943 resultados para Plasmid incompatibility


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The course of in vivo infection of five isolates of Yersinia pseudotuberculosis was followed for three weeks in Swiss mice. The strains were isolated from diarrheic and normal feces and mesenteric lymph nodes of healthy and sick stock animals. Four strains of serogroup O:3 and one of serogroup O:1a, with and without the virulence plasmid, were inoculated intragastrically and intravenously in the mice. Groups of five animals were sacrificed at 6 h and 3, 6, 10, 15, and 21 days after inoculation, and organs and tissues were checked for possible macroscopic alterations. Development of infection was monitored at these times by performing viable bacterial counts in homogenates of selected tissues. The animals were cheked daily for clinical alterations. The results of the study showed that strains with the virulence plasmid infected organs and tissues at various times and at varying intensity by both routes of infection, the strain of type O:1a being the most invasive. Moreover, clinical and pathological alterations occurred only in animals inoculated with bacteria carrying the virulence plasmid, regardless of the route of infection.

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Ribotyping and virulence markers has been used to investigate 68 Yersinia pseudotuberculosis strains of serogroups O:1a and O:3. The strains were isolated from clinical material obtained from healthy and sick animals in the Southern region of Brazil. Ribotypes were identified by double digestion of extracted DNA with the restriction endonucleases SmaI and PstI, separation by electrophoresis and hybridization with a digoxigenin-labeled cDNA probe. The presence of the chromosomal virulence marker genes inv, irp1, irp2, psn, ybtE, ybtP-ybtQ, and ybtX-ybtS, of the IS100 insertion sequence, and of the plasmid gene lcrF was detected by polymerase chain reaction. The strains were grouped into four distinct ribotypes, all of them comprising several strains. Ribotypes 1 and 4 presented distinct profiles, with 57.3% genetic similarity, ribotypes 2 and 3 presented 52.5% genetic similarity, and genetic similarity was 45% between these two groups (1/4 and 2/3). All strains possessed the inv, irp1, and irp2 genes. Additionally, strains of serogroup O:1a carried psn, ybtE, ybtP-ybtQ, ybtX-ybtS, and IS100. As expected lcrF was only detected in strains harboring the virulence plasmid. These data demonstrate the presence of Y. pseudotuberculosis strains harboring genotypic virulence markers in the livestock from Southern Brazil and that the dissemination of these bacteria may occur between herds.

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Broadly speaking, the concept of gene therapy involves the transfer of a genetic material into a cell, tissue, or organ in order to cure a disease or at least improve the clinical status of a patient. Making it simple, gene therapy consists in the insertion of functional genes into cells containing defective genes by substituting, complementing or inhibiting them. The achievement of a foreigner DNA expression into a population of cells requires its transfer to the target. Therefore, it is a key issue to create systems able to transfer and protect the DNA until it reaches the target, the vectors. The disadvantages related to the use of viral vectors have encouraged efforts to develop emulsions as non-viral vectors. In fact, they are easily produced, present controllable stability and enable transfection. The aim of this work was to develop an emulsion for gene therapy and evaluate its ability to compact nucleic acids by the development of a complex with the plasmid pIRES2-EGFP. The first step was to determine the Hydrophilic Lipophilic Balance (HLB) of the Captex® 355 (oily internal phase of the emulsion) through long and short term stability assays. Based on the results, emulsions composed of Captex® 355, Tween 20® and Span 60® with 10.7 HLB were produced by three different methods: phase inversion, spontaneous emulsification and sonication. The results showed that the lowest diameter and best stability of the emulsions were achieved by the sonication method. The cationic emulsions were made by adding DOTAP to the basic emulsion. Its association with pIRES2-EGFP was evaluated by electrophoresis. Several rates of emulsion and DNA were evaluated and the results showed that 100% of the complex was formed when the rate DOTAP/DNA(nmol/µg) was 130. In conclusion, the overall results show the ability of the proposed emulsion to compact pIRES2-EGFP, which is a requirement to a successful transfection. Therefore, such formulation may be considered a promising candidate for gene therapy

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Phenomenology is the focus of this study for its critique of the limits of positivist science, which guides most of the fields of study including Psychology. The clinical formation process in Psychology courses is especially difficult for students-interns who adopt phenomenology as their clinical framework. Such difficulty is due to the incompatibility between theory provided in Psychology courses a science traditionally based on paradigms of scientism , and the theoretical-methodological proposal adopted by the aforementioned approach. As a backdrop for our study, we carefully examined the thought of philosopher Martin Heidegger, especially the Era of Technique. This contemporary technicism society was studied so that we could understand the socio-cultural status where this formation lies. Thus, we questioned if this panorama upon which Clinical Psychology rests favors the development of a phenomenological attitude and a special look at the meanings of existence, as defined in phenomenological clinical practice. Knowing such limits, our research aimed at understanding the experience of formation of clinical psychologists who take part in internships in the field of phenomenology-existentialism. Such study was, then, a phenomenological-hermeneutic research based on Heideggerian ontology and used a semi-structured interview as access tool. Six students of the UFRN higher-degree Psychology course who were doing their supervised internship in clinical psychology and the referred approach took part in this research. The research revealed that the phenomenological-existential formation phase opens a door to discoveries on the part of the intern that transcend the dimension of the other, for they show a self disclosure while a person in the word. Despite the initial discomforts caused by the course curriculum itself and by the freedom for clinical practice, so characteristic of phenomenology, the narratives demonstrate that such difficulties may start a process of search for new meanings, which show a search for sharpening their practices and for a path in balance with the existence of the other

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The aim of this paper is to evaluate the consistency indexes among 30 Brazilian university libraries from the south and south-east regions through a specific mathematical formula. It was selected a sample of 30 university libraries that, according to the information in their official sites, have a collection consisted of more than 100.000 copies and allow the search into the on-line catalog. Searches were carried out in every university by means of their sites, requesting books that contained a certain word in its title and were printed in a certain year. The response was a list of available titles in the library, from which we chose at random a title and asked to visualize the complete record to verify the existence of a given subject. This procedure was repeated until we found the same title in five libraries with the chosen subjects. The result is 10 trials, each one consisting of one figure and one table showing the selected libraries, the subjects, the documentary languages ( tools) and the consistency indexes relaxed and rigid. These trials show great discrepancy between the values of consistency indexes with intervals between 73,3% to 34,4% in the relaxed index, and between 60% and 9,6% in the rigid one. It was revealed that the coincidence in determining the subjects is not too high remaining below 39%. It is concluded that the difference between the consistency indexes may be due to factors as: incompatibility among documentary languages; lack of updating of these languages so as to follow the knowledge evolution; absence of a well-defined indexing policy with guidelines clearly established. Procedures of indexing followed by indexers could contribute to the consistency index to be bigger in percentage, since there would be parameters for the indexing process.

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This assignment ains to prove the pertinency of using the wittgenstein´s argument against private language as a criticism to cartesian fundacionism. Therefore, I want to demonstrate in the first chaper the conceptual viability of facing the cartesian argument of cogito not as a simple silogism but as an exemple of a private experience (process of thinking). At the second chaper, the subordination of the argument against private language give us the idea that rules can only be followed by means of corrections givem by a linguistic community that is external to the private subject, in a way to be unviable the assumption that is possible to name an internal experience without searching external rules of the use of terms. At the chaper 3 the pertinency of the hypothesis raised by A. Kenny, about the overtaking of the argument against private language can be extended to the idea of epistemic and ontologic privacy that would lend validity to the fundacion present at the argument at the cartesian cogito. In oder to become evident the pertinency of use of Wittgenstein´s argument agaist Descartes´ fundation, it´s necessary, at the chaper 3, to demonstrate the impertinency of the objection to the A. Kenny´s hypothesis, based on the experiency of the thought of the brain at the recipient, to make clear the incompatibility existing between the cartesian idea of cogito and Wittgenstein´s notion that language is an activitie followed by rules, wich correction criterion may be external and intersubjective

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

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

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Deletion of reading frame YHR116W of the Saccharomyces cerevisiae nuclear genome elicits a respiratory deficiency. The encoded product, here named Cox23p, is shown to be required for the expression of cytochrome oxidase. Cox23p is homologous to Cox17p, a water-soluble copper protein previously implicated in the maturation of the Cu-A center of cytochrome oxidase. The respiratory defect of a cox23 null mutant is rescued by high concentrations of copper in the medium but only when the mutant harbors COX17 on a high copy plasmid. Overexpression of Cox17p by itself is not a sufficient condition to rescue the mutant phenotype. Cox23p, like Cox17p, is detected in the intermembrane space of mitochondria and in the postmitochondrial supernatant fraction, the latter consisting predominantly of cytosolic proteins. Because Cox23p and Cox17p are not part of a complex, the requirement of both for cytochrome oxidase assembly suggests that they function in a common pathway with Cox17p acting downstream of Cox23p.

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

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

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Repetitive DNAs have been extensively applied as physical chromosome markers on comparative studies, identification of chromosome rearrangements and sex chromosomes, chromosome evolution analysis, and applied genetics. Here we report the characterization of repetitive DNA sequences from the Nile tilapia (Oreochromis niloticus) genome by construction and screening of plasmid library enriched with repetitive DNAs, analysis of a BAC-based physical map, and hybridization to chromosomes. The physical mapping of BACs enriched with repetitive sequences and C(o)t-1 DNA (DNA enriched for highly and moderately repetitive DNA sequences) to chromosomes using FISH showed a predominant distribution of repetitive elements in the centromeric and telomeric regions and along the entire length of the largest chromosome pair (X and Y sex chromosomes) of the species. The distribution of repetitive DNAs differed significantly between the p arm of X and Y chromosomes. These findings suggest that repetitive DNAs have had an important role in the differentiation of sex chromosomes. (c) 2007 Elsevier Ltd. All rights reserved.

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Meio mineral líquido contendo benzeno (B) ou tolueno (T) ou xileno (X) a 100 mg L-1 e suas misturas de BT, BX e TX (50 + 50 mg L-1 cada mistura) e BTX (33,3 + 33,3 + 33,3 mg L-1 cada mistura) foram utilizados para avaliar a atividade de degradação de B, T e X por Pseudomonas putida CCMI 852 contendo um plasmídeo TOL. Após 18 a 24 horas de homogenização da mistura, o inoculo foi adicionado e o decréscimo da concentração dos solventes foi determinado entre 24 e 25 horas por GC. Pseudomonas putida CCMI 852 foi capaz de metabolizar T e X, mas não B. Na mistura BTX, B não foi metabolizado também e a velocidade de degradação de T e X decresceu cerca de 50% comparado com soluções contendo apenas T ou X.

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Merostachys riedeliana Rupr. é uma espécie monocárpica com floração cíclica e muito freqüente em sub-bosques de fragmentos florestais do sul do estado de Minas Gerais, Brasil. Sua biologia floral e seu sistema de reprodução foram estudados e comparados com os de outros bambus. Devido ao complexo sistema de rizomas, formam touceiras vigorosas no interior da floresta, ocorrendo a interrupção na produção de novos colmos meses antes do aparecimento das primeiras inflorescências. O início da floração maciça e da morte da população ocorreu em outubro de 1998 e maio de 1999, respectivamente, com pico de floração durante a estação quente e chuvosa (dezembro e janeiro). As inflorescências espiciformes possuem, em média, 29 espiguetas. Estas são hermafroditas com três anteras poricidas e dois estigmas plumosos que se expõem durante a antese. O pólen é abundante e facilmente liberado das anteras pelo vento ou pelos visitantes. Apis mellifera L. e Trigona spinipes (F.) foram os visitantes mais freqüentes, atuando como pilhadores de pólen e, ocasionalmente, através de movimentos vibratórios, como elementos auxiliares para a dispersão do pólen. A alta pluviosidade durante a floração e a escassez de vento no sub-bosque da floresta, podem diminuir a efetividade da anemofilia. No entanto, vários caracteres morfológicos das espiguetas, queda de folhas e o hábito espacialmente agrupado, apontam para uma polinização pelo vento. Testes de polinização controlada, mostraram que M. riedeliana é autocompatível (ISI 0,99). A auto-incompatibilidade não favorece a formação de frutos em clones vegetais, ao passo que a autocompatibilidade poderia resultar em uma elevada produção de sementes. Assim, a possível ocorrência de clones de M. riedeliana nos fragmentos florestais, originados pelo crescimento vegetativo durante os intervalos reprodutivos de 30-32 anos, poderiam explicar o alto investimento na produção de espiguetas e a formação de frutos provenientes da autocompatibilidade.