Layer‐by‐Layer Coated Gold Nanoparticles: Size‐Dependent Delivery of DNA into Cells

Because nanoparticles are finding uses in myriad biomedical applications, including the delivery of nucleic acids, a detailed knowledge of their interaction with the biological system is of utmost importance. Here the size‐dependent uptake of gold nanoparticles (AuNPs) (20, 30, 50 and 80 nm), coated with a layer‐by‐layer approach with nucleic acid and poly(ethylene imine) (PEI), into a variety of mammalian cell lines is studied. In contrast to other studies, the optimal particle diameter for cellular uptake is determined but also the number of therapeutic cargo molecules per cell. It is found that 20 nm AuNPs, with diameters of about 32 nm after the coating process and about 88 nm including the protein corona after incubation in cell culture medium, yield the highest number of nanoparticles and therapeutic DNA molecules per cell. Interestingly, PEI, which is known for its toxicity, can be applied at significantly higher concentrations than its IC₅₀ value, most likely because it is tightly bound to the AuNP surface and/or covered by a protein corona. These results are important for the future design of nanomaterials for the delivery of nucleic acids in two ways. They demonstrate that changes in the nanoparticle size can lead to significant differences in the number of therapeutic molecules delivered per cell, and they reveal that the toxicity of polyelectrolytes can be modulated by an appropriate binding to the nanoparticle surface.     

Neural Networks Approach to the Random Walk Dilemma of Financial Time Series

Predictions of financial time series often show a characteristic one step shift relative to the original data as in a random walk. This has been the cause for opposing views whether such time series do contain information that can be extracted for predictions, or are simply random walks. In this case study, we show that NNs that are capable of extracting weak low frequency periodic signals buried in a strong high frequency signal, consistently predict the next value in the series to be the current value, as in a random walk, when used for one-step-ahead predictions of the detrended S&P 500 time series. In particular for the Time Delay Feed Forward Networks and Elman Networks of various configurations, our study supports the view of the detrended S&P 500 being a random walk series. This is consistent with the long standing hypothesis that some financial time series are random walk series.     

Multi-Agent Dynamic Logics with Informational Test

This paper investigates a family of logics for reasoning about the dynamic activities and informational attitudes of agents, namely the agents' beliefs and knowledge. The logics are based on a new formalisation and semantics of the test operator of propositional dynamic logic and a representation of actions which distinguishes abstract actions from concrete actions. The new test operator, called informational test, can be used to formalise the beliefs and knowledge of particular agents as dynamic modalities. This approach is consistent with the formalisation of the agents' beliefs and knowledge as K(D)45 and S5 modalities. Properties concerning informativeness, truthfulness and preservation of beliefs are proved for a derivative of the informational test operator. It is shown that common belief and common knowledge can be expressed in the considered logics. This means, the logics are more expressive than propositional dynamic logic with an extra modality for belief or knowledge. The logics remain decidable and belong to 2EXPTIME. Versions of the considered logics express natural additional properties of beliefs or knowledge and interaction of beliefs or knowledge with actions. It is shown that a simulation of PDL can be constructed in one of these extensions.     

The WALL: participatory design workspace in support of creativity, collaboration, and socialization

A key challenge faced by organizations is to provide project teams with workspaces, information, and collaboration technologies that fosters creativity and high-performance team productivity. This requires understanding the relation between and impacts of (1) workspace, (2) activity and content that is created, and (3) social, behavioral, and cognitive aspects of work. This paper describes an exploratory study of everyday activities in the context of knowledge work in a shared workspace used by a high-tech global design team that explores future products. The study formalizes key elements for productive knowledge work as a function of tasks, context, and team. It identifies enablers, hindrances, and requirements for physical, virtual, and social work environments. The study identified, through semi-structured interviews, surveys, and on-site shadowing, a key workspace component that facilitates dynamic participation of all team members. This workspace component is a wall used as a large, public, physical display surface for project content (the WALL). The WALL acts as a mediator for individual reflection-in-action and team reflection-in-interaction. It serves as “social glue” both between individuals and between geographically distributed subgroups.     

Building common ground in global teamwork through re-representation

We explore in this paper the relation between activities, communication channels and media, and common ground building in global teams. We define re-representation as a sequence of representations of the same concept using different communication channels and media. We identified the re-representation technique to build common ground that is used by team members during multimodal and multimedia communicative events in cross-disciplinary, geographically distributed settings. Our hypotheses are as follows: (1) Significant sources of information behind decisions and request for actions are embedded within the fabric of communicative events in which participants use both informal and formal media to express their ideas. Capturing these information sources can facilitate common ground building and accelerate the execution of action requests. (2) Re-representations of concepts, i.e., sequences of representations using diverse media and communication channels, mediate and accelerate common ground building. (3) The use of intra- or interdisciplinary re-representations correlates with high team performance, i.e., effective team process and high product quality. We used AEC Global Teamwork course offered in 2008–2009 as the testbed for our study to validate our hypothesis.     

Spectrophotometric measurements of pH in-situ: laboratory and field evaluations of instrumental performance

Automated in-situ instrumentation has been developed for precise and accurate measurements of a variety of analytes in natural waters. In this work we describe the use of 'SEAS' (spectrophotometric elemental analysis system) instrumentation for measurements of solution pH. SEAS-pH incorporates a CCD-based spectrophotometer, an incandescent light source, and dual pumps for mixing natural water samples with a sulfonephthalein indicator. The SEAS-pH optical cell consists of either a liquid core waveguide (LCW, Teflon AF-2400) or a custom-made PEEK cell. Long optical path lengths allow use of indicators at low concentrations, thereby precluding large indicator-induced pH perturbations. Laboratory experiments show that pH measurements obtained using LCW and PEEK optical cells are indistinguishable from measurements obtained using conventional spectrophotometric cells and high-performance spectrophotometers. Deployments in the Equatorial Pacific and the Gulf of Mexico demonstrate that SEAS-pH instruments are capable of obtaining vertical pH profiles with high spatial resolution. SEAS-pH deployments at a fixed river-site (Hillsborough River, FL) demonstrate the capability of SEAS for observations of diel pH cycles with high temporal resolution. The in-situ precision of SEAS-pH is assessed as 0.0014 pH units, and the system's measurement frequency is approximately 0.5 Hz. This work indicates that in-situ instrumentation can be used to provide accurate, precise, and highly resolved observations of carbon-system transformations in the natural environment.     

Molecular basis of the constitutive activity and STI571 resistance of Asp816Val mutant KIT receptor tyrosine kinase

The receptor tyrosine kinase, KIT, displays activating mutations in the kinase domain, which are associated with various cancers. We have used homology modelling based on the crystal structures of the insulin receptor kinase in active and inactive conformations to predict the corresponding structures of the KIT kinase domain. We have prepared four KIT models, one each for the active and inactive conformations of the wild-type and of the Asp816Val mutant proteins. We have also placed ATP into the active conformations and the inhibitor, STI571, into the inactive conformations. All models have been fully energy minimised. The molecular modelling studies described here explain (i) why Asp816Val KIT is constitutively active, (ii) why the nature of the substituting amino acid at residue 816 is relatively unimportant, and (iii) why the Asp816Val substitution confers resistance to the KIT-inhibitory drug STI571. The models will be valuable for predicting other kinase inhibitory drugs that may be active on wild-type and mutant forms of KIT. During the course of this work, a crystal structure of the active conformation of the KIT kinase domain has been published. Our model of the active conformation of the Asp816Val mutant is strikingly similar to this crystal structure, whereas our model of the active conformation of the wild-type kinase domain of KIT differs from the crystal structure in some respects. The reasons for this apparent discrepancy are discussed.     

(Ba,Ca)TiO3 PTCR Ceramics with LaNiO3 Thin-Film Electrodes: Preparation and Characterization of the Interface

Conductive LaNiO₃ thin film electrodes were deposited by chemical solution deposition (CSD) from nitrate solutions onto polycrystalline Al₂O₃ and (Ba,Ca)TiO₃ PTCR ceramic substrates. The electrical properties of the LaNiO₃ thin film on Al₂O₃ and of the interface consisting of LaNiO₃ and the semiconductive oxide ceramic were investigated. The deposited LaNiO₃ films were about 250 nm thick and consisted of nanosized particles. The resistivity of the LaNiO₃ film was about 3 × 10^-3cm at 20°C. The PTCR ceramic consisted of m sized particles and exhibited an electronic resistivity of about 10 cm at 20°C and a steep increase of the resistivity of a few orders of magnitude above the Curie point at about 120°C. The electrical properties of the LaNiO₃/PTCR interface were dominated by the properties of a barrier layer between the PTCR ceramic and the LaNiO₃ electrode. The potential dependence of the impedance indicated that the barrier layer consisted of a depletion layer within the PTCR ceramic, when the flat band potential of LaNiO₃ on the PTCR ceramic at about –250 mV was exceeded. Additionally the formation of an insulating layer at the LaNiO₃ electrode has to be taken into account.     

How does personality matter in marriage? An examination of trait anxiety, interpersonal negativity, and marital satisfaction.

Although trait anxiety and its aliases (negative affectivity, neuroticism) have frequently been found to be associated with marital dissatisfaction, few efforts have been made to identify the processes through which trait anxiety exerts its influence. This study reports findings from a 13-year, 4-phase longitudinal study in which trait anxiety, measured when spouses were newlyweds, consistently predicted marital negativity which, in turn, was associated with partner's marital dissatisfaction. Some support was also found for effects of trait anxiety on partner's marital satisfaction, independent of marital negativity, as well as for the idea that trait anxiety is directly related to spouses' own marital satisfaction. Trait anxiety did not distinguish couples who divorced from those who remained married, and it generally did not predict declines in marital satisfaction. The disagreeable impact of trait anxiety on marriage was evident at the outset of marriage and was stable over time. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Addressing the numbers problem in directed evolution

Our previous contribution to increasing the efficiency of directed evolution is iterative saturation mutagenesis (ISM) as a systematic means of generating focused libraries for the control of substrate acceptance, enantioselectivity, or thermostability of enzymes. We have now introduced a crucial element to knowledge-guided targeted mutagenesis in general that helps to solve the numbers problem in directed evolution. We show that the choice of the amino acid (aa) alphabet, as specified by the utilized codon degeneracy, provides the experimenter with a powerful tool in designing "smarter" randomized libraries that require considerably less screening effort. A systematic comparison of two different codon degeneracies was made by examining the relative quality of the identically sized enzyme libraries in relation to the degree of oversampling required in the screening process. The specific example in our case study concerns the conventional NNK codon degeneracy (32 codons/20 aa) versus NDT (12 codons/12 aa). The model reaction is the hydrolytic kinetic resolution of a chiral trans-disubstituted epoxide, catalyzed by the epoxide hydrolase from Aspergillus niger. The NDT library proves to be of much higher quality, as measured by the dramatically higher frequency of positive variants and by the magnitude of catalyst improvement (enhanced rate and enantioselectivity). We provide a statistical analysis that constitutes a useful guide for the optimal design and generation of "smarter" focused libraries. This type of approach accelerates the process of laboratory evolution considerably and can be expected to be broadly applicable when engineering functional proteins in general.