We present unzipping force analysis of protein association (UFAPA) as a novel and versatile method for detection of the position and dynamic nature of protein-DNA interactions. A single DNA double helix was unzipped in the presence of DNA-binding proteins using a feedback-enhanced optical trap. When the unzipping fork in a DNA reached a bound protein molecule we observed a dramatic increase in the tension in the DNA, followed by a sudden tension reduction. Analysis of the unzipping force throughout an unbinding "event" revealed information about the spatial location and dynamic nature of the protein-DNA complex. The capacity of UFAPA to spatially locate protein-DNA interactions is demonstrated by noncatalytic restriction mapping on a 4-kb DNA with three restriction enzymes (BsoBI, XhoI, and EcoRI). A restriction map for a given restriction enzyme was generated with an accuracy of approximately 25 bp. UFAPA also allows direct determination of the site-specific equilibrium association constant (K(A)) for a DNA-binding protein. This capability is demonstrated by measuring the cation concentration dependence of K(A) for EcoRI binding. The measured values are in good agreement with previous measurements of K(A) over an intermediate range of cation concentration. These results demonstrate the potential utility of UFAPA for future studies of site-specific protein-DNA interactions.

In Lepidoptera, forewings and hindwings are mechanically coupled and flap in synchrony. Flight is anteromotoric, being driven primarily by action of the forewings. Here we report that lepidopterans can still fly when their hindwings are cut off, a procedure reducing their total wing surface, on average, by nearly one half. However, as we demonstrate by analysis of three-dimensional flight trajectories of a moth and a butterfly (Lymantria dispar and Pieris rapae), hindwing removal causes lepidopterans to incur a loss in both linear and turning acceleration, so that they are unable to exercise their normal flight maneuverability. Without hindwings they still are able to zigzag aerially (the ablation has no effect on their turning radius in flight) but at lesser speed and therefore less evasively. Consequently, hindwings in the expanded state in which they occur in lepidopterans seem to contribute in an essential way to lepidopteran survival. Moths in today's world, we argue, may rely on their evasive flight primarily to avoid capture by bats, whereas butterflies, which we propose advertise their evasiveness collectively through shared aposematism, may depend upon it primarily for defense against birds. Aerial agility thus may be the chief adaptive asset derived by lepidopterans from possession of oversize hindwings.aposematism ͉ coloration ͉ defense ͉ flight maneuverability ͉ mimicry

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In a naïve realist approach to reading an ontology off the models of a physical theory, the invariance of a given theory under permutations of its propertybearing objects entails the existence of distinct possible worlds from amongst which the theory cannot choose. A brand of Ontic Structural Realism (OSR) attempts to avoid this consequence by denying that objects possess primitive identity, and thus worlds with property values permuted amongst those objects are really one and the same world. Assuming that any successful ontology of objects is able to describe a universe containing a determinate number of them, I argue that no version of OSR which both retains objects and understands 'structure' in terms of relations can be successful. This follows from the fact that no set of relational facts is sufficient to fix the cardinality of the collection of objects implied by those facts. More broadly, I offer reasons to believe that no version of OSR is compatible with the existence of objects, no matter how ontologically derivative they are taken to be. Any such account would have to attribute a definite cardinality to a collection of objects while denying that those objects are possessed of a primitive identity. With no compelling reason to abandon the classical conception of cardinality, such an attribution is incoherent.