Sensor Domain of Histidine Kinase VxrA of Vibrio cholerae- A Hairpin-swapped Dimer and its Conformational Change.

VxrA and VxrB are cognate histidine kinase (HK) - response regulator (RR) pairs of a two-component signaling system (TCS) found in Vibrio cholerae, a bacterial pathogen that causes cholera. The VxrAB TCS positively regulates virulence, the Type VI Secretion System, biofilm formation, and cell wall homeostasis in V. cholerae, providing protection from environmental stresses and contributing to the transmission and virulence of the pathogen. The VxrA HK has a unique periplasmic sensor domain (SD) and, remarkably, lacks a cytoplasmic linker domain between the second transmembrane helix and the dimerization and histidine phosphotransfer (DHp) domain, indicating that this system may utilize a potentially unique signal sensing and transmission TCS mechanism. In this study, we have determined several crystal structures of VxrA-SD and its mutants. These structures reveal a novel structural fold forming an unusual ß hairpin-swapped dimer. A conformational change caused by relative rotation of the two monomers in a VxrA-SD dimer could potentially change the association of transmembrane helices and, subsequently, the pairing of cytoplasmic DHp domains. Based on the structural observation, we propose a putative scissor-like closing regulation mechanism for the VxrA HK.IMPORTANCE V. cholerae has a dynamic life cycle, which requires rapid adaptation to changing external conditions. Two-component signal transduction (TCS) systems allow V. cholerae to sense and respond to these environmental changes. The VxrAB TCS positively regulates a number of important V. cholerae phenotypes, including virulence, the Type Six Secretion System, biofilm formation, and cell wall homeostasis. Here, we provide the crystal structure of the VxrA sensor histidine kinase sensing domain and propose a mechanism for signal transduction. The cognate signal for VxrAB remains unknown, however, in this work we couple our structural analysis with functional assessments of key residues to further our understanding of this important TCS.

Emotion-word processing difficulties in abstinent alcoholics with and without lifetime externalizing disorders.

BACKGROUND: Evidence suggests that abstinent alcoholics have difficulties processing a variety of emotion-laden stimuli, and some of these difficulties may not fully resolve with long-term abstinence. The current study examined whether emotion-word processing difficulties were present in long-term abstinent alcoholics (LTAA; 18+ months of sobriety) with and without a previously diagnosed externalizing (EXT; antisocial personality disorder and/or attention-deficit/hyperactivity disorder diagnosis) disorder. METHODS: Subjects (N = 121) completed an affective go/no-go (AGNG) task with positive, negative, and neutral emotion-word stimuli, and a lexical decision-making (LDM) task with nonemotion word and nonword stimuli. Nonsubstance abusing controls (NSAC; n = 38, 50.0% women, mean age = 48 ± 7.8), LTAA with EXT (n = 32, 41% women, mean age = 47.1 ± 6.6), and LTAA without EXT (n = 51, 47% women, mean age = 49.7 ± 6.5) were compared between signal discriminability (d') and mean response times (RT) for correct responses (mcRT). RESULTS: In the LDM task, LTAA had lower (d') values and slower mcRT than NSAC. In the AGNG task, LTAA and NSAC did not differ in AGNG task mcRT. LTAA had lower (d') values than NSAC, and this effect was partially associated with group differences in LDM task (d') values. In LTAA, lower AGNG (d') values also were associated with an earlier age of first drink, greater lifetime alcohol use, and a history of EXT disorder. CONCLUSIONS: Our findings suggest that detecting the emotional content of words is impaired in LTAA, and this impairment is over and above LTAA's more general lexical processing difficulties. Results also suggest that specific emotion processing impairments in LTAA may be exacerbated by greater lifetime alcohol use burden and other comorbid EXT diagnoses.

Functional principal components analysis of workload capacity functions.

Workload capacity, an important concept in many areas of psychology, describes processing efficiency across changes in workload. The capacity coefficient is a function across time that provides a useful measure of this construct. Until now, most analyses of the capacity coefficient have focused on the magnitude of this function, and often only in terms of a qualitative comparison (greater than or less than one). This work explains how a functional extension of principal components analysis can capture the time-extended information of these functional data, using a small number of scalar values chosen to emphasize the variance between participants and conditions. This approach provides many possibilities for a more fine-grained study of differences in workload capacity across tasks and individuals.

A novel human cancer vaccine elicits cellular responses to the tumor-associated antigen, human chorionic gonadotropin beta.

PURPOSE: The oncofetal antigen, human chorionic gonadotropin beta subunit (hCGbeta), is expressed by a number of carcinomas and is a prognostic indicator in renal, colorectal, bladder, and pancreatic cancers. We describe the development of a novel antibody-based dendritic cell (DC)-targeted cancer vaccine capable of eliciting cellular immune responses directed against hCGbeta. EXPERIMENTAL DESIGN: The tumor-associated antigen hCGbeta was coupled genetically to a human anti-DC antibody (B11). The resulting fusion protein (B11-hCGbeta) was evaluated for its ability to promote tumor antigen-specific cellular immune responses in a human in vitro model. Monocyte-derived human DCs from normal donors were exposed to purified B11-hCGbeta, activated with CD40 ligand, mixed with autologous lymphocytes, and tested for their ability to promote hCGbeta-specific proliferative and cytotoxic T-lymphocyte responses. RESULTS: B11-hCGbeta was found to be a soluble, well-defined, and readily purified product that specifically recognized the human mannose receptor via the B11 antibody portion of the fusion protein. B11-hCGbeta functionally promoted the uptake and processing of tumor antigen by DCs, which led to the generation of tumor-specific HLA class I and class II-restricted T-cell responses, including CTLs capable of killing human cancer cell lines expressing hCGbeta. CONCLUSIONS: Although other hCG vaccines have been shown to be capable of eliciting antibody responses to hCGbeta, this is the first time that cellular immune responses to hCGbeta have been induced by a vaccine in a human system. This DC-targeted hCGbeta vaccine holds promise for the management of a number of cancers and merits additional clinical development.

Working memory capacity and redundant information processing efficiency.

Working memory capacity (WMC) is typically measured by the amount of task-relevant information an individual can keep in mind while resisting distraction or interference from task-irrelevant information. The current research investigated the extent to which differences in WMC were associated with performance on a novel redundant memory probes (RMP) task that systematically varied the amount of to-be-remembered (targets) and to-be-ignored (distractor) information. The RMP task was designed to both facilitate and inhibit working memory search processes, as evidenced by differences in accuracy, response time, and Linear Ballistic Accumulator (LBA) model estimates of information processing efficiency. Participants (N = 170) completed standard intelligence tests and dual-span WMC tasks, along with the RMP task. As expected, accuracy, response-time, and LBA model results indicated memory search and retrieval processes were facilitated under redundant-target conditions, but also inhibited under mixed target/distractor and redundant-distractor conditions. Repeated measures analyses also indicated that, while individuals classified as high (n = 85) and low (n = 85) WMC did not differ in the magnitude of redundancy effects, groups did differ in the efficiency of memory search and retrieval processes overall. Results suggest that redundant information reliably facilitates and inhibits the efficiency or speed of working memory search, and these effects are independent of more general limits and individual differences in the capacity or space of working memory.

An information processing/associative learning account of behavioral disinhibition in externalizing psychopathology.

Externalizing psychopathology (EXT) is associated with low executive working memory (EWM) capacity and problems with inhibitory control and decision-making; however, the specific cognitive processes underlying these problems are not well known. This study used a linear ballistic accumulator computational model of go/no-go associative-incentive learning conducted with and without a working memory (WM) load to investigate these cognitive processes in 510 young adults varying in EXT (lifetime problems with substance use, conduct disorder, ADHD, adult antisocial behavior). High scores on an EXT factor were associated with low EWM capacity and higher scores on a latent variable reflecting the cognitive processes underlying disinhibited decision-making (more false alarms, faster evidence accumulation rates for false alarms [vFA], and lower scores on a Response Precision Index [RPI] measure of information processing efficiency). The WM load increased disinhibited decision-making, decisional uncertainty, and response caution for all subjects. Higher EWM capacity was associated with lower scores on the latent disinhibited decision-making variable (lower false alarms, lower vFAs and RPI scores) in both WM load conditions. EWM capacity partially mediated the association between EXT and disinhibited decision-making under no-WM load, and completely mediated this association under WM load. The results underline the role that EWM has in associative-incentive go/no-go learning and indicate that common to numerous types of EXT are impairments in the cognitive processes associated with the evidence accumulation-evaluation-decision process.

Dimensions of disinhibited personality and their relation with alcohol use and problems.

Although alcohol use disorders (AUDs) have been associated with different aspects of disinhibited personality and antisociality, less is known about the specific relationships among different domains of disinhibited personality, antisociality, alcohol use, and alcohol problems. The current study was designed to address three goals, (i) to provide evidence of a three-factor model of disinhibited personality (comprised of impulsivity [IMP], risk taking/low harm avoidance [RTHA], excitement seeking [ES]), (ii) to test hypotheses regarding the association between each dimension and alcohol use and problems, and (iii) to test the hypothesis that antisociality (social deviance proneness [SDP]) accounts for the direct association between IMP and alcohol problems, while ES is directly related to alcohol use. Measures of disinhibited personality IMP, RTHA, ES and SDP and alcohol use and problems were assessed in a sample of young adults (N=474), which included a high proportion of individuals with AUDs. Confirmatory factor analyses supported a three-factor model of disinhibited personality reflecting IMP, RTHA, and ES. A structural equation model (SEM) showed that IMP was specifically associated with alcohol problems, while ES was specifically associated with alcohol use. In a second SEM, SDP accounted for the majority of the variance in alcohol problems associated with IMP. The results suggest that aspects of IMP associated with SDP represent a direct vulnerability to alcohol problems. In addition, the results suggest that ES reflects a specific vulnerability to excessive alcohol use, which is then associated with alcohol problems, while RTHA is not specifically associated with alcohol use or problems when controlling for IMP and ES.

Externalizing psychopathology and behavioral disinhibition: working memory mediates signal discriminability and reinforcement moderates response bias in approach-avoidance learning.

Research has suggested that reduced working memory capacity plays a key role in disinhibited patterns of behavior associated with externalizing psychopathology. In this study, participants (N = 365) completed 2 versions of a go/no-go mixed-incentive learning task that differed in the relative frequency of monetary rewards and punishments for correct and incorrect active-approach responses, respectively. Using separate structural equation models for conventional (hit and false alarm rates) and signal detection theory (signal discriminability and response bias) performance indices, distinct roles for working memory capacity and changes in payoff structure were found. Specifically, results showed that (a) working memory capacity mediated the effects of externalizing psychopathology on false alarms and discriminability of go versus no-go signals; (b) these effects were not moderated by the relative frequency of monetary rewards and punishments; (c) the relative frequency of monetary rewards and punishments moderated the effects of externalizing psychopathology on hits and response bias for go versus no-go responses; and (d) these effects were not mediated by working memory capacity. The findings implicate distinct roles for reduced working memory capacity and poorly modulated active approach and passive avoidance in the link between externalizing psychopathology and behavioral disinhibition.

Modeling Response Times in the Go/No-Go Discrimination Task.

The work presented here uses a simple stochastic model as a cognitive psychometric tool for analyzing response time data in the Go/No-Go Discrimination task with motivationally distinct conditions. The parameters of the model inform us of underlying cognitive mechanisms because they have an established psychological meaning and allow us to quantify a subjects ability and response caution. Using these model parameters, we focus on the differences between subjects with varying degrees of substance abuse and antisocial behavioral disorders and show that there are reliable differences between the decision mechanisms of these subjects. Using data from executive working memory tasks, we postulate that these differences in cognitive processes might be due to differences in working memory capacity. Ultimately, we show that formal cognitive modeling has the potential to provide valuable insights into clinical phenomena that cannot be captured by traditional data analysis techniques.

The differential sensitivity of human and rhesus macaque CCR5 to small-molecule inhibitors of human immunodeficiency virus type 1 entry is explained by a single amino acid difference and suggests a mechanism of action for these inhibitors.

AD101 and SCH-C are two chemically related small molecules that inhibit the entry of human immunodeficiency virus type 1 (HIV-1) via human CCR5. AD101 also inhibits HIV-1 entry via rhesus macaque CCR5, but SCH-C does not. Among the eight residues that differ between the human and macaque versions of the coreceptor, only one, methionine-198, accounts for the insensitivity of macaque CCR5 to inhibition by SCH-C. Thus, the macaque coreceptor engineered to contain the natural human CCR5 residue (isoleucine) at position 198 is sensitive to HIV-1 entry inhibition by SCH-C, whereas a human CCR5 mutant containing the corresponding macaque residue (methionine) is resistant. Position 198 is in CCR5 transmembrane (TM) helix 5 and is not located within the previously defined binding site for AD101 and SCH-C, which involves residues in TM helices 1, 2, 3, and 7. SCH-C binds to human CCR5 whether residue 198 is isoleucine or methionine, and it also binds to macaque CCR5. However, the binding of a conformation-dependent monoclonal antibody to human CCR5 is inhibited by SCH-C only when residue 198 is isoleucine. These observations, taken together, suggest that the antiviral effects of SCH-C and AD101 involve stabilization, or induction, of a CCR5 conformation that is not compatible with HIV-1 infection. However, SCH-C is unable to exert this effect on CCR5 conformation when residue 198 is methionine. The region of CCR5 near residue 198 has, therefore, an important influence on the conformational state of this receptor.

Mannose receptor targeting of tumor antigen pmel17 to human dendritic cells directs anti-melanoma T cell responses via multiple HLA molecules.

Targeting recycling endocytic receptors with specific Abs provides a means for introducing a variety of tumor-associated Ags into human dendritic cells (DCs), culminating in their efficient presentation to T cells. We have generated a human mAb (B11) against the mannose receptor that is rapidly internalized by DCs through receptor-mediated endocytosis. By genetically linking the melanoma Ag, pmel17, to Ab B11, we obtained the fully human fusion protein, B11-pmel17. Treatment of DCs with B11-pmel17 resulted in the presentation of pmel17 in the context of HLA class I and class II molecules. Thus, potent pmel17-specific T cells were cytotoxic toward gp100(+) HLA-matched melanoma targets, but not HLA-mismatched melanoma or gp100(-) nonmelanoma tumor lines. Importantly, competitive inhibition of lysis of an otherwise susceptible melanoma cell line by cold targets pulsed with known gp100 CD8 T cell epitopes as well as a dose-dependent proliferative response to Th epitopes demonstrates that DCs can process targeted Ag for activation of cytotoxic as well as helper arms of the immune response. Thus, the specific targeting of soluble exogenous tumor Ag to the DC mannose receptor directly contributes to the generation of multiple HLA-restricted Ag-specific T cell responses.