Targeted development of informative microsatellite (SSR) markers

We describe a novel approach, selectively amplified microsatellite (SAM) analysis, for the targeted development of informative simple sequence repeat (SSR) markers. A modified selectively amplified microsatellite polymorphic loci assay is used to generate multi-locus SSR fingerprints that provide a source of polymorphic DNA markers (SAMs) for use in genetic studies. These polymorphisms capture the repeat length variation associated with SSRs and allow their chromosomal location to be determined prior to the expense of isolating and characterising individual loci. SAMs can then be converted to locus-specific SSR markers with the design and synthesis of a single primer specific to the conserved region flanking the repeat. This approach offers a cost-efficient and rapid method for developing SSR markers for predetermined chromosomal locations and of potential informativeness. The high recovery rate of useful SSR markers makes this strategy a valuable tool for population and genetic mapping studies. The utility of SAM analysis was demonstrated by the development of SSR markers in bread wheat.

Characterization of SU1 Isoamylase, a Determinant of Storage Starch Structure in Maize1

Function of the maize (Zea mays) gene sugary1 (su1) is required for normal starch biosynthesis in endosperm. Homozygous su1- mutant endosperms accumulate a highly branched polysaccharide, phytoglycogen, at the expense of the normal branched component of starch, amylopectin. These data suggest that both branched polysaccharides share a common precursor, and that the product of the su1 gene, designated SU1, participates in kernel starch biosynthesis. SU1 is similar in sequence to α-(1→6) glucan hydrolases (starch-debranching enzymes [DBEs]). Specific antibodies were produced and used to demonstrate that SU1 is a 79-kD protein that accumulates in endosperm coincident with the time of starch biosynthesis. Nearly full-length SU1 was expressed in Escherichia coli and purified to apparent homogeneity. Two biochemical assays confirmed that SU1 hydrolyzes α-(1→6) linkages in branched polysaccharides. Determination of the specific activity of SU1 toward various substrates enabled its classification as an isoamylase. Previous studies had shown, however, that su1- mutant endosperms are deficient in a different type of DBE, a pullulanase (or R enzyme). Immunoblot analyses revealed that both SU1 and a protein detected by antibodies specific for the rice (Oryza sativa) R enzyme are missing from su1- mutant kernels. These data support the hypothesis that DBEs are directly involved in starch biosynthesis.

Bootstrap confidence levels for phylogenetic trees.

Evolutionary trees are often estimated from DNA or RNA sequence data. How much confidence should we have in the estimated trees? In 1985, Felsenstein [Felsenstein, J. (1985) Evolution 39, 783-791] suggested the use of the bootstrap to answer this question. Felsenstein's method, which in concept is a straightforward application of the bootstrap, is widely used, but has been criticized as biased in the genetics literature. This paper concerns the use of the bootstrap in the tree problem. We show that Felsenstein's method is not biased, but that it can be corrected to better agree with standard ideas of confidence levels and hypothesis testing. These corrections can be made by using the more elaborate bootstrap method presented here, at the expense of considerably more computation.

The acceleration and collimation of jets.

I will discuss several issues related to the acceleration, collimation, and propagation of jets from active galactic nuclei. Hydromagnetic stresses provide the best bet for both accelerating relativistic flows and providing a certain amount of initial collimation. However, there are limits to how much "self-collimation" can be achieved without the help of an external pressurized medium. Moreover, existing models, which postulate highly organized poloidal flux near the base of the flow, are probably unrealistic. Instead, a large fraction of the magnetic energy may reside in highly disorganized "chaotic" fields. Such a field can also accelerate the flow to relativistic speeds, in some cases with greater efficiency than highly organized fields, but at the expense of self-collimation. The observational interpretation of jet physics is still hampered by a dearth of unambiguous diagnostics. Propagating disturbances in flows, such as the oblique shocks that may constitute the kiloparsec-scale "knots" in the M87 jet, may provide a wide range of untapped diagnostics for jet properties.

Coupling the phosphotransferase system and the methyl-accepting chemotaxis protein-dependent chemotaxis signaling pathways of Escherichia coli.

Chemotactic responses in Escherichia coli are typically mediated by transmembrane receptors that monitor chemoeffector levels with periplasmic binding domains and communicate with the flagellar motors through two cytoplasmic proteins, CheA and CheY. CheA autophosphorylates and then donates its phosphate to CheY, which in turn controls flagellar rotation. E. coli also exhibits chemotactic responses to substrates that are transported by the phosphoenolpyruvate (PEP)-dependent carbohydrate phosphotransferase system (PTS). Unlike conventional chemoreception, PTS substrates are sensed during their uptake and concomitant phosphorylation by the cell. The phosphoryl groups are transferred from PEP to the carbohydrates through two common intermediates, enzyme I (EI) and phosphohistidine carrier protein (HPr), and then to sugar-specific enzymes II. We found that in mutant strains HPr-like proteins could substitute for HPr in transport but did not mediate chemotactic signaling. In in vitro assays, these proteins exhibited reduced phosphotransfer rates from EI, indicating that the phosphorylation state of EI might link the PTS phospho-relay to the flagellar signaling pathway. Tests with purified proteins revealed that unphosphorylated EI inhibited CheA autophosphorylation, whereas phosphorylated EI did not. These findings suggest the following model for signal transduction in PTS-dependent chemotaxis. During uptake of a PTS carbohydrate, EI is dephosphorylated more rapidly by HPr than it is phosphorylated at the expense of PEP. Consequently, unphosphorylated EI builds up and inhibits CheA autophosphorylation. This slows the flow of phosphates to CheY, eliciting an up-gradient swimming response by the cell.

Estudo da seletividade de catalisadores a base de cobre e paládio em transformações de hidrocarbonetos insaturados

Compreender a correlação entre as características de um catalisador particular e seu desempenho catalítico tem sido um dos principais objetos da pesquisa em catálise heterogênea a fim de usar esse conhecimento para o desenho racional de catalisadores mais ativos, seletivos e estáveis. A seletividade é um dos fatores mais importantes a ser controlado pelo desenho de catalisadores, podendo ser alcançada de diversas maneiras, levando-se em consideração mudanças do tipo estrutural, química, eletrônica, de composição, de cinética e de energia. O trabalho descrito nessa tese de doutorado compreende a síntese e caracterização de catalisadores compostos de nanopartículas de óxido de cobre, paládio e cobre-paládio e seu estudo em reações de hidrogenação e oxidação seletivas de hidrocarbonetos insaturados. Os catalisadores foram preparados através da deposição de nanopartículas dos metais cataliticamente ativos sobre suportes magneticamente recuperáveis compostos de nanopartículas de magnetita revestidas por sílica com superfícies funcionalizada com diferentes grupos orgânicos. A natureza magnética do suporte permitiu a fácil separação do catalisador do meio reacional pela simples aproximação de um ímã na parede do reator. O catalisador pôde ser completamente separado da fase líquida, fazendo com que a utilização de outros métodos de separação como filtração e centrifugação, comumente utilizados em sistemas heterogêneos líquidos, fossem completamente dispensados. Os catalisadores foram inicialmente testados em reações de hidrogenação de alquenos e alquinos. As reações de hidrogenação foram realizadas utilizando hidrogênio molecular como agente redutor, dispensando a utilização de agentes redutores mais agressivos. Os catalisadores compostos de NPs de Pd mostram excelente atividade e capacidade de reutilização na hidrogenação de cicloexeno, podendo ser utilizados em até 15 ciclos sem perda de atividade. Nas reações de hidrogenação de alquinos, os catalisadores que contêm cobre mostraram maior seletividade para a obtenção dos produtos de semi-hidrogenação, com destaque para o catalisador composto de NPs de CuPd, que não apresenta nem traços do produto de hidrogenação completa na amostra final. Esse catalisador bimetálico alia as características do paládio (elevada atividade) e do cobre (elevada seletividade) para fornecer um catalisador ativo e seletivo para a transformação desejada. Além disso, os grupos funcionais presentes na superfície do suporte catalítico mostraram influência na atividade e seletividade para a hidrogenação de alquenos e alquinos. Os catalisadores sintetizados também foram testados na reação de oxidação de cicloexeno e mostraram seletividade para a produção do composto carbonílico α,β-insaturado, cicloex-2-en-1-ona, que é um reagente de partida de grande interesse para a síntese de diversos materiais na indústria química. As reações de oxidação foram realizadas utilizando-se apenas O2 como oxidante primário, dispensando o uso de oxidantes tóxicos como cromatos, permanganatos ou compostos halogenados, que não são recomendados do ponto de vista ambiental. Os catalisadores sintetizados puderam ser reutilizados em sucessivos ciclos de oxidação, mostrando seletividade para a formação dos produtos alílicos em todos os ciclos. Os catalisadores foram estáveis sob as condições reacionais e não apresentaram problemas de lixiviação da espécie ativa para o meio reacional, que é comum na catálise heterogênea. Um estudo cinético mostrou que, mesmo no início da reação, o catalisador tem seletividade para a ocorrência de oxidação alílica em detrimento da reação de oxidação direta que dá origem ao epóxidos correspondente, e se mostrou condizente com o mecanismo proposto na literatura para a reação de oxidação de alquenos via radicalar.

Discursive constructions of social and environmental accounting in Brazil: the case of Petrobras.

In the 29 years following \"Our Common Future\" by the United Nations, there is considerable debate among governments, civil society, interest groups and business organisations about what constitutes sustainable development, which constitutes evidence for a contested discourse concerning sustainability. The purpose of this study is to understand this debate in the developing economic context of Brazil, and in particular, to understand and critique the social and environmental accounting [SEA] discursive constructions relating to the State-owned, Petrobras as well as to understand the Brazilian literature on SEA. The discourse theory [DT]-based analysis employs rhetorical redescription to analyse twenty-two reports from Petrobras from 2004-2013. I investigate the political notions by employing the methodological framework of the Logics of Critical Explanation [LCE]. LCE engenders five methodological steps: problematisation, retroduction, logics (social, political and fantasmatic), articulation and critique. The empirical discussion suggests that the hegemony of economic development operates to obfuscate, rhetorically, the development of sustainability, so as to maintain the core business of Petrobras conceived as capital accumulation. Equally, these articulations also illustrate how the constructions of SEA operate to serve the company\'s purpose with few (none) profound changes in integration of sustainability. The Brazilian literature on SEA sustains the status quo of neo-liberal market policies that operate to protect the dominant business case approach to maintain an agenda of wealth-creation in relation to social and environmental needs. The articulations of the case manifested in policies regarding, for example, corruption, which involved over-payments for contracts and unsustainable practices relating to the use of fossil fuels and demonstrated that there was antagonism between action and disclosure. The corruption scandal that emerged after SEA disclosures highlighted the rhetorical nature of disclosure when financial resources were subtracted from the company for political parties and engineering contractors hid facts through incomplete disclosures. The articulations of SEA misrepresent a broader context of the meanings associated with sustainability, which restricted the constructions of SEA to principally serve and represent the intention of the most powerful groups. The significance of SEA, then is narrowed to represent particular interests. The study argues for more critical studies as limited Brazilian literature concerning SEA kept a \'safe distance\' from substantively critiquing the constructions of SEA and its articulations in the Brazilian context. The literature review and the Petrobras\' case illustrate a variety of naming, instituting and articulatory practices that endeavour to maintain the current hegemony of development in an emerging economy, which allows Petrobras to continue to exercise significant profit at the expense of the social and environmental. The constructed idea of development in Petrobras\' discourses emphasises a rhetoric of wider development, but, in reality, these discourses were the antithesis of political, social and ethical developmental issues. These constructions aim to hide struggles between social inequalities and exploitation of natural resources and constitute excuses about a fanciful notion of rhetorical and hegemonic neo-liberal development. In summary, this thesis contributes to the prior literature in five ways: (i) the addition of DT to the analysis of SEA enhances the discussion of political elements such as hegemony, antagonism, logic of equivalence/difference, ideology and articulation; (ii) the analysis of an emerging economy such as Brazil incorporates a new perspective of the discussion of the discourses of SEA and development; (iii) this thesis includes a focus on rhetoric to discuss the maintenance of the status quo; (iv) the holistic structure of the LCE approach enlarges the understanding of the social, political and fantasmatic logics of SEA studies and; (v) this thesis combines an analysis of the literature and the case of Petrobras to characterise and critique the state of the Brazilian academy and its impacts and reflections on the significance of SEA. This thesis, therefore, argues for more critical studies in the Brazilian academy due to the persistence of idea of SEA and development that takes-for-granted deep exclusions and contradictions and provide little space for critiques.

Exploring Existentially-Based Interventions For Career Transitions in Sport

From Introduction: Career transition issues have become of increasing interest in the field of sport psychology. Confronting the end of an athletic career is an inevitable reality that every athlete will confront in his or her lifetime (Baillie, 1993), regardless of level of competition (Kerr & Dacyshyn, 2000) or the amount of free choice related to the transition. Many athletes are able to cope with the effects of the transition process effectively, and see retirement as an opportunity to pursue new ventures and identity roles in life. However, retirement from sport can be an event that often results in various adjustment difficulties for an athlete involving emotional, social, financial, and vocational conflicts. Some athletes have reported experiencing effects such as depression, eating disorders, decreased self-esteem, increased suicidality, and substance abuse (Kerr and Dacyshyn, 2000). These types of distress can be exacerbated by the fact that many athletes fail to adequately anticipate and prepare for their impending transition (Baillie, 1993), and often embark on the retirement process without any formalized support (Stier, 2007).Typically, the role of a sport psychologist has been to assist in maximizing an athlete's competitive performance during the course of their career. However, as a sport psychologist's primary responsibility is to serve active competitors and athletic organizations, this tends to come at the expense of failing to provide follow-up care for the athlete as he or she retires from sport (Taylor, Ogilvie, and Lavallee, 2006). Since the 1970's, when the efforts of professionals in European sports organizations first received attention, there has been growing interest in academic circles about career transition

Sustainable Development and its Discontents

Sustainable development (or sustainability) is a decision-making framework for maintaining and achieving human well-being, both in the present and into the future. The framework requires both consideration and achievement of environmental protection, social justice and economic development. In that framework, environmental protection must be integrated into decisions about social and economic development, and social justice and economic viability must be integrated into decisions about environmental quality. First endorsed by the world’s nations in 1992, this framework is intended to provide an effective response to the twin global challenges of growing environmental degradation and widespread extreme poverty. Sustainability provides a framework for humans to live in harmony with nature, rather than at nature’s expense. It may therefore be one of the most important ideas to come out of the 20th century. In the last two decades, the framework has become a touchstone in nearly every economic sector and at every level of government, unleashing an extraordinary range of creativity in all of those realms. Sustainable development is having a significant effect on the practice of law and on the way in which laws are written and implemented. Understanding the framework is increasingly important for law makers and lawyers. As sustainable development (or sustainability) has grown in prominence, its critics have become more numerous and more vocal. Three major lines of criticism are that the term is “too boring” to command public attention, “too vague” to provide guidance, and “too late” to address the world’s problems. Critics suggest goals such as abundance, environmental integrity, and resilience. Beginning with the international agreements that shaped the concept of sustainable development, this Article provides a functional and historical analysis of the meaning of sustainable development. It then analyzes and responds to each of these criticisms in turn. While the critics, understood constructively, suggest ways of strengthening this framework, they do not provide a compelling alternative. The challenge for lawyers, law makers, and others is to use and improve this framework to make better decisions.

La cobertura periodística de la obesidad en la prensa española (2000-2005)

Antecedentes: la obesidad es un problema de salud pública en España. Los medios de comunicación son una herramienta útil para la salud pública. Objetivo: explorar el tratamiento periodístico de la obesidad en la prensa escrita española durante 2000-2005, frecuencia de aparición, fuentes de información y enfoques, en relación con el contexto social. Materiales y métodos: análisis de contenido cuantitativo de 690 noticias publicadas en El País, El Mundo y ABC. Cálculo de frecuencias y odds ratio (OR) con intervalos de confianza (IC) del 95% y significación estadística. Resultados: incrementaron las noticias de 2000 (n=25) a 2005 (n=185). Se centraron en denuncias (36,4%) y magnitud del problema (15,7%), en detrimento de aquellas sobre iniciativas-estrategias políticas (3,8%). Destacan los hombres como fuentes informativas principales (75,5%) y las mujeres como primeras firmantes de las noticias (56,1%). Los hombres del ámbito médico-sanitario (OR=1,98;IC95%,1,11-3,57) y las mujeres del ámbito político (OR=2,54;IC95%1,46-4,42) tienen mayor probabilidad de ser la fuente informativa principal. Conclusiones: la cobertura periodística de la obesidad durante 2000-2005 aumentó, coincidiendo con el desarrollo de respuestas políticas en torno al tema. Principalmente, se denuncia el problema. Destaca la escasa cobertura periodística de iniciativas-estrategias políticas, sugiriendo incipiente interacción entre la agenda política y mediática.

Sistema Nacional de Salud español. Características y análisis

La Ley General de Sanidad 14/1986 de 25 de Abril posibilitó el tránsito del antiguo modelo de Seguridad Social al actual modelo de Sistema Nacional de Salud (SNS), financiado con impuestos y de cobertura prácticamente universal. Desde entonces se han producido profundos cambios en el sistema que culminaron en el año 2002 con la descentralización total de competencias en materia de salud en las Comunidades Autónomas. La regulación nacional de competencias en materia de salud se realiza desde el Consejo Interterritorial del Sistema Nacional de Salud, organismo que agrupa a los máximos responsables autonómicos en materia de salud de cada Comunidad Autónoma y que tiene entre otras, la responsabilidad de evitar las desigualdades en servicios sanitarios dentro del territorio nacional. La creación y competencias del Consejo Interterritorial quedan recogidas en la Ley 16/2003 de 28 de mayo de Cohesión de la calidad del Sistema Nacional de Salud. La cartera de servicios comunes del SNS se establece en el Real Decreto 1030/2006 de 15 de Septiembre, actualizando el Real Decreto 63/1995 de 20 de enero sobre Ordenación de las prestaciones sanitarias, resultando del actual marco legislativo con la descentralización de competencias y gestión de los presupuestos un horizonte de posible variabilidad en los modelos de gestión de cada CCAA, que, si bien deben garantizar la universalidad de las prestaciones, también ofrece una diversidad de modalidades de gestionar los recursos en materia de salud. En cuanto al estado de salud de los españoles, destacar que la esperanza de vida al nacer se sitúa en 79,9 años, superior a la media europea, 78,3 años, y la esperanza de vida ajustada por incapacidad fue en 2002 de 72,6 años en España respecto a los 70,8 de la UE. Según cifras del propio Ministerio de Sanidad, la percepción de la salud de los ciudadanos fue positiva para un 73% de los hombres y un 63,2 de las mujeres. Alrededor del 60% de la población tiene un peso normal y la morbilidad sitúa en los primeros lugares las enfermedades del aparato circulatorio, el cáncer y las enfermedades del aparato respiratorio (CIE-9). El gasto sanitario en España, es un capítulo presupuestario importante, al situarse en torno al 7,5 del P.I.B, y los recursos e inversiones presentan aparentes desigualdades autonómicas. Los modelos de gestión y dependencia patrimonial de los recursos, variables entre Autonomías, plantean la necesidad de monitorizar un seguimiento que permita evaluar en los próximos diez años el impacto de la descentralización de competencias del Sistema. La estructura del Sistema tiene dos niveles asistenciales mayoritarios, atención primaria y especializada, absorbiendo la atención especializada la mayor parte del presupuesto. El incremento del gasto sanitario y la universalidad de las prestaciones han condicionado en gran medida la implantación de modelos de gestión diferentes a los tradicionales. Esta situación no es exclusiva del Estado Español. En los Estados del entorno de la Unión Europea, el Consejo de Ministros de Sanidad de la UE en su sesión celebrada los días 1 y 2 de Junio de 200625 concluyeron un documento que recoge los valores y principios comunes de los sistemas sanitarios de los países de la Unión Europea, resaltando los principios y valores de los sistemas sanitarios como soporte estructural de dichos estados. Como conclusión, en este momento (2007) el Sistema Nacional de Salud Español, está inmerso en un proceso de trasformación orientado a garantizar la eficiencia de las prestaciones de manera responsable, es decir, ofertar al ciudadano la mejor calidad de servicios al mínimo coste.

Colonización y formación de un paisaje agrario. El caso de San Antonio (Requena). 1750-1920

San Antonio, actual pedanía de 2.100 hb situada a 5 km de la ciudad de Requena, es un ejemplo de transformación del paisaje por efecto de la colonización humana. En 1752 poblaban esta partida dos docenas de familias de labradores, jornaleros y pastores repartidos en una veintena de casas dispersas por la campiña. La propiedad de la tierra estaba en manos de una docena de terratenientes forasteros y del concejo municipal de Requena. La superficie cultivada ascendía a uno 600 almudes de regadío y 1.000 de secano. Cien años más tarde había 730 en riego y 3.700 en secano. Este incremento estuvo acompañado de una expansión del viñedo y los cereales, a costa de pastizales y baldíos, y de una distribución y traspaso parcial de la propiedad de la tierra a manos de los colonos que se multiplicaron por diez en poco más de un siglo. Las causas que propiciaron aquella transformación fueron básicamente dos: la desamortización y venta de bienes concejiles y eclesiásticos, y la proliferación de contratos de plantación de viña a medias, mediante los cuales decenas de jornaleros lograron hacerse con una pequeña propiedad vitícola.

Receipt signed by James Winthrop, 1772 December 10

Handwritten receipt signed by James Winthrop acknowledging money received from the Steward, also signed by Steward Jonathan Hastings. A handwritten transcription of the Corporation vote on December 10, 1772 granting money for caring for the College Library since May 1, 1772, is signed by President Samuel Locke on other side of the document.

Tutors grants for £77.8.4, 1773

Handwritten receipt signed by Joseph Willard, John Marsh, John Marsh, Andrew Eliot Jr., John Wadsworth, and S. Hall acknowledging money received between August 20th and 23rd, 1773. A handwritten transcription of the Corporation vote on September 7, 1772 that, "the Tutors be allowed in addition to their salary twenty pounds," is signed as a true copy by President Samuel Locke at the beginning of the document.

The expenses of a dinner provided at the installation of the Rev'd. Professor Williams, 1780 May 2

Handwritten one-page itemized list of costs for food and supplies purchased for the installation dinner of Samuel Williams (1743-1817) for his appointment as Hollis Professor Mathematics and Natural Philosophy at Harvard College. Signed by Caleb Gannett.