Dictionary-Learning-Based Post-Filter for HMM-Based Speech Synthesis

Oversmoothing of speech parameter trajectories is one of the causes for quality degradation of HMM-based speech synthesis. Various methods have been proposed to overcome this effect, the most recent ones being global variance (GV) and modulation-spectrum-based post-filter (MSPF). However, there is still a significant quality gap between natural and synthesized speech. In this paper, we propose a two-fold post-filtering technique to alleviate to a certain extent the oversmoothing of spectral and excitation parameter trajectories of HMM-based speech synthesis. For the spectral parameters, we propose a sparse coding-based post-filter to match the trajectories of synthetic speech to that of natural speech, and for the excitation trajectory, we introduce a perceptually motivated post-filter. Experimental evaluations show quality improvement compared with existing methods.

Hegel y la disolución de la tradición de la ley natural : el apogeo del gnosticismo moderno

Resumen: En el presente trabajo, el autor expone la disolución teórica de la doctrina tradicional sobre la ley natural y su expulsión, incluso semántica, de la Filosofía del Derecho de Hegel. Para ello, se expone una síntesis del pensamiento especulativo de Hegel y se pone de manifiesto el inmanentismo radical del filósofo suabo, que resulta ser expresión del gnosticismo moderno.

A reserva estatal de atividade econômica e a propriedade do petróleo e do gás natural

A partir de uma pesquisa histórica sobre a regulação da atividade petrolífera nacional, constatou-se que as jazidas de petróleo e de gás natural são bens públicos cuja exploração é constitucionalmente reservada ao Estado. Ademais, a delegação da exploração desses bens, por meio de concessão ou de partilha de produção, mantém a propriedade estatal desses recursos naturais.

Tropical river fisheries valuation: background papers to a global synthesis

This report is a compilation of five regional reviews that document the global status of tropical rivers and inland fisheries in three continents: Latin America, Africa and Asia. It explores the role of ‘valuation’ methods and their contribution to policy-making and river fishery management. From the compilation, the best estimate of the global value of inland fisheries for those three continents is US$ 5.58 billion (gross market value), which is equivalent to 19 percent of the current value of annual fish exports from developing countries (US$ 29 billion) for 2004. The compilation shows that there is a general shortage of information on inland fisheries, especially derived from conventional economic valuation methods, though information from economic impact assessment methods and socio-economic and livelihood analysis methods is more widely available. The status of knowledge about the impact of changes in river management on the value of tropical river fisheries is weak and patchy. Although the impacts of large dams on the hydrology, ecology and livelihood support attributes of tropical rivers are well-recognized, there have been only few valuation studies of these issues. The document highlights the need for further valuation studies of tropical river and inland fisheries in developing countries. It underlines how vital it is for policy-makers and other stakeholders to understand the importance of these natural resources in order to make appropriate decisions concerning their role in development policy and illustrates why capacity building in valuation should become a major priority for agencies concerned with fisheries management and policy-making.

Stereoselective inverse conjugate addition of nitrogen and carbon nucleophiles to allenyl phosphine oxide. Synthesis of α,β-unsaturated phosphine oxides

Issue in Honor of Prof. Paweł Kafarski

Chiral gold(I) vs chiral silver complexes as catalysts for the enantioselective synthesis of the second generation GSK-hepatitis C virus inhibitor

The synthesis of a GSK 2(nd) generation inhibitor of the hepatitis C virus, by enantioselective 1,3-dipolar cycloaddition between a leucine derived iminoester and tert-butyl acrylate, was studied. The comparison between silver(I) and gold(I) catalysts in this reaction was established by working with chiral phosphoramidites or with chiral BINAP. The best reaction conditions were used for the total synthesis of the hepatitis C virus inhibitor by a four step procedure affording this product in 99% ee and in 63% overall yield. The origin of the enantioselectivity of the chiral gold(I) catalyst was justified according to DFT calculations, the stabilizing coulombic interaction between the nitrogen atom of the thiazole moiety and one of the gold atoms being crucial.

Directed evolution of terpene synthases for non-natural substrates

Terpenes represent about half of known natural products, with terpene synthases catalyzing reactions to increase the complexity of substrates and generate cyclizations of the linear diphosphate substrates, therefore forming rings and stereocenters. With their diverse functionality, terpene synthases may be highly evolvable, with the ability to accept a wide range of non-natural compounds and with high product selectivity. Our hypothesis is that directed evolution of terpene synthases can be used to increase selectivity of the synthase on a specific substrate. In the first part of the work presented herein, three natural terpene synthases, Cop2, BcBOT2, and SSCG_02150, were tested for activity against the natural substrate and a non-natural substrate, called Surrogate 1, and the relative activities on both the natural and non-natural substrates were compared. In the second part of this work, a terpene synthase variant of BcBOT2 that has been evolved for thermostability, was used for directed evolution for increased activity and selectivity on the non-natural substrate referred to as Surrogate 2. Mutations for this evolution were introduced using random mutagenesis, with error prone polymerase chain reactions, and using site-specific saturation mutagenesis, in which an NNK library is designed with a specific active site amino acid targeted for mutation. The mutant enzymes were then screened and selected for enhancement of the desired functionality. Two neutral mutants, 19B7 W367F and 19B7 W118Q, were found to maintain activity on Surrogate 2, as measured by the screen.

Synthesis and biological activity of anticoagulant heparan sulfate glycopolymers

Heparin has been used as an anticoagulant drug for more than 70 years. The global distribution of contaminated heparin in 2007, which resulted in adverse clinical effects and over 100 deaths, emphasizes the necessity for safer alternatives to animal-sourced heparin. The structural complexity and heterogeneity of animal-sourced heparin not only impedes safe access to these biologically active molecules, but also hinders investigations on the significance of structural constituents at a molecular level. Efficient methods for preparing new synthetic heparins with targeted biological activity are necessary not only to ensure clinical safety, but to optimize derivative design to minimize potential side effects. Low molecular weight heparins have become a reliable alternative to heparin, due to their predictable dosages, long half-lives, and reduced side effects. However, heparin oligosaccharide synthesis is a challenging endeavor due to the necessity for complex protecting group manipulation and stereoselective glycosidic linkage chemistry, which often result in lengthy synthetic routes and low yields. Recently, chemoenzymatic syntheses have produced targeted ultralow molecular weight heparins with high-efficiency, but continue to be restricted by the substrate specificities of enzymes.

To address the need for access to homogeneous, complex glycosaminoglycan structures, we have synthesized novel heparan sulfate glycopolymers with well-defined carbohydrate structures and tunable chain length through ring-opening metathesis polymerization chemistry. These polymers recapitulate the key features of anticoagulant heparan sulfate by displaying the sulfation pattern responsible for heparin’s anticoagulant activity. The use of polymerization chemistry greatly simplifies the synthesis of complex glycosaminoglycan structures, providing a facile method to generate homogeneous macromolecules with tunable biological and chemical properties. Through the use of in vitro chromogenic substrate assays and ex vivo clotting assays, we found that the HS glycopolymers exhibited anticoagulant activity in a sulfation pattern and length-dependent manner. Compared to heparin standards, our short polymers did not display any activity. However, our longer polymers were able to incorporate in vitro and ex vivo characteristics of both low-molecular-weight heparin derivatives and heparin, displaying hybrid anticoagulant properties. These studies emphasize the significance of sulfation pattern specificity in specific carbohydrate-protein interactions, and demonstrate the effectiveness of multivalent molecules in recapitulating the activity of natural polysaccharides.

Palladium-catalyzed asymmetric allylic alkylation: insights, application toward cyclopentanoid and cycloheptanoid molecules, and the total synthesis of several daucane sesquiterpenes

The asymmetric construction of quaternary stereocenters is a topic of great interest in the organic chemistry community given their prevalence in natural products and biologically active molecules. Over the last decade, the Stoltz group has pursued the synthesis of this challenging motif via a palladium-catalyzed allylic alkylation using chiral phosphinooxazoline (PHOX) ligands. Recent results indicate that the alkylation of lactams and imides consistently proceeds with enantioselectivities substantially higher than any other substrate class previously examined in this system. This observation prompted exploration of the characteristics that distinguish these molecules as superior alkylation substrates, resulting in newfound insights and marked improvements in the allylic alkylation of carbocyclic compounds.

General routes to cyclopentanoid and cycloheptanoid core structures have been developed that incorporate the palladium-catalyzed allylic alkylation as a key transformation. The unique reactivity of α-quaternary vinylogous esters upon addition of hydride or organometallic reagents enables divergent access to γ-quaternary acylcyclopentenes or cycloheptenones through respective ring contraction or carbonyl transposition pathways. Derivatization of the resulting molecules provides a series of mono-, bi-, and tricyclic systems that can serve as valuable intermediates for the total synthesis of complex natural products.

The allylic alkylation and ring contraction methodology has been employed to prepare variably functionalized bicyclo[5.3.0]decane molecules and enables the enantioselective total syntheses of daucene, daucenal, epoxydaucenal B, and 14-p-anisoyloxydauc-4,8-diene. This route overcomes the challenge of accessing β-substituted acylcyclopentenes by employing a siloxyenone to effect the Grignard addition and ring opening in a single step. Subsequent ring-closing metathesis and aldol reactions form the hydroazulene core of these targets. Derivatization of a key enone intermediate allows access to either the daucane sesquiterpene or sphenobolane diterpene carbon skeletons, as well as other oxygenated scaffolds.

Enantioselective synthesis of pyrroloindolines and tryptophan derivatives by an asymmetric protonation reaction

Pyrroloindoline and unnatural tryptophan motifs are important targets for synthesis based on their incorporation into a diverse array of biologically active natural products. Both types of alkaloids have also found applications as chiral catalysts and tryptophan derivatives are commonly employed as biological probes. On account of their applications, these frameworks have inspired the development of numerous enantioselective, catalytic reactions. In particular, the past few years have witnessed an impressive number of novel approaches for pyrroloindoline formation.

The first project described herein involves our contribution to pyrroloindoline research. We have developed an (R)-BINOL•SnCl₄-catalyzed formal (3 + 2) cycloaddition reaction between 3-substituted indoles and 2-amidoacrylates that affords pyrroloindoline-2-carboxylates bearing an all-carbon quaternary center. Mechanistic studies have elucidated that the enantiodetermining step is a highly face-selective catalyst-controlled protonation reaction. The subsequent application of this asymmetric protonation strategy to the synthesis of a variety of enantioenriched tryptophan derivatives is also discussed.

Binding site size limitations of imidazole-pyrrole polyamides for recognition in the minor groove of DNA

The discovery that the three ring polyamide Im-Py-Py-Dp containing imidazole (Im) and pyrrole (Py) carboxamides binds the DNA sequence 5'-(A,T)G(A,T)C(A,T)-3' as an antiparallel dimer offers a new model for the design of ligands for specific recognition of sequences in the minor groove containing both G,C and A,T base pairs. In Chapter 2, experiments are described in which the sequential addition of five N- methylpyrrolecarboxamides to the imidazole-pyrrole polyamide Im-Py-Py-Dp affords a series of six homologous polyamides, Im-(Py)_2-7-Dp, that differ in the size of their binding site, apparent first order binding affinity, and sequence specificity. These results demonstrate that DNA sequences up to nine base pairs in length can be specifically recognized by imidazole-pyrrole polyamides containing three to seven rings by 2:1 polyamide-DNA complex formation in the minor groove. Recognition of a nine base pair site defines the new lower limit of the binding site size that can be recognized by polyamides containing exclusively imidazole and pyrrolecarboxamides. The results of this study should provide useful guidelines for the design of new polyamides that bind longer DNA sites with enhanced affinity and specificity.

In Chapter 3 the design and synthesis of the hairpin polyamide Im-Py-Im-Py-γ-Im- Py-Im-Py-Dp is described. Quantitative DNase I footprint titration experiments reveal that Im-Py-Im-Py-γ-Im-Py-Im-Py-Dp binds six base pair 5'-(A,T)GCGC(A,T)-3' sequences with 30-fold higher affinity than the unlinked polyamide Im-Py-Im-Py-Dp. The hairpin polyamide does not discriminate between A•T and T•A at the first and sixth positions of the binding site as three sites 5'-TGCGCT-3', 5'-TGCGCA-3', and 5 'AGCGCT- 3' are bound with similar affinity. However, Im-Py-Im-Py-γ-Im-Py-Im-PyDp is specific for and discriminates between G•C and C•G base pairs in the 5'-GCGC-3' core as evidenced by lower affinities for the mismatched sites 5'-AACGCA-3', 5'- TGCGTT-3', 5'-TGCGGT-3', and 5'-ACCGCT-3'.

In Chapter 4, experiments are described in which a kinetically stable hexa-aza Schiff base La³⁺ complex is covalently attached to a Tat(49-72) peptide which has been shown to bind the HIV-1 TAR RNA sequence. Although these metallo-peptides cleave TAR site-specifically in the hexanucleotide loop to afford products consistent with hydrolysis, a series of control experiments suggests that the observed cleavage is not caused by a sequence-specifically bound Tat(49-72)-La(L)³⁺ peptide.

Studies directed toward the synthesis of palau'amine and axinellamines A-D

The use of spiro [2.4]hepta-4,6-diene-1-methanol 7 as a general precursor for the synthesis of highly functionalized cyclopentyl rings is described. Diene 7 was converted to its silyl protected 4-nitrile derivative 24 in 46% overall yield. The cyclopropyl ring of 24 reacted with soft carbanionic nucleophiles to give ring opened homo-conjugate addition products 25a-h in 76-97% yield without loss of optical purity. The addition products could be further manipulated by selective mono-hydrogenation to give 1,2 substituted cyclopentenes 26a-e in 85-96% yield.

Diene 7 was used as a starting material for studies directed toward the synthesis of the stereochemically dense chloro-cyclopentyl core of palau'amine 1. Two advanced intermediates 50 and 72 were synthesized. Attempts to effect intramolecular chlorine transfer with 50 were unsuccessful. Attempted intramolecular chlorine transfer with 72 led, instead, to an oxygenated species resulting from oxygen radical trapping.

The enantioselective synthesis of the stereochemically dense chloro-cyclopenty l core of axinellamines A-D 2-5 starting from 7 is also described. The core is synthesized in 4.6% yield over 24 steps. Nakamura's radical dehalogenative hydroxylation is applied for the first time to a cyclopropyl carbonyl iodide to give the ring-opened product in 86% yield. Bolm's meso-anhydride desymmetrization is used to introduce asymmetry in a norbornene intermediate. The final step is a diastereoselective intermolecular chlorination using Barton's methodology to achieve chlorine transfer in 76% yield.

The development of an asymmetric palladium-catalyzed conjugate addition and its application toward the total syntheses of taiwaniaquinoid natural products

The asymmetric synthesis of quaternary stereocenters remains a challenging problem in organic synthesis. Past work from the Stoltz laboratory has resulted in methodology to install quaternary stereocenters α- or γ- to carbonyl compounds. Thus, the asymmetric synthesis of β-quaternary stereocenters was a desirable objective, and was accomplished by engineering the palladium-catalyzed addition of arylmetal organometallic reagents to α,β-unsaturated conjugate acceptors.

Herein, we described the rational design of a palladium-catalyzed conjugate addition reactions utilizing a catalyst derived from palladium(II) trifluoroacetate and pyridinooxazole ligands. This reaction is highly tolerant of protic solvents and oxygen atmosphere, making it a practical and operationally simple reaction. The mild conditions facilitate a remarkably high functional group tolerance, including carbonyls, halogens, and fluorinated functional groups. Furthermore, the reaction catalyzed conjugate additions with high enantioselectivity with conjugate acceptors of 5-, 6-, and 7-membered ring sizes. Extension of the methodology toward the asymmetric synthesis of flavanone products is presented, as well.

A computational and experimental investigation into the reaction mechanism provided a stereochemical model for enantioinduction, whereby the α-methylene protons adjacent the enone carbonyl clashes with the tert-butyl groups of the chiral ligand. Additionally, it was found that the addition of water and ammonium hexafluorophosphate significantly increases the reaction rate without sacrificing enantioselectivity. The synergistic effects of these additives allowed for the reaction to proceed at a lower temperature, and thus facilitated expansion of the substrate scope to sensitive functional groups such as protic amides and aryl bromides. Investigations into a scale-up synthesis of the chiral ligand (S)-tert-butylPyOx are also presented. This three-step synthetic route allowed for synthesis of the target compound of greater than 10 g scale.

Finally, the application of the newly developed conjugate addition reaction toward the synthesis of the taiwaniaquinoid class of terpenoid natural products is discussed. The conjugate addition reaction formed the key benzylic quaternary stereocenter in high enantioselectivity, joining together the majority of the carbons in the taiwaniaquinoid scaffold. Efforts toward the synthesis of the B-ring are presented.

Part I. Efforts directed towards the total synthesis of shionone. Part II. Conversion of estrone to androst-4-EN-3-ONE: a new method for activating the C-9 and C-10 positions of estrogenic steroids for substitution

Part I: An approach to the total synthesis of the triterpene shionone is described, which proceeds through the tetracyclic ketone i. The shionone side chain has been attached to this key intermediate in 5 steps, affording the olefin 2 in 29% yield. A method for the stereo-specific introduction of the angular methyl group at C-5 of shionone has been developed on a model system. The attempted utilization of this method to convert olefin 2 into shionone is described.

Part II: A method has been developed for activating the C-9 and C-10 positions of estrogenic steroids for substitution. Estrone has been converted to 4β,5β-epoxy-10β-hydroxyestr-3-one; cleavage of this epoxyketone using an Eschenmoser procedure, and subsequent modification of the product afforded 4-seco-9-estren-3,5-dione 3-ethylene acetal. This versatile intermediate, suitable for substitution at the 9 and/or 10 position, was converted to androst-4-ene-3-one by known procedures.

Fundamentos da exploração de petróleo e gás natural e análise geoeconômica da matriz energética brasileira atual e projeções futuras

A análise da matriz energética mundial, assim como a brasileira, nos mostra que o gás natural representará em breve um importante insumo energético favorecendo a balança de pagamentos nacional, visto que o gás poderá ser usado tanto para consumo interno, quanto para exportação. O aumento das reservas nacionais de hidrocarbonetos se deve ao desenvolvimento de tecnologias, que favoreceram o conhecimento das bacias brasileiras quanto ao seu potencial produtor de combustíveis fósseis, permitindo a descoberta de novas jazidas. O amadurecimento do mercado nacional quanto ao consumo de gás natural passa pela construção de uma infraestrutura robusta, eficiente e que possibilite a captação, o armazenamento e distribuição do mesmo. O Brasil tem todos os requisitos necessários para adentrar ao seleto grupo de países exportadores de hidrocarbonetos, a descoberta do Pré-sal tende a incrementar a importância do gás natural para o país. Ao final do trabalho são descritos cenários futuros (quanto o consumo de energéticos), onde se vê que o consumo de energia para os próximos anos crescerá a taxas superiores as das duas últimas décadas. Análise destes cenários permite antecipar o interesse de futuros investimentos no desenvolvimento do conhecimento geológico para áreas promissoras.