A proteomic investigation of Fusobacterium nucleatum alkaline-induced biofilms

Background: The Gram negative anaerobe Fusobacterium nucleatum has been implicated in the aetiology of periodontal diseases. Although frequently isolated from healthy dental plaque, its numbers and proportion increase in plaque associated with disease. One of the significant physico-chemical changes in the diseased gingival sulcus is increased environmental pH. When grown under controlled conditions in our laboratory, F. nucleatum subspecies polymorphum formed mono-culture biofilms when cultured at pH 8.2. Biofilm formation is a survival strategy for bacteria, often associated with altered physiology and increased virulence. A proteomic approach was used to understand the phenotypic changes in F. nucleatum cells associated with alkaline induced biofilms. The proteomic based identification of significantly altered proteins was verified where possible using additional methods including quantitative real-time PCR (qRT-PCR), enzyme assay, acidic end-product analysis, intracellular polyglucose assay and Western blotting. Results: Of 421 proteins detected on two-dimensional electrophoresis gels, spot densities of 54 proteins varied significantly (p < 0.05) in F. nucleatum cultured at pH 8.2 compared to growth at pH 7.4. Proteins that were differentially produced in biofilm cells were associated with the functional classes; metabolic enzymes, transport, stress response and hypothetical proteins. Our results suggest that biofilm cells were more metabolically efficient than planktonic cells as changes to amino acid and glucose metabolism generated additional energy needed for survival in a sub-optimal environment. The intracellular concentration of stress response proteins including heat shock protein GroEL and recombinational protein RecA increased markedly in the alkaline environment. A significant finding was the increased abundance of an adhesin, Fusobacterial outer membrane protein A (FomA). This surface protein is known for its capacity to bind to a vast number of bacterial species and human epithelial cells and its increased abundance was associated with biofilm formation. Conclusion: This investigation identified a number of proteins that were significantly altered by F. nucleatum in response to alkaline conditions similar to those reported in diseased periodontal pockets. The results provide insight into the adaptive mechanisms used by F. nucleatum biofilms in response to pH increase in the host environment.Jactty Chew, Peter S Zilm, Janet M Fuss and Neville J Gull

Valuing climate change uncertainty reductions for robust energy portfolios

Climate policy uncertainty has decisive influence on energy sector strategies. Potential stranded climate-energy investments may be enormous. Remote sensing can improve our understanding of the climate system and thus better inform climate policy and reduce associated uncertainties. We develop an integrated energy-portfolio model to value these uncertainties. The operations of individual power plants are optimized using real options given scenarios of stochastically evolving CO2 prices mimicking observation-induced climate policy uncertainty. The resulting profit distributions are used in a portfolio optimization. The optimization under imperfect information about future CO2 prices leads to substantially lower profits for a given risk level when portfolios are to be robust across all plausible scenarios. A potential uncertainty reduction associated with an improved climate modeling supported by remote sensing will thus not only lead to substantial financial efficiency gains, but will also be conducive to steering investments into the direction of higher shares of renewable energy

Permits vs. Offsets Under Investment Uncertainty

A global crediting mechanism would enable developing countries without binding emissions reduction targets to participate in the international carbon market. Linking the framework on Reducing Emissions from Deforestation and Forest Degradation (REDD) as an offset program to major cap-and-trade programs is a particularly promising approach to increase both climate finance and cost-efficiency. However, the coexistence of permits and offsets also creates a classic case of interaction effects. In this paper, we explore how the availability of multiple compliance instruments affects energy investment incentives. Alternative trading and linkage schemes are compared using a real options model of firm-level investment decisions under stochastic prices and the ability to delay investments. We first isolate the critical design factors that drive private investments in the energy sector. We then identify policy regimes that balance the different concerns in the polarized debate for and against the inclusion of forest carbon offsets

Transgender Transitioning and Change of Self-Reported Sexual Orientation

Objective: Sexual orientation is usually considered to be determined in early life and stable in the course of adulthood. In contrast, some transgender individuals report a change in sexual orientation. A common reason for this phenomenon is not known. Methods: We included 115 transsexual persons (70 male-to-female "MtF" and 45 female-to-male "FtM") patients from our endocrine outpatient clinic, who completed a questionnaire, retrospectively evaluating the history of their gender transition phase. The questionnaire focused on sexual orientation and recalled time points of changes in sexual orientation in the context of transition. Participants were further asked to provide a personal concept for a potential change in sexual orientation. Results: In total, 32.9% (n = 23) MtF reported a change in sexual orientation in contrast to 22.2% (n = 10) FtM transsexual persons (p = 0.132). Out of these patients, 39.1% (MtF) and 60% (FtM) reported a change in sexual orientation before having undergone any sex reassignment surgery. FtM that had initially been sexually oriented towards males (= androphilic), were significantly more likely to report on a change in sexual orientation than gynephilic, analloerotic or bisexual FtM (p = 0.012). Similarly, gynephilic MtF reported a change in sexual orientation more frequently than androphilic, analloerotic or bisexual MtF transsexual persons (p = 0.05). Conclusion: In line with earlier reports, we reveal that a change in self-reported sexual orientation is frequent and does not solely occur in the context of particular transition events. Transsexual persons that are attracted by individuals of the opposite biological sex are more likely to change sexual orientation. Qualitative reports suggest that the individual's biography, autogynephilic and autoandrophilic sexual arousal, confusion before and after transitioning, social and self-acceptance, as well as concept of sexual orientation itself may explain this phenomenon

Residency time of agonists does not affect the stability of GPCR–arrestin complexes

Background and purpose: The interaction of arrestins with G-protein coupled receptors (GPCRs) desensitizes agonist-dependent receptor responses and often leads to receptor internalization. GPCRs that internalize without arrestin have been classified as "class A" GPCRs whereas "class B" GPCRs co-internalize with arrestin into endosomes. The interaction of arrestins with GPCRs requires both agonist activation and receptor phosphorylation. Here, we ask the question whether agonists with very slow off-rates can cause the formation of particularly stable receptor-arrestin complexes. Experimental approach: The stability of GPCR-arrestin-3 complexes at two class A GPCRs, the β2 -adrenoceptor and the μ opioid receptor, was assessed using two different techniques, fluorescence resonance energy transfer (FRET) and fluorescence recovery after photobleaching (FRAP) employing several ligands with very different off-rates. Arrestin trafficking was determined by confocal microscopy. Key results: Upon agonist washout, GPCR-arrestin-3 complexes showed markedly different dissociation rates in single-cell FRET experiments. In FRAP experiments, however, all full agonists led to the formation of receptor-arrestin complexes of identical stability whereas the complex between the μ receptor and arrestin-3 induced by the partial agonist morphine was less stable. Agonists with very slow off-rates could not mediate the co-internalization of arrestin-3 with class A GPCRs into endosomes. Conclusions and implications: Agonist off-rates do not affect the stability of GPCR-arrestin complexes but phosphorylation patterns do. Our results imply that orthosteric agonists are not able to pharmacologically convert class A into class B GPCRs

An Integrated CVaR and Real Options Approach to Investments in the Energy Sector

The objective of this paper is to combine a real options framework with portfolio optimization techniques and to apply this new framework to investments in the electricity sector. In particular, a real options model is used to assess the adoption decision of particular technologies under uncertainty. These technologies are coal-fired power plants, biomass-fired power plants and onshore wind mills, and they are representative of technologies based on fossil fuels, biomass and renewables, respectively. The return distributions resulting from this analysis are then used as an input to a portfolio optimization, where the measure of risk is the Conditional Value-at-Risk (CVaR)

Distinguishing problematic from nonproblematic postsurgical pain: A pain trajectory analysis after total knee arthroplasty

The goal of this study was to follow a cohort of patients undergoing total knee arthroplasty over time to: (1) identify and describe the various pain trajectories beginning preoperatively and for up to 12 months after surgery, (2) identify baseline predictors of trajectory group membership, and (3) identify trajectory groups associated with poor psychosocial outcomes 12 months after surgery. One hundred seventy-three participants (female = 85 [49%]; mean age [years] = 62.9, SD = 6.8) completed pain and psychological questionnaires and functional performance tests preoperatively and 4 days, 6 weeks, and 3 and 12 months after total knee arthroplasty. Using growth mixture modeling, results showed that a 4-group model, with a quadratic slope and baseline pain data predicting trajectory group membership, best fit the data (Akaike information criterion = 2772.27). The first 3 pain trajectories represent various rates of recovery ending with relatively low levels of pain 12 months after surgery. Group 4, the constant high pain group, comprises patients who have a neutral or positive pain slope and do not show improvement in their pain experience over the first year after surgery. This model suggests that preoperative pain levels are predictive of pain trajectory group membership and moderate preoperative pain, as opposed to low or high pain, is a risk factor for a neutral or positive pain trajectory postoperatively. Consistent with previous studies, these results show that postoperative pain is not a homogeneous condition and point to the importance of examining intraindividual pain fluctuations as they relate to pain interventions and prevention strategies.M. G. Page´ is supported by a Canada Graduate Scholarship— Doctoral Award from the Canadian Institutes of Health Research (CIHR) and is a recipient of a Lillian-Wright Maternal-Child Health Scholarship from York University, a trainee member of Pain in Child Health and a CIHR Strategic Training Fellow in Pain: Molecules to Community. J. Katz is supported by a CIHR Canada Research Chair in Health Psychology at York University. H. A. Clarke is supported by a Merit Award from the Department of Anaesthesia at the University of Toronto and also supported by the STAGE Training Program in Genetic Epidemiology from the CIHR. The remaining authors have no conflicts of interest to declare