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

Plásticos reforçados a base de tecidos híbridos: efeitos da anisotropia e geometria normativa na caracterização mecânica e da fratura

As most current studies, reinforced plastics have been, in recent years, a viable alternative in building structural elements of medium and large, since the lightness accompanied by high performance possible. The design of hybrid polymer composites (combination of different types of reinforcements) may enable structural applications thereof, facing the most severe service conditions. Within this class of composite materials, reinforced the underlying tissues hybrid high performance are taking space when your application requires high load bearing and high rigidity. The objective of this research work is to study the challenges in designing these fabrics bring these materials as to its mechanical characterization and fracture mechanisms involved. Some parameters associated with the process and / or form of hybridization stand out as influential factors in the final performance of the material such as the presence of anisotropy, so the fabric weave, the process of making the same, normative geometry of the specimens, among others. This sense, four laminates were developed based hybrid reinforcement fabrics involving AS4 carbon fiber, kevlar and glass 49-E as the matrix epoxy vinyl ester resin (DERAKANE 411-350). All laminates were formed each with four layers of reinforcements. Depending on the hybrid fabric, all the influencing factors mentioned above have been studied for laminates. All laminates were manufactured industrially used being the lamination process manual (hand-lay-up). All mechanical characterization and study of the mechanism of fracture (fracture mechanics) was developed for laminates subjected to uniaxial tensile test, bending in three and uniaxial compression. The analysis of fracture mechanisms were held involving the macroscopic, optical microscopy and scanning electron microscopy

Influência da laparoscopia e da laparotomia na gasometria, leucometria diferencial e citocinas em modelo de sepse abdominal em ratos

Laparoscopic surgery is associated with reduced surgical trauma, and less acute phase response, as compared with open surgery. Cytokines are important regulators of the biological response to surgical and anesthetic stress. The aim of this study was to determine if CO2 pneumoperitoneum would change cytokine expression, gas parameters and leukocyte count in septic rats. Methods: Wistar rats were randomly assigned to five groups: control (anesthesia only), laparotomy, CO2 pneumoperitoneum, cecum ligation and puncture by laparotomy, and laparoscopic cecum ligation and puncture. After 30 min of the procedures, arterial blood samples were obtained to determine leukocytes subpopulations by hemocytometer. TNFα, IL-1β, IL-6 were determined in intraperitoneal fluid (by ELISA). Gas parameters were measured on arterial blood, intraperitoneal and subperitoneal exsudates. Results: Peritoneal TNFα, IL-1β and IL-6 concentrations were lower in pneumoperitoneum rats than in all other groups (p<0.05). TNFα, IL-1β and IL-6 expression was lower in the laparoscopic than in laparotomic sepsis (p<0.05). Rats from laparoscopic cecum ligation and puncture group developed significant hypercarbic acidosis in blood and subperitoneal fluid when compared to open procedure group. Total white blood cells and lymphocytes were significantly lower in laparoscopic cecum ligation and puncture rats than in the laparotomic (p<0.01). Nevertheless, the laparotomic cecum ligation rats had a significant increase in blood neutrophils and eosinophils when compared with controls (p<0.05). Conclusions: This study demonstrates that the CO2 pneumoperitoneum reduced the inflammatory and immune response in an animal model of peritonitis with respect to intraperitoneal cytokines, white blood cell count and clinical correlates of sepsis. The pneumoperitoneum produced hypercarbic acidosis in septic animals

Problemas e modelos que contribuíram com o desenvolvimento do cálculo diferencial e integral: dos gregos à Newton

This article refers to a research which tries to historically (re)construct the conceptual development of the Integral and Differential calculus, taking into account its constructing model feature, since the Greeks to Newton. These models were created by the problems that have been proposed by the history and were being modified by the time the new problems were put and the mathematics known advanced. In this perspective, I also show how a number of nature philosophers and mathematicians got involved by this process. Starting with the speculations over scientific and philosophical natures done by the ancient Greeks, it culminates with Newton s work in the 17th century. Moreover, I present and analyze the problems proposed (open questions), models generated (questions answered) as well as the religious, political, economic and social conditions involved. This work is divided into 6 chapters plus the final considerations. Chapter 1 shows how the research came about, given my motivation and experience. I outline the ways I have gone trough to refine the main question and present the subject of and the objectives of the research, ending the chapter showing the theoretical bases by which the research was carried out, naming such bases as Investigation Theoretical Fields (ITF). Chapter 2 presents each one of the theoretical bases, which was introduced in the chapter 1 s end. In this discuss, I try to connect the ITF to the research. The Chapter 3 discusses the methodological choices done considering the theoretical fields considered. So, the Chapters 4, 5 and 6 present the main corpus of the research, i.e., they reconstruct the calculus history under a perspective of model building (questions answered) from the problems given (open questions), analyzing since the ancient Greeks contribution (Chapter 4), pos- Greek, especially, the Romans contribution, Hindus, Arabian, and the contribution on the Medium Age (Chapter 5). I relate the European reborn and the contribution of the philosophers and scientists until culminate with the Newton s work (Chapter 6). In the final considerations, it finally gives an account on my impressions about the development of the research as well as the results reached here. By the end, I plan out a propose of curse of Differential and Integral Calculus, having by basis the last three chapters of the article

Geometria e simetria nas rendas de bilro: contribuições para matemática escolar

A tese tem como objetivo descrever e analisar características e princípios dos padrões das rendas de bilro de modo a estabelecer relações com a Matemática escolar, principalmente, no que se refere aos tópicos como Geometria, simetria, isometria, área, perímetro, entre outros. Desse modo, elaboramos atividades didáticas, com base na Matemática explorada nos padrões da criação da renda de bilro, visando concretizar um exercício investigatório nas aulas de Matemática, de modo que, sejam estabelecidas relações conceituais entre a prática investigada e os conteúdos da Matemática escolar. Para satisfazer esses objetivos, buscamos apoio metodológico na pesquisa bibliográfica, do tipo documental em catálogos como o da Professora Valdelice Girão (1984) e também o de Dawson (1984). Realizamos também a pesquisa empírica durante as visitas ao Museu do Ceará e ao Centro das Rendeiras na Prainha, em Aquiraz, no Ceará. Para realizar as atividades didáticas, apoiamo-nos em Mendes (2009). Consideramos relevante essa abordagem de ensino porque pressupõe a experiência direta do aprendiz com situações reais vivenciadas, nas quais a abordagem instrucional é centrada no aluno. Desse modo, concluímos que para o ensino de conteúdos como Geometria, simetria, isometria, relação entre perímetro e área, entre outros que são abordados na Educação Básica, os modelos decorrentes da criação renda de bilro e outros modelos já descritos na tradição cearense podem ser usados como artefato cultural na criação de atividades didáticas

Uma proposta de FSS fractal com geometria simplificada

Frequency Selective Surfaces (FSS) are periodic structures in one or two dimensions that act as spatial filters, can be formed by elements of type conductors patches or apertures, functioning as filters band-stop or band-pass respectively. The interest in the study of FSS has grown through the years, because such structures meet specific requirements as low-cost, reduced dimensions and weighs, beyond the possibility to integrate with other microwave circuits. The most varied applications for such structures have been investigated, as for example, radomes, antennas systems for airplanes, electromagnetic filters for reflective antennas, absorbers structures, etc. Several methods have been used for the analysis of FSS, among them, the Wave Method (WCIP). Are various shapes of elements that can be used in FSS, as for example, fractal type, which presents a relative geometric complexity. This work has as main objective to propose a simplification geometric procedure a fractal FSS, from the analysis of influence of details (gaps) of geometry of the same in behavior of the resonance frequency. Complementarily is shown a simple method to adjust the frequency resonance through analysis of a FSS, which uses a square basic cell, in which are inserted two reentrance and dimensions these reentrance are varied, making it possible to adjust the frequency. For this, the structures are analyzed numerically, using WCIP, and later are characterized experimentally comparing the results obtained. For the two cases is evaluated, the influence of electric and magnetic fields, the latter through the electric current density vector. Is realized a bibliographic study about the theme and are presented suggestions for the continuation of this work

Identificação em tempo real de modelo dinâmico de robô móvel com acionamento diferencial e zona morta

Several mobile robots show non-linear behavior, mainly due friction phenomena between the mechanical parts of the robot or between the robot and the ground. Linear models are efficient in some cases, but it is necessary take the robot non-linearity in consideration when precise displacement and positioning are desired. In this work a parametric model identification procedure for a mobile robot with differential drive that considers the dead-zone in the robot actuators is proposed. The method consists in dividing the system into Hammerstein systems and then uses the key-term separation principle to present the input-output relations which shows the parameters from both linear and non-linear blocks. The parameters are then simultaneously estimated through a recursive least squares algorithm. The results shows that is possible to identify the dead-zone thresholds together with the linear parameters

Controle dinâmico de robôs móveis com acionamento diferencial

This work addresses the dynamic control problem of two-wheeled differentially driven non-holonomic mobile robot. Strategies for robot positioning control and robot orientating control are presented. Such strategies just require information about the robot con¯guration (x, y and teta), which can be collected by an absolute positioning system. The strategies development is related to a change on the controlled variables for such systems, from x, y and teta to s (denoting the robot linear displacement) and teta, and makes use of the polar coordinates representation for the robot kinematic model. Thus, it is possible to obtain a linear representation for the mobile robot dynamic model and to develop such strategies. It is also presented that such strategies allow the use of linear controllers to solve the control problem. It is shown that there is flexibility to choice the linear controller (P, PI, PID, Model Matching techniques, others) to be implemented. This work presents an introduction to mobile robotics and their characteristics followed by the control strategies development and controllers design. Finally, simulated and experimental results are presented and commented

Modelagem linear e identificação do modelo dinâmico de um robô móvel com acionamento diferencial

This work presents a modelling and identification method for a wheeled mobile robot, including the actuator dynamics. Instead of the classic modelling approach, where the robot position coordinates (x,y) are utilized as state variables (resulting in a non linear model), the proposed discrete model is based on the travelled distance increment Delta_l. Thus, the resulting model is linear and time invariant and it can be identified through classical methods such as Recursive Least Mean Squares. This approach has a problem: Delta_l can not be directly measured. In this paper, this problem is solved using an estimate of Delta_l based on a second order polynomial approximation. Experimental data were colected and the proposed method was used to identify the model of a real robot

A modelagem matemática como alternativa de ensino e aprendizagem da geometria na educação de jovens e adultos

This is work itself insert in the mathematics education field of the youth and adult education to aim to practitioners of the educational action into the mathematics area performing to with this is teaching kind, adopting to as parameter the Mathematics Molding approach. The motive of the research is to draw up a application proposal of the molding mathematics as teaching and learning geometry alternative in the youth and adult education. The research it develops in three class of the third level (series 5th and 6th) of he youth and adults education in the one school municipal from the Natal outskirts. Its have qualitative nature with participating observation approach, once performing to directly in to research environment as a mathematics teacher of those same classes. We are used questionnaires, lesson notes and analyses of the officials documents as an basis of claim instruments. The results indicates that activity used the mathematic moldings were appreciated the savoir-faire of the student in to knowledge construction process, when search develop to significant learning methods, helping to student build has mathematics connections with other knowledge areas and inside mathematics himself, so much that enlarges your understanding and assist has in your participation in the other socials place, over there propitiate to change in student and teacher posture with relation to mathematic classroom dynamics

Geometria e arirmética numa visão multicultural: uma experiência pedagógica

This paper aims to build a notebook of activities that can help the teacher of elementary school mathematics. Topics covered are arithmetic and geometry and the activities proposed here were developed aiming print them a multicultural character. We take as a base line developed by Claudia Zaslavsky multiculturalism and reflected in his books "Games and activities worldwide" and "More games and activities worldwide." We structure our work around four themes: the symbol of the Olympic Games, the pyramids of Egypt, the Russian abacus abacus and Chinese. The first two themes allow you to explore basic concepts of geometry while the latter two themes allow us to explore numerical notation and arithmetic operations

Geometrias não-eucludianas como anomalias: implicações para o ensino de geometria e medidas

This present research the aim to show to the reader the Geometry non-Euclidean while anomaly indicating the pedagogical implications and then propose a sequence of activities, divided into three blocks which show the relationship of Euclidean geometry with non-Euclidean, taking the Euclidean with respect to analysis of the anomaly in non-Euclidean. PPGECNM is tied to the line of research of History, Philosophy and Sociology of Science in the Teaching of Natural Sciences and Mathematics. Treat so on Euclid of Alexandria, his most famous work The Elements and moreover, emphasize the Fifth Postulate of Euclid, particularly the difficulties (which lasted several centuries) that mathematicians have to understand him. Until the eighteenth century, three mathematicians: Lobachevsky (1793 - 1856), Bolyai (1775 - 1856) and Gauss (1777-1855) was convinced that this axiom was correct and that there was another geometry (anomalous) as consistent as the Euclid, but that did not adapt into their parameters. It is attributed to the emergence of these three non-Euclidean geometry. For the course methodology we started with some bibliographical definitions about anomalies, after we ve featured so that our definition are better understood by the readers and then only deal geometries non-Euclidean (Hyperbolic Geometry, Spherical Geometry and Taxicab Geometry) confronting them with the Euclidean to analyze the anomalies existing in non-Euclidean geometries and observe its importance to the teaching. After this characterization follows the empirical part of the proposal which consisted the application of three blocks of activities in search of pedagogical implications of anomaly. The first on parallel lines, the second on study of triangles and the third on the shortest distance between two points. These blocks offer a work with basic elements of geometry from a historical and investigative study of geometries non-Euclidean while anomaly so the concept is understood along with it s properties without necessarily be linked to the image of the geometric elements and thus expanding or adapting to other references. For example, the block applied on the second day of activities that provides extend the result of the sum of the internal angles of any triangle, to realize that is not always 180° (only when Euclid is a reference that this conclusion can be drawn)

Esperando Ulisses: o mito de Penélope à luz da comparação diferencial e discursiva

From the many weavers known, the Queen of Ithaca is certainly among the most famous.Over the years,many writers have dedicated themselves to retell the myth of Penelope in their works by their own way. According to Ute Heidmann, the modern writers recurrence to the Greek myths in order to produce their texts is a renewing discursive practice, which gives new writing and relevance to the myth. (2003, p.47). This work deals with a differential and discursive comparative analysis on the myth of Penelope linking it with two short stories from Brazilian authors: Penélope by João do Rio (1919) and Penélope by Dalton Trevisan (1959). In order to do it, we are supported by: the works of Heidmann (2003, 2006, 2008) and Maingueneau (2006). We also concentrate ourselves on the temporal trace presented in both Penelope s myth and in its modern rewriting so that we can identify how each configuration of the classical myth develops into one of the most celebrated acts of this myth: the waiting. In order to so, we seek support on the studies by Paul Ricoeur (2006), Hans Meyerhoff (1976) and Benedito Nunes (1988)