RAP74 induces promoter contacts by RNA polymerase II upstream and downstream of a DNA bend centered on the TATA box

RAP74, the large subunit of transcription factor IIF, associates with a preinitiation complex containing RNA polymerase II (pol II) and other general initiation factors. We have mapped the location of RAP74 in close proximity to promoter DNA at similar distances both upstream and downstream of a DNA bend centered on the TATA box. Binding of RAP74 induces a conformational change that affects the position of pol II relative to that of the DNA. This reorganization of the preinitiation complex minimally requires the N-terminal region of RAP74 containing both its RAP30-binding domain and another region necessary for accurate transcription in vitro. We propose a role for RAP74 in controlling the topological organization of the pol II preinitiation complex.

Inhibition of human telomerase in immortal human cells leads to progressive telomere shortening and cell death

The correlation between telomerase activity and human tumors has led to the hypothesis that tumor growth requires reactivation of telomerase and that telomerase inhibitors represent a class of chemotherapeutic agents. Herein, we examine the effects of inhibition of telomerase inside human cells. Peptide nucleic acid and 2′-O-MeRNA oligomers inhibit telomerase, leading to progressive telomere shortening and causing immortal human breast epithelial cells to undergo apoptosis with increasing frequency until no cells remain. Telomere shortening is reversible: if inhibitor addition is terminated, telomeres regain their initial lengths. Our results validate telomerase as a target for the discovery of anticancer drugs and supply general insights into the properties that successful agents will require regardless of chemical type. Chemically similar oligonucleotides are in clinical trials and have well characterized pharmacokinetics, making the inhibitors we describe practical lead compounds for testing for an antitelomerase chemotherapeutic strategy.

Linking genome and proteome by mass spectrometry: Large-scale identification of yeast proteins from two dimensional gels

The function of many of the uncharacterized open reading frames discovered by genomic sequencing can be determined at the level of expressed gene products, the proteome. However, identifying the cognate gene from minute amounts of protein has been one of the major problems in molecular biology. Using yeast as an example, we demonstrate here that mass spectrometric protein identification is a general solution to this problem given a completely sequenced genome. As a first screen, our strategy uses automated laser desorption ionization mass spectrometry of the peptide mixtures produced by in-gel tryptic digestion of a protein. Up to 90% of proteins are identified by searching sequence data bases by lists of peptide masses obtained with high accuracy. The remaining proteins are identified by partially sequencing several peptides of the unseparated mixture by nanoelectrospray tandem mass spectrometry followed by data base searching with multiple peptide sequence tags. In blind trials, the method led to unambiguous identification in all cases. In the largest individual protein identification project to date, a total of 150 gel spots—many of them at subpicomole amounts—were successfully analyzed, greatly enlarging a yeast two-dimensional gel data base. More than 32 proteins were novel and matched to previously uncharacterized open reading frames in the yeast genome. This study establishes that mass spectrometry provides the required throughput, the certainty of identification, and the general applicability to serve as the method of choice to connect genome and proteome.

Two contact regions between Stat1 and CBP/p300 in interferon γ signaling

Interferon γ (IFN-γ) induces rapid tyrosine phosphorylation of the latent cytoplasmic transcription factor, Stat1, which then forms homodimers, translocates to the nucleus and participates in IFN-γ-induced transcription. However, little is known of the interactions between Stat1 and the general transcription machinery during transcriptional activation. We show here that Stat1 can directly interact with the CREB-binding protein (CBP)/p300 family of transcriptional coactivators. Specifically, two interaction regions were identified: the amino-terminal region of Stat1 interacts with the CREB-binding domain of CBP/p300 and the carboxyl-terminal region of Stat1 interacts with the domain of CBP/p300 that binds adenovirus E1A protein. Transfection experiments suggest a role for these interactions in IFN-γ-induced transcription. Because CBP/p300-binding is required for the adenovirus E1A protein to regulate transcription of many genes during viral replication and cellular transformation, it is possible that the anti-viral effect of IFN-γ is based at least in part on direct competition by nuclear Stat1 with E1A for CBP/p300 binding.

SURVEY AND SUMMARY: Recent advances in the elucidation of the mechanisms of action of ribozymes

The cleavage of RNA can be accelerated by a number of factors. These factors include an acidic group (Lewis acid) or a basic group that aids in the deprotonation of the attacking nucleophile, in effect enhancing the nucleophilicity of the nucleophile; an acidic group that can neutralize and stabilize the leaving group; and any environment that can stabilize the pentavalent species that is either a transition state or a short-lived intermediate. The catalytic properties of ribozymes are due to factors that are derived from the complicated and specific structure of the ribozyme–substrate complex. It was postulated initially that nature had adopted a rather narrowly defined mechanism for the cleavage of RNA. However, recent findings have clearly demonstrated the diversity of the mechanisms of ribozyme-catalyzed reactions. Such mechanisms include the metal-independent cleavage that occurs in reactions catalyzed by hairpin ribozymes and the general double-metal-ion mechanism of catalysis in reactions catalyzed by the Tetrahymena group I ribozyme. Furthermore, the architecture of the complex between the substrate and the hepatitis delta virus ribozyme allows perturbation of the pKa of ring nitrogens of cytosine and adenine. The resultant perturbed ring nitrogens appear to be directly involved in acid/base catalysis. Moreover, while high concentrations of monovalent metal ions or polyamines can facilitate cleavage by hammerhead ribozymes, divalent metal ions are the most effective acid/base catalysts under physiological conditions.

A combined probabilistic framework for learning gestures and actions

In this paper we introduce a probabilistic approach to support visual supervision and gesture recognition. Task knowledge is both of geometric and visual nature and it is encoded in parametric eigenspaces. Learning processes for compute modal subspaces (eigenspaces) are the core of tracking and recognition of gestures and tasks. We describe the overall architecture of the system and detail learning processes and gesture design. Finally we show experimental results of tracking and recognition in block-world like assembling tasks and in general human gestures.

Word-formation and the translation of Marvel comic book charactonyms

This article analyzes the solutions given in Spanish translations to the morphological creativity shown in the names of Marvel comic book characters. The English versions almost invariably provide a full description of the hero (or villain) by means of a wide variety of word-formation mechanisms leading to highly expressive charactonyms. Indeed, examples shall be listed of names of comic book heroes created through compounding, derivation, including prefixation or suffixation (both classical and Anglo-Saxon but also from other origins), lexical blending, abbreviation, clipping, onomatopoeia, and borrowings from Spanish or from other languages. Early translations into Spanish seemed to be slightly less expressive than the original, even when the same word-formation mechanism was used, usually due to either problems of transparency mainly in some of the word parts or to translation constraints. In later periods, a number of factors, including the influence from other media featuring the same characters and the general trend towards globalization through English, have led translators to choose repetition as the most frequent strategy, which has almost eliminated the creative power of wordformation mechanisms in Spanish and their ability to convey the stylistic effects found in the English versions.

New galactic star clusters discovered in the VVV survey. Candidates projected on the inner disk and bulge

Context. VISTA Variables in the Vía Láctea (VVV) is one of six ESO Public Surveys using the 4 meter Visible and Infrared Survey Telescope for Astronomy (VISTA). The VVV survey covers the Milky Way bulge and an adjacent section of the disk, and one of the principal objectives is to search for new star clusters within previously unreachable obscured parts of the Galaxy. Aims. The primary motivation behind this work is to discover and analyze obscured star clusters in the direction of the inner Galactic disk and bulge. Methods. Regions of the inner disk and bulge covered by the VVV survey were visually inspected using composite JHKS color images to select new cluster candidates on the basis of apparent overdensities. DR1, DR2, CASU, and point spread function photometry of 10 × 10 arcmin fields centered on each candidate cluster were used to construct color–magnitude and color–color diagrams. Follow-up spectroscopy of the brightest members of several cluster candidates was obtained in order to clarify their nature. Results. We report the discovery of 58 new infrared cluster candidates. Fundamental parameters such as age, distance, and metallicity were determined for 20 of the most populous clusters.

The Gaia-ESO Survey: Empirical determination of the precision of stellar radial velocities and projected rotation velocities

Context. The Gaia-ESO Survey (GES) is a large public spectroscopic survey at the European Southern Observatory Very Large Telescope. Aims. A key aim is to provide precise radial velocities (RVs) and projected equatorial velocities (vsini) for representative samples of Galactic stars, which will complement information obtained by the Gaia astrometry satellite. Methods. We present an analysis to empirically quantify the size and distribution of uncertainties in RV and vsini using spectra from repeated exposures of the same stars. Results. We show that the uncertainties vary as simple scaling functions of signal-to-noise ratio (S/N) and vsini, that the uncertainties become larger with increasing photospheric temperature, but that the dependence on stellar gravity, metallicity and age is weak. The underlying uncertainty distributions have extended tails that are better represented by Student’s t-distributions than by normal distributions. Conclusions. Parametrised results are provided, which enable estimates of the RV precision for almost all GES measurements, and estimates of the vsini precision for stars in young clusters, as a function of S/N, vsini and stellar temperature. The precision of individual high S/N GES RV measurements is 0.22–0.26 km s-1, dependent on instrumental configuration.

Guarantee of financial support for student James Savage, signed by William Tudor and John Cooper, 1799 July 25

This legal agreement, a guarantee of financial support for entering student James Savage (A.B. 1803), was signed on July 25, 1799 by his two guarantors, William Tudor and John Cooper. The document was also signed by two witnesses, William Tudor's sons John Henry Tudor and Frederic Tudor. The agreement specifies that, in the event of Savage's failure to settle all financial obligations to the President and Fellows of Harvard College during the course of his studies, the two guarantors would be responsible for a payment of two hundred ounces of silver. It seems that the Tudors and Cooper were relatives of Savage, thus explaining their desire to assure his entry to Harvard by entering into this financial obligation.

Understanding Dropout and Prolonged Engagement in Adolescent Competitive Sport

Objectives: The purpose of this study is to gain understanding of training patterns and roles of significant others (i.e. coaches, parents, peers, and siblings) in adolescent swimmers’ sport participation patterns. Design: The developmental model of sport participation [Côté, J., Baker, J., & Abernethy, B. (2003). From play to practice: A developmental framework for the acquisition of expertise in team sport. In J. Starkes, & K. A. Ericsson (Eds.), Recent advances in research on sport expertise (pp. 89–114). Champaign, IL: Human Kinetics; Côté, J., & Fraser-Thomas, J. (2007). Youth involvement in sport. In P. R. E. Crocker (Ed.), Introduction to sport psychology: A Canadian perspective (pp. 266–294). Toronto: Pearson Prentice Hall] was used as a framework. Method: Ten dropout and 10 engaged swimmers, matched on key demographic variables participated in a semi-structured qualitative interview. Results: Groups had many similar experiences (e.g. early training, supportive and unsupportive coaches, involved parents). However, only dropouts spoke of early peak performances, limited one-on-one coaching, pressuring parents during adolescence, lack of swimming peers during adolescence, and sibling rivalries. In contrast, only engaged athletes spoke of clubs’ developmental philosophies, coaches’ and parents’ open communication, school friends’ support, and siblings’ general positive influences. Conclusions: Findings highlight the importance of appropriately structured programs and the fragility of athletes’ relationships with significant others during the adolescent years. Implications for sport programmers, coaches, and parents are discussed.

Transitioning Towards a Development Permit System: Recommendations for the Ministry of Municipal Affairs and Housing

Since 2007, Ontario Regulation 608/06 now provides all municipalities with the authority to establish a development permit system (DPS); however, much of this regulatory power and its functions are largely misunderstood by both professionals and the general public, which has led to a lack of widespread municipal implementation. One main contributor to the uncertainty is the lack of academic literature. Currently, the most comprehensive document has been produced by the Ministry of Municipal Affairs and Housing (MMAH), Development Permit System: A Handbook for Municipal Implementation. The study’s key objective is to identify, analyze, and evaluate the MMAH Handbook in order to effectively provide an updated set of recommendations within the context of Ontario.

European Integration and Europeanisation: Benefits and Disadvantages for Business. Bruges European Economic Policy (BEEP) Briefing 29/2013

European integration is a project of great economic importance for the 500 million consumers and 21 million companies in Europe. With the economic borders between Member States removed, Europeanisation becomes inevitable for companies. The paper proposes a framework to analyse the benefits and disadvantages for business that come with the process of European integration, structured according to the logic of the four fundamental freedoms of movement within the Internal Market (freedom of movement of goods, services, capital and people) complemented by the section on technology and innovation, and the general EU regulatory environment. Whereas the business decisions need to be taken on a case-by-case basis, taking into consideration firm’s own capabilities and resources as well as industry specificities, several recommendations for companies willing to Europeanise are made, based on an analysis of the regulatory macro-environment of the EU. Above all, any company willing to be successful in the EU has to become a learning organisation, responsive to the advancements of the macro-environment. The ability to anticipate the regulatory developments and to adjust one’s own business and corporate strategy accordingly is the key to achieving sustainable competitive advantage in the European Union.

Social Content of the International Sphere: Symbols and Meaning in Franco-German Relations, CES Germany & Europe Working Papers, No. 02.2, 2002

“The Franco-German friendship is rich in memories and gestures that are at once important and symbolic, and that characterize the exceptional nature of the relationship between our two countries,” reflects former French economics minister and European Commission President Jacques Delors. Such symbolic acts and joint memories are not primarily about cooperation in specific instances. Rather, more generally, they denote what it means to act together. They lend significance to a relationship; they signify what is “at stake,” or what it is “all about.” They are about a deeper and more general social purpose underlying specific instances of cooperation. They are about the value and intrinsic importance that social relations incorporate. Symbols contribute to the institutionalization of social meaning and social purpose in dealing with one another. In this paper I clarify the concept of “predominantly symbolic acts and practices among states,” systematically explore such acts for the bilateral Franco-German relationship between the late 1950s and the mid-1990s, and scrutinize the specific meaning and effects that these practices have helped to generate and perpetuate.

Consensus, contradiction, and conciliation of interests: the geo-economics of the Energy Union. EPC Policy Brief, 8 July 2015

European Union energy policy calls for nothing less than a profound transformation of the EU's energy system: by 2050 decarbonised electricity generation with 80-95% fewer greenhouse gas emissions, increased use of renewables, more energy efficiency, a functioning energy market and increased security of supply are to be achieved. Different EU policies (e.g., EU climate and energy package for 2020) are intended to create the political and regulatory framework for this transformation. The sectorial dynamics resulting from these EU policies already affect the systems of electricity generation, transportation and storage in Europe, and the more effective the implementation of new measures the more the structure of Europe's power system will change in the years to come. Recent initiatives such as the 2030 climate/energy package and the Energy Union are supposed to keep this dynamic up. Setting new EU targets, however, is not necessarily the same as meeting them. The impact of EU energy policy is likely to have considerable geo-economic implications for individual member states: with increasing market integration come new competitors; coal and gas power plants face new renewable challengers domestically and abroad; and diversification towards new suppliers will result in new trade routes, entry points and infrastructure. Where these implications are at odds with powerful national interests, any member state may point to Article 194, 2 of the Lisbon Treaty and argue that the EU's energy policy agenda interferes with its given right to determine the conditions for exploiting its energy resources, the choice between different energy sources and the general structure of its energy supply. The implementation of new policy initiatives therefore involves intense negotiations to conciliate contradicting interests, something that traditionally has been far from easy to achieve. In areas where this process runs into difficulties, the transfer of sovereignty to the European level is usually to be found amongst the suggested solutions. Pooling sovereignty on a new level, however, does not automatically result in a consensus, i.e., conciliate contradicting interests. Rather than focussing on the right level of decision making, European policy makers need to face the (inconvenient truth of) geo-economical frictions within the Union that make it difficult to come to an arrangement. The reminder of this text explains these latter, more structural and sector-related challenges for European energy policy in more detail, and develops some concrete steps towards a political and regulatory framework necessary to overcome them.