settings, the cost implications of performing both conventional and blocked smears on all FNA material warrants further evaluation.

Submitted by William Justo Figueiro (williamjf) on 2015-07-06T23:27:47Z No. of bitstreams: 1 09a.pdf: 771261 bytes, checksum: c8d36f36fd0e9b6396c5bad8e1ea3681 (MD5) / Made available in DSpace on 2015-07-06T23:27:47Z (GMT). No. of bitstreams: 1 09a.pdf: 771261 bytes, checksum: c8d36f36fd0e9b6396c5bad8e1ea3681 (MD5) Previous issue date: 2014 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / UNISINOS - Universidade do Vale do Rio dos Sinos / Microrganismos vivendo no interior dos tecidos das plantas sem causar efeitos prejudiciais visíveis de sua presença são chamados endofíticos. As bactérias endofíticas podem promover o crescimento das plantas de várias maneiras: através da secreção de reguladores de crescimento vegetal, pela solubilização de fosfato, entre outras. Na busca de práticas agrícolas ambientalmente sustentáveis, tem sido dada considerável atenção à fixação biológica de nitrogênio atmosférico. Assim, o objetivo do presente estudo foi avaliar, através de métodos dependentes de cultivo, a ocorrência de bactérias endofíticas em colmos e folhas de duas amostras das variedades “RG 121” e “RG 900” de arroz irrigado, num sistema de cultivo convencional. Foram coletadas plantas de arroz de duas variedades na Estação Experimental do Arroz, do Instituto Rio Grandense do Arroz (EEA-IRGA), Cachoeirinha/RS, no ano agrícola 2012/2013. As plantas coletadas em campo foram submetidas à desinfecção superficial, seguida do isolamento e cultivo das bactérias endofíticas em meios de cultura semi-sólidos, livres de nitrogênio: NFb (Azospirillum brasilense/A. lipoferum), JNFb (Herbaspirillum seropedicae/H. rubrisubalbicans) e LGI-P (Gluconacetobacter diazotrophicus). Ao final desses procedimentos, foram obtidos 190 isolados bacterianos, os quais agruparam-se, de acordo com suas características fenotípicas, em 29 morfotipos. A partir disso, o padrão de distribuição dos morfotipos entre oito condições foi avaliado por Análise de Correspondência Destendenciada (DCA). As oito condições foram: C1 = amostra (planta) 1, variedade “RG 121”, colmos; C2 = amostra 1, variedade “RG 121”, folhas; C3 = amostra 1, variedade “RG 900”, colmos; C4 = amostra 1, variedade “RG 900”, folhas; C5 = amostra (planta) 2, variedade “RG 121”, colmos; C6 = amostra 2, variedade “RG 121”, folhas; C7 = amostra 2, variedade “RG 900”, colmos; C8 = amostra 2, variedade “RG 900”, folhas. Os dois primeiros eixos da DCA explicaram 28,5 % da variância total do conjunto de dados - 23,9 % de explicação somente pelo eixo 1. Ainda, foi realizada uma Análise de Componentes Principais (PCA) na tentativa de estabelecer quais morfotipos melhor explicam as diferenças observadas entre as condições. Deste modo, os eixos 1 e 2 da PCA tiveram uma proporção de explicação acumulada correspondente a 45,61 %, sendo o eixo 1 responsável por 25,91 % destes. Por fim, em função do número de isolados obtidos por grupo fenotípico, pôde-se calcular o índice de diversidade de Shannon-Weaver (H’), que relaciona a riqueza e uniformidade de espécies (morfotipos) para cada área (condição) em estudo. Estes foram testados par-a-par pelos métodos de bootstrapping e permutação quanto a possíveis diferenças significativas. De modo que valores de H’ estatisticamente diferentes apresentados por algumas das combinações de condições puderam ser relacionados à amostra (planta), à estrutura vegetal (colmos e folhas) e à variedade, isolada ou conjuntamente. O crescimento bacteriano em meios de cultura livres de nitrogênio, em condições microaerofílicas, infere a ocorrência da capacidade de fixação biológica de nitrogênio. / Microorganisms living within the tissues of plants without causing visible detrimental effects of their presence are known as endophytes. The endophytic bacteria can promote plant growth in various ways: by secreting plant growth regulators, by phosphate solubilization, among others. In pursuit of environmentally sustainable agricultural practices, considerable attention has been given to the biological nitrogen fixation. Thus, the aim of this study was to evaluate, by culture-dependent methods, the occurrence of endophytic bacteria in stems and leaves of two samples of varieties “RG 121” e “RG 900” of irrigated rice, on a conventional cropping system. Rice plants from two varieties were collected in the Estação Experimental do Arroz, from Instituto Riograndense do Arroz (EEA-IRGA), Cachoeirinha/RS, in the agricultural year 2012/2013. Plants collected in the field were submitted to surface disinfection, followed by isolation and cultivation of endophytic bacteria in semisolid, nitrogen-free, culture media: NFb (Azospirillum brasilense/A. lipoferum), JNFb (Herbaspirillum seropedicae/H. rubrisubalbicans) and LGI-P (Gluconacetobacter diazotrophicus). At the end of these procedures, 190 bacterial isolates were obtained, which were grouped, according to their phenotypic characteristics, in 29 morphotypes. From this, the pattern of distribution of morphotypes in eight conditions was assessed by Detrended Correspondence Analysis (DCA). The eight conditions were: C1 = sample (plant) 1, variety "RG 121", stems; C2 = sample 1, variety "RG 121", leaves; C3 = sample 1, variety "RG 900", stems; C4 = sample 1, variety "RG 900", leaves; C5 = sample (plant) 2, variety "RG 121", stems; C6 = sample 2, variety "RG 121", leaves; C7 = sample 2, variety "RG 900", stems; C8 = sample 2, variety "RG 900", leaves. The first two axes of DCA explained 28.5 % of the total variance of the data set - 23.9 % of explanation only by axis 1. In addition, a Principal Components Analysis (PCA) was performed in attempt to establish which morphotypes best explain the observed differences between the conditions. Thus, the axes 1 and 2 of the PCA had a proportion of accumulated explanation corresponding to 45.61 %, being the axis 1 responsible for 25.91 % of them. Finally, according to the number of isolates obtained by phenotypic group, the Shannon-Weaver diversity index (H’) were calculated, which relates the richness and evenness of species (morphotypes) for each area (condition) in study. These were tested pair-to-pair by bootstrapping and permutation methods for possible significant differences. Thus, statistically different values of H' presented by some of the combinations of conditions could be related to the sample (plant), plant structure (stems and leaves) and variety, individually or together. The bacterial growth in nitrogen-free culture media, under microaerophilic conditions, implies the occurrence of biological nitrogen fixation capacity.

Made available in DSpace on 2014-07-29T15:07:28Z (GMT). No. of bitstreams: 1 NIVIA faria.pdf: 1723183 bytes, checksum: 8c7fa557f1d9e528a75b069ae5bd4fbf (MD5) Previous issue date: 2009-12-18 / The transmissible venereal tumor (TVT) is among the most common cancers in dogs in Brazil. This is a sexually transmitted cancer through a mechanism of transplantation of viable tumor cells, it has high incidence among dogs of both sexes in reproductive age, with high occurrence in areas of high concentration of dogs with poor control of reproduction. The diagnosis made in the routine of clinical medicine is usually based only on clinical symptoms and the chemotherapy of choice has been the vincristine sulfate. This drug has been shown in most cases a good response to treatment with complete regression of the tumor, but there are also cases where the tumor appears resistant to treatment. This study aimed to demonstrate the practicality of using fine needle aspiration cytology as a method of diagnosis and cytomorphologic classification, being effective and safe as well as measuring the response of different cytomorphologic groups forward to treatment with vincristine sulfate. Also evaluating the clinical response, hematologic and biochemical tests using serum liver function (ALT) and renal (creatinine), in the therapy instituted. Were evaluated and treated 18 dogs of both sexes, carrying TVT, which received six consecutive weeks during the administration of vincristine sulfate in the dosage of 0.03 mg / kg administered intravenously. The animals underwent weekly prior to medication, clinical assessment, haematological and biochemical. It was observed in this study that cytological examination is a simple technique, minimally invasive and painless, and a safe method that provides no risk to the patient's life and does not require the use of sophisticated equipment, sedation or anesthesia for their achievement. Animals carrying the TVT plasmacytoid type, had higher rate of tumor resistance to treatment in relation to established groups and linfocytic mixed, showing a greater resistance to treatment with this group of vincristine sulfate. The efficiency of response to treatment was presented by the linfocytic group. The vincristine sulfate showed few side effects, low hematological, hepatic and renal toxicity, it is therefore an effective option in the control of transmissible venereal tumor in not resistant cases. / O tumor venéreo transmissível (TVT) é uma das neoplasias mais freqüentes em cães no Brasil. Trata-se de uma neoplasia sexualmente transmissível, através de um mecanismo de transplantação de células tumorais viáveis, apresenta alta incidência entre cães de ambos os sexos em idade reprodutiva, tendo grande ocorrência em ambientes de alta concentração de cães errantes e com deficiente controle de reprodução. O diagnóstico realizado na rotina da clínica médica geralmente se baseia apenas na sintomatologia clínica e o quimioterápico de eleição tem sido o sulfato de vincristina. Este quimioterápico tem demonstrado na maioria dos casos boa resposta ao tratamento com regressão completa da neoplasia, mas também há casos onde o tumor se mostra resistente ao tratamento. O presente trabalho visou demonstrar a praticidade do uso da citologia aspirativa por agulha fina como método de diagnóstico e de classificação citomorfológica eficaz e seguro, assim como mensurar a resposta dos diferentes grupos citomorfológicos frente ao tratamento com sulfato de vincristina. Avaliando também a resposta clínica, hematológica e por meio de provas bioquímicas séricas a função hepática (ALT) e renal (creatinina), frente à terapia instituída. Foram avaliados e tratados 18 cães de ambos os sexos, portadores de TVT, os quais receberam durante seis semanas consecutivas a administração de sulfato de vincristina na dosagem de 0,03 mg/kg administrado via intravenosa. Os animais foram submetidos semanalmente previamente a medicação, avaliação clínica, hematológica e bioquímica. Observou-se com este estudo que o exame citológico constitui-se em uma técnica simples, minimamente invasiva e indolor, sendo um método seguro, que não proporciona riscos à vida do paciente e não requer o uso de equipamento sofisticado, sedação ou anestesia para sua realização. Os animais portadores do TVT do tipo plasmocitóide, apresentaram maior índice de resistência tumoral ao tratamento estabelecido em relação aos grupos linfocitóide e misto, demonstrando uma maior resistência deste grupo ao tratamento com sulfato de vincristina. A maior eficácia de resposta ao tratamento foi apresentada pelo grupo linfocitóide. O sulfato de vincristina apresentou poucos efeitos colaterais, baixa toxicidade hematológica, hepática e renal constituindo, portanto em opção eficaz no controle do tumor venéreo transmissível em casos não resistentes.