Antiendotoxin therapies for septic shock

Gram-negative shock is thought to result primarily from the effects of endotoxin, a component of the bacterial outer membrane. Accordingly, therapies aimed at inhibiting, neutralizing, or clearing endotoxin have been extensively explored. Despite over 30 years of research, no antiendotoxin approach to the treatment of human septic shock is of proven benefit. In recent randomized clinical trials of monoclonal antibodies against endotoxin, therapeutic efficacy was not convincingly demonstrated. This result, however, does not eliminate the possibility that other antiendotoxin therapies may be effective. The antibodies used in these clinical trials do not appear to neutralize endotoxin in vitro and are not reproducibly protective in animal models of sepsis. Newer agents with well-defined mechanisms of antiendotoxin activity may help clarify the role of endotoxin in septic shock and prove useful therapy for some patients.     

The effect of verbal elaborations on memory in young and older adults

The Stein paradigm was used to examine the circumstances under which verbal elaborations enhance memory in young and older adults. Subjects studied target adjectives that were embedded in one of three sentence contexts that varied in elaboration of the subject-adjective relationship: (1) nonelaborated base sentences; (2) base sentences with semantically consistent, but arbitrary verbal, elaborations; and (3) base sentences with explanatory verbal elaborations that clarified the significance of the subject-adjective relationship. The presence of the elaborations was varied at encoding and retrieval, and cued recall of the target adjectives was tested with incidental and intentional learning procedures. In Experiments 1A and 1B, explanatory elaborations at encoding and retrieval yielded the largest memorial facilitation for both young and older adults, and the benefit was comparable for the incidental and intentional learning measures. In Experiment 2, age-related differences in recall were minimal with explanatory elaborations at encoding and retrieval, but larger age differences occurred in the nonelaborated comparison conditions. In Experiment 3, explanatory elaborations present at encoding but not at retrieval enhanced recall when the original Stein stimuli were used, but not with the present stimuli. The implications of these results with regard to the mnemonic efficacy of verbal elaborations for young and older adults are discussed.     

The role of tyrosine-114 in the enzymatic activity of the Shiga-like toxin I A-chain

Shiga-like toxin I (SLT-I), the potent cytotoxin produced by certain pathogenic strains of Escherichia coli, is a member of a burgeoning family of ribosome-in-activating proteins (RIPs), which share common structural and mechanistic features. The prototype of the group is the plant toxin ricin. Recently we proposed a structural model for the Slt-IA active site, based in part on the known geometry of the enzymatic subunit of the ricin toxin. The model places three aromatic residues within the putative Slt-IA active site cleft: tyrosine 77, tyrosine 114, and tryptophan 203. Here we present biochemical and biophysical data regarding, the phenotypes of conservative point mutants of Slt-IA in which tyrosine 114 is altered. We used oligonucleotide-directed mutagenesis to replace tyrosine 114 with either phenylalanine (Y114F) or serine (Y114S). Periplasmic extracts of E. coli containing wild-type or mutant Slt-IA were tested for their ability to inhibit protein synthesis in vitro. Relative to wild-type, the activity of mutant Y114F was attenuated about 30-fold, while the mutant Y114S was attenuated about 500 to 1000-fold. In order to address the possibility that differential activation of the mutants rather than local effects at the active site might account for their diminished activity, we engineered the same mutations into a truncated slt-IA cassette that directs expression of a product corresponding to the activated A1 form of Slt-IA (wild-type-delta). The same general relationships held: relative to wild type-delta, Y114F-delta was attenuated about 7-fold, and Y114S-delta about 300-fold.(ABSTRACT TRUNCATED AT 250 WORDS)     

HER-2/c-erbB2 is phosphorylated by calmodulin-dependent protein kinase II on a single site in the cytoplasmic tail at threonine-1172

Calmodulin-dependent protein kinase II (Cam kinase II) is known to desensitise epidermal growth factor receptor (HER-1) tyrosine kinase activity by a process involving phosphorylation at serines 1046/47 in the cytoplasmic tail. We have developed an experimental system to investigate phosphorylation of the related HER-2/c-erbB2 proto-oncogene utilising purified Cam kinase II and recombinant glutathione-S-transferase (GST) fusion proteins. The cDNA for rat Cam kinase II-alpha was transfected into human embryonic kidney (HEK) 293 fibroblasts and the expressed protein purified to homogeneity by calmodulin-agarose affinity chromatography. A GST fusion protein comprising residues 1126-1255 of HER-2 was phosphorylated by purified Cam kinase II, in contrast to a GST protein comprising residues 1005-1125. Phosphoamino-acid analysis and site-directed mutagenesis indicated that HER-2 was phosphorylated on a single site at threonine-1172 which resides within a consensus Cam kinase II phosphorylation site (RAKT). HER-2 (threonine-1172-alanine), in the form of a ligand-inducible chimaera HER-1/2, was co-transfected into HEK-293 fibroblasts with a constitutively active form of Cam kinase II, followed by in vivo labelling of these cells with 32 P-orthophosphate. Immunoprecipitation of ligand-activated receptors followed by two-dimensional phosphopeptide mapping indicated that threonine-1172 in HER-2 is a newly identified in vivo site which can be hyper-phosphorylated by constitutively active Cam kinase II. In addition, when over-expressed in HEK-293 fibroblasts, HER-1/2 (threonine-1172-alanine) showed a defect in desensitisation and underwent a more sustained EGF-induced receptor autophosphorylation compared to wild-type HER-1/2.     

Three-parameter vs. two-parameter Weibull distribution for pultruded composite material properties

The three-parameter and two-parameter Weibull distributions are compared using 26 mechanical property data sets of fiber-reinforced polymeric (FRP) composite materials manufactured by the pultrusion process. Both strength and stiffness properties were examined. The probability distributions were compared on the basis of goodness of fit, nominal design values, and allowable load to achieve uniform reliability. It is recommended that the two-parameter Weibull distribution be used to characterize FRP composite material properties. The primary basis for this recommendation is small differences in nominal design values and small differences in allowable loads between the two-parameter and three-parameter Weibull distributions. Other supporting reasons for the recommendation are similar observed significance levels in distribution fitting, computational efficiency, and the fact that the location parameter of the three-parameter Weibull distribution is near the first order statistic.     

Developing competency-based methods for near-real-time air combat problem solving assessment

Air combat scenarios present unique problems where the solution is not obvious. An advanced networked simulation environment can be used to train high-level cognitive air combat skills such as problem solving. A problem in the development of principled and construct oriented assessment is related to determining the level of specificity of the assessment. We present a detailed discussion of the definition of critical competencies associated with combat mission problem solving performance, and we describe our approach to develop and implement an embedded performance assessment system that maps outcomes to these competencies. Finally, we discuss the implications for our approach for comprehensive assessment and discuss our goals for an evaluation of the competency-based approach to assessment.     

Do You Know What I Know? A Shared Understandings Perspective on Text‐Based Communication

This article illustrates how the hermeneutic analysis of text illuminates how shared understandings affect our interpretations of lean communication in distributed work environments. It is proposed that in contrast to the pessimistic conclusions of media richness theory that lean communication channels cannot support complex or equivocal work tasks, miscommunications are not the result of technology, but rather occur due to a lack of shared understandings among the individuals communicating. An illustrative case study based on fieldwork in franchise organizations is presented to demonstrate the possibilities for how the hermeneutic analysis of coherence, invention, intention, and reference can be used to discover how workers create and recreate shared understandings through text.     

Extracting Electron Densities in N-Type GaAs From Raman Spectra: Theory

In this paper, we present the theory for calculating Raman line shapes as functions of the Fermi energy and finite temperatures in zinc blende, n-type GaAs for donor densities between 10¹⁶ cm⁻³ and 10¹⁹ cm⁻³. Compared to other theories, this theory is unique in two respects: 1) the many-body effects are treated self-consistently and 2) the theory is valid at room temperature for arbitrary values of the ratio R = (Q²/α), where Q is the magnitude of the normalized wave vector and α is the normalized frequency used in the Raman measurements. These calculations solve the charge neutrality equation self-consistently for a two-band model of GaAs at 300 K that includes the effects of high carrier concentrations and dopant densities on the perturbed densities of states used to calculate the Fermi energy as a function of temperature. The results are then applied to obtain the carrier concentrations from Fermi energies in the context of line shapes in Raman spectra due to the coupling between longitudinal optical phonons and plasmons. Raman measurements have been proposed as a non-destructive method for wafer acceptance tests of carrier density in semiconductor epilayers. The interpretation of Raman spectra to determine the majority electron density in n-type semiconductors requires an interdisciplinary effort involving experiments, theory, and computer-based simulations and visualizations of the theoretical calculations.     

Neural correlates of sexual arousal in homosexual and heterosexual men.

Men exhibit much higher levels of genital and subjective arousal to sexual stimuli containing their preferred sex than they do to stimuli containing only the nonpreferred sex. This study used event-related functional magnetic resonance imaging to investigate how this category-specific pattern would be reflected in the brains of homosexual (n = 11) and heterosexual (n = 11) men. Comparisons of activation to preferred sexual stimuli, nonpreferred sexual stimuli, and sports stimuli revealed large networks correlated with sexual arousal, spanning multiple cortical and subcortical areas. Both homosexual and heterosexual men exhibited category-specific arousal in brain activity. Within the amygdala, greater preference-related activity was observed in homosexual men, but it is unclear whether this is a cause or a consequence of their sexuality. In a subsequent analysis of regions hypothesized to support arousal, both participant groups demonstrated widespread increases in evoked activity for preferred stimuli. Aggregate data from these regions produced significant differences between stimulus types in 16 out of 22 participants. Significant activational differences matched reported sexual orientation in 15 of these 16 participants, representing an advance in psychophysiological measures of arousal. (PsycINFO Database Record (c) 2010 APA, all rights reserved)