A public relations guide for military commanders

Thesis (M.S.)--Boston Universit

Cif Is Negatively Regulated by the TetR Family Repressor CifR

We previously reported that the novel Pseudomonas aeruginosa toxin Cif is capable of decreasing apical membrane expression of the cystic fibrosis transmembrane conductance regulator (CFTR). We further demonstrated that Cif is capable of degrading the synthetic epoxide hydrolase (EH) substrate S-NEPC [(2S,3S)-trans-3-phenyl-2-oxiranylmethyl 4-nitrophenol carbonate], suggesting that Cif may be reducing apical membrane expression of CFTR via its EH activity. Here we report that Cif is capable of degrading the xenobiotic epoxide epibromohydrin (EBH) to its vicinal diol 3-bromo-1,2-propanediol. We also demonstrate that this epoxide is a potent inducer of cif gene expression. We show that the predicted TetR family transcriptional repressor encoded by the PA2931 gene, which is immediately adjacent to and divergently transcribed from the cif-containing, three-gene operon, negatively regulates cif gene expression by binding to the promoter region immediately upstream of the cif-containing operon. Furthermore, this protein-DNA interaction is disrupted by the epoxide EBH in vitro, suggesting that the binding of EBH by the PA2931 protein product drives the disassociation from its DNA-binding site. Given its role as a repressor of cif gene expression, we have renamed PA2931 as CifR. Finally, we demonstrate that P. aeruginosa strains isolated from cystic fibrosis patient sputum with increased cif gene expression are impaired for the expression of the cifR gene

The Utilitarian Virtual Self - Using Embodied Personalized Avatars to Investigate Moral Decision-Making in Semi-Autonomous Vehicle Dilemmas

Embodied personalized avatars are a promising new tool to investigate moral decision-making by transposing the user into the &amp;#x201C;middle of the action&amp;#x201D; in moral dilemmas. Here, we tested whether avatar personalization and motor control could impact moral decision-making, physiological reactions and reaction times, as well as embodiment, presence and avatar perception. Seventeen participants, who had their personalized avatars created in a previous study, took part in a range of incongruent (i.e., harmful action led to better overall outcomes) and congruent (i.e., harmful action led to trivial outcomes) moral dilemmas as the drivers of a semi-autonomous car. They embodied four different avatars (counterbalanced - personalized motor control, personalized no motor control, generic motor control, generic no motor control). Overall, participants took a utilitarian approach by performing harmful actions only to maximize outcomes. We found increased physiological arousal (SCRs and heart rate) for personalized avatars compared to generic avatars, and increased SCRs in motor control conditions compared to no motor control. Participants had slower reaction times when they had motor control over their avatars, possibly hinting at more elaborate decision-making processes. Presence was also higher in motor control compared to no motor control conditions. Embodiment ratings were higher for personalized avatars, and generally, personalization and motor control were perceptually positive features. These findings highlight the utility of personalized avatars and open up a range of future research possibilities that could benefit from the affordances of this technology and simulate, more closely than ever, real-life action.</p

In Vitro Analysis of Tobramycin-Treated Pseudomonas aeruginosa Biofilms on Cystic Fibrosis-Derived Airway Epithelial Cells

P. aeruginosa forms biofilms in the lungs of individuals with cystic fibrosis (CF); however, there have been no effective model systems for studying biofilm formation in the CF lung. We have developed a tissue culture system for growth of P. aeruginosa biofilms on CF-derived human airway cells that promotes the formation of highly antibiotic-resistant microcolonies, which produce an extracellular polysaccharide matrix and require the known abiotic biofilm formation genes flgK and pilB. Treatment of P. aeruginosa biofilms with tobramycin reduced the virulence of the biofilms both by reducing bacterial numbers and by altering virulence gene expression. We performed microarray analysis of these biofilms on epithelial cells after treatment with tobramycin, and we compared these results with gene expression of (i) tobramycin-treated planktonic P. aeruginosa and (ii) tobramycin-treated P. aeruginosa biofilms on an abiotic surface. Despite the conservation in functions required to form a biofilm, our results show that the responses to tobramycin treatment of biofilms grown on biotic versus abiotic surfaces are different, as exemplified by downregulation of genes involved in Pseudomonas quinolone signal biosynthesis specifically in epithelial cell-grown biofilms versus plastic-grown biofilms. We also identified the gene PA0913, which is upregulated by tobramycin specifically in biofilms grown on CF airway cells and codes for a probable magnesium transporter, MgtE. Mutation of the PA0913 gene increased the bacterial virulence of biofilms on the epithelial cells, consistent with a role for the gene in the suppression of bacterial virulence. Taken together, our data show that analysis of biofilms on airway cells provides new insights into the interaction of these microbial communities with the host

What Attracts Women to the IT Field? The First Process of Occupational Socialization

Socialization in an occupation differs in important ways from organizational socialization. Entering a defined occupation is apt to involve a lifelong commitment, yet it is one that individuals often drift into gradually. Entering occupations involves five different, but overlapping processes: attraction, access, adjustment, identification and commitment (Trice & Beyer, 1993). The first of these five processes refers to the specific features of the occupation that get individuals attention and attract them to the occupation. Such features include specific members of the occupation to whom newcomers get attracted such as successful heroes and mentors. It could also be the activities distinctive of the occupational culture or the kinds of extrinsic and intrinsic rewards that the occupation appears to offer. This research study focuses on the attraction process of occupational socialization to the information technology (Ahuja, 2002) field of female students in order to understand women’s experiences and initial perceptions of the IT occupation. This study gathers empirical evidence from current female students in IT-related majors based on a qualitative approach and the use of focus groups as the elicitation technique. The goal of our research study is to contribute to a better understanding of the initial process of occupational socialization of female students in IT majors. The findings of this study, we believe, can help in improving and customizing recruitment strategies for female students that would emphasize the most attractive features of the IT occupation as perceived by women. In this research in progress we present the findings of our study based on eight focus groups conducted with students of three IT related academic majors in two academic institutions in the United States

Epoxide-Mediated Differential Packaging of Cif and Other Virulence Factors into Outer Membrane Vesicles

Pseudomonas aeruginosa produces outer membrane vesicles (OMVs) that contain a number of secreted bacterial proteins, including phospholipases, alkaline phosphatase, and the CFTR inhibitory factor (Cif). Previously, Cif, an epoxide hydrolase, was shown to be regulated at the transcriptional level by epoxides, which serve as ligands of the repressor, CifR. Here, we tested whether epoxides have an effect on Cif levels in OMVs. We showed that growth of P. aeruginosa in the presence of specific epox

Clustered Regularly Interspaced Short Palindromic Repeat-Dependent, Biofilm-Specific Death of Pseudomonas aeruginosa Mediated by Increased Expression of Phage-Related Genes

The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (CRISPR/Cas) system is an adaptive immune system present in many archaea and bacteria. CRISPR/Cas systems are incredibly diverse, and there is increasing evidence of CRISPR/Cas systems playing a role in cellular functions distinct from phage immunity. Previously, our laboratory reported one such alternate function in which the type 1-F CRISPR/Cas system of the opportunistic pathogen Pseudomonas aeruginosa strain UCBPP-PA14 (abbreviated as P. aeruginosa PA14) inhibits both biofilm formation and swarming motility when the bacterium is lysogenized by the bacteriophage DMS3. In this study, we demonstrated that the presence of just the DMS3 protospacer and the protospacer-adjacent motif (PAM) on the P. aeruginosa genome is necessary and sufficient for this CRISPR-dependent loss of these group behaviors, with no requirement of additional DMS3 sequences. We also demonstrated that the interaction of the CRISPR system with the DMS3 protospacer induces expression of SOS-regulated phage-related genes, including the well-characterized pyocin operon, through the activity of the nuclease Cas3 and subsequent RecA activation. Furthermore, our data suggest that expression of the phage-related genes results in bacterial cell death on a surface due to the inability of the CRISPR-engaged strain to downregulate phage-related gene expression, while these phage-related genes have minimal impact on growth and viability under planktonic conditions. Deletion of the phage-related genes restores biofilm formation and swarming motility while still maintaining a functional CRISPR/Cas system, demonstrating that the loss of these group behaviors is an indirect effect of CRISPR self-targeting