Le métabolisme des microorganismes est traditionnellement étudié à deux échelles: d’une part, à l’échelle locale, la description des réactions métaboliques et d’autre part, à l’échelle globale, l’étude de la physiologie de la cellule. Malgré des progrès technologiques récents facilitant les études à ces deux échelles, leur exploitation conjointe demeure complexe car le comportement physiologique de la cellule résulte de l’action coordonnée de nombreuses réactions. Les modèles mathématiques globaux du métabolisme ont toutefois récemment permis de relier ces deux échelles. Dans cette thèse, nous explorerons l’utilisation de ces modèles pour compléter la connaissance des réactions à l’aide d’une catégorie particulière de données d’échelle globale : les essentialités de gènes déterminées à partir des phénotypes de croissance de mutants de délétion. Nous nous appuierons pour cela sur la bactérie Acinetobacter baylyi ADP1. Après avoir présenté les développements effectués pour reconstruire un modèle global du métabolisme d’A. baylyi, nous montrerons que la confrontation entre phénotypes observés et phénotypes prédits permet de mettre en évidence des incohérences entre les deux échelles d’observations. Nous montrerons ensuite qu’une interprétation formelle de ces incohérences permet de corriger le modèle et d’améliorer la connaissance du métabolisme. Nous illustrerons ce propos en présentant les corrections que nous avons réalisées à l’aide de phénotypes de mutants d’A. baylyi. Enfin, dans une dernière partie, nous proposerons une méthode permettant d’automatiser la correction des incohérences causées par des erreurs d’association entre gènes et réactions. / Microbial metabolism has traditionally been investigated at two different scales: the finest involves characterizing individually each reaction occurring in the cell; the largest focuses on global cell physiology. While both scales have recently benefited from technological advances, combining them remains, however, especially complex as the global physiological behavior of a cell results from the coordinated action of a large network of reactions. Mathematical modeling approaches have yet shown recently that genome-scale metabolic models could help in linking both scales. In this thesis, we explore the use of such models to expand the knowledge of reactions with a specific type of high-level data: gene essentiality data, assessed using growth phenotypes of deletion mutants. We will use as model organism the bacterium Acinetobacter baylyi ADP1, for which a genome-wide collection of gene deletion mutants has recently been created. Following a presentation of the key steps and developments that have been required to reconstruct a global metabolic model of A. baylyi, we will show that confronting observed and predicted phenotypes highlight inconsistencies between the two scales. We will then show that a formal interpretation of these inconsistencies can guide model corrections and improvements to the knowledge of metabolism. We will illustrate this claim by presenting model corrections triggered by A. baylyi mutant phenotypes. Finally, we will introduce a method that automates the correction of inconsistencies caused by wrong associations between genes and reactions.

The phenotypic variability in human populations is partly the result of gene polymorphisms and differential gene expression. Studying the variability of gene expression across human populations is essential to understanding the molecular basis for diversity. However, key issues remain unanswered with respect to human expression variability. For example, the role of gene methylation in expression variability is uncertain, nor is it clear what role tissue-specific factors may have. Moreover, the contribution that expression variability has in aging and development is unknown. Here we classified human genes based on their expression variability in normal human breast and brain samples and identified functional aspects associated with high and low expression variability. Interestingly, both high variability and low variability gene sets are enriched for developmentally essential genes. There is limited overlap between the variably expressed genes of different tissues, indicating that tissue-specific rather than individual-specific factors are at work. We also find that methylation likely has a key role in controlling expression variability insofar as genes with low expression variability are likely to be non-methylated. Importantly, we find that genes with high population expression variability are likely to have age-, but not sex-dependent expression. Taken together, our work indicates that gene expression variability is tissue-specific, methylation-dependent, and is an important component of the natural aging process.

Renewable energy integration into diesel generation systems for remote island communities is a rapidly growing energy engineering field. Fuel supply issues are becoming more common and the disruption, instability and panic caused by fuel shortages results in inefficient and unreliable power supplies for remote island communities. This thesis develops an energy engineering approach for meeting renewable energy development, supply security, cost and sustainability objectives. The approach involves adapting proven energy engineering techniques including energy auditing, energy system modelling with basic cost analysis and demand side management. The novel aspect of this research is the development of critical load engineering in the system design, and informing this with an assessment of essentiality of energy services during the audit phase. This approach was prompted by experiences with previous fuel shortages and long term sustainability policy drivers. The methodology uses the most essential electric loads as the requirement for sizing the renewable energy capacity in the hybrid system. This approach is revolutionary because communication with the customers about availability and the need to shed non-essential loads helps to both meet cost and security requirements and to reduce levels of panic and uncertainty when fuel supply issues arise. A sustainable power generation system is a system that provides continuity of supply for electrical appliances that are considered by the residents to be essential and for which adaptability and resilience of behaviour were key design priorities over growth. The sustainable electrical energy supply should match the critical (essential) load and should have the ability to continue without major disruptions to the daily lives of the people in these communities. Essential energy end uses were identified through energy audits and surveys. The electric power system is designed so that renewable energy sources alone can meet that “essential” demand with a plant that is both economically and technically feasible. Diesel generators were supplemented to meet the short fall in meeting the unconstrained electric demand. This is to design a system that is generally competitive with the present conventional power generation. This method should be particularly suitable for handling the complexities of a modern-day energy system in terms of planning a sizable sustainable energy and electricity system, either based on wholly sustainable sources or integrating sustainable sources of energy into a conventional generation system. The final hybrid system chosen after numerous simulations for the case study (Fenfushi island in the Maldives) community has the minimum renewable energy sources to meet the essential load but uses diesel to supplement the present load. A variety of design parameters such as PV size, wind turbine sizes and numbers and battery capacity have been considered. The minimum renewable energy sources to supply the essential loads of the community were simulated with diesel generators to find the optimal supply mix for the present load (typical unconstrained demand). The final outcome has the following characteristics: NPC and COE were $1,532,340 and $0.37/kWh respectively, lower than any diesel-only systems that could supply the demand. The total annual electricity production is 386,444 units (kWh), of which 9.61% is excess electricity and the annual operating cost is $68,688. Compared to the diesel-only systems there is a fuel savings of 77,021 litres of diesel per year, which is a 66.5 % reduction. An annual carbon dioxide emission reduction of 202,824 kg was achieved, which is a reduction of 66.5%. An annual renewable energy contribution of 70% would be achieved, 34% of which would be from PV arrays and 36% from wind turbines. The selected system shows that even with 30 percent power supply from diesel generators, still the highest NPC is on diesel generation for a life of over 25 years.

Phosphoglucosamine mutase (GlmM) is an enzyme of bacterial cell wall biosynthesis. The main aim of this thesis was to find out, whether gene glmM is essential for viability of Streptococcus pneumoniae. Therefore, we prepared merodiploid strain containing two copies of glmM; the genomic gene and ectopic copy under control of zinc inducible promoter. Subsequently, depletion strain was prepared by deletion of genomic copy of glmM. This strain was further used for analysis of viability and phenotype features in the medium containing various concentrations of zinc ions, an inducer of ectopic glmM expression. We found out, that the viability of this strain was strictly dependent on the concentration of inducer and further, that depletion of GlmM resulted in remarkable morphological defects. The rescue of mutant strain was observed after addition of inducer up to the level of the control sample. These results have provided the evidence of glmM essentiality for S. pneumoniae viability. Furthermore, we analyzed, whether phosphorylation of key amino acid residues, S99 and S101, is essential for GlmM functionality. Four different strains were prepared by means of site-directed mutagenesis expressing glmM with substitutions of key serine residues for alanine or glutamic acid. Since deletion of chromosomal locus in...

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No. of bitstreams: 2 Dissertação - Rafael Carneiro Rocha - 2016.pdf: 1465045 bytes, checksum: 6a80b5f0b8f8aaa16c8024ffa1d3fe68 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-12-05 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The book Endliches und Ewiges Sein (Finite and Eternal Being), by Edith Stein, has in its own subtitle the exposition of its purpose:“Versuch eines Aufstiegs zum Sinn des Seins” (which can be translated as “An Attempt at an Ascent to the meaning of being”). Although the author's starting point for her inquiry about the sense of being is phenomenological, from the fact of the being of the ego - after all, as she herself asserts, her mother tongue is philosophically the language of Husserl - the way of the certainty of the ego (which she calls “augustinian”) occurs in the context of her work together with the way that she calls “aristotelian”, which proceeds from what is intrusive to us [Aufdrängenden] by first, namely, the world of the sensibly perceived things. Despite the apparent antagonism, the author argues that the Augustinian and the Aristotelian ways must complement each other. The philosophical project of Edith Stein, as the author herself emphasizes at the beginning of Potenz und Akt (Potency and Act), should be considered an “ontology” or a science of being, because if we do not understand what is being, we do not understand anything. On the other hand, the transcendental perspective recommends itself as an initial approach because it proposes a consideration of the origin of the meaning of all being, describing the structures of consciousness in which an by which the transcendent world is constructed from immanence. This means that without the transcendental perspective, the ontology remains inexplicable and its constitutive problems are all neglected. However, Edith Stein gives a certain preeminence to the augustinian way, which starts from the life of the ego [Ichleben]. Such preeminence s due to the fact that it is in the realm of the life of the ego that reality itself gains meaning as reality. In this way, the essentialites occupy an eminent position, because just them carry with themselves the possibility of being fully intelligible, thus transmiting intelligibility and meaning to the objects of reality. The book Endliches und Ewiges Sein (Finite and Eternal Being), by Edith Stein, has in its own subtitle the exposition of its purpose:“Versuch eines Aufstiegs zum Sinn des Seins” (which can be translated as “An Attempt at an Ascent to the meaning of being”). Although the author's starting point for her inquiry about the sense of being is phenomenological, from the fact of the being of the ego - after all, as she herself asserts, her mother tongue is philosophically the language of Husserl - the way of the certainty of the ego (which she calls “augustinian”) occurs in the context of her work together with the way that she calls “aristotelian”, which proceeds from what is intrusive to us [Aufdrängenden] by first, namely, the world of the sensibly perceived things. Despite the apparent antagonism, the author argues that the Augustinian and the Aristotelian ways must complement each other. The philosophical project of Edith Stein, as the author herself emphasizes at the beginning of Potenz und Akt (Potency and Act), should be considered an “ontology” or a science of being, because if we do not understand what is being, we do not understand anything. On the other hand, the transcendental perspective recommends itself as an initial approach because it proposes a consideration of the origin of the meaning of all being, describing the structures of consciousness in which an by which the transcendent world is constructed from immanence. This means that without the transcendental perspective, the ontology remains inexplicable and its constitutive problems are all neglected. However, Edith Stein gives a certain preeminence to the augustinian way, which starts from the life of the ego [Ichleben]. Such preeminence s due to the fact that it is in the realm of the life of the ego that reality itself gains meaning as reality. In this way, the essentialites occupy an eminent position, because just them carry with themselves the possibility of being fully intelligible, thus transmiting intelligibility and meaning to the objects of reality. / O livro Endliches und Ewiges Sein (Ser finito e Ser Eterno), de Edith Stein, tem no próprio subtítulo a exposição de seu propósito: “Versuch eines Aufstiegs zum Sinn des Seins” (que pode ser traduzido como “uma tentativa de ascensão ao sentido do ser”). Embora o ponto de partida da autora para a sua investigação sobre o sentido do ser se dê fenomenologicamente, a partir do fato do ser do eu – afinal, conforme ela mesma afirma, filosoficamente a sua língua materna é a linguagem de Husserl e dos pensadores da fenomenologia – a via da certeza do eu (que ela chama de “agostiniana”) se dá, no contexto de sua obra, em conjunto com a via que ela denomina de “aristotélica”, que procede daquilo que nos é intrusivo [Aufdrängenden] por primeiro, a saber, o mundo das coisas sensivelmente percebidas. Apesar do aparente antagonismo, a autora defende que as vias agostiniana e aristotélica devem ser complementares. O projeto filosófico de Edith Stein, como a própria autora enfatiza no início de Potência e ato, deve ser considerado, enquanto procedimento investigativo, como uma “ontologia”, ou uma ciência do ser, porque se não compreendemos o que é ser, não compreendemos nada Por outro lado, a perspectiva transcendental recomenda a si mesma como uma abordagem inicial, porque ela se propõe a uma consideração acerca da origem do sentido de todo ser, descrevendo as “estruturas de consciência em que e pela qual o mundo transcendente é construído a partir da imanência”. Isso quer dizer que sem a perspectiva transcendental, a ontologia permanece inexplicável e seus problemas constitutivos são todos eles negligenciados. Porém, Edith Stein concede uma certa preeminência à via agostiniana, que parte da “vida do eu” [Ichleben]. Tal preeminência se deve ao fato de que é no domínio da vida do eu que a própria realidade ganha sentido enquanto realidade. Dessa maneira, as essencialidades ocupam posição eminente, porque são apenas elas que carregam consigo mesmas a possibilidade de serem plenamente inteligíveis, transmitindo assim inteligibilidade e sentido aos objetos da realidade.

Regulation of gene transcription plays a major role in mediating cellular responses and physiological behavior in all known organisms. The finding that similar genes are often regulated in a similar manner (co-regulated or "co-expressed") has directed several "guilt-by-association" approaches in order to reverse-engineer the cellular transcriptional networks using gene expression data as a compass. This kind of studies has been considerably assisted in the recent years by the development of high-throughput transcript measurement platforms, specifically gene microarrays and next-generation sequencing. In this thesis, I describe several approaches for improving the extraction and interpretation of the information contained in microarray based gene expression data, through four steps: (1) microarray platform design, (2) microarray data normalization, (3) gene network reverse engineering based on expression data and (4) experimental validation of expression-based guilt-by-association inferences. In the first part test case is shown aimed at the generation of a microarray for Thellungiella salsuginea, a salt and drought resistant close relative to the model plant Arabidopsis thaliana; the transcripts of this organism are generated on the combination of publicly available ESTs and newly generated ad-hoc next-generation sequencing data. Since the design of a microarray platform requires the availability of highly reliable and non-redundant transcript models, these issues are addressed consecutively, proposing several different technical solutions. In the second part I describe how inter-array correlation artifacts are generated by the common microarray normalization methods RMA and GCRMA, together with the technical and mathematical characteristics underlying the problem. A solution is proposed in the form of a novel normalization method, called tRMA. The third part of the thesis deals with the field of expression-based gene network reverse engineering. It is shown how different centrality measures in reverse engineered gene networks can be used to distinguish specific classes of genes, in particular essential genes in Arabidopsis thaliana, and how the use of conditional correlation can add a layer of understanding over the information flow processes underlying transcript regulation. Furthermore, several network reverse engineering approaches are compared, with a particular focus on the LASSO, a linear regression derivative rarely applied before in global gene network reconstruction, despite its theoretical advantages in robustness and interpretability over more standard methods. The performance of LASSO is assessed through several in silico analyses dealing with the reliability of the inferred gene networks. In the final part, LASSO and other reverse engineering methods are used to experimentally identify novel genes involved in two independent scenarios: the seed coat mucilage pathway in Arabidopsis thaliana and the hypoxic tuber development in Solanum tuberosum. In both cases an interesting method complementarity is shown, which strongly suggests a general use of hybrid approaches for transcript expression-based inferences. In conclusion, this work has helped to improve our understanding of gene transcription regulation through a better interpretation of high-throughput expression data. Part of the network reverse engineering methods described in this thesis have been included in a tool (CorTo) for gene network reverse engineering and annotated visualization from custom transcription datasets. / Die Regulation der Gentranskription spielt eine wichtige Rolle bei der Steuerung des physiologischen Verhaltens in allen Organismen. Dass ähnliche Gene oft in gleicher Weise reguliert werden (koreguliert oder koexpimiert), hat zu diversen „guilt-by-association“-Ansätzen zur Rekonstruktion von zellulären Transkriptionsnetzwerken geführt, die Genexpressionsdaten zur Orientierung nutzen. Studien dieser Art wurden in den letzten Jahren durch die Entwicklung von Hochdurchsatzmessungen von Transkriptmengen mittels Mikroarrays und ‚Next Generation‘ Sequenziertechniken stark gefördert. In der vorliegenden Arbeit werden verschiedene Ansätze zur Verbesserung der Extraktion und Interpretation von Mikroarray-basierten Genexpressionsdaten in vier Schritten beschrieben: (1) Mikroarray-Sonden-Design, (2) Mikroarray Datennormalisierung, (3) Rekonstruktion von Gennetzwerken unter Verwendung von Expressionsdaten und (4) experimentelle Überprüfung von expressionsbasierten „guilt-by-association“ Schlussfolgerungen. Im ersten Teil wird ein Beispiel zur Erstellung eines Mikroarrays für Thelungiella salsuginea gezeigt, einem salz- und trockenresistenten Verwandten von Arabidopsis thaliana. Zur Rekonstruktion der Transkripte wurden sowohl öffentliche ESTs (‚expressed sequence tags‘) als auch neu erzeugte ‚Next Generation‘ Sequenzierdaten genutzt. Da das Design von Mikroarrays speziesspezifische, nicht-redundante Transkriptmodelle erfordert, werden diese Aufgaben nacheinander abgearbeitet und verschiedene technische Lösungsmöglichkeiten aufgezeigt. Im zweiten Teil wird beschrieben, wie übliche Mikroarray-Normalisierungsverfahren wie RMA und GCRMA zu Korrelationsartefakten führen können. Technische sowie mathematische Hintergründe werden erläutert und zur Lösung des Problems wird mit tRMA eine neue Normalisierungsmethode vorgestellt. Der dritte Teil der Arbeit beschäftigt sich der expressionsbasierten Rekonstruktion von Gennetzwerken. Es wird demonstriert, wie dabei verschiedene „Zentralitäten“ bei zur Unterscheidung von spezifischen Genklassen, hier beispielhaft essentielle Gene von Arabidopsis thaliana, genutzt werden können und wie die Verwendung von konditioneller Korrelation tieferes Verständnis des der Transkriptionsregulation zugrundeliegenden Informationsflusses ermöglicht. Weiterhin werden Ansätze zur Netzwerkrekonstruktion verglichen. Besonderes Augenmerk liegt dabei auf der LASSO Technik, einer Art linearer Regression, die trotz ihren theoretischen Vorteilen in Robustheit und Interpretierbarkeit gegenüber Standardmethoden bisher selten zur Rekonstruktion von globalen Gennetzwerken genutzt wurde. Die Leistungsfähigkeit von LASSO wird durch in silico Analysen der Zuverlässigkeit der erstellten Gennetzwerke gemessen. Im letzten Teil der Arbeit wurden LASSO und andere Rekonstruktionsmethoden genutzt um experimentell neue Gene der folgenden zwei Szenarien zu identifizieren: im Samenschleim von Arabidopsis thaliana und während der Knollenentwicklung von Solanum tuberosum unter Sauerstoffmangel. In beiden Fällen wird eine interessante Methodenkomplementarität gezeigt, nach welcher eine Mischung mehrerer Ansätze zu empfehlen ist um Schlüsse aufgrund von Transkriptexpression zu ziehen. Zusammenfassend zielt diese Arbeit darauf ab, das Verständnis der Regulation von Gentranskriptionsnetzwerken durch bessere Interpretation von Hochdurchsatzexpressionsdaten zu verbessern. Ein Teil der in dieser Arbeit beschriebenen Methoden wurden im Programm CorTo zur Gennetzwerkrekonstruktion und annotierten Visualisierung von benutzerdefinierten Transkriptionsdaten verarbeitet.

Made available in DSpace on 2014-06-11T19:32:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-08-12Bitstream added on 2014-06-13T20:04:44Z : No. of bitstreams: 1 neves_rcf_dr_botfmvz.pdf: 810484 bytes, checksum: 4c35f4860011be79141d0a439a5abd04 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / O objetivo do presente trabalho foi determinar a concentração de cobre em amostras de filé da tilápia do Nilo (Oreochromis niloticus) coletadas no comércio da cidade de Botucatu-SP-Brasil e amostras fornecidas pelo Laboratório de Nutrição Organismos Aquáticos/FMVZ-UNEST/Botucatu-SP/Brasil. Na etapa de preparação, as amostras de filé foram liofilizadas e moídas criogenicamente até obtenção de partículas menores que 60 mm e a extração do cobre foi feita por ultra-som utilizando-se HCl 0,10 mol L-1 como solução extratora. As determinações de cobre foram feitas por espectrometria de Absorção Atômica em Forno de Grafite (GFAAS) utilizando temperaturas de secagem de 100 oC - 250oC, temperatura de pirólise de 1400 oC, temperatura de atomização de 2400 oC e temperatura de limpeza de 2800 oC. Foi utilizado como modificador químico nitrato de paládio injetado junto com as amostras e tungstênio como modificador permanente. Entre as seis amostras de filé de tilápia coletadas foram encontradas concentrações de cobre da ordem de 0,70 a 1,60 mg kg-1, valores estes considerados de acordo com a legislação brasileira, ( concentrações de cobre em amostras de pescados de até 30,00 mg kg-1). A exatidão e precisão das concentrações de cobre determinadas neste estudo foram avaliadas utilizando-se padrão certificado Lake Michigan Fish Tissue – NIST SRM 1947 / The aim of this work was to determine the copper concentrations in filet samples of Nile tilapia (Oreochromis niloticus) collected in the Botucatu City - Brazil. In the samples preparation step, the samples were lyophilised and the steak was cryogenically to produce particles smaller than 60 mm and the copper extractant was carried out by ultrasound using HCl 0.10 mol L-1 as extracting solution. The copper determination were carried out by GFAAS, whit a drying temperature of 100oC - 250oC, pyrolysis temperature of 1400oC, atomization temperature of 2400oC, and cleaning temperature of 2800oC. Palladium nitrate was used as a chemical modifier coinjected with the samples, and tungsten was use permanent modifier. Among the six fillet samples of Nile tilapia collected were found copper concentrations of 0.70 to 1.60 mg kg-1, these values are in agreement with Brazilian law, (copper concentrations in fish samples up to 30.00 mg kg-1). The accuracy and precision of the copper concentrations determined in this study were evaluated using certified standard Lake Michigan Fish Tissue - NIST SRM 1947

No presente trabalho será apresentado um estudo sobre a seletividade em função da essencialidade no Imposto sobre Circulação de Mercadorias e Serviços(ICMS). O ICMS é o tributo responsável pela maior parte da arrecadação dos Estados brasileiros, e a norma da seletividade é fator decisivo na configuração dessa arrecadação. A Constituição de 1988, ao instituir o ICMS, trouxe para este tributo a seletividade em função da essencialidade das mercadorias e dos serviços tributados. A partir da verificação dos textos das Constituições das legislações estaduais sobre o imposto será demonstrada a adoção da seletividade por todos os Estados da Federação. A verificação de como a tributação é aplicada pelos Estados e a comparação das alíquotas de alguns produtos irá expor o desrespeito ao critério constitucionalmente eleito para a seleção, a essencialidade dos produtos e serviços tributados. A pesquisa da jurisprudência dos Tribunais Estaduais, do Superior Tribunal de Justiça e do Supremo Tribunal Federal demonstrará que o tema não vem tendo a atenção que deveria em razão de sua importância. / In the present work it will be presented a study regarding the selectivity based on the essentiality in the State Valu e Added Tax (ICMS). The ICMS tax is responsible for most of the collection of Brazilian states, and the rule of selectivity is a decisive factor in the configuration of this collection. The 1988 Constitution, by introducing the ICMS, brought to this tax th e selectivity based on the essentiality of the goods and services that are taxed. From the analysis of the texts of the Constitutions and state legislations that rule the tax, it will be showed the adoption of selectivity by all states of Brazil. The verification of how the tax is applied by States and the comparison of the rates of some products will expose the disrespect to the criterion that was constitutionally elected for the selection: the essentiality of the goods and services taxed. The research of the jurisprudence of the State Courts, of the Superior Court of Justice and of the Supreme Court will demonstrate that the issue is not receiving the deserved attention that should receive due to its importance.

Orientador: Pedro Magalhães Padilha / Banca: José Pedro de Serra Valente / Banca: Luciana Francisco Fleuri / Banca: Lincoln Carlos Silva de Oliveira / Banca: Gustavo Rocha de Castro / Resumo: O objetivo do presente trabalho foi determinar a concentração de cobre em amostras de filé da tilápia do Nilo (Oreochromis niloticus) coletadas no comércio da cidade de Botucatu-SP-Brasil e amostras fornecidas pelo Laboratório de Nutrição Organismos Aquáticos/FMVZ-UNEST/Botucatu-SP/Brasil. Na etapa de preparação, as amostras de filé foram liofilizadas e moídas criogenicamente até obtenção de partículas menores que 60 mm e a extração do cobre foi feita por ultra-som utilizando-se HCl 0,10 mol L-1 como solução extratora. As determinações de cobre foram feitas por espectrometria de Absorção Atômica em Forno de Grafite (GFAAS) utilizando temperaturas de secagem de 100 oC - 250oC, temperatura de pirólise de 1400 oC, temperatura de atomização de 2400 oC e temperatura de limpeza de 2800 oC. Foi utilizado como modificador químico nitrato de paládio injetado junto com as amostras e tungstênio como modificador permanente. Entre as seis amostras de filé de tilápia coletadas foram encontradas concentrações de cobre da ordem de 0,70 a 1,60 mg kg-1, valores estes considerados de acordo com a legislação brasileira, ( concentrações de cobre em amostras de pescados de até 30,00 mg kg-1). A exatidão e precisão das concentrações de cobre determinadas neste estudo foram avaliadas utilizando-se padrão certificado Lake Michigan Fish Tissue - NIST SRM 1947 / Abstract: The aim of this work was to determine the copper concentrations in filet samples of Nile tilapia (Oreochromis niloticus) collected in the Botucatu City - Brazil. In the samples preparation step, the samples were lyophilised and the steak was cryogenically to produce particles smaller than 60 mm and the copper extractant was carried out by ultrasound using HCl 0.10 mol L-1 as extracting solution. The copper determination were carried out by GFAAS, whit a drying temperature of 100oC - 250oC, pyrolysis temperature of 1400oC, atomization temperature of 2400oC, and cleaning temperature of 2800oC. Palladium nitrate was used as a chemical modifier coinjected with the samples, and tungsten was use permanent modifier. Among the six fillet samples of Nile tilapia collected were found copper concentrations of 0.70 to 1.60 mg kg-1, these values are in agreement with Brazilian law, (copper concentrations in fish samples up to 30.00 mg kg-1). The accuracy and precision of the copper concentrations determined in this study were evaluated using certified standard Lake Michigan Fish Tissue - NIST SRM 1947 / Doutor

No presente trabalho será apresentado um estudo sobre a seletividade em função da essencialidade no Imposto sobre Circulação de Mercadorias e Serviços(ICMS). O ICMS é o tributo responsável pela maior parte da arrecadação dos Estados brasileiros, e a norma da seletividade é fator decisivo na configuração dessa arrecadação. A Constituição de 1988, ao instituir o ICMS, trouxe para este tributo a seletividade em função da essencialidade das mercadorias e dos serviços tributados. A partir da verificação dos textos das Constituições das legislações estaduais sobre o imposto será demonstrada a adoção da seletividade por todos os Estados da Federação. A verificação de como a tributação é aplicada pelos Estados e a comparação das alíquotas de alguns produtos irá expor o desrespeito ao critério constitucionalmente eleito para a seleção, a essencialidade dos produtos e serviços tributados. A pesquisa da jurisprudência dos Tribunais Estaduais, do Superior Tribunal de Justiça e do Supremo Tribunal Federal demonstrará que o tema não vem tendo a atenção que deveria em razão de sua importância. / In the present work it will be presented a study regarding the selectivity based on the essentiality in the State Valu e Added Tax (ICMS). The ICMS tax is responsible for most of the collection of Brazilian states, and the rule of selectivity is a decisive factor in the configuration of this collection. The 1988 Constitution, by introducing the ICMS, brought to this tax th e selectivity based on the essentiality of the goods and services that are taxed. From the analysis of the texts of the Constitutions and state legislations that rule the tax, it will be showed the adoption of selectivity by all states of Brazil. The verification of how the tax is applied by States and the comparison of the rates of some products will expose the disrespect to the criterion that was constitutionally elected for the selection: the essentiality of the goods and services taxed. The research of the jurisprudence of the State Courts, of the Superior Court of Justice and of the Supreme Court will demonstrate that the issue is not receiving the deserved attention that should receive due to its importance.