Currently, the application of bioremediation requires extensive and costly monitoring due to limited understanding of the terminal electron accepting processes (TEAPs) that control biodegradation, which impairs the accurate quantification of contaminant mass loss. The measurement of redox conditions and evaluation of TEAPs are critical in assessing the capacity for bioremediation at any site. A series of batch microcosm experiments, using sediment collected from a gasoline-contaminated aquifer at Fort McCoy, Wisconsin, was designed to: 1) evaluate the role of Fe(III) in the development of TEAPs during biodegradation of benzene, toluene, ethylbenzene, and the xylenes (BTEX); 2) examine the biodegradation potential in different portions of the plume; and 3) compare methods of TEAP characterization. In general, the presence of Fe-oxides in microcosms inhibited methanogenesis. Although Fe-reducers did not actively degrade BTEX, Fe-reduction did occur, and most probable number (MPN) counts showed that added Fe(III) increased numbers of Fe-reducers in the microcosms. Methane production in microcosms constructed from sediment near the source area was ~5 times lower than levels produced by the mid-plume sediment. No Fe-reduction occurred in microcosms containing sediment from the source area. These results suggest that the source area has much lower biological activity than the mid-plume. TEAP characterization was conducted using a variety of methods, including geochemical indicators, redox dyes, MPN, and hydrogen concentrations. Monitoring of CH4 concentration yielded useful information in delineation of redox processes; Fe(II) monitoring was unreliable as a geochemical indicator. Redox dyes supplied basic information on reducing environments. MPN counts estimated microbial populations in lieu of faulty geochemical indicators, i.e., Fe(II). The measurement of H2 proved to be one of the more simple and reliable methods for TEAP identification. Results of this study indicate that TEAP characterization should include use of multiple methods; relying on geochemical indicators alone is not sufficient. / Master of Science

La ribonucléase III humaine Dicer est responsable de la biosynthèse des microARN et représente un pilier de la régulation génique, et donc de l’homéostasie cellulaire. Les microARN sont de petits éléments régulateurs de 19 à 24 nt qui régulent ~60% des gènes chez l’humain et sont impliqués dans la majorité des processus cellulaires. De ce fait, il n’est pas étonnant qu’il y ait, dans la majorité des cancers, une diminution globale du niveau des microARN et que, dans plusieurs cas, un changement d’expression ou de localisation de la protéine Dicer ait été noté. Malgré le rôle important que joue Dicer dans la cellule, sa régulation n’est pas très bien connue, ni même les complexes de nature protéique au sein duquel il se trouve. Pour pallier à ce manque, nous avons criblé une librairie d’ADN complémentaires par double-hybride chez la levure, ce qui nous a permis de découvrir, et de caractériser, de nouvelles protéines pouvant interagir avec Dicer. Les résultats obtenus démontrent que Dicer est stabilisé par son interaction avec la protéine résidente du réticulum endoplasmique (RE) cytoskeleton-linking endoplasmic réticulum (ER) membrane protein of 63 kDa (CLIMP-63). La protéine Dicer forme un complexe avec CLIMP-63 peu de temps après sa traduction de novo. Les protéines semblent interagir à l’intérieur du RE et mener à l’export de la protéine Dicer hors de la cellule. Durant la mitose, où les niveaux de protéines Dicer sont réduits, Dicer peut être acétylé par l’acétyl-transférase General Control of Amino-acid Synthesis 5 (GCN5), une modification qui semble stabiliser Dicer. La localisation nucléaire de Dicer peut être causée par la protéine Mitogen-activated protein kinase upstream kinase-binding inhibitory protein (MBIP), au cours d’un processus nécessitant une lysine acétylable à la position 301 du signal de localisation nucléaire de MBIP. Finalement, la cytokine (TWEAK) pourrait moduler l’activité de clivage de Dicer via une interaction directe entre les deux protéines. L’utilisation du double-hybride chez la levure a permis de jeter un éclairage nouveau sur les protéines pouvant réguler la fonction et la localisation de l’enzyme Dicer dans les cellules humaines, au-delà de sa simple activité catalytique.

Flyktingströmmarna över Medelhavet har de senaste åren ökat dramatiskt. Dessa migranter och flyktingar har oftast färdats i undermåliga farkoster, vilket i flera fall lett till att de hamnat i sjönöd. Detta har i sin tur resulterat i att ett stort antal fartyg engagerats i att undsätta dessa människor. Denna studie har genom intervjuer med svenska sjöfarare och genom en litteraturstudie undersökt vilka skillnader och likheter det finns mellan gällande riktlinjer för sjöräddning och räddningsinsatser i Medelhavet. Syftet med studien var att undersöka om riktlinjerna identifierar samma problem och risker som personer som deltagit i räddningsinsatser av nödställda migranter i Medelhavet gjort. Av intervjuerna framkom att det inte fanns några större skillnader mellan de internationella riktlinjerna och respondenternas upplevelse. Dock framkom det av intervjuerna att det stora antalet människor ombord utgjorde en säkerhetsrisk. Detta är något som inte togs i beaktande i de riktlinjer som undersökts. Studien poängterar vikten av att undvika panik i alla situationer, ha tydlig ansvarsfördelning samt att vara väl medveten om att omhändertagandet är resurskrävande och innebär en stor arbetsinsats. Den mest centrala rekommendationen för att uppnå ett framgångsrikt resultat ansågs vara att planera väl före en sjöräddningsinsats påbörjades. / The stream of refugees across the Mediterranean Sea has in recent years increased dramatically. This study has through interviews with Swedish seafarers and through a literature study, examined what differences there are between international guidelines and actual efforts made in the Mediterranean Sea. The purpose of the study was to enquire if these guidelines identified the same problems and risks during a SAR-operation as the people involved in the rescue of distressed migrants in the Mediterranean Sea. The interviews revealed that there were no major differences between the guidelines and the experience of the people involved. It emerged however from the interviews that the large number of people on board constituted a security risk. This is something that was not taken into account in the guidelines that were examined. The study showed the pressing need to avoid panic in any situation, have clear responsibilities and be aware that the care of the distressed migrants is resource-intensive and involves a lot of work. The most important recommendation to achieve a successful outcome was considered to plan well before a search and rescue effort began.

published_or_final_version / Chemistry / Doctoral / Doctor of Philosophy

Par l'utilisation de techniques statistiques poussées, l'approche de population rend l'estimation des propriétés pharmacocinétiques (étude des phénomènes d'absorption, distribution, métabolisme et élimination du médicament dans l'organisme) possible chez les patients inclus dans les études de Phase III, en dépit d'un nombre de prélèvements sanguins réduit. Elle permet également d'évaluer, parmi les caractéristiques des patients, les sources de variabilité interindividuelle (covariables) pouvant expliquer les différences potentielles observées dans telle ou telle sous-population.Après un bref rappel bibliographique de la technique, nous présentons les résultats des analyses effectuées sur trois candidats-médicaments en fin de développement clinique. Le premier exemple détaille le processus d'une analyse de population réalisée chez plus de 3000 patients ainsi que les résultats d'une analyse pharmacocinétique/pharmacodynamique. Dans la deuxième partie, nous insistons sur une technique particulière de validation (le bootstrap) et détaillons l'étape de simulation mise en place pour appuyer les conclusions de l'analyse. La dernière analyse présente enfin un modèle pharmacocinétique de population combinant les données d'un produit parent et de son métabolite actif, l'autre originalité de l'analyse étant l'utilisation du randomization test afin de vérifier l'inclusion de covariables dans le modèle.Les conclusions de ces analyses permettent de mieux appréhender la cinétique des trois molécules dans leur population cible par la mise en évidence d'éventuelles sous-populations à risque et, ainsi, de veiller à leur efficacité et à leur absence de toxicité. / Using sophisticated statistical techniques, the population approach allows estimating the pharmacokinetic properties (study of the absorption, distribution, metabolism and elimination phenomena of a drug into the body) in patients included in Phase III studies, despite a low number of blood samples collected. It also allows evaluating, from the patients' characteristics, the sources of interindividual variability (covariates) that may explain the potential differences observed in a given subpopulation.After a brief review of the technical aspects of the population approach, the results of the analyses performed on three drugs in the end of their clinical development will be presented. The first example details the process of a population analysis carried out in more than 3000 patients, together with the results of a pharmacokinetic/pharmacodynamic analysis. In a second part, we underline a validation technique (the bootstrap) and the simulation study performed to emphasize the conclusions of the study. The last example presents a population pharmacokinetic model applied to the simultaneous fit of data from a parent drug and its active metabolite, the other originality of the analysis being the use of the randomization test to verify the covariates inclusion in the model.The conclusions of these analyses allow a better understanding of the kinetics of the three drugs in their target population by emphasizing potential subpopulations at risk and, hence, participate to verify their efficacy and their absence of toxicity.

En 2010, Haurie et al. ont identifié deux isoformes différentes de la Pol III humaine : Pol IIIα, quin’est présente que dans les cellules souches embryonnaires et dans celles tumorales, et Pol IIIβ,exprimée constitutivement.L’expression ectopique de Pol IIIα affecte profondément l’équilibre de cellules différentiées, jusqu’àen induire la transformation oncogénique. Ceci n’est pas observé dans le cas de l’expression ectopiquede Pol IIIβ.A fin de définir les causes moléculaires de la transformation cellulaire induite par Pol IIIα, nousavons décidé d’étudier son transcriptome en comparaison avec celui de Pol IIIβ, à la recherche desgènes qui pourraient soutenir l’oncogenèse observée. Dans notre étude, nous avons adopté latechnique du ChIP-Seq, une approche innovatrice et puissante qui permet de cartographier tous lessites de liaison d’une protéine sur le génome. Elle comporte la purification de la protéine d’intérêtcomplexe avec ces cibles génomiques et le séquençage de ces dernières de manière massive.Le premier but de cette thèse a été représenté par la mise a point d’un protocole de ChIP-Seqoptimisé spécifiquement pour Pol IIIα et Pol IIIβ. Une fois obtenue une préparation de chromatine dequalité adéquate aux standards du ChIP-Seq, nous avons effectué les séquençages massifs etcartographié sur le génome tous les loci contactés par les deux isoformes de polymérase.De cette manière, nous avons identifié 1287 loci occupés par Pol IIIα et 1281 par Pol IIIβ. Les deuxpolymérases se co-localisent sur la plupart de ces sites, mais montrent aussi une occupationpréférentielle et spécifique sur une fraction de loci Pol III. Cette disproportion pourrait contribuer auphénomène cellulaire observé par Haurie et al.Le manuscrit de thèse détaille les résultats de cette analyse et les conclusions qui en dérivent.Nous y avons ajouté une synthèse des travaux réalisés antérieurement et concertants la caractérisationdes promoteurs de gène qui codent pour les snoARN chez la levure S.cerevisiae, positionnée en toutefin de document. / In eukaryotes, transcription is carried out by DNA-dependent RNA polymerases I, II and III (or I-V inplants). These RNA polymerases are specialized in the transcription of specific groups of genes.Human RNA polymerase III (Pol III) transcribes small noncoding RNAs involved in the regulation oftranscription (7SK RNA), RNA processing (U6 RNA, RNAse P, RNAse MRP), translation (tRNAs,5S RNA) or other cellular processes (vault RNAs [multidrug resistance], adenoviral RNAs [VA-I,VA-II], Epstein-Barr virus RNAs [EBER1, EBER2]). It has furthermore been reported that somemicroRNAs of viral or cellular origin may also be transcribed by Pol III.Interestingly, increased Pol III transcription levels accompany or cause cell transformation. Themechanisms underlying this phenomenon are still largely unknown. Recently, two distinct isoforms ofhuman Pol III have been discovered (Haurie et al., 2010). RPC32β-containing Pol IIIβ is ubiquitouslyexpressed and essential for growth and survival of human cells. In contrast, RPC32α-containing PolIIIα is dispensable for cell survival and its expression is restricted to undifferentiated embryonic stem(ES) cells and to tumor cells.The distinct effects of Pol IIIα and Pol III β on cell growth and transformation may be explained bythe transcription of isoform-specific target genes. To identify such isoform-specific target genes, wespecifically targeted RPC32α and of RPC32β subunits in chromatin immunoprecipitation (ChIP)experiments and analyzed the co-precipitated DNA sequences by high throughput sequencing (ChIPseq.).Genome wide localization of RPC32α and RPC32β subunits of Pol III revealed the presence of bothsubunits on many of the known Pol III-transcribed genes, suggesting redundant activities of bothisoforms of Pol III in transcription of these genes. We also found that some of the genes known to betranscribed by Pol III are only occupied by either RPC32α or by RPC32β, suggesting that these genesare exclusively transcribed by Pol IIIα or by Pol IIIβ, respectively.RPC32α and RPC32β ChIP-seq. results furthermore led to the identification of novel Pol III candidategenes in HeLa cells. Moreover, we found high levels of Pol IIIα or Pol III β at some of the annotatedtRNA pseudogenes, implicating that these genes may be transcribed. The functions of RNAstranscribed from novel putative Pol III genes or from tRNA pseudogenes remain to be determined.

This work concentrates on the issue of capital adequacy of banks. Inadequate capital base of the banking sector is often referred to as one of the causes of the latest financial crisis. Such topic is currently very actual. Even banking regulators pay high attention to this topic. In my thesis I focus on legal and economic aspects of this topic. It is typical for the banking sector that banks operate with much more leverage effect than enterprises belonging to other sectors. Therefore, I focus on the root cause of this phenomenon in the third chapter. I assess the bank indebtedness using the theory of optimal capital structure and I also address the relations between economic and regulatory capital. In the fourth chapter I focus on the concept of capital adequacy and its comparison with leverage indicator. The next chapter concentrates on the development of regulatory minimum capital adequacy standards developed by the Basel Committee on Banking Supervision, known as Basel I and Basel II. The last but one chapter focuses on the latest regulatory rules Basel III governing the capital adequacy of banks and their legal implementation. Final chapter analyzes the impact of Basel III rules on the capital structure of banks in the European Union and the Czech Republic. A separate part of the chapter is...

Orientador: Olga Maria Piazentin Rolim Rodrigues / Resumo: Compreender os processos envolvidos desde o início do desenvolvimento da linguagem tem sido o desafio de múltiplas áreas do conhecimento devido a sua complexidade e a gama de variáveis que podem estar correlacionadas. Para alguns estudiosos, as interações iniciais que o ser humano é exposto, desempenha papel fundamental no desenvolvimento de habilidades linguísticas. Culturalmente, a mãe está no cenário das primeiras relações com maior frequência e, por isso, acaba viabilizando mais oportunidades de interação com o bebê. Considerando assim, o presente estudo propôs-se a descrever e relacionar aspectos da interação mãe-bebê no desenvolvimento da linguagem expressiva e receptiva de bebês de três meses. Participaram deste estudo trinta mães e seus bebês com três meses que frequentam um serviço de acompanhamento do desenvolvimento infantil no primeiro ano de vida, em uma universidade pública do interior do estado de São Paulo. Para a avaliação da interação mãe-bebê, a díade foi filmada em situação estruturada em um episódio de três minutos. Para a avaliação das habilidades comunicativas foi utilizada a Escala de Desenvolvimento Bayley-III, contemplando apenas as áreas de Linguagem Expressiva e Linguagem Receptiva. As filmagens foram analisadas através da cotação de comportamentos interativos maternos e do bebê por segundo, utilizando o protocolo Interadíade. Os resultados demonstraram que, tanto as mães quanto os bebês, apresentaram maior frequência de comportamentos positivos na... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Understanding the processes involved since the beginning of language development has been the challenge of multiple areas of knowledge due to their complexity and range of variables that may be correlated. For some studies, the initial interactions that the human being is exposed plays a fundamental role in the development of language skills. Culturally, the mother is in the scene of the first relationships with more frequency and for this reason it ends up making viable more opportunities of interaction with the baby. Considering this, the present study aimed to describe and relate aspects of the mother-baby interaction in the development of expressive and receptive language of three-month-old babies. Thirty mothers and their three-month-old infants attending a child development follow-up service in the first year of life at a university in the interior of the state of São Paulo participated in this study. For the evaluation of the mother-baby interaction, the dyad was filmed in a structured situation in a three-minute episode. For the evaluation of communicative abilities, the Development Scale Bayley-III was used, considering only the areas of Expressive Language and Receptive Language. Filming was analyzed through the quotation of interactive maternal and baby behaviors per second. The results showed that both mothers and infants had a higher frequency of positive behaviors in the dyad. Babies had more mother-to-be-behaving behaviors with neutral or positive sounds, and m... (Complete abstract click electronic access below) / Mestre

Esta tese apresenta um estudo sobre a contração de anel de olefinas e cetonas cíclicas promovida por sais de tálio(III). A reação de uma série de cicloexanonas e cis e trans-2-decalonas com TTN em CH2Cl2 levou aos correspondentes produtos de contração em rendimentos muito bons, desde que não houvesse um grupo metila em α-carbonila. A reação de 3- e 4-alquilcicloexanonas, bem como de trans-2-decalonas, ocorreu com alto grau de seletividade. As diastereosseletividades observadas estão de acordo com o mecanismo proposto por McKillop. O sistema indânico, resultante da contração de um dos anéis do sistema bicíclico[4.4.0], foi construído de três maneiras diferentes. A reação de 1-tetralonas com TTN/K-10 em pentano forneceu 1-indanocarboxilatos de metila em rendimentos razoáveis, enquanto que o tratamento de 1,2-diidronaftalenos com TTN em MeOH levou aos correspondentes produtos de contração em bons rendimentos, quando não havia grupos alquílicos ligados à dupla ligação. Finalmente, a reação de dois álcoois homoalílicos, contendo a dupla ligação endocíclica, com TTN em uma mistura 1:1 de AcOH e H2O, levou às correspondentes 1-(2,3-diidro-1H-1-indanil)-3-hidroxi-1- propanonas em rendimentos excelentes. / This thesis presents studies toward the ring contraction of ketones and olefins promoted by thallium(III) The reaction of alkylcyclohexanones and cis and trans-2-decalones with TTN in CH2Cl2 led to the corresponding ring contraction products in very good yields, providing there is no methyl group at α-carbonyl position. The reaction of 3- and 4-alkylcyclohexanones, as well as trans-2-decalones, occurred with high degree of selectivity. The observed diastereoselectivities agree with the McKillop\'s mechanism. The indan ring system was constructed by three different protocols. The reaction of 1-tetralones with TTN/K-10 in pentane afforded methyl 1-indanecarboxylates in reasonable yields, while treatment of 1,2-dihydronaphathlenes with TTN in MeOH furnished the corresponding ring contraction products in good yields, as long as there is no alkyl group at the double bond. Finally, the reaction of two homoallylic alcohols, bearing an endocyclic double bond, with TTN in a 1:1 mixture of AcOH and H2O, led to 1-(2,3-dihydro-1H-1-indenyl)-3-hydroxypropan-1-ones in excellent yields.

Este trabalho apresenta a síntese de complexos de ouro(III) com ligante (dietilaminotiocarbonil)benzimidoíla-morfolinil-tiossemicarbazona e co-ligante L variando entre Cl-, CN-, SCN-, C3H5OS2- e C9H13N. Para a caracterização destes complexos foram envolvidas diversas técnicas, como espectroscopia na região do infravermelho e do UV-Vis, ponto de fusão, condutividade, espectroscopia de ressonância magnética nuclear 1H e 13C, espectrometria de massas ESI(+)-MS e análise elementar (C,H,N). O estudo consiste na avaliação da influência do ligante L frente a atividade antitumoral e anti-Trypanossoma cruzi, assim como na avaliação de sua influência em interações com possíveis alvos biológicos (DNA e albumina). Nos experimentos biológicos foi verificada a potencial atividade antitumoral destes complexos em células HepG2, HeLa, DU-145 e MDA-MB-231, dando destaque aos resultados obtidos para os complexos do tipo [AuCl(dmstc)] (IC50 = 8,05 ?molL-1) e [Au(xant)(dmstc)] (IC50 = 28,5 ?molL-1) frente à célula MDA-MB-231, ao complexo [AuCl(dmstc)] (IC50 = 8,37 ?molL-1) frente à célula DU-145 e ao complexo [AuSCN(dmstc)] o qual apresentou resultados promissores para todas as linhagens celulares tumorais avaliadas. Os ensaios in vitro contra cepas Tulahuen LacZ de Trypanossoma cruzi apresentaram resultados promissores, demonstrando uma elevada atividade contra o parasita e citotoxicidade geral negligenciável frente à células de baço de camundongo Swiss para todos os complexos avaliados, com exceção do complexo [Au(xant)(dmstc)]. Através do estudo de interação destes complexos com DNA-ct e albumina humana (HSA), é possível descartar o DNA como principal alvo biológico responsável pelas atividades observadas e sugerir uma adequada distribuição in vivo por meio da HSA. / This work describes the synthesis of gold(III) complexes with the ligand (diethylaminocarbonyl)benzimidoyl-morpholinyl thiosemicarbazone and the co-ligand L varying between Cl-, CN-, SCN-, C3H5OS2- and C9H13N. The complexes characterization involved various techniques such as melting point, conductivity, infrared, UV-Vis, 1H and 13C NMR spectroscopies, ESI(+)-MS mass spectrometry and elemental analysis (C, H, N). The study consists on the evaluation of the influence of the ligand L front anti-tumor and anti-Trypanosoma cruzi activities, as well as in assessing their influence on interactions with possible biological targets (DNA and albumin). The biological experiments showed potential antitumor activity for the complexes against HepG2, HeLa, DU-145 and MDA-MB-231 cells, especially the results obtained for the complexes of the type [AuCl(dmstc)] (IC50 = 8,05 ?molL-1) and [Au(xant)(dmstc)] (IC50 = 28,5 ?molL-1) against the MDA-MB-231 cell, the complex [AuCl(dmstc)] (IC50 = 8,37 ?molL-1) against the DU-145 cell, and the complex [AuSCN(dmstc)] which showed promising results for all tumor cells lines evaluated. In vitro assays against LacZ Tulahuen Trypanosoma cruzi presented promising results, showing high activity against the parasite with negligible general cytotoxicity against the Swiss mouse spleen cells in all evaluating complexes, except for complex [Au(xant)(dmstc)]. Through the studies of interaction of the complexes with DNA-ct and human albumin (HSA) can discard the DNA as the primary biological target responsible for the observed activities and suggest a proper distribution in vivo through the HSA.