Bacterial mercury resistance from atoms to ecosystems

Bacterial resistance to inorganic and organic mercury compounds (HgR) is one of the most widely observed phenotypes in eubacteria. Loci conferring HgR in Gram-positive or Gram-negative bacteria typically have at minimum a mercuric reductase enzyme (MerA) that reduces reactive ionic Hg(II) to volatile, relatively inert, monoatomic Hg(0) vapor and a membrane-bound protein (MerT) for uptake of Hg(II) arranged in an operon under control of MerR, a novel metal-responsive regulator. Many HgR loci encode an additional enzyme, MerB, that degrades organomercurials by protonolysis, and one or more additional proteins apparently involved in transport. Genes conferring HgR occur on chromosomes, plasmids, and transposons and their operon arrangements can be quite diverse, frequently involving duplications of the above noted structural genes, several of which are modular themselves. How this very mobile and plastic suite of proteins protects host cells from this pervasive toxic metal, what roles it has in the biogeochemical cycling of Hg, and how it has been employed in ameliorating environmental contamination are the subjects of this review.     

Numerical methods to determine calcium release flux from calcium transients in muscle cells.

Several methods are currently in use to estimate the rate of depolarization-induced calcium release in muscle cells from measured calcium transients. One approach first characterizes calcium removal of the cell. This is done by determining parameters of a reaction scheme from a fit to the decay of elevated calcium after the depolarizing stimulus. In a second step, the release rate during depolarization is estimated based on the fitted model. Using simulated calcium transients with known underlying release rates, we tested the fidelity of this analysis in determining the time course of calcium release under different conditions. The analysis reproduced in a satisfactory way the characteristics of the input release rate, even when the assumption that release had ended before the start of the fitting interval was severely violated. Equally good reconstructions of the release rate time course could be obtained when the model used for the analysis differed in structure from the one used for simulating the data. We tested the application of a new strategy (multiple shooting) for fitting parameters in nonlinear differential equation systems. This procedure rendered the analysis less sensitive to ill-chosen initial guesses of the parameters and to noise. A locally adaptive kernel estimator for calculating numerical derivatives allowed good reconstructions of the original release rate time course from noisy calcium transients when other methods failed.     

Visual methods for the nanomolar detection of electrophilic reagents

A series of highly colored nitrophenolates and nitrothiophenolates has been tested as spray reagents for the detection of electrophilic species of the types commonly used in peptide and protein chemistry. Sensitive TLC detection of agents for alkylation, acylation, sulfonylation and phosphorylation was demonstrated. In addition, the thiophenolate sprays were sensitive for oxidizing agents in nanomolar quantities. Selective TLC detection of acylating and phosphorylating agents was accomplished by subsequent alkali treatment resulting in the restoration of color.     

Microscale methods to assemble mammalian cells into tissue-like structures

Different cell types make up tissues and organs hierarchically and communicate within a complex, three-dimensional (3D) environment. The in vitro recapitulation of tissue-like structures is meaningful, not only for fundamental cell biology research, but also for tissue engineering (TE). Currently, TE research adopts either the top-down or bottom-up approach. The top-down approach involves defining the macroscopic tissue features using biomaterial scaffolds and seeding cells into these scaffolds. Conversely, the bottom-up approach aims at crafting small tissue building blocks with precision-engineered structural and functional microscale features, using physical and/or chemical approaches. The bottom-up strategy takes advantage of the repeating structural and functional units that facilitate cell-cell interactions and cultures multiple cells together as a functional unit of tissue. In this review, we focus on currently available microscale methods that can control mammalian cells to assemble into 3D tissue-like structures.     

Visual data analysis and decision support methods for non-deterministic LCA

Principal components analysis coupled with non-parametric bootstrapping is introduced with an example as a powerful tool to help visualise, analyse and decide on comparative or single non-deterministic LCI/LCIA results. Decision support is provided by adding non-parametric bootstrapping (NPB) to the GA1A plane, which is a special case of principle comments analysis (PCA) built around the Promethée multicriteria decision aid model. In addition to an easy to analyse visual presentation of otherwise complex and cluttered numerical results, PCA with NPB is equally at ease will all data uncertainty formats used to date in LCA.     

Projection of Orthogonal Tiling from the Retina to the Visual Cortex

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A Direct Projection from Mouse Primary Visual Cortex to Dorsomedial Striatum

The mammalian striatum receives inputs from many cortical areas, but the existence of a direct axonal projection from the primary visual cortex (V1) is controversial. In this study we use anterograde and retrograde tracing techniques to demonstrate that V1 directly innervates a topographically defined longitudinal strip of dorsomedial striatum in mice. We find that this projection forms functional excitatory synapses with direct and indirect pathway striatal projection neurons (SPNs) and engages feed-forward inhibition onto these cells. Importantly, stimulation of V1 afferents is sufficient to evoke phasic firing in SPNs. These findings therefore identify a striatal region that is functionally innervated by V1 and suggest that early visual processing may play an important role in striatal-based behaviors.     

Improved methods to detect GTP-binding proteins from plants

Improved methods are described for the detection of G1P-binding proteins (G-proteins) in the protonema of mossFunaria hygrometrica and coleoptiles of corn(Zea mays) and sorghum(Sorghum vulgare). We optimized conditions for the transfer of proteins to nitrocellulose, production of high titer polyclonal anti-Gα (common) antibodies and finally the detection of G-proteins by amplification. In addition to the α-subunit of heterotrimeric G-proteins (M _r 41–43 kDa), a small molecular weight class (< 30 kDa) was also detected by anti-Gα (common) antibodies. An easy, reliable and efficient filter assay is also described to quantify the toxin catalyzed ADP-ribosylation. The apparentK _m of the NAD has been determined to be approximately 1.5μM for the microsomal fraction of moss. Inclusion of G1P stimulated ADP-ribosylation by 2–27-fold. One to three polypeptides representing the α-subunit of heterotrimeric G-proteins of (M_r 37–43 kDa) were ADP-ribosylated in all three plants. The anti-Gβ (C-terminus) antibody cross-reacted strongly with 39 and 34 kDa polypeptide in moss and corn respectively. By employing improved methods two classes of G-proteins have been shown to be present in three plant species.     

Visual census methods underestimate density and diversity of cryptic reef fishes

The diversity and density of small, benthic reef fishes were estimated using visual census and enclosed rotenone stations. Visual census underestimated the number of species present and the density of common species by up to 91%.     

The Visual N1 Is Sensitive to Deviations from Natural Texture Appearance

Disruptions of natural texture appearance are known to negatively impact performance in texture discrimination tasks, for example, such that contrast-negated textures, synthetic textures, and textures depicting abstract art are processed less efficiently than natural textures. Presently, we examined how visual ERP responses (the P1 and the N1 in particular) were affected by violations of natural texture appearance. We presented participants with images depicting either natural textures or synthetic textures made from the original stimuli. Both stimulus types were additionally rendered either in positive or negative contrast. These appearance manipulations (negation and texture synthesis) preserve a range of low-level features, but also disrupt higher-order aspects of texture appearance. We recorded continuous EEG while participants completed a same/different image discrimination task using these images and measured both the P1 and N1 components over occipital recording sites. While the P1 exhibited no sensitivity to either contrast polarity or real/synthetic appearance, the N1 was sensitive to both deviations from natural appearance. Polarity reversal and synthetic appearance affected the N1 latency differently, however, suggesting a differential impact on processing. Our results suggest that stages of visual processing indexed by the P1 and N1 are sensitive to high-order statistical regularities in natural textures and also suggest that distinct violations of natural appearance impact neural responses differently.     

A technique for identifying atoms from a screen image

Improving the interfaces in molecular graphics applications, making them more natural and easy to use, is an important task, given the current complexity of the displayed objects and of modeling operations. Clicking near an atom center is the usual method of atom selection. However, this method has certain disadvantages when working with images composed of different atomic representations such as sticks, CPK, or dotted surfaces. We propose another technique allowing the user to obtain the correct answer when he or she clicks on any element of the atom image.     

Possible transitions from molecules to cells

In view of the available data the present state of biogenesis research will be described. A reconstruction of the primordial cell evolution is carried through the Multistep hypothesis. The modular principle is an important criterion as a macro-mechanism of the evolution.