Characterization of the Saccharomyces cerevisiae ERG27 gene encoding the 3-keto reductase involved in C-4 sterol demethylation

The last unidentified gene encoding an enzyme involved in ergosterol biosynthesis in Saccharomyces cerevisiae has been cloned. This gene, designated ERG27, encodes the 3-keto sterol reductase, which, in concert with the C-4 sterol methyloxidase (ERG25) and the C-3 sterol dehydrogenase (ERG26), catalyzes the sequential removal of the two methyl groups at the sterol C-4 position. We developed a strategy to isolate a mutant deficient in converting 3-keto to 3-hydroxy-sterols. An ergosterol auxotroph unable to synthesize sterol or grow without sterol supplementation was mutagenized. Colonies were then selected that were nystatin-resistant in the presence of 3-ketoergostadiene and cholesterol. A new ergosterol auxotroph unable to grow on 3-ketosterols without the addition of cholesterol was isolated. The gene (YLR100w) was identified by complementation. Segregants containing the YLR100w disruption failed to grow on various types of 3-keto sterol substrates. Surprisingly, when erg27 was grown on cholesterol- or ergosterol-supplemented media, the endogenous compounds that accumulated were noncyclic sterol intermediates (squalene, squalene epoxide, and squalene dioxide), and there was little or no accumulation of lanosterol or 3-ketosterols. Feeding experiments in which erg27 strains were supplemented with lanosterol (an upstream intermediate of the C-4 demethylation process) and cholesterol (an end-product sterol) demonstrated accumulation of four types of 3-keto sterols identified by GC/MS and chromatographic properties: 4-methyl-zymosterone, zymosterone, 4-methyl-fecosterone, and ergosta-7,24 (28)-dien-3-one. In addition, a fifth intermediate was isolated and identified by 1H NMR as a 4-methyl-24,25-epoxy-cholesta-7-en-3-one. Implications of these results are discussed.

Dynamics of maize endosperm development and DNA endoreduplication.

Endosperm development in Zea mays is characterized by a period of intense mitotic activity followed by a period in which mitosis is essentially eliminated and the cell cycle becomes one of alternating S and G phases, leading to endoreduplication of the nuclear DNA. The endosperm represents a significant contribution to the grain yield of maize; thus, methods that facilitate the study of cellular kinetics may be useful in discerning cellular and molecular components of grain yield. Two mathematical models have been developed to describe the kinetics of endosperm growth. The first describes the kinetics of mitosis during endosperm development; the second describes the kinetics of DNA endoreduplication during endosperm development. The mitotic model is a modification of standard growth curves. The endoreduplication model is composed of six differential equations that represent the progression of nuclei from one DNA content to another during the endoreduplication process. Total nuclei number per endosperm and the number of 3C, 6C, 12C, 24C, 48C, and 96C nuclei per endosperm (C is the haploid DNA content per nucleus) for inbred W64A from 8 to 18 days after pollination were determined by flow cytometry. The results indicate that the change in number of nuclei expressed as a function of the number of days after pollination is the same from one yearly crop to another. These data were used in the model to determine the endosperm growth rate, the maximum nuclei number per endosperm, and transition rates from one C value to the next higher C value. The kinetics of endosperm development are reasonably well represented by the models. Thus, the models provide a means to quantify the complex pattern of endosperm development.

Social Citizenship and Institution Building: EU-Enlargement and the Restructuring of Welfare States in East Central Europe. CES Germany & Europe Working Paper, No. 01.2, April 2001

With the EU-enlargement process well underway, this paper focuses on social citizenship as a conceptual frame for analyzing the restructuring of social institutions in applicant countries in East Central Europe. So far, comparative welfare state analysis has concentrated mainly on the developed economies of the OECD-countries; there is little systematic analytical work on the transitions in post-communist Europe. Theoretically, this paper builds on comparative welfare state analysis as well as on new institutionalism. The initial hypothesis is built on the assumption that emerging patterns of social support and social security diverge from the typology described in the comparative welfare state literature inasmuch as the transformation of postcommunist societies is distinctly different from the building of welfare states in Europe. The paper argues that institutionbuilding is shaped by and embedded in the process of European integration and part of governance in the EU. Anticipating full membership in the European Union, the applicant countries have to adapt to the rules and regulations of the EU, including the "social acquis." Therefore, framing becomes an important feature of institutional changes. The paper seeks to identify distinct patterns and problems of the institutionalization of social citizenship.

COACHING E MENTORING COMO PRÁTICAS DE COMPARTILHAMENTO DO CONHECIMENTO: ESTUDOS DE CASOS EM EMPRESAS PÚBLICAS PAULISTAS

A pesquisa teve como objetivo geral analisar as principais razões pelas quais as empresas públicas paulistas utilizam coaching e mentoring como práticas de compartilhamento de conhecimento. No ano de 2009, foi instituído pelo governador do Estado de São Paulo, o decreto nº 53.963 que instituiu a Política de Gestão do Conhecimento e Inovação para as empresas públicas. Kuniyoshi e Santos (2007) realizaram uma pesquisa, na qual identificaram práticas e iniciativas de gestão do conhecimento adotadas por algumas empresas, dentre elas, coaching e mentoring. As práticas são processos que necessitam de investimento não somente financeiro, mas de tempo e pessoas adequadas, por serem processos mais complexos, instigam a investigação de ações no contexto organizacional de empresas públicas. Este estudo busca contribuir para o desenvolvimento de estudos na área pública. O método utilizado neste estudo de abordagem qualitativa é do tipo exploratória. O objeto desta pesquisa foram as empresas públicas paulistas, que, atualmente, somam 21. Foi realizado estudo de caso, com entrevista e análise documental em duas destas empresas, A Sabesp, empresa do segmento de saneamento de água e esgoto, teve como objetivo analisar a prática de coaching e, o Instituto de Pesquisa Tecnológicas (IPT), referência nacional em metrologia, teve como objetivo analisar a prática de mentoring. Uma vez que não existem práticas exclusivas à Gestão do Conhecimento, e o sucesso de uma prática está relacionado ao contexto na qual está inserida. No caso da Sabesp, a prática de coaching é utilizada como uma das atividades dentro de dois programas, visando desenvolver o capital humano como força competitiva. O IPT teve como objetivo da aplicação do programa de mentoring, especificamente, o compartilhar conhecimento tácito. Foi constatado que as práticas de coaching e mentoring podem ser utilizadas como recurso capaz de tornar a empresa singular perante as demais, mesmo empresas públicas não tendo foco em competitividade, mas utilizam o conhecimento de forma estratégica para melhorar a qualidade de atendimento à sociedade.

Effects of oral vitamin C on monocyte:Endothelial cell adhesion in healthy subjects

Monocyte recruitment and retention in the vasculature is influenced by oxidative stress and is involved in cardiovascular disease (CVD). Individuals with low plasma ascorbate are at elevated risk of CVD. It is unknown whether vitamin C supplementation affects monocyte adhesion to endothelial cells (ECs) in healthy non-smokers. In a randomised double-blind crossover study the effect of vitamin C supplementation (six weeks, 250 mg/day) was determined in subjects with normal (HIC) and below average (LOC) plasma vitamin C concentration at baseline (mean = 67μM, n = 20, mean = 32μM, n = 20, respectively). LOC subjects showed 30% greater monocyte adhesion to ECs. This was significantly reduced by 37% (P < 0.02) following vitamin C supplementation to levels of HIC monocyte adhesion. No differences in plasma malondialdehyde concentrations were observed between groups or after supplementation. In conclusion, vitamin C supplementation normalises monocyte adhesion in subjects with low plasma vitamin C (LOC). This process may be related to a direct effect on monocytes, independent of lipid peroxidation. © 2002 Elsevier Science (USA). All rights reserved.

Outsourcing in the innovation process: locational and strategic determinants

There is now substantial evidence that locational and agglomeration influences can have a significant positive effect on innovation performance. Networking and boundary-spanning activities are also increasingly recognised as important contributors to innovation success. In this article we attempt to discover whether these factors are associated: in particular, is there any link between plant location, agglomeration effects and the extent of outsourcing in the innovation process? Using data for a large sample of UK and German manufacturing plants, we find that organisational and strategic factors play a much greater and more consistent role than locational influences in shaping the level of outsourcing in the innovation process. Strategic approaches to outsourcing may also benefit plants in obtaining economies of scope in the management or governance of outsourcing within the innovation process.

Modelling the innovation value chain

Innovation events – the introduction of new products or processes – represent the end of a process of knowledge sourcing and transformation. They also represent the beginning of a process of exploitation which may result in an improvement in the performance of the innovating business. This recursive process of knowledge sourcing, transformation and exploitation comprises the innovation value chain. Modelling the innovation value chain for a large group of manufacturing firms in Ireland and Northern Ireland highlights the drivers of innovation, productivity and firm growth. In terms of knowledge sourcing,we find strong complementarity between horizontal, forwards, backwards, public and internal knowledge sourcing activities. Each of these forms of knowledge sourcing also makes a positive contribution to innovation in both products and processes although public knowledge sources have only an indirect effect on innovation outputs. In the exploitation phase, innovation in both products and processes contribute positively tocompany growth, with product innovation having a short-term ‘disruption’ effect on labour productivity. Modelling the complete innovation value chain highlights the structure and complexity of the process of translating knowledge into business value and emphasises the role of skills, capital investment and firms’ other resources in the value creation process.

The innovation value chain

Innovation events - the introduction of new products or processes - represent the end of a process of knowledge sourcing and transformation. They also represent the beginning of a process of exploitation which may result in an improvement in the performance of the innovating business. This recursive process of knowledge sourcing, transformation and exploitation we call the innovation value chain. Modelling the innovation value chain for a large group of manufacturing firms in Ireland and Northern Ireland highlights the drivers of innovation, productivity and firm growth. In terms of knowledge sourcing, we find strong complementarity between horizontal, forwards, backwards, public and internal knowledge sourcing activities. Each of these forms of knowledge sourcing also makes a positive contribution to innovation in both products and processes although public knowledge sources have only an indirect effect on innovation outputs. In the exploitation phase, innovation in both products and processes contribute positively to company growth, with product innovation having a short-term ‘disruption’ effect on labour productivity. Modelling the complete innovation value chain highlights the structure and complexity of the process of translating knowledge into business value and emphasises the role of skills, capital investment and firms’ other resources in the value creation process.

An experimental analysis of operating conditions in cold roll-forming

A detailed literature survey confirmed cold roll-forming to be a complex and little understood process. In spite of its growing value, the process remains largely un-automated with few principles used in set-up of the rolling mill. This work concentrates on experimental investigations of operating conditions in order to gain a scientific understanding of the process. The operating conditions are; inter-pass distance, roll load, roll speed, horizontal roll alignment. Fifty tests have been carried out under varied operating conditions, measuring section quality and longitudinal straining to give a picture of bending. A channel section was chosen for its simplicity and compatibility with previous work. Quality measurements were measured in terms of vertical bow, twist and cross-sectional geometric accuracy, and a complete method of classifying quality has been devised. The longitudinal strain profile was recorded, by the use of strain gauges attached to the strip surface at five locations. Parameter control is shown to be important in allowing consistency in section quality. At present rolling mills are constructed with large tolerances on operating conditions. By reduction of the variability in parameters, section consistency is maintained and mill down-time is reduced. Roll load, alignment and differential roll speed are all shown to affect quality, and can be used to control quality. Set-up time is reduced by improving the design of the mill so that parameter values can be measured and set, without the need for judgment by eye. Values of parameters can be guided by models of the process, although elements of experience are still unavoidable. Despite increased parameter control, section quality is variable, if only due to variability in strip material properties. Parameters must therefore be changed during rolling. Ideally this can take place by closed-loop feedback control. Future work lies in overcoming the problems connected with this control.

Value creation in a reregulated market : Divestments, firms and dynamic processes

This thesis makes use of the unique reregulation of pharmaceutical monopoly in Sweden to critically examine intraindustry firm heterogeneity. It contributes to existing divestiture research as it studies the dynamism in between reconfigurations of value constellations and its effects on value creation of divested pharmacies. Because the findings showed that the predominant theory of intraindustry firm heterogeneity could not explain firm performance, the value constellation concept was applied as it captured the phenomena. A patterned finding informed how reconfigurations of value constellations in a reregulated market characterized by strict rules, regulations, and high competition did not generate additional value for firms on short term. My study unveils that value creation is hampered in situations where rules and regulations significantly affect firms’ ability to reconfigure their value constellations. The key practical implication is an alternative perspective on fundamental aspects of the reregulation and how policy-makers may impede firm performance and the intended creation of new value for not only firms but for society as a whole.

I film therefore I am: process and participation, networks and knowledge – examples from Scottish community media projects.

This chapter examines community media projects in Scotland as social processes that nurture knowledge through participation in production. A visual and media anthropology framework (Ginsburg, 2005) with an emphasis on the social context of media production informs the analysis of community media. Drawing on community media projects in the Govan area of Glasgow and the Isle of Bute, the techniques of production foreground “the relational aspects of filmmaking” (Grimshaw and Ravetz, 2005: 7) and act as a catalyst for knowledge and networks of relations embedded in time and place. Community media is defined here as a creative social process, characterised by an approach to production that is multi-authored, collaborative and informed by the lives of participants, and which recognises the relevance of networks of relations to that practice (Caines, 2007: 2). As a networked process, community media production is recognised as existing in collaboration between a director or producer, such as myself, and organisations, institutions and participants, who are connected through a range of identities, practices and place. These relations born of the production process reflect a complex area of practice and participation that brings together “parallel and overlapping public spheres” (Meadows et al., 2002: 3). This relates to broader concerns with networks (Carpentier, Servaes and Lie, 2003; Rodríguez, 2001), both revealed during the process of production and enhanced by it, and how they can be described with reference to the knowledge practice of community media.

Novel method for measuring nanofriction by atomic force microscope

The authors describe a novel approach to the measurement of nanofriction, and demonstrate the application of the method by measurement of the coefficient of friction for diamondlike carbon (DLC) on DLC, Si on DLC, and Si on Si surfaces. The technique employs an atomic force microscope in a mode in which the tip moves only in the z (vertical) direction and the sample surface is sloped. As the tip moves vertically on the sloped surface, lateral tip slipping occurs, allowing the cantilever vertical deflection and the frictional (lateral) force to be monitored as a function of tip vertical deflection. The advantage of the approach is that cantilever calibration to obtain its spring constants is not necessary. Using this method, the authors have measured friction coefficients, for load range 0 < L M 6 mu N, of 0.047 +/- 0.002 for Si on Si, 0.0173 +/- 0.0009 for Si on DLC, and 0.0080 +/- 0.0005 for DLC on DLC. For load range 9 < L < 13 mu N, the DLC on DLC coefficient of friction increased to 0.051 +/- 0.003. (C) 2008 American Vacuum Society.

Gene expression analysis of Paracoccidioides brasiliensis transition from conidium to yeast cell

Paracoccidioides brasiliensis infectious process relies on the initial expression of virulence faactors that are assumed to be controlled by molecular mechanisms through which the conidia and/or mycelial fragments convert to yeast cells. In order to analyze the profile of the thermally-induced dimorphic gene expression, 48 h C-L transition cultures which had been incubated at 36 degrees C were studied. By this time approximately 50% of the conidial population had already reverted to yeast form cells. At this transition time, an EST-Orestes library was constructed and characterized. As a result, 79 sequences were obtained, of which 39 (49.4%) had not been described previously in other libraries of this fungus and which could represent novel exclusive C-Y transition genes. Two of these sequences are, among others, cholestanol delta-isomerase, and electron transfer flavoprotein-ubiquinoneoxidoreductase (ETF-QO). The other 40 (50.6%) sequences were shared with Mycelia (M), Yeast (Y) or Mycelia to yest transition (M-Y) libraries. An important component of this group of sequences is a putative response regulator receiver SKN7, a protein of high importance in stress adaptation and a regulator of virulence in some bacteria and fungi. This is the first report identifying genes expressed during the C-Y transition process, the initial step required to understand the natural history of P brasiliensis conidia induced infection.

The perception of Executives and other Employees on the answer to the Sarbanes-Oxley: case study of a subsidary of the American auto industry established in Brazil

To attend and obtain the systems and. internal controls mechanisms proposed by Sarbanes-Oxley certifications is actually a big challenge,for most of the multinational companies registered in SEC (US Securities and Exchange Commission). This work has the objective of contributing to the analysis of this methodology, not only to attend the law but to reduce cost and generate value through the strengthen of the internal control systems, turning them into animating value generation process mechanisms. So, the idea is to identify the main gaps in the theory through the literature revision and a case study in order to put a question to the main deficiencies, strong points or contributions through the evaluation of the noticed practices. Finally, we can say that a a result of the research and the analyses made in. this case, the vast majority of executives and other employees recognize the benefit that Sarbanes-Oxley Act has brought to the company searched. Also recognize that, although there is still necessity for systemic adequacy and infrastructure, it helps and reinforce reducing and controlling the risks. the system of internal controls in all areas of expertise. They approach and understand that there is the need for a change in the other employees` culture to be inserted in the day-today routine as internal controls, attention to Sarbanes-Oxley and Corporate Governance, making the control cost smaller when compared to the benefits generated.