640 resultados para Pull


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Aquesta tesi doctoral s'engloba dins d'un projecte general d'estudi de gens implicats en l'embriogènesi del blat de moro. L'embriogènesi del blat de moro, i en general la de totes les plantes superiors, es dóna en tres etapes: una primera etapa on es diferencien tots els diversos teixits que formaran l'embrió, una segona etapa on l'embrió acumula productes de reserva i un tercer període, la dormància, que finalitza quan les condicions ambientals són les idònies per a la germinació. En el laboratori estàvem interessats, concretament, en l'estudi de gens implicats en la primera etapa morfogenètica, on els diferents teixits i estructures embrionàries queden definides. Per tal d'estudiar gens que s'expressaven en aquest període, una de les estratègies que es va realitzar fou un crivellat diferencial entre teixit embrionari i teixit de planta adulta. D'entre els diferents clons obtinguts, un corresponia a un clon parcial que presentava similitud amb receptors quinasa i que fou objecte d'estudi. A partir d'aquest clon es va obtenir el clon complet i es va anomenar MARK (per Maize Atypical Receptor Kinase). MARK presenta una estructura típica d'un receptor quinasa amb un domini extracel.lular, que conté 6 còpies imperfectes de LRR (Leucine- Rich Repeats), un únic domini transmembrana i un domini quinasa intracel.lular. El domini quinasa de MARK presenta, però, algunes variacions en els residus aminoacídics que es consideren claus per a la funció catalítica dels dominis quinasa. En concret cinc dels aminoàcids considerats essencials per a la fosforilació es troben substituits en el domini quinasa de MARK (DK-MARK). Els experiments de fosforilació in vitro que es van realitzar al laboratori, van mostrar com MARK era incapaç de fosforilar in vitro. Aquesta característica no és, però, exclusiva de MARK. Una búsqueda en les bases de dades ens van permetre identificar altres seqüències que també presentaven els mateixos o altres canvis en aquestes posicions aminoacídiques. En les bases de dades de plantes es van identificar un conjunt de seqüències genòmiques o ESTs amb aquestes característiques i només una d'elles, la proteïna TMKL1 d'Arabidopsis, ha sigut descrita com un receptor quinasa incapaç de fosforilar in vitro. Respecte a la búsqueda de receptors similars a MARK en les bases de dades d'animals, es van identificar també un conjunt de proteïnes que, en alguns casos, s'ha descrit que no tenen activitat quinasa in vivo. Per exemple, un dels casos més ben estudiats és el del receptor erbB3 que forma part de la família de receptors del EGF (Epidermal Growth Factor). Aquesta família de receptors està formada per 4 receptors: erbB1, erbB2, erbB3 i erbB4, dels quals només l'erbB3 no presenta activitat catalítica. S'ha descrit que erbB3 és capaç, tot i no fosforilar in vivo, de participar activament en la transducció del senyal formant heterodímers amb els altres membres de la família. Així, erbB3 és fosforilat pel seu partner i pot iniciar la cascada de transducció del senyal. La participació d'erbB3 en la transducció del senyal és essencial ja que embrions de ratolí knock-out pel gen erbB3 són inviables. Així doncs, el fet que receptors quinasa catalíticament inactius participin en les cascades de transducció del senyal, suggereix l'existència de nous mecanismes d'acció per a la transducció del senyal. Per tant, l'objectiu d'aquest treball fou l'estudi del mecanisme d'acció de MARK mitjançant la caracterització les proteïnes capaces d'interaccionar amb el seu domini quinasa. Per tal d'assolir aquest objectiu, es va realitzar un crivellat de doble-híbrid amb una llibreria de cDNA d'embrions de blat de moro de 7 DAP. D'aquest crivellat es va obtenir un conjunt de possibles clons positius que foren seqüenciats i entre els quals es van escollir per un estudi més detallat aquells que s'havien obtingut més vegades com a clons independents. Aquests clons codificaven per: una SAMDC (S-Adenosil Descarboxilasa), una eIF5 (Eukaryotic translation initiation), una hypothetical protein, una unknown protein, una gamma-adaptina i una MAP4K. Amb aquests 6 clons es van fer estudis in vitro i in vivo per tal de confirmar al seva interacció amb DK-MARK. Els estudis in vivo es van realitzar amb la soca de llevat AH109, una soca més astringent que la utilitzada en el crivellat, ja que presenta tres gens marcadors: Histidina, Adenina i Lacz. Els resultats obtinguts van mostrar que els clons codificants per SAMDC i eIF5 no van créixer en un medi selectiu per His i Ade i, per tant o es tracta de falsos positius del sistema o la seva interacció amb DK-MARK és dèbil. D'altra banda, la resta dels clons analitzats (proteïna hipotètica, una proteïna de funció desconeguda, la gamma-adaptina i una MAP4K) van créixer en medis en absència de Histidina i Adenina. Els assatjos de b-galactosidasa van ser tots positius a excepció de la proteïna hipotètica suggerint que potser aquesta interacció sigui més feble. D'altra banda també es van realitzar estudis in vitro amb la tècnica del pull-down. Els resultats obtinguts amb aquesta tècnica van recolzar els obtinguts en cèl.lules de llevat, ja que tots els clons analitzats a excepció dels codificants per SAMDC i eIF5 van donar un resultat d'interacció amb KD-MARK in vitro positiu. Davant aquests resultats ens vam centrar en l'estudi de la proteïna similar a MAP4K, doncs algunes proteïnes de la seva família s'han relacionat amb receptors de membrana. Els clons que es va obtenir del crivellat codificaven per una proteïna similar amb el domini C-terminal a les proteïnes BnMAP4Ka1 i a2 de Brassica napus. Aquestes proteïnes presenten una forta similitud de seqüència amb proteïnes de la família GCK/SPS1 que formen part d'un grup particular de MAPK relacionades amb la proteïna Ste20 (sterile 20 protein) de llevat. Ste20p activa la MAP3K de llevat Ste11 directament per fosforilació, transduint d'aquesta manera el senyal del receptor de feromones de creuament de les cèl.lules de llevat i es pot, doncs, considerar com una proteïna del tipus MAP4K (mitogen-activated protein kinase kinase kinase kinase). En els darrers anys, s'han identificat un gran nombre de proteïnes similars a Ste20: fins a una trentena en mamífers, en Drosophila, en Caenorhabditis elegans i en altres organismes. Segons la seva estructura aminoacídica, la família Ste20 s'ha classificat en dues subfamílies: les proteïnes STE20/PAK (p21-activated kinases) i la subfamília GCK/SPS1 (germinal center kinases). Les dues subfamílies estan formades per proteïnes que contenen un domini quinasa i un domini regulador, però, mentre que les proteïnes PAK presenten el domini quinasa en la part C-terminal, les GCKs el presenten en la regió N terminal. Les proteïnes GCK presenten una elevada diversitat estructural en el domini regulador permetent la seva classificació en 6 subfamílies. Mitjançant la tècnica del RACE es va obtenir el clon de cDNA complet que es va anomenar MIK (MARK Interacting Kinase). Amb la tècnica del Southern blot es va poder determinar que el gen MIK és un gen de còpia única en el genoma de blat de moro. Per tal d'analitzar la possible interacció entre DK-MARK i MIK, es va estudiar tant el patró d'expressió d'ambdós gens com el seu patró d'acumulació d'ambdues proteïnes durant l'embriogènesi del blat de moro. El patró d'expressió, analitzat per Northen blot va mostrar uns patrons coincidents al llarg de l'embriogènesi des del seu inici fins als 20 DAP amb una acumulació màxima de mRNA en embrions de 15 DAP. D'altra banda per tal d'estudiar el patró d'acumulació de la proteïna MIK així com per comparar-lo amb el de MARK, es van realitzar estudis de Westerns blot. Els resultats també van mostrar una coincidència en el temps de l'acumulació de les proteïnes MARK i MIK durant l'embriogènesi de blat de moro amb una major acumulació en embrions de 15 i 20 DAP. Es van dur a terme també estudis d'immunolocalitzacions sobre embrions de blat de moro de 15 DAP per tal d'estudiar en quins teixits s'acumulaven ambdues proteïnes. Les immunolocalitzacions van mostrar una major acumulació tant de MARK com de MIK en les zones meristemàtiques i en el teixit vascular sobretot del coleòptil on s'aprecia una forta co-localització de MARK i MIK. Totes aquestes dades són compatibles, doncs, amb una possible interacció de les proteïnes MARK i MIK, tot i que no la demostren. Per tal de demostrar la interacció es van realitzar experiments d'immunoprecipitació in vivo a partir d'extractes d'embrions. Malauradament, els resultats no són clars i en aquests moments en el laboratori s'estan posant a punt aquests experiments. També es van realitzar estudis comparatius de seqüència amb diferents proteïnes de la família GCK, mostrant una major similitud amb les proteïnes de la subfamília GCK-III. La subfamília GCK-III ha estat molt poc estudiada i en formen part un conjunt de proteïnes amb funcions molt diverses des de l'apoptosi, la citoquinesi o l'anòxia cel.lular. Per tant, la similitud de seqüència possiblement fa referència a una conservació en el mecanisme d'acció més que no pas a una conservació funcional. La possible interacció de MARK amb el domini C-terminal de MIK (el domini regulador) podria activar aquesta última iniciant una cascada de transducció del senyal en un model en el que una proteïna del tipus GCK-III faria de lligam directa entre un receptor de membrana i una cascada de senyalització intracel.lular. Aquest tipus de lligam entre un recepctor de membrana i mòduls intracel.lulars de senyalització s'ha descrit per a altres proteïnes GCK, si bé no directament sinó a través de proteïnes adaptadores. D'altra banda, la interacció directa de MARK, un receptor quinasa atípic que no té activitat catalítica, amb MIK suggereix un mecanisme on receptors atípics podrien interaccionar en la transducció del senyal activant la via de les MAPK.

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El uso de barras de materiales compuestos (FRP) se propone como una alternativa efectiva para las tradicionales estructuras de hormigón armadas con acero que sufren corrosión en ambientes agresivos. La aceptación de estos materiales en el mundo de la construcción está condicionada a la compresión de su comportamiento estructural. Este trabajo estudia el comportamiento adherente entre barras de FRP y hormigón mediante dos programas experimentales. El primero incluye la caracterización de la adherencia entre barras de FRP y hormigón mediante ensayos de pull-out y el segundo estudia el proceso de fisuración de tirantes de hormigón reforzados con barras de GFRP mediante ensayo a tracción directa. El trabajo se concluye con el desarrollo de un modelo numérico para la simulación del comportamiento de elementos de hormigón reforzado bajo cargas de tracción. La flexibilidad del modelo lo convierte en una herramienta flexible para la realización de un estudio paramétrico sobre las variables que influyen en el proceso de fisuración.

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Smooth flow of production in construction is hampered by disparity between individual trade teams' goals and the goals of stable production flow for the project as a whole. This is exacerbated by the difficulty of visualizing the flow of work in a construction project. While the addresses some of the issues in Building information modeling provides a powerful platform for visualizing work flow in control systems that also enable pull flow and deeper collaboration between teams on and off site. The requirements for implementation of a BIM-enabled pull flow construction management software system based on the Last Planner System™, called ‘KanBIM’, have been specified, and a set of functional mock-ups of the proposed system has been implemented and evaluated in a series of three focus group workshops. The requirements cover the areas of maintenance of work flow stability, enabling negotiation and commitment between teams, lean production planning with sophisticated pull flow control, and effective communication and visualization of flow. The evaluation results show that the system holds the potential to improve work flow and reduce waste by providing both process and product visualization at the work face.

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The use of semiochemicals for manipulation of the pollen beetle Meligethes aeneus (Fabricius) (Coleoptera: Nitidulidae) is being investigated for potential incorporation into a push-pull control strategy for this pest, which damages oilseed rape, Brassica napus L. (Brassicaceae), throughout Europe. The response of M. aeneus to non-host plant volatiles was investigated in laboratory assays to establish whether they have any effect on host plant location behaviour. Two approaches were used. First a novel, moving-air bioassay using air funnels was developed to compare the response of M. aeneus to several non-host plant essential oils. The beetles avoided the host plant flowers in the presence of non-host volatiles, suggesting that M. aeneus uses olfactory cues in host location and/or acceptance. The results were expressed as 'repellency values' in order to compare the effects of the different oils tested. Lavender (Lavendula angustifolia Miller) (Lamiaceae) essential oil gave the highest repellency value. In addition, a four-arm olfactometer was used to investigate olfactory responses, as this technique eliminated the influence of host plant visual and contact cues. The attraction to host plant volatiles was reduced by the addition of non-host plant volatiles, but in addition to masking the host plant volatiles, the non-host volatiles were avoided when these were presented alone. This is encouraging for the potential use of non-host plants within a push-pull strategy to reduce the pest colonisation of crops. Further testing in more realistic semi-field and field trials is underway.

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We argue the case for a new branch of mathematics and its applications: Mathematics for the Digital Society. There is a challenge for mathematics, a strong “pull” from new and emerging commercial and public activities; and a need to train and inspire a generation of quantitative scientists who will seek careers within the associated sectors. Although now going through an early phase of boiling up, prior to scholarly distillation, we discuss how data rich activities and applications may benefit from a wide range of continuous and discrete models, methods, analysis and inference. In ten years time such applications will be common place and associated courses may be embedded within the undergraduate curriculum.

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Coronavirus nucleoproteins (N proteins) localize to the cytoplasm and the nucleolus, a subnuclear structure, in both virus-infected primary cells and in cells transfected with plasmids that express N protein. The nucleolus is the site of ribosome biogenesis and sequesters cell cycle regulatory complexes. Two of the major components of the nucleolus are fibrillarin and nucleolin. These proteins are involved in nucleolar assembly and ribosome biogenesis and act as chaperones for the import of proteins into the nucleolus. We have found that fibrillarin is reorganized in primary cells infected with the avian coronavirus infectious bronchitis virus (IBV) and in continuous cell lines that express either IBV or mouse hepatitis virus N protein. Both N protein and a fibrillarin-green fluorescent protein fusion protein colocalized to the perinuclear region and the nucleolus. Pull-down assays demonstrated that IBV N protein interacted with nucleolin and therefore provided a possible explanation as to how coronavirus N proteins localize to the nucleolus. Nucleoli, and proteins that localize to the nucleolus, have been implicated in cell growth-cell cycle regulation. Comparison of cells expressing IBV N protein with controls indicated that cells expressing N protein had delayed cellular growth. This result could not to be attributed to apoptosis. Morphological analysis of these cells indicated that cytokinesis was disrupted, an observation subsequently found in primary cells infected with IBV. Coronaviruses might therefore delay the cell cycle in interphase, where maximum translation of viral mRNAs can occur.

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Another Proof of the Preceding Theory was produced as part of a residency run by Artists in Archeology in conjunction with the Stonehenge Riverside project. The film explores the relationship between science, work and ritual, imagining archaeology as a future cult. As two robed disciples stray off from the dig, they are drawn to the drone of the stones and proceed to play the henge like a gigantic Theremin. Just as a Theremin is played with the hand interfering in an electric circuit and producing sound without contact, so the stones respond to the choreographed bodily proximity. Finally, one of the two continues alone to the avenue at Avebury, where the magnetic pull of the stones reaches its climax. Shot on VHS, the film features a score by Zuzushi Monkey, with percussion and theremin sounds mirroring the action. The performers are mostly artists and archeologists from the art and archaeology teams. The archeologists were encouraged to perform their normal work in the robes, in an attempt to explore the meeting points of science and ritual and interrogate our relationship to an ultimately unknowable prehistoric past where activities we do not understand are relegated to the realm of religion. Stonehenge has unique acoustic properties, it’s large sarsen stones are finely worked on the inside, left rough on the outside, intensifying sound waves within the inner horseshoe, but since their real use, having been built over centuries, remains ambiguous, the film proposes that our attempts to decode them may themselves become encoded in their cumulative meaning for future researchers.

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Os tripanossomatídeos são caracterizados por processos moleculares diferenciados como a transcrição policistrônica e regulação pós-transcricional da expressão gênica. Em mamíferos, a tradução se inicia com a ligação do complexo eIF4F (formado pelos eIF4A, eIF4E e eIF4G) a extremidade 5' dos mRNAs, o que facilita seu reconhecimento pelo ribossomo. A atividade do eIF4F é reforçada pela proteína de ligação a cauda poli-A (PABP), na extremidade 3' dos mRNAs, que interage com o eIF4G. Dois complexos do tipo eIF4F foram identificados em tripanossomatídeos: o primeiro formado pelos EIF4G3, EIF4E4 e EIF4AI com a PABP1; e um outro baseado na interação do EIF4G4 com o EIF4E3 e o EIF4A1. Este trabalho buscou caracterizar as interações entre as subunidades destes complexos e sua associação com PABPs de Leishmania, avaliando o efeito de mutações em motivos específicos. Proteínas recombinantes foram geradas fusionadas a GST e avaliadas quanto a sua habilidade de interagir com parceiros marcados radioativamente em ensaios do tipo pull-down. Para o EIF4G3, mutações individuais em dois resíduos vizinhos (I8A e R9A), afetaram a interação com o EIF4E4 e a mutação de ambos os resíduos equivalentes do EIF4G4 (IL25-26AA) também impediu sua ligação ao EIF4E3, sugerindo um motivo comum para a ligação aos seus parceiros. As proteínas EIF4E3 e EIF4E4 foram avaliadas quanto à capacidade de interagir com a PABP2 e PABP1 respectivamente, e mutações em motivos conservados nas regiões N-terminais dos EIF4E (Boxes A, B e C) aboliram sua interação com os homólogos da PABP. Para identificar que regiões da PABP1 estão relacionadas às interações com o parceiro EIF4E4, foram obtidas proteínas PABP1 mutantes em motivos conservados e observou-se que a mutação no motivo TGM, C-terminal, aboliu sua interação com o EIF4E4. Com estas abordagens conseguiu-se avançar na definição das interações entre as referidas subunidades do eIF4F e PABP, identificando-se diferenças relevantes em relação a outros eucariotos

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A semiochemical based push-pull strategy for control of oilseed rape pests is being developed at Rothamsted Research. This strategy uses insect and plant derived semiochemicals to manipulate pests and their natural enemies. An important element within this strategy is an understanding of the importance of non-host plant cues for pest insects and how such signals could be used to manipulate their behaviour. Previous studies using a range of non-host plants have shown that, for the pollen beetle Meligethes aeneus (Coleoptera: Nitidulidae), the essential oil of lavender, Lavandula angustifolia (Lamiaceae), was the most repellent. The aim of this study was to identify the active components in L. angustifolia oil, and to investigate the behaviour of M. aeneus to these chemicals, to establish the most effective use of repellent stimuli to disrupt colonisation of oilseed rape crops. Coupled gas chromatography-electroantennography (GC-EAG) and gas chromatography-mass spectrometry (GC-MS) resulted in the identification of seven active compounds which were tested for behavioural activity using a 4-way olfactometer. Repellent responses were observed with (±)-linalool and (±)-linalyl acetate. The use of these chemicals within a push-pull pest control strategy is discussed.

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Haptic computer interfaces provide users with feedback through the sense of touch, thereby allowing users to feel a graphical user interface. Force feedback gravity wells, i.e. attractive basins that can pull the cursor toward a target, are one type of haptic effect that have been shown to provide improvements in "point and click" tasks. For motion-impaired users, gravity wells could improve times by as much as 50%. It has been reported that the presentation of information to multiple sensory modalities, e.g. haptics and vision, can provide performance benefits. However, previous studies investigating the use of force feedback gravity wells have generally not provided visual representations of the haptic effect. Where force fields extend beyond clickable targets, the addition of visual cues may affect performance. This paper investigates how the performance of motion-impaired computer users is affected by having visual representations of force feedback gravity wells presented on-screen. Results indicate that the visual representation does not affect times and errors in a "point and click" task involving multiple targets.

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Gossip (or Epidemic) protocols have emerged as a communication and computation paradigm for large-scale networked systems. These protocols are based on randomised communication, which provides probabilistic guarantees on convergence speed and accuracy. They also provide robustness, scalability, computational and communication efficiency and high stability under disruption. This work presents a novel Gossip protocol named Symmetric Push-Sum Protocol for the computation of global aggregates (e.g., average) in decentralised and asynchronous systems. The proposed approach combines the simplicity of the push-based approach and the efficiency of the push-pull schemes. The push-pull schemes cannot be directly employed in asynchronous systems as they require synchronous paired communication operations to guarantee their accuracy. Although push schemes guarantee accuracy even with asynchronous communication, they suffer from a slower and unstable convergence. Symmetric Push- Sum Protocol does not require synchronous communication and achieves a convergence speed similar to the push-pull schemes, while keeping the accuracy stability of the push scheme. In the experimental analysis, we focus on computing the global average as an important class of node aggregation problems. The results have confirmed that the proposed method inherits the advantages of both other schemes and outperforms well-known state of the art protocols for decentralized Gossip-based aggregation.

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Purpose: Malawi’s current extension policy supports pluralism and advocates responsiveness to farmer demand. We investigate whether smallholder farmers’ experience supports the assumption that access to multiple service providers leads to extension and advisory services that respond to the needs of farmers. Design/methodology/approach: Within a case study approach, two villages were purposively selected for in-depth qualitative analysis of available services and farmers’ experiences. Focus group discussions were held separately with male and female farmers in each village, followed by semi-structured interviews with 12 key informants selected through snowball sampling. Transcripts were analysed by themes and summaries of themes were made from cross case analysis. Findings: Farmers appreciate having access to a variety of sources of technical advice and enterprise specific technology. However, most service providers continue to dominate and dictate what they will offer. Market access remains a challenge, as providers still emphasize pushing a particular technology to increase farm productivity rather than addressing farmers’ expressed needs. Although farmers work in groups, providers do not seek to strengthen these to enable active interaction and to link them to input and produce markets. This limits farmers’ capacity to continue with innovations after service providers pull out. Poor coordination between providers limits exploitation of potential synergies amongst actors. Practical implications: Services providers can adapt their approach to engage farmers in discussion of their needs and work collaboratively to address them. At a system level, institutions that have a coordination function can play a more dynamic role in brokering interaction between providers and farmers to ensure coverage and responsiveness. Originality/value: The study provides a new farmer perspective on the implementation of extension reforms.

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The use of semiochemicals for the manipulation of the pollen beetle, Meliethes aeneus (Fabricius) (Coleoptera: Nitidulidae), is being investigated for potential incorporation into a push-pull strategy for this pest, which damages oilseed rape, Brassica napus L. (Brassicaceae), throughout Europe. Previous laboratory behavioural studies using volatiles from non-host plants showed that M. aeneus is repelled by the odour of lavender, Lavendula angustifolia Mill. (Lamiaceae), essential oil. This article reports on semi-field and field trials to investigate this behaviour under more realistic conditions. Semi-field experiments were conducted to assess the relative importance of olfaction at different points in host location behaviour by M. aeneus. The results showed that oilseed rape plants treated with lavender odour were less colonised by M. aeneus in comparison with an untreated control, but that the treatment effect was much reduced if the lavender odour was applied after colonisation. The field experiment demonstrated that lavender odour caused a significant reduction in the number of adultM. aeneus infesting the oilseed rape plants in the treatment plots compared to the control plots. Overall, these findings are very encouraging for the future development of a push-pull pest control system.

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Consistent with a negativity bias account, neuroscientific and behavioral evidence demonstrates modulation of even early sensory processes by unpleasant, potentially threat-relevant information. The aim of this research is to assess the extent to which pleasant and unpleasant visual stimuli presented extrafoveally capture attention and impact eye movement control. We report an experiment examining deviations in saccade metrics in the presence of emotional image distractors that are close to a nonemotional target. We additionally manipulate the saccade latency to test when the emotional distractor has its biggest impact on oculomotor control. The results demonstrate that saccade landing position was pulled toward unpleasant distractors, and that this pull was due to the quick saccade responses. Overall, these findings support a negativity bias account of early attentional control and call for the need to consider the time course of motivated attention when affect is implicit

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The City of London is a major financial centre where the property market is heavily dependent on the space requirements of office occupiers. Technological change, however, is changing the way in which organisations conduct their business and is challenging the continued locational pull of the City. This research presents findings from a major survey of City occupiers carried out by The College research team during 2002. The research examines how the shape and form of the office property market in the City will be affected by process change in eBusiness and related technology over the next 5 years. The report, which includes an extensive literature review, examines the implications of increased homeworking and other trends for future office supply in the City. The research was funded by the Worshipful Company of Chartered Surveyors Trust (Toby Sutton Research Award), and part of the literature review by the Small Business Support Research Unit of DTI. It is essential reading for surveyors, planners, facilities managers and all those interested in the technology and property interface.