Improving outcome after stroke: overcoming the translational roadblock

Stroke poses a massive burden of disease, yet we have few effective therapies. The paucity of therapeutic options stands contrary to intensive research efforts. The failure of these past investments demands a thorough re-examination of the pathophysiology of ischaemic brain injury. Several critical areas hold the key to overcoming the translational roadblock: (1) vascular occlusion: current recanalization strategies have limited effectiveness and may have serious side effects; (2) complexity of stroke pathobiology: therapy must acknowledge the 'Janus-faced' nature of many stroke targets and must identify endogenous neuroprotective and repair mechanisms; (3) inflammation and brain-immune-system interaction: inflammation contributes to lesion expansion, but is also instrumental in lesion containment and repair; stroke outcome is modulated by the interaction of the injured brain with the immune system; (4) regeneration: the potential of the brain for reorganization, plasticity and repair after injury is much greater than previously thought; (5) confounding factors, long-term outcome and predictive modelling. These 5 areas are linked on all levels and therefore need to be tackled by an integrative approach and innovative therapeutic strategies.

Government initiative and start-up firms’ eco-innovation : A case study through the ecological modernisation lens

Government programs to finance small firms or start-ups have attracted a little empirical attention. From an economical perspective, the effect of government grants is evaluated by a measure of innovation or firm productivity. Yet, this paper takes a different approach from economical view aiming to address the research question “How do start ups firms view the relationship between government grants and their co-efficient innovation effort?” Semi-structured interviews with grant recipients (start-up business owners revealed that the grants assist firms to leverage their resource limitations but at the same time the grants also act as a major roadblock for their product development success.

Standardisation of digital human models

Digital human models (DHM) have evolved as useful tools for ergonomic workplace design and product development, and found in various industries and education. DHM systems which dominate the market were developed for specific purposes and differ significantly, which is not only reflected in non-compatible results of DHM simulations, but also provoking misunderstanding of how DHM simulations relate to real world problems. While DHM developers are restricted by uncertainty about the user need and lack of model data related standards, users are confined to one specific product and cannot exchange results, or upgrade to another DHM system, as their previous results would be rendered worthless. Furthermore, origin and validity of anthropometric and biomechanical data is not transparent to the user. The lack of standardisation in DHM systems has become a major roadblock in further system development, affecting all stakeholders in the DHM industry. Evidently, a framework for standardising digital human models is necessary to overcome current obstructions.

Jet and flash imprint defectivity - Assessment and reduction for semiconductor applications

Defectivity has been historically identified as a leading technical roadblock to the implementation of nanoimprint lithography for semiconductor high volume manufacturing. The lack of confidence in nanoimprint's ability to meet defect requirements originates in part from the industry's past experiences with 1 × lithography and the shortage in enduser generated defect data. SEMATECH has therefore initiated a defect assessment aimed at addressing these concerns. The goal is to determine whether nanoimprint, specifically Jet and Flash Imprint Lithography from Molecular Imprints, is capable of meeting semiconductor industry defect requirements. At this time, several cycles of learning have been completed in SEMATECH's defect assessment, with promising results. J-FIL process random defectivity of < 0.1 def/cm2 has been demonstrated using a 120nm half-pitch template, providing proof of concept that a low defect nanoimprint process is possible. Template defectivity has also improved significantly as shown by a pre-production grade template at 80nm pitch. Cycles of learning continue on feature sizes down to 22nm. © 2011 SPIE.

Type IA topoisomerase inhibition by clamp closure

Bacterial DNA topoisomerase I (topoI) catalyzes relaxation of negatively supercoiled DNA. The enzyme alters DNA topology through protein-operated DNA gate, switching between open and closed conformations during its reaction. We describe the mechanism of inhibition of Mycobacterium smegmatis and Mycobacterium tuberculosis topoI by monoclonal antibodies (mAbs) that bind with high affinity and inhibit at 10-50 nM concentration. Unlike other inhibitors of topoisomerases, the mAbs inhibited several steps of relaxation reaction, namely DNA binding, cleavage, strand passage, and enzyme-DNA dissociation. The enhanced religation of the cleaved DNA in presence of the mAb indicated closing of the enzyme DNA gate. The formation of enzyme-DNA heterocatenane in the presence of the mAbs as a result of closing the gate could be inferred by the salt resistance of the complex, visualized by atomic force microscopy and confirmed by fluorescence measurements. Locking the enzyme-DNA complex as a closed clamp restricted the movements of the DNA gate, affecting all of the major steps of the relaxation reaction. Enzyme trapped on DNA in closed clamp conformation formed roadblock for the elongating DNA polymerase. The unusual multistep inhibition of mycobacterial topoisomerases may facilitate lead molecule development, and the mAbs would also serve as valuable tools to probe the enzyme mechanism.

Strategies for adding features to CAD models in order to optimize performance

This paper presents an approach which enables new parameters to be added to a CAD model for optimization purposes. It aims to remove a common roadblock to CAD based optimization, where the parameterization of the model does not offer the shape sufficient flexibility for a truly optimized shape to be created. A technique has been developed which uses adjoint based sensitivity maps to predict
the sensitivity of performance to the addition to a model of four different feature types, allowing the feature providing the greatest benefit to be selected. The optimum position to add the feature is also discussed. It is anticipated that the approach could be used to iteratively add features to a model, providing greater flexibility to the shape of the model, and allowing the newly-added parameters to be used as design variables in a subsequent shape optimization.

Cryptic species in the cosmopolitan Bugulan eritina complex (Bryozoa,Cheilostomata)

Previous analyses of the mitochondrial gene cytochrome c oxidase subunit 1 (COI) and γ-proteobacterial endosymbiont diversity have suggested that the marine bryozoan Bugula neritina is a complex of three cryptic species, namely Types S, D and N. Types D and N were previously reported to have restricted distributions along California (western USA) and Delaware and Connecticut (eastern USA), respectively, whereas Type S is considered widespread in tropical, subtropical and temperate regions due to anthropogenic transport. Here, Bayesian species delimitation analysis of a data set composed of two mitochondrial (COI and large ribosomal RNA subunit [16S]) and two nuclear genes (dynein light chain roadblock type-2 protein [DYN] and voltage-dependent anion-selective channel protein [VDAC]) demonstrated that Types S, D and N correspond to three biological species. This finding was significantly supported, in spite of the combinations of priors applied for ancestral population size and root age. Furthermore, COI sequences were used to assess the introduction patterns of the cosmopolitan Type S species. Two COI haplotypes of Type S (S1a and S1d) were found occurring at a global scale. Mantel tests showed correlation between these haplotypes and local sea surface temperature tolerance. Accordingly, the distributions of Type S haplotypes may reflect intraspecific temperature tolerance variation, in addition to the role of introduction vectors. Finally, we show that the Type N may also have been introduced widely, as this species was found for the first time in Central California and north-eastern Australia.

Consequences of DNA damage for gene transcription : direct effects of modified nucleobases and the role of base excision repair = Folgen von DNA-Schäden für die Gentranskription

The presence of damaged nucleobases in DNA can negatively influence transcription of genes. One of the mechanisms by which DNA damage interferes with reading of genetic information is a direct blockage of the elongating RNA polymerase complexes – an effect well described for bulky adducts induced by several chemical substances and UV-irradiation. However, other mechanisms must exist as well because many of the endogenously occurring non-bulky DNA base modifications have transcription-inhibitory properties in cells, whilstrnnot constituting a roadblock for RNA polymerases under cell free conditions. The inhibition of transcription by non-blocking DNA damage was investigated in this work by employing the reporter gene-based assays. Comparison between various types of DNA damage (UV-induced pyrimidine photoproducts, oxidative purine modifications induced by photosensitisation, defined synthetic modified bases such as 8-oxoguanine and uracil, and sequence-specific single-strand breaks) showed that distinct mechanisms of inhibition of transcription can be engaged, and that DNA repair can influence transcription of the affectedrngenes in several different ways.rnQuantitative expression analyses of reporter genes damaged either by the exposure of cells to UV or delivered into cells by transient transfection supported the earlier evidence that transcription arrest at the damage sites is the major mechanism for the inhibition of transcription by this kind of DNA lesions and that recovery of transcription requires a functional nucleotide excision repair gene Csb (ERCC6) in mouse cells. In contrast, oxidisedrnpurines generated by photosensitisation do not cause transcriptional blockage by a direct mechanism, but rather lead to transcriptional repression of the damaged gene which is associated with altered histone acetylation in the promoter region. The whole chain of events leading to transcriptional silencing in response to DNA damage remains to be uncovered. Yet, the data presented here identify repair-induced single-strand breaks – which arise from excision of damaged bases by the DNA repair glycosylases or endonucleases – as arnputative initiatory factor in this process. Such an indirect mechanism was supported by requirement of the 8-oxoguanine DNA glycosylase (OGG1) for the inhibition of transcription by synthetic 8-oxodG incorporated into a reporter gene and by the delays observed for the inhibition of transcription caused by structurally unrelated base modifications (8-oxoguanine and uracil). It is thereby hypothesized that excision of the modified bases could be a generalrnmechanism for inhibition of transcription by DNA damage which is processed by the base excision repair (BER) pathway. Further gene expression analyses of plasmids containing single-strand breaks or abasic sites in the transcribed sequences revealed strong transcription inhibitory potentials of these lesions, in agreement with the presumption that BER intermediates are largely responsible for the observed effects. Experiments with synthetic base modifications positioned within the defined DNA sequences showed thatrninhibition of transcription did not require the localisation of the lesion in the transcribed DNA strand; therefore the damage sensing mechanism has to be different from the direct encounters of transcribing RNA polymerase complexes with DNA damage.rnAltogether, this work provides new evidence that processing of various DNA basernmodifications by BER can perturb transcription of damaged genes by triggering a gene silencing mechanism. As gene expression can be influenced even by a single DNA damage event, this mechanism could have relevance for the endogenous DNA damage induced in cells under normal physiological conditions, with a possible link to gene silencing in general.

Representaciones del conflicto y medios masivos : la pueblada de Salta en noviembre de 2000 a través de Clarín y El Tribuno

En el presente artículo pretendemos indagar sobre las diversas maneras en que fue mencionado e interpretado el episodio de protesta mediante corte de rutas acontecido en noviembre de 2000 en el norte de la provincia de Salta, en el marco de un extenso proceso de luchas que comienzan a mediados de la década de 1990. El estudio se basa en las representaciones que fueron divulgadas por dos medios masivos de información (Clarín y El Tribuno). Reconstruiremos, mediante la lectura crítica de las noticias, las caracterizaciones sobre los protagonistas de la protesta, quiénes fueron sus emisores, los momentos en que fueron realizadas, etc. En tal sentido, el objetivo será elucidar el modo en que enfrentamientos físicos y simbólicos, lejos de estar divorciados, se condicionan mutuamente.

Improving the Manufacturability of Active Microfluidic Devices using Stereolithography (SL)

Thesis (Master's)--University of Washington, 2016-06

Weathering characteristics and lithology of samples from the Rauer Group, Antarctica, obtained in February/March 2007, and conducted with field support of POLARSTERN cruise ANT-XXIII/9

The presence of glacial sediments across the Rauer Group indicates that the East Antarctic ice sheet formerly covered the entire archipelago and has since retreated at least 15 km from its maximum extent. The degree of weathering of these glacial sediments suggests that ice retreat from this maximum position occurred sometime during the latter half of the last glacial cycle. Following this phase of retreat, the ice sheet margin has not expanded more than ~1 km seaward of its present position. This pattern of ice sheet change matches that recorded in Vestfold Hills, providing further evidence that the diminutive Marine Isotope Stage 2 ice sheet advance in the nearby Larsemann Hills may have been influenced by local factors rather than a regional ice-sheet response to climate and sea-level change.

IGF-1R inhibition sensitizes breast cancer cells to ATM-Related Kinase (ATR) inhibitor and cisplatin

The complexity of the IGF-1 signalling axis is clearly a roadblock in targeting this receptor in cancer therapy. Here, we sought to identify mediators of resistance, and potential co-targets for IGF-1R inhibition. By using an siRNA functional screen with the IGF-1R tyrosine kinase inhibitor (TKI) BMS-754807 in MCF-7 cells we identified several genes encoding components of the DNA damage response (DDR) pathways as mediators of resistance to IGF-1R kinase inhibition. These included ATM and Ataxia Telangiectasia and RAD3-related kinase (ATR). We also observed a clear induction of DDR in cells that were exposed to IGF-1R TKIs (BMS-754807 and OSI-906) as indicated by accumulation of γ-H2AX, and phosphorylated Chk1. Combination of the IGF-1R/IR TKIs with an ATR kinase inhibitor VE-821 resulted in additive to synergistic cytotoxicity compared to either drug alone. In MCF-7 cells with stably acquired resistance to the IGF-1R TKI (MCF-7-R), DNA damage was also observed, and again, dual inhibition of the ATR kinase and IGF-1R/IR kinase resulted in synergistic cytotoxicity. Interestingly, dual inhibition of ATR and IGF-1R was more effective in MCF-7-R cells than parental cells. IGF-1R TKIs also potentiated the effects of cisplatin in a panel of breast cancer cell lines. Overall, our findings identify induction of DDR by IGF-1R kinase inhibition as a rationale for co-targeting the IGF-1R with ATR kinase inhibitors or cisplatin, particularly in cells with acquired resistance to TKIs.