Enhancing the catalytic repertoire of nucleic acids: a systematic study of linker length and rigidity

The incorporation of potentially catalytic groups in DNA is of interest for the in vitro selection of novel deoxyribozymes, A series of 10 C5-modified analogues of 2'-deoxyuridine triphosphate have been synthesised that possess side chains of differing flexibility and bearing a primary amino or imidazole functionality, For each series of nucleotide analogues differing degrees of flexibility of the C5 side chain was achieved through the use of alkynyl, alkenyl and alkyl moieties, The imidazole function was conjugated to these CS-amino-modified nucleotides using either imidazole 4-acetic acid or imidazole 4-acrylic acid (urocanic acid), The substrate properties of the nucleotides (fully replacing dTTP) with Taq polymerase during PCR have been investigated in order to evaluate their potential applications for in vitro selection experiments, 5-(3-Aminopropynyl)dUTP and 5-(E-3-aminopropenyl)dUTP and their imidazole 4-acetic acid- and urocanic acid-modified conjugates were found to be substrates, In contrast, C5-amino-modified dUTPs with alkane or Z-alkene linkers and their corresponding conjugates were not substrates, The incorporation of these analogues during PCR has been confirmed by inhibition of restriction enzyme digestion using XbaI and by mass spectrometry of the PCR products.

Fibroblast growth factor 2 maintains the neurogenic capacity of embryonic neural progenitor cells in vitro but changes their neuronal subtype specification

Many in vitro systems used to examine multipotential neural progenitor cells (NPCs) rely on mitogens including fibroblast growth factor 2 (FGF2) for their continued expansion. However, FGF2 has also been shown to alter the expression of transcription factors (TFs) that determine cell fate. Here, we report that NPCs from the embryonic telencephalon grown without FGF2 retain many of their in vivo characteristics, making them a good model for investigating molecular mechanisms involved in cell fate specification and differentiation. However, exposure of cortical NPCs to FGF2 results in a profound change in the types of neurons generated, switching them from a glutamatergic to a GABAergic phenotype. This change closely correlates with the dramatic upregulation of TFs more characteristic of ventral telencephalic NPCs. In addition, exposure of cortical NPCs to FGF2 maintains their neurogenic potential in vitro, and NPCs spontaneously undergo differentiation following FGF2 withdrawal. These results highlight the importance of TFs in determining the types of neurons generated by NPCs in vitro. In addition, they show that FGF2, as well as acting as a mitogen, changes the developmental capabilities of NPCs. These findings have implications for the cell fate specification of in vitro-expanded NPCs and their ability to generate specific cell types for therapeutic applications. Disclosure of potential conflicts of interest is found at the end of this article.

Ligand dissociation from estrogen receptor is mediated by receptor dimerization: Evidence from molecular dynamics simulations

Estrogen Receptor (ER) is an important target for pharmaceutical design. Like other ligand-dependent transcription factors, hormone binding regulates ER transcriptional activity. Nevertheless, the mechanisms by which ligands enter and leave ERs and other nuclear receptors remain poorly understood. Here, we report results of locally enhanced sampling molecular dynamics simulations to identify dissociation pathways of two ER ligands [the natural hormone 17 beta-estradiol (E-2) and the selective ER modulator raloxifene (RAL)] from the human ER alpha ligand-binding domain in monomeric and dimeric forms. E-2 dissociation occurs via three different pathways in ER monomers. One resembles the mousetrap mechanism (Path I), involving repositioning of helix 12 (H12), others involve the separation of H8 and H11 (Path II), and a variant of this pathway at the bottom of the ligand-binding domain (Path II`). RAL leaves the receptor through Path I and a Path I variant in which the ligand leaves the receptor through the loop region between H11 and H12 (Path I`). Remarkably, ER dimerization strongly suppresses Paths II and II` for E-2 dissociation and modifies RAL escape routes. We propose that differences in ligand release pathways detected in the simulations for ER monomers and dimers provide an explanation for previously observed effects of ER quaternary state on ligand dissociation rates and suggest that dimerization may play an important, and hitherto unexpected, role in regulation of ligand dissociation rates throughout the nuclear receptor family.

Evidences for charge-transfer complex formation in the benzene adsorption on sulfated TiO2-a resonance Raman spectroscopy investigation

Benzene adsorbed on highly acidic sulfated TiO2 (S-TiO2) shows an intriguing resonance Raman (RR) effect, with excitation in the blue-violet region. There are very interesting spectral features: the preferential enhancement of the e(2g) mode (1595 cm(-1)) in relation to the a(1g) mode (ring-breathing mode at 995 cm(-1)) and the appearance of bands at 1565 and 1514 cm(-1). The band at 1565 cm(-1) is probably one of the components of the e(2g) split band, originally a doubly degenerate mode (8a, 8b) in neat benzene, and the band at 1514 cm(-1) is assigned to the 19a mode, an inactive mode in neat benzene. These facts indicate a lowering of symmetry in adsorbed benzene, which may be caused by a strong interaction between S-TiO2 and the benzene molecule with formation of a benzene to Ti (IV) charge transfer (CT) complex or by the formation of a benzene radical cation species. However, the RR spectra of the adsorbed benzene cannot be assigned to the benzene radical cation because the observed wavenumber of the ring-breathing mode does not have the value expected for this species. Moreover, it was found by ESR measurements that the amount of radicals was very low, and so it was concluded that a CT complex is the species that originates the RR spectra. The most favorable intensification of the band at 1595 cm(-1) in the RR spectra of benzene/S-TiO2 at higher excitation energy corroborates this hypothesis, as an absorption band in this energy range, assigned to a CT transition, is observed. Copyright (C) 2008 John Wiley & Sons, Ltd.

Avaliação dos parâmetros genéticos da população de arroz irrigado CNA 11 e da divergência genética entre os genitores

Os avanços nas práticas de manejo e no melhoramento das cultivares de arroz irrigado têm permitido consolidar o Rio Grande do Sul como maior produtor deste cereal no Brasil. A seleção recorrente é um método alternativo de melhoramento que permite a recombinação cíclica de genótipos selecionados em uma população geneticamente divergente. Os objetivos deste trabalho foram avaliar o potencial da população CNA 11 para fins de melhoramento para tolerância ao frio, estudar a divergência genética dos seus genitores e quantificar as alterações nas freqüências alélicas como resultado do processo de seleção. Para isso foram conduzidos dois ensaios de campo compostos de 140 famílias S0:2 derivadas da população CNA 11 e quatro cultivares testemunhas, no delineamento látice 12 x 12 e em dois locais: Cachoeirinha (RS) e Santa Vitória do Palmar (RS), onde foram avaliados dez caracteres fenotípicos. O DNA dos genitores da população CNA 11 e de 133 famílias S0:2 foi analisado com marcadores moleculares do tipo microssatélites. A população CNA 11 apresentou alto potencial para fins de melhoramento, com ampla variabilidade genética em todos os dez caracteres avaliados. Os caracteres estatura de planta, esterilidade de espiguetas e número de grãos por panícula apresentaram altas correlações genotípicas com rendimento de grãos, podendo ser usados como critérios de seleção indireta. A esterilidade de espiguetas mostrou ser um critério eficiente de seleção para identificação de genótipos de arroz com tolerância ao frio no estádio reprodutivo. Os genitores da população CNA 11 apresentaram ampla divergência genética e, de modo geral, as freqüências dos alelos de microssatélites característicos dos genótipos da subespécie índica aumentaram e as da subespécie japônica diminuíram à medida que a intensidade de seleção aumentou. As melhores famílias S0:2 mostraram maior similaridade genética com os genitores da subespécie índica.

Enforcing international financial standards in Brazil: limits and possibilities for adoption of IOSCO principles

Foreign capital and institutional investors play a key role in the Brazilian capital and financial markets. Internationally promoted regulatory patterns, especially IOSCO principles, have been increasingly influencing administrative rule making by the Brazilian Securities and Exchange Commission (CVM) as well as the adoption of transnational rules in Brazil by means of self-regulatory activity. Even though there is a certain level of convergence of market regulatory standards at the transnational level, implementation and enforcement of rules remains essentially domestic. We analyze two case studies regarding the transposition of international standards into the Brazilian legal system, which illustrate this tension between the transnational and domestic dimensions of financial markets regulation. The first case concerns a CVM rule on disclosure of executive compensation and the its interpretation by local courts. The second case refers to the adoption of suitability rules.

Dodd Frank act and the Brazilian capital market: extraterritorial effects of regulation to the over-the-counter derivatives market

This paper aims to describe the chief alterations proposed by the Dodd Frank Act to the American over-the-counter derivatives market and, at the same time, understand the extraterritorial reach of this law compared to the regulatory framework of the Brazilian derivative market. In order to do so, I will study the extraterritorial effects of the law, particularly in reference to the international nature of Title II of the Dodd Frank, which deals with the over-the-counter derivatives, in order to evaluate its reach to foreign markets, especially the Brazilian market.

Applying negotiation skills in the design of public policies: analysis of the city of São Paulo’s invoice program

The author argues that by applying problem-solving negotiation skills in the design of public policies, public administrators benefit from more effective and wide-ranging outcomes in the realization of their goals. In order to demonstrate this idea, the author analyzes how negotiation skills – such as identifying key actors and their interests, recognizing hardbargaining tactics and changing the players, knowing your best alternative, creating value and building trust – permeated and contributed to the success of the City of São Paulo’s Invoice Program (“Programa Nota Fiscal Paulistana”), a public policy aimed at combating tax evasion of service tax in the City of São Paulo.

Vida e morte no direito brasileiro : uma reflexão sobre a relação entre dogmática jurídica e estrutura dos tribunais a partir da decisão da ADPF n. 54

This paper analyzes the Brazilian Supreme Court (STF) Decision (ADPF n. 54) in which the court, by majority of votes, stated that the termination of the pregnancy of an anencephalic fetus does not constitute a crime of abortion. The paper analyzes the arguments used by the judges in their opinions to show that they agree on the conclusion, but strongly disagree on their reasons, with the result that the court remains deeply divided on the legal concept of life. The paper then points to the fact that the possibility of different majority positions regarding the decision and the arguments that justify it is not accidental, but the result of the court’s structure, which does not guarantee the collective rationality of the court’s decisions.

Main problems with the taxation of civil society organizations in Brazil: certifications and impact on payroll

The objective of this paper is to identify and analyze the main problems in the taxation—regarding both taxes themselves and compliance costs of taxation—of civil society organizations in Brazil. This study is qualitative descriptive research. A multiple case study with 26 organizations was performed. The results show that the problems mainly affect organizations with lower revenue and that do not work in the areas of education, health or social care. The main problems involve the taxation of the payroll and the difficulties related to obtaining and maintaining certifications. The study concludes with suggestions for the improvement of the regulatory framework.

Beyond ‘best practices': the international regulation of capital markets

It is widely acknowledged that there is considerable international pressure for international ‘best practices’ to be adopted via national legislation. This would occur either by means of model laws or through the passing of country specific legislation that closely replicates foreign legal formats, administrative rules, and or regulation. These attempts to spread the implementation of ‘best practices’ have gained importance in the international debate due to the liberalization of international capital flows. The oversight, country reports, and technical assistance carried out by international organizations along with the growing internationalization of investors have also contributed to this growing pressure. In this respect, due to the constant evolution of transactions and the end objective of making sure that capital markets are developed with just rules, structures, and methods, this article looks to analyze the adoption of standardized models of capital market regulation. Furthermore it looks to examine the motivation and interest of states and other ‘stakeholders’ at the international level.

Teixeira de Freitas leitor de Savigny

The international circulation and reception of the works of Friedrich Carl von Savigny (1779-1861) constitute one of the main features of the first globalization of legal thought. Reaching law professors and practitioners from Brazil to Japan, readings of Savigny’s books offer a promising perspective for understanding how legal cultures around the world coped with the challenges of modernity. By focusing empirically on the circulation of books and analytically on the adaptation of texts to local contexts, the approach forwarded here tries to capture the creative aspects of the diffusion of knowledge. For this purpose, it concentrates on Brazilian readers of Savigny in the 19th Century, especially on the celebrated lawyer Augusto Teixeira de Freitas (1816-1883). It argues that Savigny’s works provided a decisive argumentative framework for some of the main issues discussed in Brazilian jurisprudence of the time. Freitas’ work documents a productive reading of Savigny that shaped his views on both normative and methodological issues, from slavery to codification. By tracing Freitas’ selective appropriation of Savigny’s texts, the article concludes that he was able both to reproduce and to subvert Savigny’s conception of private law, whenever the local context and his personal convictions demanded him to.

Alavanca

Ilustração componente do jogo “Escola Mágica (http://www.loa.sead.ufscar.br/escola-magica.html)” desenvolvido pela equipe do Laboratório de Objetos de Aprendizagem da Universidade Federal de São Carlos (LOA/UFSCar).

Parâmetros eletrocardiográficos e valores séricos e urinários de sódio e potássio em cães tratados com furosemida

The serum and urine Na+ and K+ levels and their relationship with electrocardiography characteristics,were studied in 15 male adult mongrel dogs, treated twice-a-day,with 2, 4, 8mg/kg of live weight doses of furosemide and furosemide associated with KCl during 35 days. Urine samples were analyzed weekly in order to determine the Na+ and K+ concentrations and to evaluate the urinary excretion of both cations. Electrocardiography was performed simultaneously, using limb derivations, speed 50mm/sec and calibration of 1 cm corresponding to 1mV. Data showed decreased seric concentrations of Na+ and Ki, increase in Na+ fractional excretion between days 7 and 21 of the treatment, slower heart rate and longer PR, QRS and QT intervals in the ECG. Briefly 2mg/kg furosemide associated with KCl was the most suitable treatment since it induced lesser side effects.