Individual Palmitoyl Residues Serve Distinct Roles in H-ras Trafficking, Microlocalization, and Signaling

H-ras is anchored to the plasma membrane by two palmitoylated cysteine residues, Cys181 and Cys184, operating in concert with a C-terminal S-farnesyl cysteine carboxymethylester. Here we demonstrate that the two palmitates serve distinct biological roles. Monopalmitoylation of Cys181 is required and sufficient for efficient trafficking of H-ras to the plasma membrane, whereas monopallmitoylation of Cys184 does not permit efficient trafficking beyond the Golgi apparatus. However, once at the plasma membrane, monopalmitoylation of Cys184 supports correct GTP-regulated lateral segregation of H-ras between cbolesterol-dependent and cholesterol-independent microdomains. In contrast, monopallmitoylation of Cys181 dramatically reverses H-ras lateral segregation, driving GTP-loaded H-ras into cholesterol-dependent microdomains. Intriguingly, the Cys181 monopalmitoylated H-ras anchor emulates the GTP-regulated microdomain interactions of N-ras. These results identify N-ras as the Ras isoform that normally signals from lipid rafts but also reveal that spacing between palmitate and prenyl groups influences anchor interactions with the lipid bilayer. This concept is further supported by the different plasma membrane affinities of the monopalmitoylated anchors: Cys181-palmitate is equivalent to the dually palmitoylated wild-type anchor, whereas Cys184-pahnitate is weaker. Thus, membrane affinity of a pallmitoylated anchor is a function both of the hydrophobicity of the lipid moieties and their spatial organization. Finally we show that the plasma membrane affinity of monopahnitoylated anchors is absolutely dependent on cholesterol, identifying a new role for cholesterol in promoting interactions with the raft and nonraft plasma membrane.

A versatile synthetic approach to peptidyl privileged structures using a "safety-catch" linker

Peptidyl privileged structures have been widely used by many groups to discover biologically active molecules. In this context, privileged substructures are used as hydrophobic anchors, to which peptide functionality is appended to gain specificity. Utilization of this concept has led to the discovery of many different active compounds at a wide range of biological receptors. A synthetic approach to these compounds has been developed on a safety-catch linker that allows rapid preparation of large libraries of these molecules. Importantly, amide bond formation/cleavage through treatment with amines is the final step; it is a linker strategy that allows significant diversification to be easily incorporated, and it only requires the inclusion of an amide bond. In addition, chemistry has been developed that permits the urea moiety to be inserted at the N-terminus of the peptide, allowing the same set of amines (either privileged substructures or amino acid analogues) to be used at both the N- and C-termini of the molecule. To show the robustness of this approach, a small library of peptidyl privileged structures were synthesized, illustrating that large combinatorial libraries can be synthesized using these technologies.

Hypermethylation of the 5 ' CpG island of the gene encoding the serine protease Testisin promotes its loss in testicular tumorigenesis

The Testisin gene (PRSS21) encodes a glycosylphosphatidylinositol (GPI)-linked serine protease that exhibits testis tissue-specific expression. Loss of Testisin has been implicated in testicular tumorigenesis, but its role in testis biology and tumorigenesis is not known. Here we have investigated the role of CpG methylation in Testisin gene inactivation and tested the hypothesis that Testisin may act as a tumour suppressor for testicular tumorigenesis. Using sequence analysis of bisulphite-treated genomic DNA, we find a strong relationship between hypermethylation of a 385 bp 50 CpG rich island of the Testisin gene, and silencing of the Testisin gene in a range of human tumour cell lines and in 100% (eight/eight) of testicular germ cell tumours. We show that treatment of Testisin-negative cell lines with demethylating agents and/or a histone deacetylase inhibitor results in reactivation of Testisin gene expression, implicating hypermethylation in Testisin gene silencing. Stable expression of Testisin in the Testisin-negative Tera-2 testicular cancer line suppressed tumorigenicity as revealed by inhibition of both anchorage-dependent cell growth and tumour formation in an SCID mouse model of testicular tumorigenesis. Together, these data show that loss of Testisin is caused, at least in part, by DNA hypermethylation and histone deacetylation, and suggest a tumour suppressor role for Testisin in testicular tumorigenesis.

Mechanisms of transfer film formation on data recording heads

The increasing demand for high capacity data storage requires decreasing the head-to-tape gap and reducing the track width. A problem very often encountered is the development of adhesive debris on the heads at low humidity and high temperatures that can lead to an increase of space between the head and media, and thus a decrease in the playback signal. The influence of stains on the playback signal of reading heads is studied using RAW (Read After Write) tests and their influence on the wear of the heads by using indentation technique. The playback signal has been found to vary and the errors to increase as stains form a patchy pattern and grow in size to form a continuous layer. The indentation technique shows that stains reduce the wear rate of the heads. In addition, the wear tends to be more pronounced at the leading edge of the head compared to the trailing one. Chemical analysis of the stains using ferrite samples in conjunction with MP (metal particulate) tapes shows that stains contain iron particles and polymeric binder transferred from the MP tape. The chemical anchors in the binder used to grip the iron particles now react with the ferrite surface to create strong chemical bonds. At high humidity, a thin layer of iron oxyhydroxide forms on the surface of the ferrite. This soft material increases the wear rate and so reduces the amount of stain present on the heads. The stability of the binder under high humidity and under high temperature as well as the chemical reactions that might occur on the ferrite poles of the heads influences the dynamic behaviour of stains. A model of stain formation taking into account the channels of binder degradation and evolution upon different environmental conditions is proposed.

Effectiveness of performance appraisal:an integrated framework

Based on a robust analysis of the existing literature on performance appraisal (PA), this paper makes a case for an integrated framework of effectiveness of performance appraisal (EPA). To achieve this, it draws on the expanded view of measurement criteria of EPA, i.e. purposefulness, fairness and accuracy, and identifies their relationships with ratee reactions. The analysis reveals that the expanded view of purposefulness includes more theoretical anchors for the purposes of PA and relates to various aspects of human resource functions, e.g. feedback and goal orientation. The expansion in the PA fairness criterion suggests certain newly established nomological networks, which were ignored in the past, e.g. the relationship between distributive fairness and organization-referenced outcomes. Further, refinements in PA accuracy reveal a more comprehensive categorization of rating biases. Coherence among measurement criteria has resulted in a ratee reactions-based integrated framework, which should be useful for both researchers and practitioners.

Selective oxidation of cyclohexene through gold functionalized silica monolith microreactors

Two simple, reproducible methods of preparing evenly distributed Au nanoparticle containing mesoporous silica monoliths are investigated. These Au nanoparticle containing monoliths are subsequently investigated as flow reactors for the selective oxidation of cyclohexene. In the first strategy, the silica monolith was directly impregnated with Au nanoparticles during the formation of the monolith. The second approach was to pre-functionalize the monolith with thiol groups tethered within the silica mesostructure. These can act as evenly distributed anchors for the Au nanoparticles to be incorporated by flowing a Au nanoparticle solution through the thiol functionalized monolith. Both methods led to successfully achieving even distribution of Au nanoparticles along the length of the monolith as demonstrated by ICP-OES. However, the impregnation method led to strong agglomeration of the Au nanoparticles during subsequent heating steps while the thiol anchoring procedure maintained the nanoparticles in the range of 6.8 ± 1.4 nm. Both Au nanoparticle containing monoliths as well as samples with no Au incorporated were tested for the selective oxidation of cyclohexene under constant flow at 30 °C. The Au free materials were found to be catalytically inactive with Au being the minimum necessary requirement for the reaction to proceed. The impregnated Au-containing monolith was found to be less active than the thiol functionalized Au-containing material, attributable to the low metal surface area of the Au nanoparticles. The reaction on the thiol functionalized Au-containing monolith was found to depend strongly on the type of oxidant used: tert-butyl hydroperoxide (TBHP) was more active than H2O2, likely due to the thiol induced hydrophobicity in the monolith.

Modelagem numérica de uma estrutura de contenção de estacas espaçadas atirantadas em areia

Retaining walls design involves factors such as plastification, loading and unloading, pre-stressing, excessive displacements and earth and water thrust. Furthermore, the interaction between the retained soil and the structure is rather complex and hard to predict. Despite the advances in numerical simulation techniques and monitoring of forces and displacements with field instrumentation, design projects are still based on classical methods, whose simplifying assumptions may overestimate structural elements of the retaining wall. This dissertation involves a three-dimensional numerical study on the behavior of a retaining wall using the finite element method (FEM). The retaining wall structure is a contiguous bored pile wall with tie-back anchors. The numerical results were compared to data obtained from field instrumentation. The influence of the position of one or two layers of anchors and the effects of the construction of a slab bounded at the top of the retaining wall was evaluated. Furthermore, this study aimed at investigating the phenomenon of arching in the soil behind the wall. Arching was evaluated by analyzing the effects of pile spacing on horizontal stresses and displacements. Parametric analysis with one layers of anchors showed that the smallest horizontal displacements of the structure were achieved for between 0.3 and 0.5 times the excavation depth. Parametric analyses with two anchor layers showed that the smallest horizontal displacements were achieve for anchors positioned in depths of 0.4H and 0.7H. The construction of a slab at the top of the retaining wall decreased the horizontal displacements by 0.14% times the excavation depth as compared to analyses without the slab. With regard to the arching , analyzes showed an optimal range of spacing between the faces of the piles between 0.4 and 0.6 times the diameter of the pile

Minicontos: uma prática de letramento emergente na escola

Like numerous other mini ficçion genres, the Flash fiction is configured as a concise and brief genre, but it can still preserve some elements of the traditional tale, such as: narrator, space-temporal unit, characters and plot. For some authors, the emergence of this genre is strictly connected to the speed and diversity of forms of communication in the current social context, that require the reader other skills in addition to reading and writing. Considering the importance of these skills in a literate society and recognizing this genre as potentially effective tool for the development of literacy, this research has as objective to describe the development and implications of didactic sequence directed the literacy practices with the Flash fiction genre. Thus, the research becomes effective in an elementary school vespertine class in 9th grade from Escola Estadual Dr. José Gonçalves de Medeiros, located in the city of Acari-RN. As theoretical assumptions, it will be used fundamentals from Literacy Studies as social practice, especially that ones discussed by the writings Kleiman (1995), Hamilton (2000), Soares (2012), Mortatti (2004) and Rojo (2012). To subsidize the literary literacy, it will be used the theoretical contibutions of Borges (2000), Candido (2004), Pinheiro (2001) and Silva e Silveira (2013). In what relates to the study about the storytellers, it was taken by support the postulates of Gotlib (2006), Bosi (1997), Cortázar (1999), Talbot (2002), Poe (s/d), Luzia de Maria (2004), as well as studies by researchers and writers as Spalding (2007; 2008; 2011), George (2007), Freire (2004) and Trevisan (1994), among others who are dedicated to study Flash fiction. In the methodological field, the proposed perspective anchors itself in an action-researching guidelines (MOREIRA; CALEFFE, 2006) and in the approach of a qualitative data (BOGDAN; BIKLEN, 1994), since it talks to interpretation of diagnosed situation, implementation of intervention actions organized in didactic sequence (SCHNEUWLY; DOLZ, 2004; BARROS; RIVERS-registry, 2014), including description and evaluation of results achieved from the implementation of such actions. The discussions generated indicate that the work with Flash ficition, developed by means of didactic sequences, contributes to the reflection on teaching strategies of Portuguese, used to prepare the students to meet the demands of reading and writing of contemporary society.

Performance improvements of mooring systems for wave energy converters

In the development of wave energy converters, the mooring system is a key component for a safe station-keeping and an important factor in the cost of the wave energy production. Generally, when designing a mooring system for a wave energy converter, two important conditions must be considered: (i) that the mooring system must be strong enough to limit the drifting motions, even in extreme waves, tidal and wind conditions and (ii) it must be compliant enough so that the impact on wave energy production can be minimised. It is frequently found that these two conditions are contradictory. The existing solutions mainly include the use of heavy chains, which create a catenary shaped mooring configuration, allowing limited flexibility within the mooring system, and hence very large forces may still be present on mooring lines and thus on anchors. This solution is normally quite expensive if the costs of the materials and installation are included. This paper presents a new solution to the mooring system for wave energy converters within the FP7 project, ‘GeoWAVE’, which is a project aiming to develop a new generation of the moorings system for minimising the loads on mooring lines and anchors, the impact on the device motions for power conversion, and the footprint if it is applicable, and meanwhile the new types of anchors are also addressed within the project. However this paper will focus on the new mooring system by presenting the wave tank test results of the Pelamis wave energy converter model and the new developed mooring system. It can be seen that the new generation of mooring system can significantly reduce the loads on mooring lines and anchors, and reduce the device excursions as a result of the new mooring system when compare to the conventional catenary mooring.

Dynamic effects of anchor positional tolerance on tension moored floating wind turbine

For water depths greater than 60m floating wind turbines will become the most economical option for generating offshore wind energy. Tension mooring stabilised units are one type of platform being considered by the offshore wind energy industry. The complex mooring arrangement used by this type of platform means that the dynamics are greatly effected by offsets in the positioning of the anchors. This paper examines the issue of tendon anchor position tolerances. The dynamic effects of three positional tolerances are analysed in survival state using the time domain FASTLink. The severe impact of worst case anchor positional offsets on platform and turbine survivability is shown. The worst anchor misposition combinations are highlighted and should be strongly avoided. Novel methods to mitigate this issue are presented.

The Interaction Between Monetary and Fiscal Policies (Working Paper 30)

One can view the period since 1970 as one in which the authorities struggled to establish appropriate medium-term anchors for both monetary and fiscal policies. During this time, they learned about the appropriate interaction between those two policies in the context of economic stabilization and growth under a flexible exchange rate regime. This lecture deals with four interrelated topics: the appropriate goals for fiscal and monetary policy, building policy credibility, the appropriate stabilization role for the two policies, and policy cooperation. The transparent medium term frameworks that have been established by the authorities will be extremely helpful in meeting the challenges that the future is sure to bring. These frameworks mean that the required adjustments in the economy will take place against a relatively stable background. Thank you for the invitation to give the Gow Lecture for 2002. Donald Gow had a great interest in public administration and in budgetary reform in the federal government (Gow 1973). He was one in a long line of

Les métamorphoses du rapport politique à la mort (recherche de pivots structurant la réflexion en médecine palliative)

Notre recherche explore quelques moments forts des métamorphoses du rapport politique à la mortalité sous examen des thèmes de l’interdit, de la dignité, de l’autonomie et de l’altérité. Nous dégageons des ancrages propices à nourrir la pensée actuelle en médecine palliative. Ainsi, nous livrons une enquête philosophique, appréciant pour nous Occidentaux, les influences marquantes des pensées gréco-romaine, chrétienne et moderne. Ces bases, édifiant notre monde politique, ont suscité l’émergence de la médecine palliative. C’est pourquoi, nous tentons de caractériser et de comprendre les problématiques nouvelles, dans leurs aspects politique et éthique, envisagées à l’aune des formes contemporaines d’accompagnement des mourants. Notre effort tente de discerner les aspirations et les impasses. L’étude de la métamorphose des repères fait ressortir une dissociation accentuée au fil du temps. En effet, au fur et à mesure, notre entreprise d’interprétation du fondement de ces questions politiques dévoilait : une reconnaissance universelle de l’interdit d’homicide, mais accusant une perte du lien moral au profit d’une visée amorale ; un aval unanime du respect de la dignité, mais manifestant une confusion et une division ostensible entre conceptions intrinsèque et extrinsèque ; une affirmation péremptoire de l’autonomie, mais avec une distanciation marquée au regard de la façon d’envisager la part de l’autre ; une déclinaison de liens humains reconnus de tous, mais exacerbés dans une tension artificielle entre individualisme et altruisme. Au surplus, en constatant la distance et la dislocation entre le public et le privé, entre la réclamation de fraternité et la recherche d’amicalité signifiante, nous avons envisagé la communauté palliative comme un lieu de résistance à cette décomposition menaçante au sein de la communauté politique. À terme de l’analyse, nous avons fondé les concepts « d’allonomie » et de « suspension éthique ». Il s’agit de contributions originales destinées à donner à la philosophie toute sa dimension sapientielle au service de l’accompagnement palliatif.

Teaching Sight-Reading to Undergraduate Choral Ensemble Singers: Lessons from Successful Learners

This study was designed to investigate professional choral singers’ training, perceptions on the importance of sight-reading skill in their work, and thoughts on effective pedagogy for teaching sight-reading to undergraduate choral ensemble singers. Participants in this study (N=48) included self-selected professional singers and choral conductors from the Summer 2015 Oregon Bach Festival’s Berwick Chorus and conducting Master Class. Data were gathered from questionnaire responses and audio recorded focus group sessions. Focus group data showed that the majority of participants developed proficiency in their sight-reading skills from instrumental study, aural skills classes, and through on-the-job training at a church job or other professional choral singing employment. While participants brought up a number of important job skills, sightreading was listed as perhaps the single most important skill that a professional choral singer could develop. When reading music during the rehearsal process, the data revealed two main strategies that professional singers used to interpret the pitches in their musical line: an intervallic approach and a harmonic approach. Participants marked their scores systematically to identify problem spots and leave reminders to aid with future readings, such as marking intervals, solfege syllables, or rhythmic counts. Participants reported using a variety of skills other than score marking to try to accurately find their pitches, such as looking at other vocal or instrumental lines, looking ahead, and using knowledge about a musical style or time period to make more intuitive “guesses” when sight-reading. Participants described using additional approaches when sight-reading in an audition situation, including scanning for anchors or anomalies and positive self-talk. Singers learned these sight-reading techniques from a variety of sources. Participants had many different ideas about how best to teach sight-reading in the undergraduate choral ensemble rehearsal. The top response was that sight-reading needed to be practiced consistently in order for students to improve. Other responses included developing personal accountability, empowering students, combining different teaching methods, and discussing real-life applications of becoming strong sight-readers. There was discussion about the ultimate purpose of choir at the university level and whether it is to teach musicianship skills or produce excellent performances.

Dimensionamento de uma estrutura de contenção de terras flexível com vários níveis de apoio

O presente trabalho, no âmbito de projeto final de curso de metrado em Engenharia da Construção, teve como objetivo o estudo do comportamento de estruturas de suporte de terras flexíveis multi-apoiadas (com diferentes tipos de apoio) para dois tipos solos homogéneos. Recorreu-se às teorias clássicas, como a de Rankine, desenvolvidas para estruturas de suporte de terras rígidas. Às teorias semi-empíricas de Terzaghi & Peck que culminaram nos diagramas de Terzaghi & Peck. Apesar de os digramas de Terzaghi & Peck serem diagramas de pressões de terras a usar em estruturas de suporte de terras flexíveis, apresentam algumas limitações importantes, como a sua aplicação apenas em solos heterogéneos, com presença ou não de níveis freáticos, e sem fornecer distribuição das pressões de terras na zona passiva (zona enterrada). Como na atualidade os modelos de elementos finitos permitem simular de modo muito mais rigoroso os problemas da engenharia. O presente trabalho esteve focado em analisar um caso prático em diferentes solos e com diferentes tipos de apoios. Será estudado mediante os métodos analíticos usando as teorias clássicas e posteriormente métodos numéricos (com diferentes programas de cálculo). Finalmente serão comparados os resultados obtidos mediante os diferentes métodos usados. As estruturas foram inicialmente pré-dimensionadas usando os métodos clássicos. Assim foram usados os diagramas de pressões de terras de Terzaghi & Peck para a zona ativa (zona em escavação) e a teoria de Rankine para conhecer as pressões de terras na zona enterrada da cortina (parede moldada) e recorrendo ao software Ftool para a obtenção dos parâmetros de dimensionamento de estruturas de suporte de terras objeto de estudo. Posteriormente utilizaram-se os programas de cálculo automático CYPE 2015 k, e o programa de cálculo de elementos finitos PLAXIS Introductory 2010. Estes programas permitem simular o faseamento construtivo do muro. Para estudar a influência de algúns parâmetros no comportamento da Resumo IV cortina o estudo foi realizado com dois solos distintos, um solo argiloso mole e um solo arenoso denso. Assim como para dois tipos de apoios distintos, ancoragens ativas e escoras passivas. Foram analisados diferentes parâmetros na estrutura de suporte; pressões horizontais das terras, deslocamentos horizontais, esforço axial, transverso e momento fletor.

Carreiras em transformação e seus paradoxais reflexos nos indivíduos: Metáforas de carreira e de competências

Este artigo analisa a evolução da carreira e seus diversos significados e apresenta um estudo realizado com profissionais de Administração de empresas, suas autopercepções e aspirações (âncoras) de carreira e as exigências em termos de competências profissionais. Foram realizadas entrevistas em profundidade, conjugadas com a aplicação do Inventário de Âncoras de Carreira de Edgar Schein, como etapa inicial e exploratória de um estudo mais amplo. As trajetórias estudadas, e as metáforas e âncoras encontradas, revelaramse exemplares de tendências atuais, como a busca pela ampliação das competências e a carreira em ziguezague, com características de autodirecionamento da carreira proteana, defrontandose com diversas alternativas e evidenciando uma postura mais próativa. Essa metodologia permitiu o aprofundamento de questões internas dos sujeitos pesquisados, em relação ao trabalho e à carreira, possibilitando a manifestação de seus sentimentos, angústias e aspirações, assim como o repensar de suas escolhas e trajetórias profissionais, de forma livre e criativa.