Toward controlling gene expression at will: Specific regulation of the erbB-2/HER-2 promoter by using polydactyl zinc finger proteins constructed from modular building blocks

To create a universal system for the control of gene expression, we have studied methods for the construction of novel polydactyl zinc finger proteins that recognize extended DNA sequences. Elsewhere we have described the generation of zinc finger domains recognizing sequences of the 5′-GNN-3′ subset of a 64-member zinc finger alphabet. Here we report on the use of these domains as modular building blocks for the construction of polydactyl proteins specifically recognizing 9- or 18-bp sequences. A rapid PCR assembly method was developed that, together with this predefined set of zinc finger domains, provides ready access to 17 million novel proteins that bind the 5′-(GNN)6-3′ family of 18-bp DNA sites. To examine the efficacy of this strategy in gene control, the human erbB-2 gene was chosen as a model. A polydactyl protein specifically recognizing an 18-bp sequence in the 5′-untranslated region of this gene was converted into a transcriptional repressor by fusion with Krüppel-associated box (KRAB), ERD, or SID repressor domains. Transcriptional activators were generated by fusion with the herpes simplex VP16 activation domain or with a tetrameric repeat of VP16’s minimal activation domain, termed VP64. We demonstrate that both gene repression and activation can be achieved by targeting designed proteins to a single site within the transcribed region of a gene. We anticipate that gene-specific transcriptional regulators of the type described here will find diverse applications in gene therapy, functional genomics, and the generation of transgenic organisms.

Supramolecular coordination complexes as building blocks for porous and magnetic materials

In this work, new coordination polymers based on two different classes of synthons are presented. In addition, manganese-based metallacrowns of magnetic interest are studied, both in the solid state and in solution. Firstly, functionalized bispyrazolylmethane derivatives are employed as bridging ligands for the establishment of silver-based coordination polymers; the influence of the substituent groups and of the counterions on the supramolecular packing is also investigated. Secondly, the use of metallacrown (MC) complexes as building blocks for porous coordination polymers is discussed. The design of a new metallacrown species is presented, which shows the tendency of aggregating in the solid state to form coordination polymers. Two new coordination polymers are indeed reported, of which one is the first MC-based permanently porous coordination network ever presented. The solid resists solvent evacuation and exhibits gas uptake ability. Furthermore, the isolation and characterization of a new metallacryptate species based on manganese ions is described. The metal-rich structure comprises nine Mn(II)/Mn(III) ions and presents an inverse metallacrown core subunit that binds a μ3-O2- ion. The metallacryptate is isolated in high yields and stable in solution. Lastly, a family of 3d-4f heterometallic metallacrowns is characterized in solution by means of UV-Vis spectrophotometry and of paramagnetically shifted 1H-NMR. The lanthanide-induced shifts observed in the spectra are employed to describe the molecules behaviour in solution and are qualitatively related to the magnetic properties of the compounds.

Intramolecular Bromoamination of Conjugated N-Tosylaminodienes Using N-Bromosuccinimide. Synthesis of Bromoallyl-Substituted N-Heterocycles and Their Applications as Building Blocks

The bromonium-promoted cyclization of conjugated aminodienes is described. The reaction proceeds smoothly in the presence of N-bromosuccinimide as halonium promoter, and using N-tosyl-protected aminodienes as substrates, to give the corresponding halocyclization products in high yields and with high diastereoselectivities. It can be envisaged that the formation of these products is the result of an SN2′-type ring-opening of a terminal bromonium intermediate in a 5-exo-trig or 6-exo-trig cyclization mode. The presence of an allyl bromide moiety in the haloamination products makes these molecules highly attractive from a synthetic point of view, as it opens the way for further transformations. Thus, allylic substitution reactions with different nucleophiles (acetate, azide, cyanide, and malonate), palladium-catalysed Suzuki coupling, and silver-mediated bromine displacement reactions were carried out successfully.

TMO revisited : an evaluation report : toward a multicultural organization : building blocks to meet 21st century challenges.

"FS-631"--P. [4] of cover.

The sugar cane biofactory - building blocks for the future

The sugarcane plant, with its enormous genetic capacity to accumulate carbon and manufacture and store sucrose, also has the potential to accumulate carbon and metabolically create a wide range of new molecules for industrial and other commercial uses. The extent to which this change can be developed and realised commercially is a function of the technical competence of the industry's R&D capacity, the reality of the commercial drivers which support this global agenda, and the determination of the industry to achieve such goals. The outcomes of existing R&D work already strongly support the technical challenges of this opportunity in sugarcane. The current challenge remains the commercialisation of the technology in a global market in which the current business structures and systems for the manufacture and distribution of existing (competitive) products makes the development of new product lines a higher risk than might otherwise be the case. This is despite all the claims that global markets are expecting and (in some cases) legislating the creation of more sustainable production systems. The options and issues for the development of a sugarcane biofactory system are discussed.

Selective functionalisation of Re6 cluster anionic units:from hexa-hydroxo [Re6Q8(OH)6]4− (Q = S, Se) to neutral trans-[Re6Q8L4L′2] hybrid building blocks

The reaction of [Re6Q8(OH)6]4- (Q = S, Se) with p-tertbutylpyridine (TBP) in water leads to neutral trans-[Re6Q8(TBP)4(OH)2] whose hydroxyl reactivity with carboxylic acid and TBP exchange reaction with functional pyridine have been investigated.

Synthesis and characterization of electron deficient heterocycles as building blocks for electronic materials

This thesis describes the synthesis of a new electropolymerizable viologen derivative. A reasonably high-yielding route is reported, and a preliminary investigation of its polymerisation is described. The viologen and its precursors were examined by 1H NMR, MS, IR and elemental analysis. The energies of the band gap for the materials have been calculated using UV-vis spectroscopy, and cyclic voltammetry was also used to estimate the oxidation and the reduction potentials and to calculate the HOMO and LUMO energies. Theoretical calculations were performed using DFT. The attempted synthesis of a new flavin-functionalised phenanthroline derivative is described. Unfortunately, the protocol used failed to provide the desired compounds.

Which Reality? Interactions at the southern border of Mexico as fieldwork of creative practice

El siguiente ensayo sintetiza y describe el proyecto de investigación creación ¿Cuál realidad? cuyo objetivo es la creación y análisis de una práctica artística como forma de entender y generar conocimiento a partir de las manifestaciones y acontecimientos sociales y culturales en diversos lugares de la frontera sur de México. Lo que se busca es reflexionar acerca de las fronteras, no sólo como lo que delimita el fin o el principio de los territorios de estados nacionales o geográficos, sino como productoras simbólicas de las diferencias entre las personas, recalcando todo aquello en lo que no son iguales por encima de lo que tienen en común. Los espacios geopolíticos que delimitan el territorio de un estado-nación son el repertorio palpable de la movilidad de personas, cosas y acontecimientos. Estas prácticas son efectuadas de manera más o menos visible y demuestran que existen y han estado ahí constantemente redes de comunicación y formas de acción común, que tienen como fin procurar bienestar y elevar la calidad de vida de los partícipes. El proyecto ¿Cuál realidad? en su materialidad y visibilidad fue planteado como una serie de intervenciones en sitio, tales como esculturas públicas, instalaciones, acciones participativas, fotografía y video, a partir de la interacción en algunas ciudades fronterizas en los estados de Chiapas, Tabasco y Quintana Roo y su colindancia con los países de Guatemala y Belice. La producción a girado entorno a las siguientes tres líneas de búsqueda temática: La línea fronteriza, el espacio físico; las personas y grupos que confluyen en estos territorios; el contrabando de cosas y el trasiego de las personas entre los países o en el interior del territorio nacional (entre ciudades del mismo estado, inmigrantes rurales etc.). Los lugares seleccionados son significativos de la frontera sur y tanto las esculturas como las intervenciones han sido construidas con objetos y/o procesos que funcionen simbólicamente para los grupos o habitantes de la zona y en colaboración con ellos: objetos útiles para el trabajo, procesos de transformación de materiales, técnicas de fabricación artesanal, ropa, donaciones, entre muchas otras construcciones.

El trabajo en equipo en el Proceso Administrativo y de Compras en OPL Carga de Bucaramanga

El proceso administrativo y de compras de OPL Carga tiene algunas falencias entre ellas: fallas en la Comunicación entre el personal operativo, no se realizan llamadas internas usando con frecuencia el email, produciendo la saturación de solicitudes las cuales terminan sin ser resueltas en cuanto a roles se refiere, no hay enfoque de procesos en vista que no se tiene claras las tareas de cada cargo, adicionalmente no hay claridad en los subprocesos, perjudicando el proceso con el aumento de costos, pérdida de tiempo, las responsabilidades de los funcionario no todas las veces se ejecutan en el tiempo asignado, el liderazgo compartido presenta ambigüedades. Objetivos: Definir el trabajo en equipo en el proceso administrativo y de compras en OPL carga de Bucaramanga. La investigación que a realizar es de tipo descriptivo, busca descubrir las falencias o características que permiten diseñar y desarrollar un modelo de solución para los problemas del equipo de OPL Carga S.A.S. Materiales y métodos: La investigación efectuada es de tipo descriptivo, el objetivo es definir el modelo del trabajo en equipo y describir las falencias en el proceso administrativo y de compras en OPL carga de Bucaramanga, que permitan obtener un diagnóstico integral que conlleve a la implementación de estrategias de solución. Resultados: Se identificaron las falencias en los siguientes aspectos: Variable comunicación, rendimiento, destrezas complementarias, propósito significativo y meta específicas de los funcionarios en OPL carga sección administrativa. Conclusiones: El modelo de trabajo en equipo que OPL aplica es jerárquico, en el que se ofrece estabilidad, seguridad, se toman decisiones en forma piramidal, mediante la planeación de tareas, la colaboración, igualdad y respeto por los miembros, trabajando en pro de la solución de problemas. Se construyó un plano conceptual que permitió exponer la interpretación que la estudiante tiene de las teorías, investigaciones y antecedentes válidos para la comprensión del problema investigado. Área comunicacional: Coordinar acciones tendientes para que los funcionarios respondan a tiempo los emails atenientes a su trabajo. Área condiciones de trabajo: Clarificar y diseñar las reglas de comportamiento al interior de los equipos de trabajo que redunden en el mejoramiento del mismo y la búsqueda de soluciones oportunas. Área metas específicas: Procurar mediante auditorías el cumplimiento de las metas y objetivos propuestos por cada equipo de trabajo.

An educational review of cartilage repair: precepts & practice - myths & misconceptions - progress & prospects.

OBJECTIVE The repair of cartilaginous lesions within synovial joints is still an unresolved and weighty clinical problem. Although research activity in this area has been indefatigably sustained, no significant progress has been made during the past decade. The aim of this educational review is to heighten the awareness amongst students and scientists of the basic issues that must be tackled and resolved before we can hope to escape from the whirlpool of stagnation into which we have fallen: cartilage repair redivivus! DESIGN Articular-cartilage lesions may be induced traumatically (e.g., by sports injuries and occupational accidents) or pathologically during the course of a degenerative disease (e.g., osteoarthritis). This review addresses the biological basis of cartilage repair and surveys current trends in treatment strategies, focussing on those that are most widely adopted by orthopaedic surgeons [viz., abrasive chondroplasty, microfracturing/microdrilling, osteochondral grafting and autologous-chondrocyte implantation (ACI)]. Also described are current research activities in the field of cartilage-tissue engineering, which, as a therapeutic principle, holds more promise for success than any other experimental approach. RESULTS AND CONCLUSIONS Tissue engineering aims to reconstitute a tissue both structurally and functionally. This process can be conducted entirely in vitro, initially in vitro and then in vivo (in situ), or entirely in vivo. Three key constituents usually form the building blocks of such an approach: a matrix scaffold, cells, and signalling molecules. Of the proposed approaches, none have yet advanced beyond the phase of experimental development to the level of clinical induction. The hurdles that need to be surmounted for ultimate success are discussed.

The manufacture of concrete blocks and their use in building construction /

Includes index.

Developing Innovative TEL Practice in Teacher Education

This interactive symposium will focus on the use of different technologies in developing innovative practice in teacher education at one university in England. Technology Enhanced Learning (TEL) is a field of educational policy and practice that has the power to ignite diametrically opposing views and reactions amongst teachers and teacher educators, ranging across a spectrum from immense enthusiasm to untold terror. In a field where the skills and experience of individuals vary from those of digital natives (Prensky 2001) to lags and lurkers in digital spaces, the challenges of harnessing the potential of TEL are complex. The challenges include developing the IT skills of trainees and educators and the creative application of these skills to pedagogy in all areas of the curriculum. The symposium draws on examples from primary, secondary and post-compulsory teacher education to discuss issues and approaches to developing research capacity and innovative practice using different etools, many of which are freely available. The first paper offers theoretical and policy perspectives on finding spaces in busy professional lives to engage in research and develop research-informed practice. It draws on notions of teachers as researchers, practitioner research and evidenc-ebased practice to argue that engagement in research is integral to teacher education and an empowering source of creative professional learning for teachers and teacher educators. Whilst acknowledging the challenges of this stance, examples from our own research practice illustrate how e-tools can assist us in building the capacity and confidence of staff and students in researching and enhancing teaching, learning and assessment practice. The second paper discusses IT skills development through the TEL pathway for trainee teachers in secondary education across different curriculum subjects. The lead tutor for the TEL pathway will use examples of activities developed with trainee teachers and university subject tutors to enhance their skills in using e-tools, such as QR codes, Kahoot, Padlet, Pinterest and cloud based learning. The paper will also focus on how these skills and tools can be used for action Discussant - the wider use of technologies in a university centre for teacher education; course management, recruitment and mentor training. research, evaluation and feedback and for marking and administrative tasks. The discussion will finish with thoughts on widening trainee teachers’ horizons into the future direction of educational technology. The third paper considers institutional policies and strategies for promoting and embedding TEL, including an initiative called ‘The Learning Conversation’, which aims ‘to share, highlight, celebrate, discuss, problematise, find things out...’ about TEL through an online space. The lead for ‘The Learning Conversation’ will offer reflections on this and other initiatives across the institution involving trainee teachers, university subject tutors, librarians and staff in student support services who are using TEL to engage, enthuse and support students on campus and during placements in schools. The fourth paper reflects on the use of TEL to engage with trainee teachers in post-compulsory education. This sector of education and training is more fragmented than primary and secondary schools sectors and so the challenges of building a community of practice that can support the development of innovative practice are greater.