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41.
Serine hydrolases have diverse intracellular substrates, biological functions, and structural plasticity, and are thus important for biocatalyst design. Amongst serine hydrolases, the recently described ybfF enzyme family are promising novel biocatalysts with an unusual bifurcated substrate‐binding cleft and the ability to recognize commercially relevant substrates. We characterized in detail the substrate selectivity of a novel ybfF enzyme from Vibrio cholerae (Vc‐ybfF) by using a 21‐member library of fluorogenic ester substrates. We assigned the roles of the two substrate‐binding clefts in controlling the substrate selectivity and folded stability of Vc‐ybfF by comprehensive substitution analysis. The overall substrate preference of Vc‐ybfF was for short polar chains, but it retained significant activity with a range of cyclic and extended esters. This broad substrate specificity combined with the substitutional analysis demonstrates that the larger binding cleft controls the substrate specificity of Vc‐ybfF. Key selectivity residues (Tyr116, Arg120, Tyr209) are also located at the larger binding pocket and control the substrate specificity profile. In the structure of ybfF the narrower binding cleft contains water molecules prepositioned for hydrolysis, but based on substitution this cleft showed only minimal contribution to catalysis. Instead, the residues surrounding the narrow binding cleft and at the entrance to the binding pocket contributed significantly to the folded stability of Vc‐ybfF. The relative contributions of each cleft of the binding pocket to the catalytic activity and folded stability of Vc‐ybfF provide a valuable map for designing future biocatalysts based on the ybfF scaffold.  相似文献   
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Independent components of magnetoencephalography: localization   总被引:5,自引:0,他引:5  
We applied second-order blind identification (SOBI), an independent component analysis method, to MEG data collected during cognitive tasks. We explored SOBI's ability to help isolate underlying neuronal sources with relatively poor signal-to-noise ratios, allowing their identification and localization. We compare localization of the SOBI-separated components to localization from unprocessed sensor signals, using an equivalent current dipole modeling method. For visual and somatosensory modalities, SOBI preprocessing resulted in components that can be localized to physiologically and anatomically meaningful locations. Furthermore, this preprocessing allowed the detection of neuronal source activations that were otherwise undetectable. This increased probability of neuronal source detection and localization can be particularly beneficial for MEG studies of higher-level cognitive functions, which often have greater signal variability and degraded signal-to-noise ratios than sensory activation tasks.  相似文献   
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Experimentally manipulated the conditions of beverage content and expectancy for alcohol in 29 heavy social-drinking (as determined by the Drinking Practices Questionnaire) male undergraduates. After completing a pencil-and-paper task, Ss found themselves in a situation where graffiti could apparently be written in and around the immediate surroundings with impunity. Regardless of expectations, Ss who actually drank alcohol were significantly more likely to write graffiti in this situation than were Ss who did not drink alcohol. This finding supports the importance of alcohol rather than expectancy as a causal influence on this form of disinhibited behavior. (12 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)  相似文献   
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Neuronal cell networks have been reconstructed on planar microelectrode arrays (MEAs) from dissociated hippocampal pyramidal neurons. Microcontact printing (microCP) and a photoresist-liftoff method were used to selectively localize poly-L-lysine (PLL) on the surface of MEAs. Haptotaxis led to the organization of the neurons into networks localized adjacent to microelectrodes. Various grids of PLL with 2-25-microm-wide lines spaced by 50-200 microm with 15-25-microm nodes at intersection points were used to guide cell body attachment and neurite outgrowth. Bursting activity with spike amplitude attenuation was observed, and multichannel recordings detected instances of coincident firing activity. Finally, we present here an extracellular recording from a approximately 2 microm bundle of guided neurites.  相似文献   
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In the present study, we investigated the induction of HSP70 proteins in Chironomus tentans in response to copper exposure. Larvae were exposed for 24 h to 0, 0.25, 0.5, 1.0 and 2.0 mgl(-1) copper, concentrations that were previously shown to cause growth reduction or mortality in 10 d bioassays. The induction of HSP70 was evaluated by Western blotting, and we observed a significant increase in HSP70 proteins over controls at all tested concentrations. The results suggest that short-term laboratory exposures to chronically toxic but acutely sublethal copper concentrations induce HSP70 proteins in C. tentans larvae.  相似文献   
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Humans have a remarkable capability to respond efficiently to a stimulus of interest despite other stimuli competing for neural resources. The current study investigated how the human system copes with distracting stimuli. During each trial, participants viewed 2 sequential stimuli that were each associated with a specific action based on an arbitrary mapping. The 1st stimulus served as a distractor, and the 2nd stimulus required a response (target). When the distractor preceded the target by more than a few hundred milliseconds, response latencies were slower when the 2 stimuli were associated with the same response. The authors propose that this negative compatibility effect stemmed from an inhibitory mechanism that the human system utilizes to prevent the distractor from eliciting an unwanted response. (PsycINFO Database Record (c) 2010 APA, all rights reserved)  相似文献   
50.
Today, people can easily select media outlets sharing their political predispositions, a behavior known as partisan selective exposure. Additional research is needed, however, to better understand the causes and consequences of partisan selective exposure. This study investigates the relationship between partisan selective exposure and political polarization using data from the National Annenberg Election Survey. Cross‐sectional results show strong evidence that partisan selective exposure is related to polarization. Over‐time analyses document that partisan selective exposure leads to polarization. Some evidence supports the reverse causal direction, namely that polarization leads to partisan selective exposure. Implications for the study of media effects and normative implications—both positive and negative—are discussed.  相似文献   
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