Outer-Membrane Protease (OmpT) Based E. coli Sensing with Anionic Polythiophene and Unlabeled Peptide Substrate.

E. coli and Salmonella are two of the most common bacterial pathogens involved in foodborne and waterborne related deaths. Hence, it is critical to develop rapid and sensitive detection strategies for near-outbreak applications. Reported is a simple and specific assay to detect as low as 1 CFU mL-1 of E. coli in water within 6 hours by targeting the bacteria's surface protease activity. The assay relies on polythiophene acetic acid (PTAA) as an optical reporter and a short unlabeled peptide (LL37FRRV ) previously optimized as a substrate for OmpT, an outer-membrane protease on E. coli. LL37FRRV interacts with PTAA to enhance its fluorescence while also inducing the formation of a helical PTAA-LL37FRRV construct, as confirmed by circular dichroism. However, in the presence of E. coli LL37FRRV is cleaved and can no longer affect the conformations and optical properties of PTAA. This ability to distinguish between an intact and cleaved peptide was investigated in detail using LL37FRRV sequence variants.

A Bottom-Up Proteomic Approach to Identify Substrate Specificity of Outer-Membrane Protease OmpT.

Identifying peptide substrates that are efficiently cleaved by proteases gives insights into substrate recognition and specificity, guides development of inhibitors, and improves assay sensitivity. Peptide arrays and SAMDI mass spectrometry were used to identify a tetrapeptide substrate exhibiting high activity for the bacterial outer-membrane protease (OmpT). Analysis of protease activity for the preferred residues at the cleavage site (P1, P1') and nearest-neighbor positions (P2, P2') and their positional interdependence revealed FRRV as the optimal peptide with the highest OmpT activity. Substituting FRRV into a fragment of LL37, a natural substrate of OmpT, led to a greater than 400-fold improvement in OmpT catalytic efficiency, with a kcat /Km value of 6.1×106  L mol-1 s-1 . Wild-type and mutant OmpT displayed significant differences in their substrate specificities, demonstrating that even modest mutants may not be suitable substitutes for the native enzyme.

On the road to self-perception: interpretation of self-behaviors can be altered by priming.

In 3 experiments, some participants read a story describing ambiguously mean behaviors performed by another person. Other participants read the story and imagined that they performed the behaviors. Results showed that (a) exposure to a conceptual priming manipulation caused assimilation effects in actor meanness judgments, regardless of whether the actor was self or other, (b) tasks designed neither to heighten self-concept accessibility nor to threaten the self moderated the effects of conceptual meanness primes on self-meanness judgments, and (c) this lack of moderation occurred despite considerable evidence of self-enhancement effects elsewhere in self-judgments. A fourth experiment examined the extent to which priming affected interpretations of real self or other behavior. Results were consistent with the idea that priming altered event interpretation and subsequent judgments but also suggested that judgments were influenced by self-enhancement motivation. Implications of the results for theorizing in personality and self-knowledge acquisition are discussed.

YscP and YscU switch the substrate specificity of the Yersinia type III secretion system by regulating export of the inner rod protein YscI.

Pathogenic yersiniae utilize a type III secretion system to inject antihost factors, called Yops, directly into the cytosol of eukaryotic cells. The Yops are injected via a needle-like structure, comprising the YscF protein, on the bacterial surface. While the needle is being assembled, Yops cannot be secreted. YscP and YscU switch the substrate specificity of the secretion system to enable Yop export once the needle attains its proper length. Here, we demonstrate that the inner rod protein YscI plays a critical role in substrate specificity switching. We show that YscI is secreted by the type III secretion system and that YscI secretion by a yscP mutant is abnormally elevated. Furthermore, we show that mutations in the cytoplasmic domain of YscU reduce YscI secretion by the yscP null strain. We also demonstrate that mutants expressing one of three forms of YscI (those with mutations Q84A, L87A, and L96A) secrete substantial amounts of Yops yet exhibit severe defects in needle formation. In the absence of YscP, mutants with the same changes in YscI assemble needles but are unable to secrete Yops. Together, these results suggest that the formation of the inner rod, not the needle, is critical for substrate specificity switching and that YscP and YscU exert their effects on substrate export by controlling the secretion of YscI.

Sex differences in jealousy: the (lack of) influence of researcher theoretical perspective.

The sex difference in jealousy is an effect that has generated significant controversy in the academic literature (resulting in two meta-analyses that reached different conclusions on the presence or absence of the effect). In this study, we had a team of researchers from different theoretical perspectives use identical protocols to test whether the sex difference in jealousy would occur across many different samples (while testing whether mate value would moderate the effect). In our samples, we found the sex difference in jealousy to occur using both forced choice and continuous measures, this effect appeared in several different settings, and, we found that mate value moderated participant responses. The results are discussed in light of the controversy surrounding the presence of the effect.

A gene expression-based comparison of cell adhesion to extracellular matrix and RGD-terminated monolayers.

This work uses global gene expression analysis to compare the extent to which model substrates presenting peptide adhesion motifs mimic the use of conventional extracellular matrix protein coated substrates for cell culture. We compared the transcriptional activities of genes in cells that were cultured on matrix-coated substrates with those cultured on self-assembled monolayers presenting either a linear or cyclic RGD peptide. Cells adherent to cyclic RGD were most similar to those cultured on native ECM, while cells cultured on monolayers presenting the linear RGD peptide had transcriptional activities that were more similar to cells cultured on the uncoated substrates. This study suggests that biomaterials presenting the cyclic RGD peptide are substantially better mimics of extracellular matrix than are uncoated materials or materials presenting the common linear RGD peptide.

Comparing two perceived characteristics of autobiographical memory: Memory detail and accessibility.

Four samples of participants recalled autobiographical memories. While some evidence emerged from regression analyses suggesting that judgements of the amount of detail contained in each memory and judgements of the ease with which events could be recalled were partially independent, the analyses generally showed that these judgements were similarly predicted by various event characteristics (age, typicality, self-importance, emotional intensity at event occurrence, rehearsal types). Co-occurrence frequency data yielded similar conclusions, showing that while ease ratings and detail ratings occasionally diverged, they were more often consistent with each other. Finally, the data also suggested that events that prompted emotional ambivalence were not judged to be more easily recalled, or to contain more detail, than non-ambivalent events.