Phosphorothioate backbone modifications of nucleotide-based drugs are potent platelet activators

Nucleotide-based drug candidates such as antisense oligonucleotides, aptamers, immunoreceptor-activating nucleotides, or (anti)microRNAs hold great therapeutic promise for many human diseases. Phosphorothioate (PS) backbone modification of nucleotide-based drugs is common practice to protect these promising drug candidates from rapid degradation by plasma and intracellular nucleases. Effects of the changes in physicochemical properties associated with PS modification on platelets have not been elucidated so far. Here we report the unexpected binding of PS-modified oligonucleotides to platelets eliciting strong platelet activation, signaling, reactive oxygen species generation, adhesion, spreading, aggregation, and thrombus formation in vitro and in vivo. Mechanistically, the platelet-specific receptor glycoprotein VI (GPVI) mediates these platelet-activating effects. Notably, platelets from GPVI function-deficient patients do not exhibit binding of PS-modified oligonucleotides, and platelet activation is fully abolished. Our data demonstrate a novel, unexpected, PS backbone-dependent, platelet-activating effect of nucleotide-based drug candidates mediated by GPVI. This unforeseen effect should be considered in the ongoing development programs for the broad range of upcoming and promising DNA/RNA therapeutics.

Elucidating the structure of chiral molecules by using amplified vibrational circular dichroism: from theory to experimental realization

Recent experimental observations of enhanced vibrational circular dichroism (VCD) in molecular systems with low-lying electronically excited states suggest interesting new applications of VCD spectroscopy. The theory describing VCD enhancement through vibronic coupling schemes was derived by Nafie in 1983, but only recently experimental evidence of VCD amplification has demonstrated the extent to which this effect can be exploited as a structure elucidation tool to probe local structure. In this Concept paper, we give an overview of the physics behind vibrational circular dichroism, in particular the equations governing the VCD amplification effect, and review the latest experimental developments with a prospective view on the application of amplified VCD to locally probe biomolecular structure.

FORCED CONVECTION TO LAMINAR FLOW OF LIQUID EGG YOLK IN CIRCULAR AND ANNULAR DUCTS

The steady-state heat transfer in laminar flow of liquid egg yolk - an important pseudoplastic fluid food - in circular and concentric annular ducts was experimentally investigated. The average convection heat transfer coefficients, determined by measuring temperatures before and after heating sections with constant temperatures at the tube wall, were used to obtain simple new empirical expressions to estimate the Nusselt numbers for fully established flows at the thermal entrance of the considered geometries. The comparisons with existing correlations for Newtonian and non-Newtonian fluids resulted in excellent agreement. The main contribution of this work is to supply practical and easily applicable correlations, which are, especially for the case of annulus, rather scarce and extensively required in the design of heat transfer operations dealing with similar shear-thinning products. In addition, the experimental results may support existing theoretical analyses.

Selectivity in the mechanism of action of antimicrobial mastoparan peptide Polybia-MP1

Many potent antimicrobial peptides also present hemolytic activity, an undesired collateral effect for the therapeutic application. Unlike other mastoparan peptides, Polybia-MP1 (IDWKKLLDAAKQIL), obtained from the venom of the social wasp Polybia paulista, is highly selective of bacterial cells. The study of its mechanism of action demonstrated that it permeates vesicles at a greater rate of leakage on the anionic over the zwitterionic, impaired by the presence of cholesterol or cardiolipin; its lytic activity is characterized by a threshold peptide to lipid molar ratio that depends on the phospholipid composition of the vesicles. At these particular threshold concentrations, the apparent average pore number is distinctive between anionic and zwitterionic vesicles, suggesting that pores are similarly formed depending on the ionic character of the bilayer. To prospect the molecular reasons for the strengthened selectivity in Polybia-MP1 and its absence in Mastoparan-X, MD simulations were carried out. Both peptides presented amphipathic alpha-helical structures, as previously observed in Circular Dichroism spectra, with important differences in the extension and stability of the helix; their backbone solvation analysis also indicate a different profile, suggesting that the selectivity of Polybia-MP1 is a consequence of the distribution of the charged and polar residues along the peptide helix, and on how the solvent molecules orient themselves according to these electrostatic interactions. We suggest that the lack of hemolytic activity of Polybia-MP1 is due to the presence and position of Asp residues that enable the equilibrium of electrostatic interactions and favor the preference for the more hydrophilic environment.

gamma-zein secondary structure in solution by circular dichroism

The proline-rich N-terminal domain of gamma-zein has been reported in relevant process, which include its ability to cross the cell membranes. Evidences indicate that synthetic hexapeptide (PPPVHL), naturally found in N-terminal portion of gamma-zein, can adopt the polyproline II (PPII) conformation in aqueous solution. The secondary structure of gamma-zein in maize protein bodies had been analyzed by solid state Fourier transform infrared and nuclear magnetic resonance spectroscopies. However, it was not possible to measure PPII content in physiological environment since the beta-sheet and PPII signals overlap in both solid state techniques. Here, the secondary structure of gamma-zein has been analyzed by circular dichroism in SDS aqueous solution with and without ditiothreitol (DTT), and in 60% of 2-propanol and water with DTT The results show that gamma-zein has high helical content in all solutions. The PPII conformation was present at about 7% only in water/DTT solution. (c) 2007 Wiley Periodicals, Inc.

Conformational Properties of Angiotensin II and Its Active and Inactive TOAC-Labeled Analogs in the Presence of Micelles. Electron Paramagnetic Resonance, Fluorescence, and Circular Dichroism Studies

The interaction between angiotensin II (AII, DRVYIHPF) and its analogs carrying 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC) and detergents-negatively charged sodium dodecyl sulfate (SDS) and zwitterionic N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS)-was examined by means of EPR, CD, and fluorescence. EPR spectra of partially active TOAC(1)-AII and inactive TOAC(3)-AII in aqueous solution indicated fast tumbling, the freedom of motion being greater at the N-terminus. Line broadening occurred upon interaction with micelles. Below SDS critical micelle concentration, broader lines indicated complex formation with tighter molecular packing than in micelles. Small changes in hyperfine splittings evinced TOAC location at the micelle-water interface. The interaction with anionic micelles was more effective than with zwitterionic micelles. Peptide-micelle interaction caused fluorescence increase. The TOAC-promoted intramolecular fluorescence quenching was more, pronounced for TOAC(3)-AII because of the proximity between the nitroxide and Tyr(4). CD spectra showed that although both AII and TOAC(1)-AII presented flexible conformations in water, TOAC(3)-AII displayed conformational restriction because of the TOAC-imposed bend (Schreier et al., Biopolymers 2004, 74, 389). In HPS, conformational changes were observed for the labeled peptides at neutral and basic pH. In SDS, all peptides underwent pH-dependent conformational changes. Although the spectra suggested similar folds for All and TOAC(1)-AII, different conformations were acquired by TOAC(3)-AII. The membrane environment has been hypothesized to shift conformational equilibria so as to stabilize the receptor-bound conformation of ligands. The fact that TOAC(3)-AII is unable to acquire conformations similar to those of native AII and partially active TOAC(1)-AII is probably the explanation for its lack of biological activity. (C) 2009 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 92: 525-537, 2009.

Single-mode acceleration of matter waves in circular waveguides

Ultracold gases in ring geometries hold promise for significant improvements of gyroscopic sensitivity. Recent experiments have realized atomic and molecular storage rings with radii in the centimeter range, sizes whose practical use in inertial sensors requires velocities significantly in excess of typical recoil velocities. We use a combination of analytical and numerical techniques to study the coherent acceleration of matter waves in circular waveguides, with particular emphasis on its impact on single-mode propagation. In the simplest case we find that single-mode propagation is best maintained by the application of time-dependent acceleration force with the temporal profile of a Blackmann pulse. We also assess the impact of classical noise on the acceleration process.

Thomas-Fermi ground state of dipolar fermions in a circular storage ring

Recent developments in the field of ultracold gases has led to the production of degenerate samples of polar molecules. These have large static electric-dipole moments, which in turn causes the molecules to interact strongly. We investigate the interaction of polar particles in waveguide geometries subject to an applied polarizing field. For circular waveguides, tilting the direction of the polarizing field creates a periodic inhomogeneity of the interparticle interaction. We explore the consequences of geometry and interaction for stability of the ground state within the Thomas-Fermi model. Certain combinations of tilt angles and interaction strengths are found to preclude the existence of a stable Thomas-Fermi ground state. The system is shown to exhibit different behavior for quasi-one-dimensional and three-dimensional trapping geometries.

O papel dos valores e crenças africanas no tempo de trabalho circular, nas estratégias, na negociação das diferenças entre operários e dirigentes no corredor de Maputo

A categoria tempo circular ou indiferenciado mostra que o trabalho formal é vivido de acordo com a subjetividade dos seus participantes, com uma multiplicidade de significados e de configurações sempre variáveis. Neste estudo, identificou-se a existência de uma relação entre o tempo circular e as estratégias elaboradas pelos trabalhadores na conquista de uma maior autonomia nos espaços de trabalho. Além disso, observou-se a convivência entre trabalhadores, empresários e dirigentes governamentais e sindicais presentes num projeto de modernização no sul de Moçambique, denominado Corredor de Desenvolvimento de Maputo e constatou-se o modo como é realizada a negociação das diferenças. Realizou-se um levantamento das principais estratégias levadas a cabo por empresários nacionais, estrangeiros, governo e sindicatos na gestão da força de trabalho, formação, qualificação e o uso do tempo nos espaços de trabalho, bem como as perspectivas de desenvolvimento humano trazido por este projeto na região de Maputo. Por enquanto, ainda não foi possível detectar um interesse real dos grupos empresariais e governamentais em delinear caminhos tendentes a encontrar equilíbrios possíveis entre a modernização e a tradição, que sejam compatíveis com a satisfação das necessidades, valores e crenças e o respeito ao imaginário dos grupos envolvidos na construção da modernização tecnológica do país. Parecem evidentes estratégias em que a responsabilidade social está ausente e a estratégia de abertura das Zonas Francas Industriais ao investimento externo carece de uma verdadeira negociação entre “parceiros” em que sejam distribuídas de forma equitativa as vantagens e desvantagens inerentes aos projetos de industrialização a serem implantados localmente. Os testemunhos obtidos junto dos protagonistas moçambicanos expressaram uma grande frustração e desencantamento quanto a resultados que possam vir a ser promissores para as populações, trabalhadores e empresários nacionais do projeto em curso. Portanto, este projeto mostra ser um processo exógeno introduzido à revelia de seus principais protagonistas nesta fase de implantação em território moçambicano e onde os aspectos culturais foram desrespeitados, além de serem usados como estratégia para tirar vantagens adicionais.

Análise experimental do sistema "vórtice ferradura" no escoamento ao redor de um cilindro circular

Este trabalho apresenta o estudo de um sistema hidrodinâmico encontrada em muitas situações dentro da mecânica dos fluidos: o Vórtice Ferradura. Esta estrutura possui características bastante complexas e seu comportamento ainda é pouco compreendido e explicado. Assim, o escoamento ao redor do cilindro circular, em leito fixo, foi investigado experimentalmente, com o propósito de tentar caracterizar o comportamento hidrodinâmico do Vórtice Ferradura através da visualização de escoamento e medições de pressão. Para o trabalho, um cilindro foi montado. As visualizações do Vórtice foram feitas através de uma janela de visualização na seção de testes do canal. A filmagem foi feita através de uma câmera de vídeo VHS, de uso doméstico, e injeção de corante. As medições de pressão foram feitas através do emprego de transdutores de pressão, e divididas em dois grupos: (i) medições de pressão feitas no leito do canal, na linha de simetria, na região de escoamento próxima ao Vórtice; (ii) medições de pressão do escoamento na superfície do cilindro. As imagens, assim como os dados de transdutor, foram transportados para o meio digital para posterior manipulação e análise. Os resultados confirmam algumas descrições fornecidas pela literatura, bem como, também levantam algumas considerações sobre o assunto: as imagens permitiram a visualização das etapas de formação do Vórtice, descrever modificações em seu diâmetro ao longo dos ensaios, reconhecer a variabilidade espacial de sua posição e detectar a presença de um vórtice secundário. Os dados de transdutor descreveram o campo de pressões no leito do canal, modificado pela presença do cilindro e, ao redor da superfície do mesmo, bem como, o comportamento das flutuações de pressão, correlações, autocorrelações e espectros das flutuações de pressão dos transdutores, para estas situações.

Memória de um tempo circular no território de Carlos Fuentes

Ao contrário da concepção linear do pensamento ocidental, o tempo mexicano, situado na tradição pré-hispânica, indica uma multiplicidade de tempos, possibilitando que variadas e distantes épocas sejam realidades presentes. Nesse enfoque, diferentes tempos da história mexicana incorporam a narrativa de Carlos Fuentes, onde é comum passado, presente e futuro estarem imbricados em circularidades que resgatam com a imaginação e a linguagem uma memória encoberta pelo discurso totalizador. Na sua obra, Carlos Fuentes percorre mitos, tempos e espaços através de uma escritura que traduz uma cultura mestiça construída por vertentes americanas e européias. No entrelaçamento da pluralidade dos mundos, descobre os acontecimentos do passado, introduzindo significações aos silêncios relegados à margem da história. Através das análises de “Las dos orillas” – conto do livro El naranjo; do romance La muerte de Artemio Cruz e de personagens que transitam nos contos de La frontera de cristal, são investigados três momentos da história mexicana: Conquista espanhola, Revolução mexicana e migrações da atualidade na fronteira. Desse modo, são exploradas possibilidades históricas disseminadas na produção de Carlos Fuentes

Ampliação de movimentos na dança circular com pessoas com deficiência visual

This paper aims at studying how circular dance can afford to sight-disabled peoples movement and how they can learn to cope with the deep movement of relation, consciousness, appropriation and communion with the world. Inside circular dance, a cosmic metaphor, is inscribed the movement of the world, which tells and changes amorously the human history. In the works of Paulo Freire and Maurice Merleau-Ponty one can find the necessary support to discuss, as long as possible, movement and existence. Research-action is used as a methodological approach whose empirical center is placed on the Institute of Education and Rehabilitation of Blind, in Natal, which shelters eight sightdisabled adults. The research s data reveal that the practice of circular dance concurs to enlarge the movement of the research s subjects, to develop a more accurate perception of their selves and of their own capacities, as well as improve the relations Me/Others, Me/World, which require a context of differences. The study has revealed that the practice of dance develops a better perception of the limits and surpasses as a human condition and, in consequence, the discovery of one s own body and the other s body as a resource of lessons and representations of the self and of the world. It lets out the development of a new way of thinking and coping with discrimination surrounding the disabled persons. In movement, in circular dance, the barrier between sight disablement and vision loses force.

Proposta de integração das redes de comunicação da COSERN como uma solução de contigêncvia em caso de falhas nos enlaces do backbone de automação

T'his dissertation proposes alternative models to allow the interconnectioin of the data communication networks of COSERN Companhia Energética do Rio Grande do Norte. These networks comprise the oorporative data network, based on TCP/IP architecture, and the automation system linking remote electric energy distribution substations to the main Operatin Centre, based on digital radio links and using the IEC 60870-5-101 protoco1s. The envisaged interconnection aims to provide automation data originated from substations with a contingent route to the Operation Center, in moments of failure or maintenance of the digital radio links. Among the presented models, the one chosen for development consists of a computational prototype based on a standard personal computer, working under LINUX operational system and running na application, developesd in C language, wich functions as a Gateway between the protocols of the TCP/IP stack and the IEC 60870-5-101 suite. So, it is described this model analysis, implementation and tests of functionality and performance. During the test phase it was basically verified the delay introduced by the TCP/IP network when transporting automation data, in order to guarantee that it was cionsistent with the time periods present on the automation network. Besides , additional modules are suggested to the prototype, in order to handle other issues such as security and prioriz\ation of the automation system data, whenever they are travesing the TCP/IP network. Finally, a study hás been done aiming to integrate, in more complete way, the two considered networks. It uses IP platform as a solution of convergence to the communication subsystem of na unified network, as the most recente market tendencies for supervisory and other automation systems indicate

Antenas monopolo planar com patch em anel circular para sistemas UWB

The microstrip antennas are largely used in wireless communication systems due to their low cost, weight, less complex construction and manufacturing, in addition to its versatility. UWB systems have emerged as an alternative to wireless communications over short distances because they offer of higher capacity and lower multipath distortion than other systems with the same purpose. Combining the advantages of microstrip antennas to the characteristics of UWB, it is possible to develop more and more smaller devices, with diverse geometries to operate satisfactorily in these systems. This paper aims to propose alternatives to microstrip antennas for UWB systems operate in the range between 3.1 and 10.6 GHz, with a patch on circular ring. Some techniques are analyzed and employed to increase the bandwidth of proposed antenna: the insertion of a parasitic elements and a rectangular slit in the displaced ground plane. For this, key issues are presented as the basic principles of UWB systems, the fundamental theory of antennas and microstrip antennas. The simulations and experimental characterization of constructed antennas are presented, as well as analysis of parameters such as bandwidth and radiation pattern

Expression, purification, and circular dichroism analysis of human CDK9

The human cyclin-dependent kinase 9 (CDK9) protein was expressed in E coli BL21 using the pET23a vector at 30 degrees C. Several milligrams of protein were purified from soluble fraction using ionic exchange and ATP-affinity chromatography. The structural quality of recombinant CDK9 and the estimation of its secondary structure were obtained by circular dichroism. Structural models of CDK9 presented 26% of helices in agreement with the spectra by circular dichroism analysis. This is the first report on human CDK9 expression in Escherichia coli and structure analysis and provides the first step for the development of CDK9 inhibitors. (c) 2006 Elsevier B.V. All rights reserved.