Benzene induces gene-duplicating but not gene-inactivating mutations at the glycophorin A locus in exposed humans.

Occupational exposure to benzene is known to cause leukemia, but the mechanism remains unclear. Unlike most other carcinogens, benzene and its metabolites are weakly or nonmutagenic in most simple gene mutation assays. Benzene and its metabolites do, however, produce chromosomal damage in a variety of systems. Here, we have used the glycophorin A (GPA) gene loss mutation assay to evaluate the nature of DNA damage produced by benzene in 24 workers heavily exposed to benzene and 23 matched control individuals in Shanghai, China. The GPA assay identifies stem cell or precursor erythroid cell mutations expressed in peripheral erythrocytes of MN-heterozygous subjects, distinguishing the NN and N phi mutant variants. A significant increase in the NN GPA variant cell frequency (Vf) was found in benzene-exposed workers as compared with unexposed control individuals (mean +/- SEM, 13.9 +/- 1.7 per million cells vs. 7.4 +/- 1.1 per million cells in control individuals; P = 0.0002). In contrast, no significant difference existed between the two groups for the N phi Vf (9.1 +/- 0.9 vs. 8.8 +/- 1.8 per million cells; P = 0.21). Further, lifetime cumulative occupational exposure to benzene was associated with the NN Vf (P = 0.005) but not with the N phi Vf (P = 0.31), suggesting that NN mutations occur in longer-lived bone marrow stem cells. NN variants result from loss of the GPA M allele and duplication of the N allele, presumably through recombination mechanisms, whereas NO variants arise from gene inactivation, presumably due to point mutations and deletions. Thus, these results suggest that benzene produces gene-duplicating mutations but does not produce gene-inactivating mutations at the GPA locus in bone marrow cells of humans exposed to high benzene levels. This finding is consistent with data on the genetic toxicology of benzene and its metabolites and adds further weight to the hypothesis that chromosome damage and mitotic recombination are important in benzene-induced leukemia.

Evolution of the chalcone synthase gene family in the genus Ipomoea.

The evolution of the chalcone synthase [CHS; malonyl-CoA:4-coumaroyl-CoA malonyltransferase (cyclizing), EC 2.3.1.74] multigene family in the genus Ipomoea is explored. Thirteen CHS genes from seven Ipomoea species (family Convolvulaceae) were sequenced--three from genomic clones and the remainder from PCR amplification with primers designed from the 5' flanking region and the end of the 3' coding region of Ipomoea purpurea Roth. Analysis of the data indicates a duplication of CHS that predates the divergence of the Ipomoea species in this study. The Ipomoea CHS genes are among the most rapidly evolving of the CHS genes sequenced to date. The CHS genes in this study are most closely related to the Petunia CHS-B gene, which is also rapidly evolving and highly divergent from the rest of the Petunia CHS sequences.

EIA-RIMA insuficiente como avaliação de impactos ambientais enquanto busca de sustentabilidade urbana: o caso de São Sebastião

O presente trabalho comprova que o atual instrumento EIA-RIMA - Estudo de Impacto Ambiental e seu respectivo Relatório de Impacto Ambiental, é insuficiente enquanto busca de sustentabilidade urbana no Litoral Norte do Estado de São Paulo, especificamente no município de São Sebastião. O método de analise dos EIA-RIMA sempre enfatizam os ganhos econômicos e atenuam os impactos negativos, apesar das fragilidades naturais, estruturais e restrições legais incidentes. A tese comprova que os instrumentos utilizados no âmbito do processo de licenciamento ambiental dos empreendimentos, como a ampliação do porto comercial de São Sebastião, estão pouco comprometidos com o real desenvolvimento sustentável da cidade e região. As especificidades geográficas, sobretudo as facilidades de interface do território terrestre com o marítimo, proporcionadas pelas características naturais do Canal de São Sebastião, sempre ofereceram condições privilegiadas para o abrigo de embarcações, característica decisiva na escolha do lugar como cidade portuária, vinculada aos mais importantes ciclos econômicos do país. Agora, em 2015, está prestes a entrar em um novo ciclo, principalmente com a ampliação do porto comercial, duplicação da rodovia dos Tamoios e da SP-055 e a exploração da camada pré-sal na Bacia de Santos. A região já apresenta todos os problemas de uma ocupação acelerada em um território ambientalmente frágil, localizado em estreita faixa litorânea, composta de um lado pela Serra do Mar e do outro pelo Oceano Atlântico. Se não houver um planejamento urbano e regional que considere as características e capacidade de suporte deste território, a zona costeira entrará em colapso. Saber dosar o uso de suas potencialidades sem esgotá-las será o desafio de nossa geração. Os Estudos de Impactos Ambientais precisam ser aperfeiçoados para que ações mitigadoras alcancem muito mais do que o mínimo, alinhadas a investimentos, tecnologias inovadoras, infraestruturas necessárias e obrigatórias que sejam pensadas de maneira conjunta e cumulativa nas diversas esferas do poder, com efetiva participação pública, caso contrário, o crescente interesse econômico e político tenderá à destruição irreversível do patrimônio histórico e natural deste território. O EIA está se transformando em um poderoso instrumento político de ordenamento territorial que acaba por definir critérios e parâmetros de uso e ocupação, desempenhando uma função que não lhe compete, além de aprofundar o desencontro entre uma vida urbana de qualidade e um ambiente equilibrado em nome do desenvolvimento econômico e a serviço dos interesses privados.

Evolução do veneno em cnidários baseada em dados de genomas e proteomas

A evolução do veneno, uma das misturas mais complexas da natureza, tem sustentado o sucesso da diversificação de inúmeras linhagens de animais. Serpentes deslizantes ou medusas flutuantes utilizam o veneno, um coquetel de peptídeos farmacologicamente ativos, sais e moléculas orgânicas. Esses animais surpreendentes têm provocado grande fascínio ao longo da história humana. Nesta dissertação propomos um estudo da evolução dos venenos no filo Cnidaria, englobando dados proteômicos e genômicos. Este projeto teve como objetivos: (1) caracterizar e elucidar a evolução da composição do veneno em Cnidaria por meio da comparação de listas de proteínas; (2) testar a hipótese de que a variação na família de toxinas específica de cnidários tem sido o resultado de um regime de seleção positiva; e (3) determinar a extensão em que a duplicação de genes pode ser considerada como a principal razão para a diversificação de toxinas em Cnidaria. O capítulo \"Comparative proteomics reveals common components of a powerful arsenal in the earliest animal venomous lineage, the cnidarians\" propõe o estudo comparado mais completo sobre a composição do veneno de cnidários e uma hipótese sobre a montagem evolutiva do complexo arsenal bioquímico de cnidários e do veneno ancestral desse grupo basal. Vinte e oito famílias de proteínas foram identificadas. Destas, 13 famílias foram registradas pela primeira vez no proteoma de Cnidaria. Pelo menos 15 famílias de toxinas foram recrutadas no proteoma de veneno de cnidários antes da diversificação dos grupos Anthozoa e Medusozoa. Nos capítulos \"Evidence of episodic positive selection in the evolution of jellyfish toxins of the cnidarian venom\" e \"Gene duplications are extensive and contribute significantly to the toxic proteome of nematocysts isolated from Acropora digitifera (Cnidaria: Anthozoa: Scleractinia)\", nossas análises demonstram que as famílias de toxinas nos cnidários se diversificam amplamente mediante a duplicação de genes. Além disso, em contraste com as famílias de toxinas do veneno na maioria das linhagens animais; nós identificamos um padrão diferente na família de toxinas específica de cnidários, em que há uma seleção purificadora por longos períodos seguindo longos tempos de diversificação ou vice-versa

Análise de frequências alélicas de 15 marcadores STR em alunos da Faculdade de Odontologia de Bauru

Entre as muitas aplicações das tecnologias de identificação biológica humana, estão as finalidades forenses. O objetivo desta pesquisa foi verificar frequências alélicas de Short Tandem Repeat (STR) e os parâmetros estatísticos de interesse em genética de populações e forense para desenvolver o primeiro banco de dados populacional de DNA na Faculdade de Odontologia de Bauru, Universidade de São Paulo, (FOB/USP) para futuros usos forenses. Frequências alélicas de 15 locos autossômicos e do marcador de gênero amelogenina foram determinadas utilizando amostras de 200 μL de saliva doados por 296 alunos de graduação da FOB/USP, com idade ≥ 18 anos, após aprovação ética. Os testes laboratoriais foram feitos com kits comerciais. Resultados e parâmetros estatísticos foram obtidos por meio de programas clássicos: GeneMapper-ID-X, MS Excel 2002 versão 10.6871.6870, GenAlEx 6.5 e Arlequin 3.5, comparando quatro populações (brasileira, portuguesa, norte-americana e a população deste estudo). Os locos mais polimórficos foram D18S51 (17 alelos) e FGA (15 alelos), seguidos pelo D21S11 (13 alelos) e os menos polimórficos foram D16S539 e TH01 (7 alelos cada). A análise comparativa com amostra da população brasileira proveniente de estudos anteriores (n > 100.000) pelo teste goodness of fit X2 index não mostrou diferenças significativas entre estes grupos (p = 0,9999). Outros parâmetros estatísticos foram calculados comparando as populações: local (deste estudo), portuguesa e norte-americana. A análise de variância molecular (AMOVA) entre as três populações, entre as pessoas da mesma população e para cada pessoa de cada população mostrou que existe uma elevada variância individual (99%), que esta variância é mantida uniformemente entre as pessoas da mesma amostra/região (1%) e entre as três populações estudadas (0%). O estudo confirmou o elevado grau de polimorfismo e a alta heterozigosidade (96,5%) da população. Houve diferença significativa quanto ao gênero (79,7% mulheres) quando comparado à população brasileira em geral (50,4%), explicada pelas características do corpo discente da FOB/USP composto por 80,6% de pessoas do gênero feminino. Interessante foi a observação de uma microvariante alélica no loco D18S51, fora da escada padrão e da escala de abrangência do kit, correspondente ao alelo 29, ainda não definida na base de dados internacional (STRBase, atualizada em 07/08/2015). Esta microvariante deverá ser confirmada por testes familiares e sequenciamento de DNA para verificar a possibilidade de outra ocorrência familiar ou duplicação de nucleotídeos. No futuro, os dados obtidos neste estudo devem ser incorporados ao banco de dados da população brasileira e podem ser considerados como referência genética da população regional, ajudando a elucidar casos forenses. Após a confirmação, a potencial nova microvariante alélica contribuirá para a base de dados internacional STRBase.

Diffraction-managed superlensing using metallodielectric heterostructures

We show that subwavelength diffracted wave fields may be managed inside multilayered plasmonic devices to achieve ultra-resolving lensing. For that purpose we first transform both homogeneous waves and a broad band of evanescent waves into propagating Bloch modes by means of a metal/dielectric (MD) superlattice. Beam spreading is subsequently compensated by means of negative refraction in a plasmon-induced anisotropic effective-medium that is cemented behind. A precise design of the superlens doublet may lead to nearly aberration-free images with subwavelength resolution in spite of using optical paths longer than a wavelength.

Substantial enlargement of angular existence range for Dyakonov-like surface waves at semi-infinite metal-dielectric superlattice

We investigated surface waves guided by the boundary of a semi-infinite layered metal-dielectric nanostructure cut normally to the layers and a semi-infinite dielectric material. Using the Floquet-Bloch formalism, we found that Dyakonov-like surface waves with hybrid polarization can propagate in dramatically enhanced angular range compared to conventional birefringent materials. Our numerical simulations for an Ag-GaAs stack in contact with glass show a low to moderate influence of losses.

Use of Costas arrays in subpixel metrology

Subpixel methods increase the accuracy and efficiency of image detectors, processing units, and algorithms and provide very cost-effective systems for object tracking. A recently proposed method permits micropixel and submicropixel accuracies providing certain design constraints on the target are met. In this paper, we explore the use of Costas arrays - permutation matrices with ideal auto-ambiguity properties - for the design of such targets.

Innovative education networking aimed at multimedia tools for geometrical optics learning

We present a purposeful initiative to open new grounds for teaching Geometrical Optics. It is based on the creation of an innovative education networking involving academic staff from three Spanish universities linked together around Optics. Nowadays, students demand online resources such as innovative multimedia tools for complementing the understanding of their studies. Geometrical Optics relies on basics of light phenomena like reflection and refraction and the use of simple optical elements such as mirrors, prisms, lenses, and fibers. The mathematical treatment is simple and the equations are not too complicated. But from our long time experience in teaching to undergraduate students, we realize that important concepts are missed by these students because they do not work ray tracing as they should do. Moreover, Geometrical Optics laboratory is crucial by providing many short Optics experiments and thus stimulating students interest in the study of such a topic. Multimedia applications help teachers to cover those student demands. In that sense, our educational networking shares and develops online materials based on 1) video-tutorials of laboratory experiences and of ray tracing exercises, 2) different online platforms for student self-examinations and 3) computer assisted geometrical optics exercises. That will result in interesting educational synergies and promote student autonomy for learning Optics.

Chitosan enhances parasitism of Meloidogyne javanica eggs by the nematophagous fungus Pochonia chlamydosporia

Pochonia chlamydosporia (Pc), a nematophagous fungus and root endophyte, uses appressoria and extracellular enzymes, principally proteases, to infect the eggs of plant parasitic nematodes (PPN). Unlike other fungi, Pc is resistant to chitosan, a deacetylated form of chitin, used in agriculture as a biopesticide to control plant pathogens. In the present work, we show that chitosan increases Meloidogyne javanica egg parasitism by P. chlamydosporia. Using antibodies specific to the Pc enzymes VCP1 (a subtilisin), and SCP1 (a serine carboxypeptidase), we demonstrate chitosan elicitation of the fungal proteases during the parasitic process. Chitosan increases VCP1 immuno-labelling in the cell wall of Pc conidia, hyphal tips of germinating spores, and in appressoria on infected M. javanica eggs. These results support the role of proteases in egg parasitism by the fungus and their activation by chitosan. Phylogenetic analysis of the Pc genome reveals a large diversity of subtilisins (S8) and serine carboxypeptidases (S10). The VCP1 group in the S8 tree shows evidence of gene duplication indicating recent adaptations to nutrient sources. Our results demonstrate that chitosan enhances Pc infectivity of nematode eggs through increased proteolytic activities and appressoria formation and might be used to improve the efficacy of M. javanica biocontrol.

Política pública en el desarrollo de la juventud rural

La política pública es fundamental para cumplir con el propósito de atender los problemas colectivos a través de la negociación colectiva. En el medio rural se expresa en toda su amplitud, como agenda de formulación, de ejecución y de evaluación ajena a los actores municipales. De esta forma, este artículo tiene como objetivo comprobar los procesos, condiciones y efectos que genera en la población que las recibe, las acepta y se adapta a ellas. Desde el enfoque territorial, se encontró que la política pública y algunos programas dirigidos a los jóvenes, se centran en la juventud urbana; mientras que los jóvenes rurales desconocen las políticas públicas y los programas que son creados o aplicados a ellos. Las instancias de gobierno municipal, registran una falta de atención a los jóvenes, por la escases de estructura administrativa y de recursos económicos, que profundizan la ausencia de coordinación y comunicación entre los tres niveles de gobierno y la sociedad civil; causando que la duplicidad de programas con recursos limitados e insuficientes se sumen a los demás factores que limitan la atención de los jóvenes de la Sierra Norte de Puebla.

The white gene and locomotor recovery from anoxia in Drosophila melanogaster

Locomotor recovery from anoxia is complicated and little is known about the molecular and cellular mechanisms regulating anoxic recovery in Drosophila. For this thesis I established a protocol for large-scale analysis of locomotor activity in adult flies with exposure to a transient anoxia. Using this protocol I observed that wild-type Canton-S flies recovered faster and more consistently from anoxia than the white-eyed mutant w1118, which carries a null allele of w1118 in an isogenic genetic background. Both Canton-S and w1118 are commonly used controls in the Drosophila community. Genetic analysis including serial backcrossing, RNAi knockdown, w+ duplication to Y chromosome as well as gene mutation revealed a strong association between the white gene and the timing of locomotor recovery. I also found that the locomotor recovery phenotype is independent of white-associated eye pigmentation, that heterozygous w+ allele was haplo-insufficient to induce fast and consistent locomotor recovery from anoxia in female flies, and that mini-white is insufficient to promote fast and consistent locomotor recovery. Moreover, locomotor recovery was delayed in flies with RNAi knockdown of white in subsets of serotonin neurons in the central nervous system. I further demonstrated that mutations of phosphodiesterase genes (PDE) displayed wild-type-like fast and consistent locomotor recovery, and that locomotor recovery was light-sensitive in the night in w1118. The delayed locomotor recovery and the light sensitivity were eliminated in PDE mutants that were dual-specific or cyclic guanosine monophosphate (cGMP)-specific. Up-regulation of cGMP using multiple approaches including PDE mutation, sildenafil feeding or specific expression of an atypical soluble guanylyl cyclase (Gyc88E) was sufficient to suppress w-RNAi induced delay of locomotor recovery. Taken together, these data strongly support the hypothesis that White transports cGMP and promotes fast and consistent locomotor recovery from anoxia.

The Dynamics of Decentralization in Italy: Towards a Federal Solution? European Diversity and Autonomy Papers, EDAP/04 2013

This paper analyses the recent process of state decentralisation in Italy from the perspectives of political science and constitutional law. It considers the conflicting pressures and partisan opportunism of the decentralising process, and how these have adversely affected the consistency and completeness of the new constitutional framework. The paper evaluates the major institutional reforms affecting state decentralisation, including the 2001 constitutional reform and the more recent legislation on fiscal federalism. It argues that while the legal framework for decentralisation remains unclear and contradictory in parts, the Constitutional Court has performed a key role in interpreting the provisions and giving life to the decentralised system, in which regional governments now perform a much more prominent role. This new system of more decentralised multi-level government must nevertheless contend with a political culture and party system that remains highly centralised, while the administrative apparatus has undergone no comparable shift to take account of state decentralisation, leading to the duplication of bureaucracy at all territorial levels and continuing conflicts over policy jurisdiction. Unlike in federal systems these conflicts cannot be resolved in Italy through mechanisms of “shared rule”, since formal inter-governmental coordination structure are weak and entirely consultative.

Greater TTIP Ambition in Chemicals: Why and how. CEPS Special Report, 28 July 2015. Paper No. 10 in the CEPS-CTR project ‘TTIP in the Balance’ and CEPS Special Report No. 114/July 2015.

To date, the negotiations over chemicals in the Translatlantic Trade and Investment Partnership (TTIP) have not shown sufficient ambition. The talks have focused too much on the differences in the two ‘systems’, rather than on the actual levels of health and environmental protection for substances regulated by both the US and the EU. Given the accomplishments within the OECD and the UN Globally Harmonised System of Classification and Labelling of Chemicals (GHS), the question is whether TTIP can be any more ambitious in the area of chemicals? We find that there is no detailed or systematic knowledge about how the two levels of protection in chemicals compare, although caricatures and stereotypes abound. This is partly due to an obsessive focus on a single US federal law, the Toxic Subtances Control Act (TSCA), whereas in practice US protection depends on many statutes and regulations, as well as on voluntary withdrawals (under pressure from the Environmental Protection Agency) and severe common law liability. This paper makes the economic case for firmly addressing the regulatory barriers, discusses the EU’s proposals, finds that the European Parliament’s Resolution on TTIP of July 2015 lacks a rationale (for chemicals), argues that both TSCA and REACH ought to be improved (based on ‘better regulation’), discusses the link with a global regime, advocates significant improvement of market access where equivalence of health and environmental objectives is agreed and, finally, proposes to lower the costs for companies selling in both markets by allowing them to opt into the other party’s more stringent rules, thereby avoiding duplication while racing-to-the-top. The ‘living agreement’ on chemicals ought to be led by a new TTIP institution authorised to establish the level of health and environmental protection on both sides of the Atlantic for substances regulated on both sides. These findings will lay the foundation for a highly beneficial lowering of trading costs without in any way affecting the level of protection. Indeed, this is exactly what TTIP is, or should be, all about.This paper is the 10th in a series produced in the context of the “TTIP in the Balance” project, jointly organised by CEPS and the Center for Transatlantic Relations (CTR) in Washington, D.C. It is published simultaneously on the CEPS (www.ceps.eu) and CTR websites (http://transatlantic.sais-jhu.edu).

Rule-Makers or Rule-Takers? Exploring the Transatlantic Trade and Investment Partnership

The Transatlantic Trade and Investment Partnership (TTIP) is an effort by the United States and the European Union to reposition themselves for a world of diffuse economic power and intensified global competition. It is a next-generation economic negotiation that breaks the mould of traditional trade agreements. At the heart of the ongoing talks is the question whether and in which areas the two major democratic actors in the global economy can address costly frictions generated by their deep commercial integration by aligning rules and other instruments. The aim is to reduce duplication in various ways in areas where levels of regulatory protection are equivalent as well as to foster wide-ranging regulatory cooperation and set a benchmark for high-quality global norms. In this volume, European and American experts explain the economic context of TTIP and its geopolitical implications, and then explore the challenges and consequences of US-EU negotiations across numerous sensitive areas, ranging from food safety and public procurement to economic and regulatory assessments of technical barriers to trade, automotive, chemicals, energy, services, investor-state dispute settlement mechanisms and regulatory cooperation. Their insights cut through the confusion and tremendous public controversies now swirling around TTIP, and help decision-makers understand how the United States and the European Union can remain rule-makers rather than rule-takers in a globalising world in which their relative influence is waning.