Preparo de lâminas fixadas e coradas: técnicas de coloração de Gram

Apresenta a descrição de técnicas de coloração para preparação de amostra de material biológico para ser observada ao microscópio óptico, com destaque para a coloração diferencial de Gram. Todas as etapas do procedimento são reproduzidas detalhadamente: desde os cuidados para a fixação dos microrganismos nas lâminas, a preparação do esfregaço a partir de cultura crescida em meio líquido e em meio sólido até a técnica de coloração de Gram propriamente dita. Após a descrição dos procedimentos apresentam-se exemplos de morfologia e coloração e a descrição detalhada do que ocorre com a célula durante cada etapa do processo de coloração.

Detecção de Curtobacterium flaccumfaciens pv. flaccumfaciens em sementes de feijoeiro produzidas em Santa Catarina

Foi avaliada a presença de Curtobacterium flaccumfaciens pv. flaccumfaciens em 37 amostras de sementes de feijoeiro, dos grupos preto e carioca, produzidas em municípios do Estado de Santa Catarina, nas safras 2004/2005 e 2005/2006. Para a detecção, as sementes foram maceradas e alíquotas de sua suspensão foram transferidas para o meio de cultura semi-seletivo MSCFF. A identidade dos isolados obtidos foi comprovada por meio da observação da morfologia celular, coloração diferencial de Gram, tolerância a NaCl a 7% e patogenicidade em cultivares de feijoeiro suscetíveis. Detectou-se a presença de Curtobacterium flaccumfaciens pv. flaccumfaciens em 23 amostras (62,2%), indicando a importância das sementes como fonte de inoculo inicial.

Transmissão planta - semente de Curtobacterium flaccumfaciens pv. flacumfaciens em cultivares de feijoeiro

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Características físico-químicas de amidos modificados com permanganato de potássio/ácido lático e hipoclorito de sódio/ácido lático

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Marcadores citológicos no cariótipo de espécies de Leptodactylus (Amphibia, Anura, Leptodactylidae) analisado com técnicas de citogenética clássica e molecular

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Identificação e caracterização de Xanthomonas euvesicatoria de pimentão no Brasil

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Estímulo da síntese da glutationa na maturação in vitro de oócitos bovinos e sua influência no desenvolvimento embrionário

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aplicabilidade taxonômica de variações anatômicas em fungos liquenizados

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Nephila clavipes e Nephila sexpunctata (Araneae, Araneomorphae, Nephilidae): cariótipo, heterocromatina constitutiva e região organizadora de nucléolo

As espécies de Nephila estão incluídas na família Nephilidae que pertence ao grupo das aranhas Entelegynae, o qual é considerado filogeneticamente derivado em relação aos outros grupos da ordem Araneae. Análises citogenéticas realizadas nas aranhas Entelegynae com técnicas de coloração convencional, têm mostrado que a maioria das espécies possui uniformidade cariotípica principalmente em relação a morfologia telo-acrocêntrica dos cromossomos e sistema cromossômico sexual (SCS) do tipo X1X20/X1X1X2X2. Além disso, estas análises têm demonstrado que algumas famílias de Entelegynae também possuem uniformidade cariotípica concernente ao número diplóide de cromossomos na maioria das suas espécies. Por outro lado, o emprego adicional de técnicas de coloração diferencial de regiões cromossômicas específicas em alguns representantes de Entelegynae têm revelado características que podem ser usadas para determinar os mecanismos envolvidos na evolução cromossômica e na diferenciação cariotípica de espécies relacionadas. Contudo, poucas espécies de Entelegynae tiveram seus cariótipos analisados com técnicas de coloração diferencial. O Brasil possui aproximadamente 4.000 espécies de Araneae descritas taxonomicamente; entretanto apenas cerca de 20 destas foram analisadas do ponto de vista citogenético. Considerando estas informações, o objetivo deste trabalho foi investigar o cariótipo de duas espécies de Nephila, Nephila clavipes e Nephila sexpunctata, com técnicas de coloração convencional e diferencial para determinar o número diplóide, a morfologia cromossômica, o tipo de SCS, e o padrão de distribuição da heterocromatina constitutiva (bandas C) e das regiões organizadoras de nucléolo (RONs) ativas, e comparar os dados obtidos com aqueles de espécies relacionadas, para estabelecer os mecanismos de evolução cromossômica...(Resumo completo, clicar acesso eletrônico abaixo)

Caracterização cromossômica por coloração convencional e diferencial de três populações de grilos do gênero Zucchiella Mello, 1990 (Trigonidiidae, nemobiinae)

Pós-graduação em Ciências Biológicas (Zoologia) - IBRC

Análise proteômica de Chromobacterium violaceum: acúmulo estacionário e diferencial após exposição à luz UVC

Chromobacterium violaceum is a free-living bacillus, Gram-negative commonly found in water and sand of tropical and subtropical regions. One of its main characteristic it's the ability to produce the purple pigment named violacein, that shows countless biological activities. In 2003, the genome of this organism was totally sequenced and revealed important informations about the physiology of this bacteria. However, few post-genomics studies had been accomplished. This work evaluated the protein profile of C. violaceum cultivated in LB medium at 28ºC that allowed the identification and characterization of proteins related to a possible secretion system that wasn't identified and characterized yet in C. violaceum, to the quorum sensing system, to regulatory process of transcription and translation, stress adaptation and biotechnological potential. Moreover, the response of the bacteria to UVC radiation was evaluated. The comparison of the protein profile, analyzed through 2-D electrophoresis, of the control group versus the treatment group allowed the identification of 52 proteins that arose after stress induction. The obtained results enable the elaboration of a stress response pathway in C. violaceum generated by the UVC light. This pathway, that seems to be a general stress response, involves the expression of proteins related to cellular division, purine and pirimidine metabolism, heat chock or chaperones, energy supply, regulation of biofilm formation, transport, regulation of lytic cycle of bacteriophages, besides proteins that show undefined function. Despite the response present similarities with the classic SOS response of E. coli, we still cannot assert that C. violaceum shows a SOS-like response, mainly due to the absence of characterization of a LexA-like protein in this organism

Proteômica diferencial da Chromobacterium violaceum em resposta ao estresse oxidativo induzido por peróxido de hidrogênio

The β-proteobacterium Chromobacterium violaceum is a Gram-negative, free-living, saprophytic and opportunistic pathogen that inhabits tropical and subtropical ecosystems among them, in soil and water of the Amazon. It has great biotechnological potential, and because of this potential, its genome was completely sequenced in 2003. Genome analysis showed that this bacterium has several genes with functions related to the ability to survive under different kinds of environmental stresses. In order to understand the physiological response of C. violaceum under oxidative stress, we applied the tool of shotgun proteomics. Thus, colonies of C. violaceum ATCC 12472 were grown in the presence and absence of 8 mM H2O2 for two hours, total proteins were extracted from bacteria, subjected to SDS-PAGE, stained and hydrolysed. The tryptic peptides generated were subjected to a linear-liquid chromatography (LC) followed by mass spectrometer (LTQ-XL-Orbitrap) to obtain quantitative and qualitative data. A shotgun proteomics allows to compare directly in complex samples, differential expression of proteins and found that in C. Violaceum, 131 proteins are expressed exclusively in the control condition, 177 proteins began to be expressed under oxidative stress and 1175 proteins have expression in both conditions. The results showed that, under the condition of oxidative stress, this bacterium changes its metabolism by increasing the expression of proteins capable of combating oxidative stress and decreasing the expression of proteins related processes bacterial growth and catabolism (transcription, translation, carbon metabolism and fatty acids). A tool with of proteomics as an approach of integrative biology provided an overview of the metabolic pathways involved in the response of C. violaceum to oxidative stress, as well as significantly amplified understanding physiological response to environmental stress. Biochemical and "in silico" assays with the hypothetical ORF CV_0868 found that this is part of an operon. Phylogenetic analysis of superoxide dismutase, protein belonging to the operon also showed that the gene is duplicated in genome of C. violaceum and the second copy was acquired through a horizontal transfer event. Possibly, not only the SOD gene but also all genes comprising this operon were obtained in the same manner. It was concluded that C. violaceum has complex, efficient and versatile mechanisms in oxidative stress response

Indução diferencial das vias MyD88 e TRIF-dependentes em leucócitos totais de equinos estimulados com LPS de E. coli

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Técnica para segmentação automática de imagens microscópicas de componentes sanguíneos e classificação diferencial de leucócitos baseada em lógica fuzzy

Automatic detection of blood components is an important topic in the field of hematology. The segmentation is an important stage because it allows components to be grouped into common areas and processed separately and leukocyte differential classification enables them to be analyzed separately. With the auto-segmentation and differential classification, this work is contributing to the analysis process of blood components by providing tools that reduce the manual labor and increasing its accuracy and efficiency. Using techniques of digital image processing associated with a generic and automatic fuzzy approach, this work proposes two Fuzzy Inference Systems, defined as I and II, for autosegmentation of blood components and leukocyte differential classification, respectively, in microscopic images smears. Using the Fuzzy Inference System I, the proposed technique performs the segmentation of the image in four regions: the leukocyte’s nucleus and cytoplasm, erythrocyte and plasma area and using the Fuzzy Inference System II and the segmented leukocyte (nucleus and cytoplasm) classify them differentially in five types: basophils, eosinophils, lymphocytes, monocytes and neutrophils. Were used for testing 530 images containing microscopic samples of blood smears with different methods. The images were processed and its accuracy indices and Gold Standards were calculated and compared with the manual results and other results found at literature for the same problems. Regarding segmentation, a technique developed showed percentages of accuracy of 97.31% for leukocytes, 95.39% to erythrocytes and 95.06% for blood plasma. As for the differential classification, the percentage varied between 92.98% and 98.39% for the different leukocyte types. In addition to promoting auto-segmentation and differential classification, the proposed technique also contributes to the definition of new descriptors and the construction of an image database using various processes hematological staining

Análise proteômica de Chromobacterium violaceum: acúmulo estacionário e diferencial após exposição à luz UVC

Chromobacterium violaceum is a free-living bacillus, Gram-negative commonly found in water and sand of tropical and subtropical regions. One of its main characteristic it's the ability to produce the purple pigment named violacein, that shows countless biological activities. In 2003, the genome of this organism was totally sequenced and revealed important informations about the physiology of this bacteria. However, few post-genomics studies had been accomplished. This work evaluated the protein profile of C. violaceum cultivated in LB medium at 28ºC that allowed the identification and characterization of proteins related to a possible secretion system that wasn't identified and characterized yet in C. violaceum, to the quorum sensing system, to regulatory process of transcription and translation, stress adaptation and biotechnological potential. Moreover, the response of the bacteria to UVC radiation was evaluated. The comparison of the protein profile, analyzed through 2-D electrophoresis, of the control group versus the treatment group allowed the identification of 52 proteins that arose after stress induction. The obtained results enable the elaboration of a stress response pathway in C. violaceum generated by the UVC light. This pathway, that seems to be a general stress response, involves the expression of proteins related to cellular division, purine and pirimidine metabolism, heat chock or chaperones, energy supply, regulation of biofilm formation, transport, regulation of lytic cycle of bacteriophages, besides proteins that show undefined function. Despite the response present similarities with the classic SOS response of E. coli, we still cannot assert that C. violaceum shows a SOS-like response, mainly due to the absence of characterization of a LexA-like protein in this organism