La construction d'un quotient, en topologie, est relativement simple; si $G$ est un groupe topologique agissant sur un espace topologique $X$, on peut considérer l'application naturelle de $X$ dans $X/G$, l'espace d'orbites muni de la topologie quotient. En géométrie algébrique, malheureusement, il n'est généralement pas possible de munir l'espace d'orbites d'une structure de variété. Dans le cas de l'action d'un groupe linéairement réductif $G$ sur une variété projective $X$, la théorie géométrique des invariants nous permet toutefois de construire un morphisme de variété d'un ouvert $U$ de $X$ vers une variété projective $X//U$, se rapprochant autant que possible d'une application quotient, au sens topologique du terme. Considérons par exemple $X\subseteq P^{n}$, une $k$-variété projective sur laquelle agit un groupe linéairement réductif $G$ et supposons que cette action soit induite par une action linéaire de $G$ sur $A^{n+1}$. Soit $\widehat{X}\subseteq A^{n+1}$, le cône affine au dessus de $\X$. Par un théorème de la théorie classique des invariants, il existe alors des invariants homogènes $f_{1},...,f_{r}\in C[\widehat{X}]^{G}$ tels que $$C[\widehat{X}]^{G}= C[f_{1},...,f_{r}].$$ On appellera le nilcone, que l'on notera $N$, la sous-variété de $\X$ définie par le locus des invariants $f_{1},...,f_{r}$. Soit $Proj(C[\widehat{X}]^{G})$, le spectre projectif de l'anneau des invariants. L'application rationnelle $$\pi:X\dashrightarrow Proj(C[f_{1},...,f_{r}])$$ induite par l'inclusion de $C[\widehat{X}]^{G}$ dans $C[\widehat{X}]$ est alors surjective, constante sur les orbites et sépare les orbites autant qu'il est possible de le faire; plus précisément, chaque fibre contient exactement une orbite fermée. Pour obtenir une application régulière satisfaisant les mêmes propriétés, il est nécessaire de jeter les points du nilcone. On obtient alors l'application quotient $$\pi:X\backslash N\rightarrow Proj(C[f_{1},...,f_{r}]).$$ Le critère de Hilbert-Mumford, dû à Hilbert et repris par Mumford près d'un demi-siècle plus tard, permet de décrire $N$ sans connaître les $f_{1},...,f_{r}$. Ce critère est d'autant plus utile que les générateurs de l'anneau des invariants ne sont connus que dans certains cas particuliers. Malgré les applications concrètes de ce théorème en géométrie algébrique classique, les démonstrations que l'on en trouve dans la littérature sont généralement données dans le cadre peu accessible des schémas. L'objectif de ce mémoire sera, entre autres, de donner une démonstration de ce critère en utilisant autant que possible les outils de la géométrie algébrique classique et de l'algèbre commutative. La version que nous démontrerons est un peu plus générale que la version originale de Hilbert \cite{hilbert} et se retrouve, par exemple, dans \cite{kempf}. Notre preuve est valide sur $C$ mais pourrait être généralisée à un corps $k$ de caractéristique nulle, pas nécessairement algébriquement clos. Dans la seconde partie de ce mémoire, nous étudierons la relation entre la construction précédente et celle obtenue en incluant les covariants en plus des invariants. Nous démontrerons dans ce cas un critère analogue au critère de Hilbert-Mumford (Théorème 6.3.2). C'est un théorème de Brion pour lequel nous donnerons une version un peu plus générale. Cette version, de même qu'une preuve simplifiée d'un théorème de Grosshans (Théorème 6.1.7), sont les éléments de ce mémoire que l'on ne retrouve pas dans la littérature. / The topological notion of a quotient is fairly simple. Given a topological group $G$ acting on a topological space $X$, one gets the natural application from $X$ to the quotient space $X/G$. In algebraic geometry, unfortunately, it is generally not possible to give the orbit space the structure of an algebraic variety. In the special case of a linearly reductive group acting on a projective variety $X$, the geometric invariant theory allows us to get a morphism of variety from an open $U$ of $X$ to a projective variety $X//G$, which is as close as possible to a quotient map, from a topological point of view. As an example, let $ X\subseteq P^{n}$ be a $k$-projective variety on which acts a linearly reductive group $G$. Suppose further that this action is induced by a linear action of $G$ on $A^{n+1}$ and let $\widehat{X}\subseteq A^{n +1}$ be the affine cone over $X$. By an important theorem of the classical invariants theory, there exist homogeneous invariants $f_{1},..., f_{r}\in C[\widehat{X}]^{G}$ such as $$\C[\widehat{X}]^{G}=\C[f_{1},...,f_{r}].$$ The locus in $X$ of $f_{1},...,f_{r}$ is called the nullcone, noted $N$. Let $Proj(C[\widehat{X}]^{G})$ be the projective spectrum of the invariants ring. The rational map $$\pi:X\dashrightarrow Proj(C[f_{1},...,f_{r}])$$ induced by the inclusion of $C[\widehat{X}]^{G}$ in $C[\widehat{X}] $ is then surjective, constant on the orbits and separates orbits as much as possible, that is, the fibres contains exactly one closed orbit. A regular map is obtained by removing the nullcone; we then get a regular map $$\pi:X \backslash N\rightarrow Proj(C[f_{1},...,f_{r}])$$ which still satisfy the preceding properties. The Hilbert-Mumford criterion, due to Hilbert and revisited by Mumford nearly half-century later, can be used to describe $N$ without knowing the generators of the invariants ring. Since those are rarely known, this criterion had proved to be quite useful. Despite the important applications of this criterion in classical algebraic geometry, the demonstrations found in the literature are usually given trough the difficult theory of schemes. The aim of this master thesis is therefore, among others, to provide a demonstration of this criterion using classical algebraic geometry and of commutative algebra. The version that we demonstrate is somewhat wider than the original version of Hilbert \cite{hilbert}; a schematic proof of this general version is given in \cite{kempf}. Finally, the proof given here is valid for $C$ but could be generalised to a field $k$ of characteristic zero, not necessarily algebraically closed. In the second part of this thesis, we study the relationship between the preceding constructions and those obtained by including covariants in addition to the invariants. We give a Hilbert-Mumford criterion for covariants (Theorem 6.3.2) which is a theorem from Brion for which we prove a slightly more general version. This theorem, together with a simplified proof of a theorem of Grosshans (Theorem 6.1.7), are the elements of this thesis that can't be found in the literature.

The purpose of this qualitative explorative study is to explain individualized lived experiences of registered nurses working and participating in clinical supervision for nursing students within the clinical environment at selected hospitals. The study explores the challenges faced by registered nurses on a daily basis on how to structure clinical activities for the nursing students and provide high quality care to patients.

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Dans ce travail, nous exploitons des propriétés déjà connues pour les systèmes de poids des représentations afin de les définir pour les orbites des groupes de Weyl des algèbres de Lie simples, traitées individuellement, et nous étendons certaines de ces propriétés aux orbites des groupes de Coxeter non cristallographiques. D'abord, nous considérons les points d'une orbite d'un groupe de Coxeter fini G comme les sommets d'un polytope (G-polytope) centré à l'origine d'un espace euclidien réel à n dimensions. Nous introduisons les produits et les puissances symétrisées de G-polytopes et nous en décrivons la décomposition en des sommes de G-polytopes. Plusieurs invariants des G-polytopes sont présentés. Ensuite, les orbites des groupes de Weyl des algèbres de Lie simples de tous types sont réduites en l'union d'orbites des groupes de Weyl des sous-algèbres réductives maximales de l'algèbre. Nous listons les matrices qui transforment les points des orbites de l'algèbre en des points des orbites des sous-algèbres pour tous les cas n<=8 ainsi que pour plusieurs séries infinies des paires d'algèbre-sous-algèbre. De nombreux exemples de règles de branchement sont présentés. Finalement, nous fournissons une nouvelle description, uniforme et complète, des centralisateurs des sous-groupes réguliers maximaux des groupes de Lie simples de tous types et de tous rangs. Nous présentons des formules explicites pour l'action de tels centralisateurs sur les représentations irréductibles des algèbres de Lie simples et montrons qu'elles peuvent être utilisées dans le calcul des règles de branchement impliquant ces sous-algèbres. / In this work, we exploit properties well known for weight systems of representations to define them for individual orbits of the Weyl groups of simple Lie algebras, and we extend some of these properties to orbits of non-crystallographic Coxeter groups. Points of an orbit of a finite Coxeter group G are considered as vertices of a polytope (G-polytope) centered at the origin of a real n-dimensional Euclidean space. Products and symmetrized powers of G-polytopes are introduced and their decomposition into the sums of G-polytopes is described. Several invariants of G-polytopes are found. The orbits of Weyl groups of simple Lie algebras of all types are reduced to the union of orbits of the Weyl groups of maximal reductive subalgebras of the algebra. Matrices transforming points of the orbits of the algebra into points of subalgebra orbits are listed for all cases n<=8 and for many infinite series of algebra-subalgebra pairs. Numerous examples of branching rules are shown. Finally, we present a new, uniform and comprehensive description of centralizers of the maximal regular subgroups in compact simple Lie groups of all types and ranks. Explicit formulas for the action of such centralizers on irreducible representations of the simple Lie algebras are given and shown to have application to computation of the branching rules with respect to these subalgebras.

L'acétylation des protéines constitue l’une des plus importantes modifications post-traductionnelles (PTMs). Elle intervient dans de multiples processus bologiques et physiopathologiques tels que, l’activité transcriptionnelle, l'apoptose, la régulation des voies métaboliques, les cancers, les maladies inflammatoires et cardiovasculaires. Face à l’importance de l’acétylation des protéines, il apparaît donc indispensable de bien comprendre les mécanismes qui y sont associés, et donc, de pouvoir identifier et quantifier les protéines acétylées à partir du protéome complet d’échantillons complexes tels que des extraits cellulaires ou tissulaires. La spectrométrie de masse est une technique de choix pour de telles études, car elle permet d’identifier les protéines et les sites d’acétylation, mais aussi de les quantifier en l’associant à des techniques de quantification (label free, SILAC, iTRAQ/TMT, AQUA). Malheureusement, ces méthodes ne ciblent pas particulièrement les acétylations et requièrent l’utilisation de techniques d’enrichissement ou de fractionnement qui ne sont dédiées qu’à certains types d’acetylation : les N-ter et K-acetylation. Aucun enrichissement n’est disponible pour les O- acétylations et ces méthodes d’enrichissement ne sont pas toujours compatibles avec les techniques de quantification citées ci-dessus. Pour améliorer la détection et la quantification des acétylations, nous proposons la méthode RAQIAT (Relatif Absolute Quantification Isobaric Affinity Tag) qui se résume en trois grandes étapes: i) Le blocage des fonctions amines libres à l'aide de la di-méthylation réductrice, ceci empêchera ces dernières de réagir avec le réactif RAQIAT, ii) La désacétylation des lysines acétylées pour permettre une quantification sélective des acétylations, iii) Le marquage des amines primaires précédemment désacétylées dans l’étape 2 par le réactif RAQIAT pour permettre leurs identifications et quantifications. Ce manuscrit a porté en partie sur les deux premières étapes de la méthode RAQIAT.Dans la première étape, les échantillons de protéines de levure ont été digérés puis di-méthylés et fractionnés par OFFGEL en 24 fractions. Ensuite, chacune de ces 24 fractions OFFGEL a été soumise à un fractionnement nano-RPLC et analysée par MALDI TOF/TOF (4800 MALDI-TOF/TOF, Sciex). En parallèle, la même expérience a été réalisée, cette fois-ci sans di-méthylation. L'analyse des données a été réalisée en utilisant le logiciel Mascot comme moteur de recherche.L’efficacité de la réaction de di-méthylation démontrée, nous avons montré que sans réaliser la di-méthylation réductrice 164 sites acétylés ont pu été identifiés alors que 385 sites acétylés distincts ont été identifiés avec la di-méthylation réductrice. De plus, l'amélioration de la détection de l'acétylation en utilisant la méthode de di-méthylation a été observée pour chacune des différents types acétylations: N-ter, K- et O-acétylation.Dans la deuxième étape, nous avons présenté des résultats préliminaires de déacétylation par la sirtuine 1 en présence du peptide de la p53 (Ac-Arg-His-Lys-Lys-(Ac)-AMC) connu comme étant un substrat de cette enzyme. Nous avons observé la formation d’un peptide non acétylé, suggérant une déacétylation de ce peptide acétylé de p53. Cependant, la formation de cet ion étant très faible et l’ion acétylé étant fortement préservé, nous en avons conclu que l’efficacité de la déacétylation du peptide de p53 n’était pas suffisante pour l’intégrer à la méthode RAQIAT. / Protein acetylation is one of the most widespread post-translational modifications which is involved in many cellular physiologies and pathologies such as cancers. Regarding the important biological effect of protein acetylation and a non-negligible number of proteins bearing this PTM, several methods emerged last decade to investigate such PTM. But the detection of acetylations and their quantification are still limited and enrichment method allowing a better detection of acetylation target mostly one kind of acetylation (K-acetylation). To improve the detection of the three kind of acetylation (N-ter, K, and O-) and their quantification, we propose the RAQIAT method (Relative Absolute Quantification Isobaric Affinity Tag), based on protein digestion followed by 3 steps : i) a protection of free primary amines at N-ter, lysine (i.e. primary amine not bearing PTM) based on a reductive di-methylation strategy ii) a deacetylation of acetylated residues to obtain free primary amine corresponding to peptides previously acetylated iii) a RAQIAT labeling on the free primary amine obtained in the previous step to allow the enrichment of peptides previously acetylated and their quantifications. Herein, we present the investigation of the two first steps of RAQIAT method.In the first step, we evidenced that the reductive di-methylation strategy improved the detection of the three kind of acetylation: N-ter, K- and O- acetylations. Yeast protein samples were digested with trypsin prior di-methylation of resulting peptide mixture. Then, di-methylated peptide mixtures were fractionated by OFFGEL and reverse phase liquid chromatography followed by MALDI-TOF/TOF mass spectrometry analysis. Data analysis was performed by using Mascot as search engines.Our results showed that OFFGEL fractionation is a useful step to increase detection of acetylations. Moreover, we showed that our di-methylation treatment improved significantly detection of acetylation. Indeed, after di-methylation treatment, 385 unique acetylated sites were identified while 164 unique acetylated peptides were detected without di-methylation treatment. The improvement of acetylation detection using our di-methylation strategy is observed for each of acetylations: N-ter, K- and O-acetylations. Thus, this new proteomic method is promising to enhance N-ter, K- and O-acetylation detection.In the second step, we presented preliminary results of deacetylation by sirtuin 1 in the presence of p53 peptide (Ac-Arg-His-Lys-Lys- (Ac) –AMC. However, the low deacetylation efficiency of the p53 peptide observed, conclude that is not suitable to applicate into RAQIAT Method

O presente trabalho pretende analisar alguns reflexos da incorporação dos influxos da nova ordem globalizada e do neoliberalismo na ordem jurídica nacional. Será demonstrado que os valores sugeridos ainda que essa sugestão tenha caráter quase cogente pelo Banco Mundial, por meio de sua atividade paranormativa, são incorporados pela ordem legal brasileira através de reformas processuais que priorizam a justiça quantitativa. Essas reformas, somadas a outros fatores tais como o aumento das demandas levadas ao Poder Judiciário, a influência da economia na análise do Direito, a utilização equivocada, irrefletida e mecânica de discursos de fundamentação prévia, a pobreza do ensino jurídico e a submissão dos juízes ao que ditam os tribunais, resultam em um patamar de jurisdição padronizada. Paralelamente a esse processo, a população carcerária brasileira cresce em acelerado ritmo, o que pode ser relacionado com o avanço dos valores neoliberais e da retração do estado do bem estar social.Diante desse quadro, é proposto, como forma de aproximação da teoria jurídica com a prática forense e de maneira a proporcionar abertura do direito penal ao mundo dos fatos, a dogmática funcional redutora, de modo que o Direito Penal atue como dique de contenção do estado de polícia que subjaz a cada estado de direito. Ademais, propõe-se que essa adoção se dê nas salas de aula dos cursos de direito, utilizando-se, tanto quanto possível, de diversos recursos didáticos para tornar mais palpáveis e inteligíveis os conceitos e ideias propostos. / This study aims to analyze some consequences of the new global order and the neoliberalism influence inBrazilian legal system.It will be demonstrated that the values suggested even though this suggestion is, in fact, an obligation by the World Bank, through his paranormative activity, are incorporated by the Brazilian legal system through changes in procedure law, which focus only in quantitative justice. These reforms, added to others factors, such as the growth in the number of cases led to the judiciary, the economy influence in law analysis, the equivocate, unreflective and mechanic comprehension and use of the previously set foundation of judicial decisions, the poor quality law education and the judges submission to the Courts, result in a standardized jurisdiction. Simultaneously, the encarcerated population has grown swiftly, phenomenon that can be related to neoliberalism values widespread and to the weakening of the welfare state.Considering this scenario, as a way to approximate theory and practice of the law and to allow the opening of penal law to exterior facts,it is proposed reductive functional penal law dogmatic, so that penal law can act like a dike that holds the police state that hibernate within every constitutional state. Moreover, it is proposed the use of this theory initiates in law courses, using, as much as possible, several didactics resources to make easier to understand these concepts and ideas.

Soit K un corps algébriquement clos de caractéristique nulle. Il est bien connu, d'après un résultat de Duflo, Khalgui et Torasso, qu'une algèbre de Lie algébrique quasi-réductive (définie sur K) est stable. La réciproque est fausse en général. Se pose la question de savoir, si pour certaines classes particulières d'algèbres de Lie non réductives, il y a équivalence entre ces deux notions. Plus généralement, les sous-algèbres biparaboliques forment une classe très intéressante (incluant la classe des sous-algèbres paraboliques et de Levi) d'algèbres de Lie qui ne sont pas toutes réductives. Panyushev conjecture que si une sous-algèbre biparabolique est stable, alors son stabilisateur générique est un tore. Cette conjecture peut être reformulée ainsi : une sous-algèbre de Lie biparabolique est stable si et seulement si elle est quasi-réductive. Compte tenu des résultats obtenus par ce dernier pour le cas des sous-algèbres paraboliques d'une algèbre de Lie simple de type A et C, on donne dans cette thèse une réponse positive à cette conjecture pour la classe des sous-algèbres paraboliques d'une algèbre de Lie simple. Au passage, nous montrons également qu'une sous-algèbre de Lie de gl(n, K) qui stabilise une forme bilinéaire alternée de rang maximal et un drapeau en position générique est stable si et seulement si elle est quasi-réductive. / Let K be an algebraically closed field of characteristic 0. It is well known by work of Duflo, Khalgui and Torasso that any quasi-reductive algebraic Lie algebra (defined over K) is stable. However, there are stable Lie algebras which are not quasi-reductive. This raises the question, if for some particular class of non-reductive Lie algebras, there is equivalence between stability and quasi-reductivity. More generally, biparabolic subalgebras form a very interesting class (including the class of parabolic subalgebras and of Levi subalgebras) of non-reductive Lie algebras. It was conjectured by Panyushev that these two notions are equivalent for biparabolic subalgebras of a reductive Lie algebra. In this thesis, we give by considering the results of Panyushev for parabolic subalgerbras of simple Lie algebra of type A and C a positive answer to this conjecture in the case of parabolic subalgebras. In passing, we prove that these two notions are equivalent for certain subalgebras of gl(n,K) which stabilize an alternating bilinear form of maximal rank and a flag in generic position.

O presente trabalho pretende analisar alguns reflexos da incorporação dos influxos da nova ordem globalizada e do neoliberalismo na ordem jurídica nacional. Será demonstrado que os valores sugeridos ainda que essa sugestão tenha caráter quase cogente pelo Banco Mundial, por meio de sua atividade paranormativa, são incorporados pela ordem legal brasileira através de reformas processuais que priorizam a justiça quantitativa. Essas reformas, somadas a outros fatores tais como o aumento das demandas levadas ao Poder Judiciário, a influência da economia na análise do Direito, a utilização equivocada, irrefletida e mecânica de discursos de fundamentação prévia, a pobreza do ensino jurídico e a submissão dos juízes ao que ditam os tribunais, resultam em um patamar de jurisdição padronizada. Paralelamente a esse processo, a população carcerária brasileira cresce em acelerado ritmo, o que pode ser relacionado com o avanço dos valores neoliberais e da retração do estado do bem estar social.Diante desse quadro, é proposto, como forma de aproximação da teoria jurídica com a prática forense e de maneira a proporcionar abertura do direito penal ao mundo dos fatos, a dogmática funcional redutora, de modo que o Direito Penal atue como dique de contenção do estado de polícia que subjaz a cada estado de direito. Ademais, propõe-se que essa adoção se dê nas salas de aula dos cursos de direito, utilizando-se, tanto quanto possível, de diversos recursos didáticos para tornar mais palpáveis e inteligíveis os conceitos e ideias propostos. / This study aims to analyze some consequences of the new global order and the neoliberalism influence inBrazilian legal system.It will be demonstrated that the values suggested even though this suggestion is, in fact, an obligation by the World Bank, through his paranormative activity, are incorporated by the Brazilian legal system through changes in procedure law, which focus only in quantitative justice. These reforms, added to others factors, such as the growth in the number of cases led to the judiciary, the economy influence in law analysis, the equivocate, unreflective and mechanic comprehension and use of the previously set foundation of judicial decisions, the poor quality law education and the judges submission to the Courts, result in a standardized jurisdiction. Simultaneously, the encarcerated population has grown swiftly, phenomenon that can be related to neoliberalism values widespread and to the weakening of the welfare state.Considering this scenario, as a way to approximate theory and practice of the law and to allow the opening of penal law to exterior facts,it is proposed reductive functional penal law dogmatic, so that penal law can act like a dike that holds the police state that hibernate within every constitutional state. Moreover, it is proposed the use of this theory initiates in law courses, using, as much as possible, several didactics resources to make easier to understand these concepts and ideas.

Neste trabalho estudou-se o uso do ferro de valência zero em meio redutivo e oxidativo, a fim de desenvolver um processo que promova tanto a degradação quanto a mineralização de substâncias orgânicas com centros deficientes de elétrons, como, por exemplo, espécies que contêm o grupo nitro ligado ao anel aromático. O processo de degradação empregando-se ferro metálico, em ambos os meios estudados, apresentou grande eficiência no tratamento do p-nitrofenol, utilizado como poluente modelo. Para soluções contendo concentração inicial de p-nitrofenol igual a 100 mg L-1, tanto para o meio redutivo quanto para o oxidativo, em condições ótimas, as taxas de degradação obtidas foram superiores a 98% ao final de 15 minutos de tratamento. Para o meio com atmosfera oxidante, foi possível observar a geração de H2O2 in situ (cerca de 0,030 mmol L-1, em 45 minutos), proporcionando uma reação do tipo Fenton. Dessa forma, foi possível obter uma redução do teor de carbono orgânico dissolvido de cerca de 20%, em 60 minutos de reação. A fim de elevar a taxa de mineralização do p-nitrofenol, realizou-se a integração dos processos redutivo e oxidativo. O emprego deste sistema de tratamento proporcionou um aumento de 100% na redução do teor de carbono orgânico dissolvido, para o mesmo tempo de tratamento. Assim, pode-se inferir que a associação dos processos estudados levou à formação de substâncias mais susceptíveis ao ataque de agentes oxidantes. Por fim, avaliou-se o emprego de ferro de valência zero sintetizado via redução de íons Fe2+ por boridreto de sódio. Utilizando-se o sistema de tratamento integrado, este material proporcionou uma taxa de mineralização de cerca de 50%, demonstrando-se como uma boa alternativa para a obtenção do ferro de valência zero. Dessa forma, pode-se concluir que o sistema integrado apresenta-se como uma alternativa viável e eficiente, com grande potencial de aplicabilidade para o tratamento de substâncias recalcitrantes aos processos de tratamento convencionais, uma vez que apresentou bons resultados na degradação/mineralização da substância modelo. / In this work, we studied the use of zero-valent iron under reductive and oxidative medium, in order to develop a process that promotes the degradation and the mineralization of organic substances with deficient electron centers, such as nitroaromatic compounds. In both mediums, the degradation process promoted by the metallic iron showed good efficiency to degrade p-itrophenol, used as pollutant model. For solutions of p-nitrophenol with initial concentration of 100 mg L-1, degradation percentages were above 98% after 15 minutes of the treatment in reductive and oxidative mediums (optimal conditions). Under oxidant atmosphere, the in situ generation of H2O2 (about 0,030 mmol L-1, in 45 minutes) leaded to a Fenton-like reaction. So, it provided a reduction of total organic carbon levels up to 20% in 60 minutes. In order to raise the mineralization of p-nitrophenol, the reductive and oxidative processes were integrated. The used of this treatment system provided an increase of 100% in the reduction of the dissolved organic carbon, for the same treatment time. Thus, we can in infer that the association of the two processes led to the formation of more susceptible substances to the attack of oxidant agents. Finally, it was evaluated the use zero-valent iron synthesized through reduction of Fe2+ ions by NaBH4. The obtained material provided about 50% of p-nitrophenol mineralization, showing to be a good source of zero-valent iron. So, it can be concluded that the use of the integrated system is a viable and efficient alternative, with a great potential of applicability for the treatment of recalcitrants substances to the conventional treatment processes, because it presented good results in the degradation/mineralization of studied pollutant model.

O estudo que ora se apresenta integrou o conjunto de pesquisas do sub-projeto - Diversidade de Bactérias Associadas à Degradação de Compostos Recalcitrantes, do projeto temático BIOTA FAPESP - Ecologia Molecular e Taxonomia Polifásica de Bactérias de Interesse Ambiental e Agro-Industrial. Apresenta caráter inovador, na medida em que procurou avaliar o potencial de aplicação biotecnológica de microrganismos anaeróbios de uma área severamente contaminada no Brasil, o estuário de Santos-São Vicente, em degradar o pentaclorofenol (PCP). A pergunta fundamental a ser respondida pelos resultados experimentais realizados foi: são os microrganismos autóctones do estuário capazes de servirem de inóculo para degradar o pentaclorofenol em biorreatores sob condições metanogênicas? Dois grupos de amostras foram avaliados, o primeiro, uma parcela composta por vários sedimentos coletados no estuário e, o segundo, sedimentos coletados na região do Largo de Canéus e na frente da Companhia Siderúrgica Paulista (COSIPA). O estabelecimento da determinação cromatográfica do PCP e congêneres menos clorados para o monitoramento experimental mostrou que na análise da presença dos clorofenóis nos sedimentos, o método de extração por ultrassom com posterior metilação dos analitos foi adequado para concentração mínima de 200 'mü'g clorofenóis/Kg sedimento para 2,3; 2,6 diclorofenóis; 2,4,6 e 2,3,6 triclorofenóis. Contudo, não foi adequado para a determinação do PCP e 2,3,4 triclorofenol. Para o meio de cultivo, o método de extração do PCP por agitação em vórtex e acetilação dos analitos mostrou-se adequado para todos os clorofenóis e com limite de quantificação de 0,1 mg/L. A avaliação do potencial metanogênico foi realizada com as amostras compostas do estuário enriquecidas sob condições halofílicas. O valor estável de metano no biogás de 50% foi obtido nos primeiros 20 dias de incubação. O sedimento nessas condições foi utilizado como inóculo para fins de isolamento de culturas metanogênicas, redutoras do íon sulfato e degradadoras de PCP. Não foram obtidas culturas desalogenadoras; porém foram isoladas arquéias metanogênicas, cultivadas em metanol, acetato e formiato de sódio, bem como bactérias cultivadas em lactato de sódio na presença e ausência de sulfato de sódio. Ensaios fisiológicos aliados aos métodos moleculares FISH e DGGE permitiram identificar arquéias metanogênicas do gênero Methanosarcina sp. e microrganismos do domínio Bacteria. Os sedimentos individualmente estudados foram coletados com maior controle de anaerobiose empregando-se amostrador do tipo corer. O enriquecimento destes sedimentos, inicialmente sob concentração de 2,5 mg PCP/g STV e com adições periódicas de 50% da concentração inicial do clorofenol e 1,25 a 2,5 g/L de glicose por 13 meses a 30 graus Celsius, resultou na obtenção de culturas degradadoras do PCP sob anaerobiose estrita. Nos reatores controles sem PCP, já primeiros 20 dias de incubação 70% de metano foi determinado no biogás. Nos reatores com PCP, a produção do metano (20%) iniciou após 100 dias. A adsorção foi o principal mecanismo de remoção de 50% do PCP nas primeiras 12 horas de incubação dos enriquecimentos. Posteriormente, a redução do PCP no meio de 77% para a amostra de Canéus e 70% para a da COSIPA foi relacionada a mecanismos de biodegradação anaeróbia, como a desalogenação redutiva. Exames microscópicos mostraram a seleção de microrganismos na presença de PCP, com predomínio de bacilos formadores de esporos semelhantes ao gênero Clostridium sp., filamentos diversos e filamentos semelhantes as arquéias metanogênicas acetoclásticas relacionadas ao gênero Methanosaeta sp. O enriquecimento foi realizado sob condições halofílicas, segunda a salinidade de cada amostra estudada no estuário. Não houve perda do clorofenol por volatilização. Os resultados obtidos no enriquecimento anterior viabilizaram o estudo seguro do potencial anaeróbio dos microrganismos oriundos do sedimento da COSIPA em metabolizar o PCP em reator contínuo do tipo RAHLF. Assim, sedimento enriquecido sob condições metanogênicas e halofílicas foi inoculado no RAHLF, controlando-se rigorosamente a anaerobiose. O reator preenchido com cubos de espuma de poliuretana foi operado por 126 dias e tempo de detenção hidráulica de 18 horas, com meio de cultura salino contendo glicose (1 g/L) como principal fonte de carbono e PCP-Na nas concentrações de 5, 13, 15 e 21 mg/L. O desempenho do RAHLF foi estável com boa eficiência durante toda a operação. A redução dos níveis de matéria orgânica medida em DQO variou de 70 a 100% e a de PCP foi de 99%, com detecção de intermediários menos clorados e sob teores de metano no biogás de 20%. Da massa de 1111,73 mg de PCP aplicada no reator, 286,9 mg ficou retida no sistema pelo processo de adsorção nas biopartículas e 824,83 mg foi biodegradada. As análises morfológicas dos tipos celulares, em conjunto com as técnicas moleculares DGGE e FISH, revelaram a presença de grupos microbianos do domínio Archaea pertencente à família Methanosarcinacea e grupos do domínio Bacteria. A participação de grupos de domínio Bacteria, cuja estrutura dos tipos microbianos na comunidade variou ao longo do RAHLF e das concentrações de PCP, e dos organismos metanogênicos da família Methanosarcinacea, possibilitou responder a questão inicialmente formulada, uma vez que se pode afirmar que os microrganismos autóctones foram capazes de degradar o PCP sob condições metanogênicas e halofílicas, com eficiência adequada. A prática para a seleção dos microrganismos retirados do ambiente estuarino, de interesse para biotecnologia anaeróbia aplicada ao saneamento ambiental, que empregou o reator do tipo RAHLF, parece promissora para avanços da engenharia na remediação biológica de uma área cuja relevância é indiscutível para o estado de São Paulo / This study has an innovative character, once aimed to evaluate the biotechnological application of anaerobic microorganisms degrading pentachlorophenol (PCP) at a severely polluted area in Brazil, estuary of Santos-São Vicente, São Paulo state. This study integrated a group of researches of the sub-project - Diversity of Bacteria Associated to the Degradation of Recalcitrant Compounds, supported by major project BIOTA FAPESP which theme is Molecular Ecology and Polyphasic Taxonomy of Bacteria of Environmental and Agriculture-Industrial Interest. The fundamental question to be answered by the accomplished experimental results was: are the autochthonus microorganisms of the estuary capable of serving as inoculum to degrade the PCP in bioreactors under methanogenic conditions? In order to evaluate this hypothesis, two groups of samples were collected: sediments taken from several sites of the estuary and sediments taken from two sites, one severely contaminated with organochlorine compounds, in front of the São Paulo Metallurgical Company (COSIPA) and other, less contaminated, in the area of Canéus. The establishment of protocol for the PCP and less chlorinated compounds chromatographic determination showed that the extraction method by ultrosonication followed by metilation of the chlorinated compounds was appropriated to evaluate concentrations of 200 ug chlorophenol/Kg of sediment, 2,3; 2,6 dichlorophenols; 2,4,6 and 2,3,6 trichlorophenols. However, it was not appropriated for evaluation of PCP and 2,3,4 trichlorophenol. For the culture medium, the extraction method by vortex agitation followed by acetilation was appropriated for all of the chlorophenols compounds. The quantification limit was 0,1 mg/L. The evaluation of the methanogenic potential and PCP biodegradation was accomplished with the samples of the estuary enriched under halophylic conditions. In the experiments without PCP was possible to obtain cultures of Methanosarcina sp, identified by FISH technique and cells of domain Bacteria, identified by DGGE. In the experiment with PCP dehalogenated cultures were not obtained. To evaluate PCP anaerobic biodegradation by sediments of COSIPA and Caneus sites, sediments collected under anaerobic conditions by a corer device were enriched in a halophylic brine medium with glucose and PCP at concentration of 2.5 mg PCP/g VTS. Periodic additions of 50% of the initial concentration of PCP and 1.25 to 2.5 g/L of glucose were done. With this strategy was possible to obtain PCP dehalogenated cultures. The adsorption was the main mechanism of 50% of PCP removal in the first 12 hours of incubation. The PCP reduction of 77% for Caneus reactor and 70% for COSIPA reactor was related to anaerobic biodegradation. Microscopic exams showed selection of microorganisms, with predominance of cells related to Clostridium sp., and filaments related to methanogenic acetoclastic archaea Methanosaeta sp. chlorophenols volatilization was not observed. The biotechnological application of HAIB reactor in PCP biodegradation was evaluated using sediment of COSIPA site previously enriched with glucose. The reactor was operated by 126 days and hydraulic detention time of 18 hours, with saline brine medium containing glucose (1 g/L) and NaPCP in concentrations of 5, 13, 15 and 21 mg/L. PCP amendments did not affect the overall performance and functional stability of the process. COD and PCP reduction was close to 80% and 100%, respectively with detection of trichlorophenols and dichlorophenols. Percentage of methane in the biogas closed to 30%. Adsorption analyses demonstrated that 287 mg of PCP was removed by adsorption in the biofilm and 825 mg was removed by biodegradation. Biofilm DGGE-profiling showed the presence of specific bands of Bacteria and Archaea domains when PCP was amended. The appearance of new bands of Bacteria showed that this organisms had a direct influence at PCP dehalogentation. Archaea organisms of Methanosarcinaceae family had an indirect influence in this metabolism. This thesis demonstrated that HAIB reactors, using autochthonous microorganisms under halophylic and methanogenic conditions, are a potential alternative for organochlorines bioremediation