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.

Active Transport of Glutamate in Leishmania (Leishmania) amazonensis

Leishmania spp. are the causative agents of leishmaniasis, a complex of diseases with a broad spectrum of clinical manifestations. Leishmania (Leishmania) amazonensis is a main etiological agent of diffuse cutaneous leishmaniasis. Leishmania spp., as other trypanosomatids, possess a metabolism based significantly on the consumption of amino acids. However, the transport of amino acids in these organisms remains poorly understood with few exceptions. Glutamate transport is an important biological process in many organisms. In the present work, the transport of glutamate is characterized. This process is performed by a single kinetic system (K-m=0.59 +/- 0.04 mM, V-max=0.123 +/- 0.003 nmol/min per 20 x 10(6) cells) showing an energy of activation of 52.38 +/- 4.7 kJ/mol and was shown to be partially inhibited by analogues, such as glutamine, aspartate, alpha-ketoglutarate and oxaloacetate, methionine, and alanine. The transport activity was sensitive to the extracellular concentration of H+ but not to Na+ or K+. However, unlike other amino acid transporters presently characterized, the treatment with specific ionophores confirmed the participation of a K+, and not H+ membrane gradient in the transport process.

Over-expression of COQ10 in Saccharomyces cerevisiae inhibits mitochondrial respiration

COQ10 deletion in Saccharomyces cerevisiae elicits a defect in mitochondrial respiration correctable by addition of coenzyme Q(2). Rescue of respiration by Q(2) is a characteristic of mutants blocked in coenzyme Q(6) synthesis. Unlike Q(6) deficient mutants, mitochondria of the coq10 null mutant have wild-type concentrations Of Q(6). The physiological significance of earlier observations that purified Coq10p contains bound Q(6) was examined in the present study by testing the in vivo effect of over-expression of Coq10p on respiration. Mitochondria with elevated levels of Coq10p display reduced respiration in the bc1 span of the electron transport chain, which can be restored with exogenous Q(2). This suggests that in vivo binding of Q(6) by excess Coq10p reduces the pool of this redox carrier available for its normal function in providing electrons to the bc1 complex. This is confirmed by observing that extra Coq8p relieves the inhibitory effect of excess Coq10p. Coq8p is a putative kinase, and a high-copy suppressor of the coq10 null mutant. As shown here, when over-produced in coq mutants, Coq8p counteracts turnover of Coq3p and Coq4p subunits of the Q-biosynthetic complex. This can account for the observed rescue by COQ8 of the respiratory defect in strains over-producing Coq10p. (C) 2010 Elsevier Inc. All rights reserved.

Quantitative analysis of indexed publications on seventeen model organisms in nine countries, from 1974 to 2006

Developed countries have an even distribution of published papers on the seventeen model organisms. Developing countries have biased preferences for a few model organisms which are associated with endemic human diseases. A variant of the Hirsch-index, that we call the mean (mo)h-index (""model organism h-index""), shows an exponential relationship with the amount of papers published in each country on the selected model organisms. Developing countries cluster together with low mean (mo)h-indexes, even those with high number of publications. The growth curves of publications on the recent model Caenorhabditis elegans in developed countries shows different formats. We also analyzed the growth curves of indexed publications originating from developing countries. Brazil and South Korea were selected for this comparison. The most prevalent model organisms in those countries show different growth curves when compared to a global analysis, reflecting the size and composition of their research communities.

Ultrasensitive Simultaneous Quantification of 1,N(2)-Etheno-2 `-deoxyguanosine and 1,N(2-)Propano-2 `-deoxyguanosine in DNA by an Online Liquid Chromatography-Electrospray Tandem Mass Spectrometry Assay

Exocyclic DNA adducts produced by exogenous and endogenous compounds are emerging as potential tools to study a variety of human diseases and air pollution exposure. A highly sensitive method involving online reverse-phase high performance liquid chromatography with electrospray tandem mass spectrometry detection in the multiple reaction monitoring mode and employing stable isotope-labeled internal standards was developed for the simultaneous quantification of 1,N(2)-etheno-2`-deoxyguanosine (1,N(2)-epsilon dGuo) and 1,N(2)-propano-2`-deoxyguanosine (1,N(2)-propanodGuo) in DNA. This methodology permits direct online quantification of 2`-deoxyguanosine and ca. 500 amol of adducts in 100 mu g of hydrolyzed DNA M the same analysis. Using the newly developed technique, accurate determinations of 1,N(2)-etheno-2`-deoxyguanosine and 1,N2-propano-2`-deoxyguanosine levels in DNA extracts of human cultured cells (4.01 +/- 0.32 1,N(2)-epsilon dGuo/10(8) dGuo and 3.43 +/- 0.33 1,N(2)-propanodGuo/10(8) dGuo) and rat tissue (liver, 2.47 +/- 0.61 1,N(2)-epsilon dGuo/10(8) dGuo and 4.61 +/- 0.69 1,N(2)-propanodGuo/108 dGuo; brain, 2.96 +/- 1.43,N(2)-epsilon dGuo/10(8) dGuo and 5.66 +/- 3.70 1,N(2)-propanoclGuo/10(8) dGuo; and lung, 0,87 +/- 0.34 1,N(2)-edGuo/ 10(8) dGuo and 2.25 +/- 1.72 1,N(2)-propanodGuo/10(8) dGuo) were performed. The method described herein can be used to study the biological significance of exocyclic DNA adducts through the quantification of different adducts in humans and experimental an with pathological conditions and after air pollution exposure.

Avaliação do potencial de mutagenicidade e toxidade da lectina hipoglicemiante de folha de Bauhinia monandra (pata-de-vaca)

Medicinal plants have been used since antiquity to treat various human diseases. The leaves of Bauhinia monandra are widely used in Brazil as herbal remedies in the treatment of Diabetes Mellitus. From the leaves of B. monandra was purified a galactose-specific lectin, called BmoLL, which also showed a significant hypoglycemic capacity. Following the proposed rules by decree No 116 of 1996/08/08 of the Ministry of Health of Brazil, the study aimed to evaluate the potential for toxicity and mutagenicity of BmoLL from the use of tests with Escherichia coli strain CC104 (Forward mutagenesis assay) with Salmonella typhimurium strain TA (Kado test), with plasmid pBCKS (Break occurrences in plasmid DNA) and enzyme exonuclease III (Search of abasic sites). The results demonstrated that the lectin was unable to increase the frequency of reverse mutation of strains of S. typhimurium, with and without metabolic activity. However, a significant decrease in the frequency of spontaneous mutation was observed in strains of E. coli, especially in poor repair (CC104mutMmutY), indicating an antioxidant potential of the lectin. BmoLL is unable to generate genotoxic and cytotoxic damage, based on the concentrations and the tests performed

Bandas de música, escolas de vida

The focus of this dissertation is to understand the child and youth musical learning trajectory by using two philharmonic orchestras from Seridó, an area in the interior of Rio Grande do Norte. One is called the Hermann Gmeiner Philharmonic Orchestra of Aldeias Infantis SOS Project from Caicó, and the other is the 24 de Outubro Philharmonic Orchestra from Cruzeta. It emphasizes the singular relationship between the band s master and his students, based on a permanent interaction with tradition, creativity and change, resulting from a shared educational process. The idea that unites the work of these two communities is the gamble about the individual's formation as a complex human being, which it is not just concerned with the learning of a musical instrument, but is, more importantly, an educational process capable of facilitating the discovery of the self, others and society. The history and the course of the musical education of these two experiences were facilitated using a base of semi-structured interviews of the bands masters, students and people from two municipal districts. Their history is understood through analysis of the band s pictures and the theoretical-social approach is governed by the complexity idea expressed by Edgar Morin. For the author, complex means to `weave together`; therefore it is multidimensional, and it creates a dialogue about several domains and aspects that occur on the same phenomenon. The complex always supposes creativity and the relationship between the subject s reason and affectivity

Doses e horário de aplicação do diquat no controle de Eichhornia crassipes

O aguapé (Eichhornia crassipes) é uma das mais importantes macrófitas aquáticas que colonizam reservatórios e corpos hídricos nas regiões tropicais e subtropicais do mundo, causando uma série de interferências no uso múltiplo da água e do corpo hídrico e no controle de vetores de doenças humanas. Para evitar prejuízos econômicos, sociais, estéticos e ao meio ambiente, essas plantas são controladas por vários métodos, incluindo o controle químico com o uso de herbicidas. Com o objetivo de avaliar a eficácia do herbicida diquat no controle dessa macrófita aquática, foram instalados dois ensaios em casa de vegetação. No primeiro, foram comparadas duas formulações (Reward® e Reglone®) aplicadas em dois horários (diurno e noturno) e em quatro doses (0,1, 0,2, 0,3 e 0,4 kg ha-1), num esquema fatorial 2 x 2 x 4, com seis repetições. Os resultados mostraram que a formulação Reward® foi mais eficiente no controle dessa macrófita, agindo mais rapidamente, sobretudo em menores doses, e que as aplicações realizadas no período noturno apresentavam melhor desempenho no controle em relação às realizadas no período diurno. Assim, um segundo ensaio foi realizado utilizando apenas a formulação Reward®, comparando a aplicação no período noturno e no diurno e as doses de 0,1, 0,2, 0,3 e 0,4 kg ha-1, obedecendo a um esquema fatorial 2 x 4. As plantas de aguapé foram cultivadas em caixas-d'água, e o delineamento experimental adotado foi em blocos ao acaso com quatro repetições, considerando a posição das caixas dentro da casa de vegetação. Os resultados confirmaram que o diquat é eficiente no controle do aguapé, principalmente nas doses de 0,3 e 0,4 kg ha¹ em aplicações noturnas. Doses baixas e aplicações diurnas não impediram intensas rebrotas das plantas afetadas.

Sensibilidade interoceptiva e respostas psicofisiológicas ao exercício submaximo

The complex human behavior related to exercise involves cognitive, physical and emotional processing. The recent theories about exercise s intensity regulation have highlighted the role played by psychophysics aspects in controlling exercise s intensity. In this regard, recent evidences have shown that there is variability in human capacity in perceiving interoceptives clues. Thus, subjects more sensitive show higher physiological arousal to physical and/or emotional stress, and sensations with higher intensity. In fact, studies have evidenced that interoceptive feedback modifies behavior in exercise with free load. However, exercise recommendations are based in a constant load standard. Therefore, we aimed to analyze the influence of interoceptive sensibility on psychophysics responses during dynamic exercise performed with constant load. Twenty-four adult males were allocated into two groups accordingly with their interoceptive sensibility: high sensibility (n=11) and low sensibility (13). They underwent to an incremental test (IT) and then randomly to two sections of moderate and severe exercise intensity for 20 minutes. Heart rate (HR), rating of perceived exertion (RPE), affective feelings (AF), alert state (AS), and percentage of associative thoughts were collect during exercise. A two-way ANOVA with repeated measures was used to assess differences between psychophysics responses. There were differences between group in RPE, AF, and AS in moderate intensity. There was no difference in any measure in severe intensity. We conclude that subjects with high interoceptive sensibility feel dynamic moderate exercise more intense than the subjecs with low interoceptive sensibility

Tuberculose: conhecimentos, representações sociais e experiências da doença na visão do portador

Tuberculosis is considered one of the most ancient human diseases, cases were registered 3900 years before Christ, and it is currently regarded as a serious public health problem in the world due to several factors such as income mismanagement, precarious standard of life and some sort of prejudice comprised by the word tuberculosis. Taking this into consideration, it was developed a descriptive and exploratory study aiming at analyzing the social representations of tuberculosis made by its patient from the Unidades de Saúde da Família (Family Health Units a public health program) in Campina Grande City PB, in relation to the decentralization of the policies that administrate the disease. It was interviewed 34 tuberculosis patient that were being treated from 2007 to 2008. The age group of the interviewees varied from 10 to 60 years old, but most of them were between 36 and 60 years old (58,8%, n=20), some were young adult and adult (21 35 years old), with 11 (32,3%) respondents, and, less frequent, children and teenagers (11 20 years old), with 03 (8,8%) participants. Data was collected through semi-structured interview. The questions that guided the research were elaborated based on the operational recommendations of DOTS strategy; that is: access to laboratory examinations; medication guarantee; directly observed treatment. Besides that, the experiences of the patient were considered in their relation with the family and the different social groups. The analysis of the discursive material was submitted to the Analyse Lexicale par Context d un Ensemble de Segments de Texte software - ALCESTE 4.7. Data interpretation showed five categories for the social representations of the tuberculosis patient that participated in DOTS strategy: 1) the accessibility of the health assistance service; 2) the patient perspective of the disease; 3) the change in the operation of the productive life; 4) the signals and symptoms of the tuberculosis disease; 5) the rearrangement and mechanisms used to face the disease. The Central Nucleus reveals that tuberculosis is a transmissible disease that can be prevented by people through educational practices, health promotion, active search for symptomatic respiratory and control of the carriers communication; these mechanisms should be incorporated to the routine of all participants of the family health groups. The Intermediate Elements, based on quotidian life, as well as the individual experiences of the tuberculosis patient, reveals prejudiced attitude and beliefs that lead to isolation and restriction of interpersonal relationship. Peripheral Elements were constituted by themes that showed the patient feelings of indignation because of the social barriers they had to face in the Family Health Units during the treatment. These elements demonstrate a negative perspective of the representation concerning the accessibility, i.e. inadequate structure of the health service; long distance to the Health Centre, this factormakes it difficult for the patient to continue the treatment; scheduling delay; and limited service regarding other requests (doctor, dentist etc). One expects to contribute for the construction of a new perspective of the health question between the different agents who make the assistencial institutions and formation of professionals, either in central or local scope

Association between MUC1 gene polymorphism and expected progeny differences in Nelore cattle (Bos primigenius indicus)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Virulence characteristics and epidemiology of Yersinia enterocolitica and Yersiniae other than Y-pseudotuberculosis and Y-pestis isolated from water and sewage

Aims: To determine the species, bio-sero-phagetypes, antimicrobial drug resistance and also the pathogenic potential of 144 strains of Yersinia spp. isolated from water sources and sewage in Brazil.Methods and Results: the 144 Yersinia strains were characterized biochemically, serologically and had their antibiotic resistance and phenotypic virulence markers determined by microbiological and serological standard techniques. The Y. enterocolitica strains related to human diseases were also tested for the presence of virulence genes, by the PCR technique. The isolates were classified as Y. enterocolitica, Y. intermedia, Y. frederiksenii, Y. kristensenii and Yersinia biochemically atypical. The 144 isolates belonged to various bio-serogroups. Half of the strains showed resistance to three or more drugs. The Y. enterocolitica strains related to human diseases exhibited phenotypic virulence characteristics and virulence genes.Conclusions: Water from various sources and sewage are contaminated with Yersinia spp. in Brasil. Among these bacteria, virulent strains of Y. enterocolitica were found, with biotypes and serogroups related to human diseases.Significance and Impact of the Study: This is the first documented description of the occurrence of pathogenic Y. enterocolitica in water sources and sewage in Brazil. The occurrence of virulence strains of Y. enterocolitica shows that the environment is a potential source of human infection by this species in this country.

Measurement of the relative sweetness of stevia extract, aspartame and cyclamate/saccharin blend as compared to sucrose at different concentrations

Special diets are used to mitigate many human diseases. When these diets require changes in carbohydrate content, then sweetness becomes an important characteristic. The range of low-calorie sweeteners available to the food industry is expanding. It is essential to have an exact knowledge of the relative sweetness of various sweeteners in relation to different sucrose concentrations. The objective of this study was to determine the variation on the relative sweetness of aspartame (APM), stevia [Stevia rebaudiana (Bert.) Bertoni] leaf extract (SrB) and the mixture cyclamate/saccharin - two parts of cyclamate and one part of saccharin - (C/S) with the increase in their concentrations, and in neutral and acid pH in equi-sweet concentration to 10% sucrose, using magnitude estimation. Sweetness equivalence of SrB in relation to sucrose concentrations of 20% or higher and of APM and C/S to sucrose concentrations of 40% or higher could not be determined, because a bitter taste predominated. The potency of all sweeteners decreased as the level of sweetner increased. In equi-sweet concentration of sucrose at 10%, with pH 7.0 and pH 3.0, the potency was practically the same for all sweeteners evaluated.

Turismo religioso na cidade da santa: a percepção da comunidade sobre a construção do Complexo Turístico e Religioso Alto de Santa Rita, Santa Cruz/RN

The general objective of this study was to analyze the city of Santa Cruz cope with the impacts of the changes caused by the insertion of religious tourism with the construction of the Tourist Complex Religious Alto de Santa Rita, and specific objectives: to identify the economic activities from building the Tourist Complex Religious Alto de Santa Rita with the monument of Santa Rita; describe the research scenario to respect the traditions, rituals and spatial organization of the city and to evaluate the impacts through construction of movement by Religious Tourism Complex human capital and economic development fostered by Religious Tourism. To this end, we used the methods of application of Questionnaires and interviews, as well as the method was conducted participant observation, individual and systematic, having exploratory held through a correlational analysis regarding the impacts by inserting Tourism religious elsewhere. Was performed also discourse analysis of speeches Stakeholder city of Santa Cruz / RN for understanding the local reality. Literature search was performed readings of books, articles, theses and dissertations dealing with the issue, it was necessary to use data from websites. As for the media research field was qualitative. At the end of the study, it was concluded that the city of Santa Cruz suffered socio-economic and cultural, as a result, especially the lack of a previously developed tourism planning

Isolation of Shiga toxigenic Escherichia coli from butcheries in Taquaritinga city, State of São Paulo, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)