Pull-Down of Metalloproteins in Their Native States by Using Desthiobiotin-Based Probes

One-third of all proteins are estimated to require metals for structural stability and/or catalytic activity. Desthiobiotin probes containing metal binding groups can be used to capture metalloproteins with exposed active-site metals under mild conditions so as to prevent changes in metallation state. The proof-of-concept was demonstrated with carbonic anhydrase (CA), an open active site, Zn²⁺-containing protein. CA was targeted by using sulfonamide derivatives. Linkers of various lengths and structures were screened to determine the optimal structure for capture of the native protein. The optimized probes could selectively pull down CA from red blood cell lysate and other protein mixtures. Pull-down of differently metallated CAs was also investigated.     

Optimization of Pathogen Capture in Flowing Fluids with Magnetic Nanoparticles

Magnetic nanoparticles have been employed to capture pathogens for many biological applications; however, optimal particle sizes have been determined empirically in specific capturing protocols. Here, a theoretical model that simulates capture of bacteria is described and used to calculate bacterial collision frequencies and magnetophoretic properties for a range of particle sizes. The model predicts that particles with a diameter of 460 nm should produce optimal separation of bacteria in buffer flowing at 1 L h⁻¹. Validating the predictive power of the model, Staphylococcus aureus is separated from buffer and blood flowing through magnetic capture devices using six different sizes of magnetic particles. Experimental magnetic separation in buffer conditions confirms that particles with a diameter closest to the predicted optimal particle size provide the most effective capture. Modeling the capturing process in plasma and blood by introducing empirical constants (c_e), which integrate the interfering effects of biological components on the binding kinetics of magnetic beads to bacteria, smaller beads with 50 nm diameters are predicted that exhibit maximum magnetic separation of bacteria from blood and experimentally validated this trend. The predictive power of the model suggests its utility for the future design of magnetic separation for diagnostic and therapeutic applications.     

Sorting Second-Grade Students: Differentiating Those Retained From Those Promoted.

A cohort of 2nd-grade students provided comparisons of academic and social competence based on school retention/promotion decisions. Sample groups were (a) retained, (b) at risk for retention, (c) special education, and (d) promoted. Findings suggested most children with academic deficiencies are identified by schools early and are sorted into educational treatments differing in intensity that represent a continuum of competence. The authors provide empirical evidence counter to the assumptions that retained students have the requisite ability to catch up and have more problem behaviors than other low-achieving students. The relevance of high-stakes test scores for promotion/retention decisions and the parallels between schools' implementation of retention policy and implementation of regulations for identifying children with disabilities are included in the discussion. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Molecular correlations in sheared thermotropic copolyesters showing banded textures

Banded textures produced in a thermotropic liquid crystal polymer by shearing between glass slides are examined by using both transmission electron and polarized light microscopy. The periodic variation in director orientation about the shear axis, as measured by light microscopy, is shown to be distinctly different from that indicated by electron diffraction. Measurements of birefringence and observation of Zernicke phase contrast indicate periodic variations in optical properties of the polymer, in step with the bands. Such effects are accounted for in terms of a synchronous rotation of the planar aromatic groups about the molecular chain axes. Evidence for an out-of-plane component of molecular orientation is also presented.     

Silver impregnation applied to striated muscle: The sarcoplasmic reticulum

The classical black reaction developed by Camillo Golgi is shown to impregnate the tubules and fenestrations of the sarcoplasmic reticulum (SR) in striated muscle. This is a double impregnation of chromate and silver, which usually fills extracellular spaces. The method is difficult insofar as long incubation times are required, and location of the successfully “stained” SR in plastic-embedded tissue blocks is unpredictable. The light microscope is absolutely necessary to find the good regions which can then be cut from the blocks in 1-μm-thick sections and examined in the electron microscope. Stereo pairs give the best results since these resolve overlap problems common to thick sections. A variety of artifacts are illustrated which can help avoid erroneous interpretations. The Golgi-“stained” SR shows this elusive network with unsurpassed contrast and should benefit the morphological studies of muscle-membrane enthusiasts.     

The effect of temperature on crazing mechanisms in polystyrene

At room temperature scission is the dominant mechanism for the modification of the entanglement network required for craze formation in polystyrene, but as the temperature is increased towardsT _g, there is the possibility that disentanglement processes may contribute. These will be most important for short chains. If disentanglement can occur, a molecular weight dependence of the crazing stress as a function of temperature will result. This prediction is tested by straining thin films of a range of monodisperse samples of polystyrene at temperatures between 40 and 90° C. The nature of the ensuing deformation has been characterized by transmission electron microscopy. It is observed that whereas only crazing occurs over the entire temperature range for the lowest molecular weight sample, shear processes become important for higher molecular weight materials. For the longest chains, crazing is almost entirely suppressed at 80° C, with the preferential formation of shear deformation zones occurring. These observations are consistent with the idea that disentanglement is playing a significant role in craze formation at sufficiently high temperatures.     

The social consequences of expressive suppression.

At times, people keep their emotions from showing during social interactions. The authors' analysis suggests that such expressive suppression should disrupt communication and increase stress levels. To test this hypothesis, the authors conducted 2 studies in which unacquainted pairs of women discussed an upsetting topic. In Study 1, one member of each pair was randomly assigned to (a) suppress her emotional behavior, (b) respond naturally, or (c) cognitively reappraise in a way that reduced emotional responding. Suppression alone disrupted communication and magnified blood pressure responses in the suppressors' partners. In Study 2, suppression had a negative impact on the regulators' emotional experience and increased blood pressure in both regulators and their partners. Suppression also reduced rapport and inhibited relationship formation. (PsycINFO Database Record (c) 2010 APA, all rights reserved)