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

O ciclo indiano de Marguerite Duras à luz da comparação diferencial: a reconfiguração de um personagem

La présente recherche a pour objectif analyser la reconfiguration d'un personnage dans deux romans et dans un texte dramatique de l’écrivaine française Marguerite Duras, qui font partie du Cicle indien. Ils sont : Le Ravissement de Lol V. Stein (1964), Le vice-consul (1965) e India Song (1975). Adoptant la perspective de la Comparaison Différentielle comme approche littéraire, proposée par Ute Heidmann (2008, 2010, 2012), professeur-docteur de l'Université de Lausanne (Suisse), ce travail cherche étudier le personnage Anne-Marie Stretter présent dans ces trois œuvres. On cherche comprendre de quelle manière se produit la réécriture du personnage à partir de trois axes d'analyse : les modalités de l'énonciation, en considérant l'analyse et la comparaison de l'œuvre littéraire comme un événement énonciatif, partant du concept de la scène d'énonciation, proposé par Dominique Maingueneau (2010) ; les modalités d'inscription générique, étroitement liées aux modalités énonciatives, avec un fondement théorique en Todorov (1980) et Heidmann (2012) ; et les modalités de dialogisme intertextuel et interdiscursif, considérant dans la relation dialogique des nouveaux et différentes propositions de sens. Para la compréhension de cela nous partons de Bakhtine (2010), Kristeva (1974), Todorov (1981) et de l'idée de dialogisme et intertextualité, pour arriver à l'idée d'interdiscours proposée par Heidmann (2010, 2012). À partir de cette étude, nous pouvons observer dans l'œuvre d'un même auteur différentes manières de construction d'un personnage, chacun avec ses spécificités et complexités, variant dans le genre et espaces discursifs. L'intérêt de la comparaison différentielle dans cette étude, partant d'un trait commun entre les trois œuvres, est de rendre visible les différences épistémologiques et les nouvelles propositions de sens.

Semantikos: proposta de interfaces para aplicativo mediador de técnica de diferencial semântico em mobile tablet

This research aims to systematize a proposal of developing a mobile tablet application in order to help implementing the Semantic Differential technique – SD, under the approach of Participatory Design. In 1975, Osgood et al. created the Semantic Differential technique. Since then, many experiments use it to measure the affective perception of individuals concerning objects and concepts by means of compounded scales of bipolar adjectives, based on the theoretical models that support the technique: the conductible, spatial and metric models. During the application of the technique with potential users, the researcher must simultaneously manage several contexts, that is, audio recorder, when authorized, and observe and record spontaneous reports of the respondent. It is noticeable that often occurs a cognitive overload during this event. Thus, the use of a single application whose interface is assigned to its users and respondents could assist researchers in applying the SD technique. This research aimed to understand the processes inherent to the task of implementing the Semantic Differential technique and obeyed the following steps: a) training of users, b) background questionnaire c) interview with Focus Group, and d) cooperative evaluation. Besides these procedures, one can also observe the degrees of facilitation or difficulty concerning the use of the conventional model, which is the development and application of scales with the aid of printed material, pencil, pens, clipboards, and recorder software for editing the document and data analysis. This paper comprises reactions and impressions from the experiences of users of SD technique. Considering the data recollected from the user’s observation, the hypothesis of the experiment proved to be right. It means that the development of the application for mobile tablet employing the technique of Semantic Differential is viable, since it assembles all the steps in one only tool, increases the accomplishment of the task between user/researcher and user/respondent resulting in their mutual satisfaction.

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

Transcriptoma diferencial entre células-tronco mesenquimais humanas jovens e senescentes

Human mesenchymal stem cells (MSC) are powerful sources for cell therapy in regenerative medicine. The long time cultivation can result in replicative senescence or can be related to the emergence of chromosomal alterations responsible for the acquisition of tumorigenesis features in vitro. In this study, for the first time, the expression profile of MSC with a paracentric chromosomal inversion (MSC/inv) was compared to normal karyotype (MSC/n) in early and late passages. Furthermore, we compared the transcriptome of each MSC in early passages with late passages. MSC used in this study were obtained from the umbilical vein of three donors, two MSC/n and one MSC/inv. After their cryopreservation, they have been expanded in vitro until reached senescence. Total RNA was extracted using the RNeasy mini kit (Qiagen) and marked with the GeneChip ® 3 IVT Express Kit (Affymetrix Inc.). Subsequently, the fragmented aRNA was hybridized on the microarranjo Affymetrix Human Genome U133 Plus 2.0 arrays (Affymetrix Inc.). The statistical analysis of differential gene expression was performed between groups MSC by the Partek Genomic Suite software, version 6.4 (Partek Inc.). Was considered statistically significant differences in expression to p-value Bonferroni correction ˂.01. Only signals with fold change ˃ 3.0 were included in the list of differentially expressed. Differences in gene expression data obtained from microarrays were confirmed by Real Time RT-PCR. For the interpretation of biological expression data were used: IPA (Ingenuity Systems) for analysis enrichment functions, the STRING 9.0 for construction of network interactions; Cytoscape 2.8 to the network visualization and analysis bottlenecks with the aid of the GraphPad Prism 5.0 software. BiNGO Cytoscape pluggin was used to access overrepresentation of Gene Ontology categories in Biological Networks. The comparison between senescent and young at each group of MSC has shown that there is a difference in the expression parttern, being higher in the senescent MSC/inv group. The results also showed difference in expression profiles between the MSC/inv versus MSC/n, being greater when they are senescent. New networks were identified for genes related to the response of two of MSC over cultivation time. Were also identified genes that can coordinate functional categories over represented at networks, such as CXCL12, SFRP1, xvi EGF, SPP1, MMP1 e THBS1. The biological interpretation of these data suggests that the population of MSC/inv has different constitutional characteristics, related to their potential for differentiation, proliferation and response to stimuli, responsible for a distinct process of replicative senescence in MSC/inv compared to MSC/n. The genes identified in this study are candidates for biomarkers of cellular senescence in MSC, but their functional relevance in this process should be evaluated in additional in vitro and/or in vivo assays