In this commentary, the author expands on Schlinger's 2015 contribution to the research topic, “Can an Emphasis on the Single Subject Provide Novel Neuroscientific Insights? Where Neuroscience Meets Behavior Analysis.” Whereas Schlinger suggested behavior analysis affords an experimental and theoretical model useful for neuroscientists, this commentary specifies areas of intersection that are currently rich with possibility. Concurrent challenges and opportunities are present for researchers and practitioners interested in better understanding the stimulus control of complex behavioral events or “cognitive processes” through a single-subject informed lens. It is suggested that neuroscience-informed behavior analysis is poised to improve the technology of both measurement and behavior change at the individual level, and that such a relationship could inform research and clinical practices of both behavior analysts and neuroscientists.How and where can behavioral neuroscientists incorporate single subject design in their existing repertoires? Behavioral neuroscience already employs approximations to incorporating single-subject designs. Czerniawski and Guzowski  used a dIn addition to the potential for experimental contributions, collaboration between behavior analysis and neuroscience is poised to inform more individualized clinical treatment and training for health care providers. Behavior analysis informs the evidence-based treatment of psychiatric, genetic, and behavioral disorders and co-occurring challenges. For instance, consequential contingencies often play an important role in hallucinatory behavior (e.g., Layng and Andronis, Strengthening connections between human neuroscience and behavior analysis holds theoretical implications for both fields. There is an ethical need to incorporate more effective measurement of the changes experienced by an individual exposed to long-term self-harm or other challenging behavior, drug side effects, and autism treatment. These areas hold a dangerous amount of applied significance, as widespread changes may be taking place in the brains, behavior streams, and behavioral environments of thousands of children prescribed antipsychotics, mood stabilizers, and ADHD medications with insufficiently researched side effects. Further, recent policy driven funding opportunities for children to receive tens of weekly hours of behavior analytic therapy for autism spectrum disorders may pose a new potential intersection between human neuroscientists and behavior analysts. It is consistent with Schlinger's  suggestiIn closing, this article suggests that consistent with Schlinger's position Schlinger , it is pThe author confirms being the sole contributor of this work and approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The cough reflex is modulated throughout growth and development. Cough—but not expiration reflex—appears to be absent at birth, but increases with maturation. Thus, acute cough is the most frequent respiratory symptom during the first few years of life. Later on, the pubertal development seems to play a significant role in changing of the cough threshold during childhood and adolescence resulting in sex-related differences in cough reflex sensitivity in adulthood. Asthma is the major cause of chronic cough in children. Prolonged acute cough is usually related to the long-lasting effects of a previous viral airway infection or to the particular entity called protracted bacterial bronchitis. Cough pointers and type may orient toward specific etiologies, such as barking cough in croup or tracheomalacia, paroxystic whooping cough in Pertussis. Cough is productive in protracted bacterial bronchitis, sinusitis or bronchiectasis. Cough is usually associated with wheeze or dyspnea on exertion in asthma; however, it may be the sole symptom in cough variant asthma. Thus, pediatric cough has particularities differentiating it from adult cough, so the approach and management should be developmentally specific. Cough and expiration reflex are vital airway defensive mechanisms. Cough is traditionally defined as a 3-phase motor act characterized by an inspiratory effort (inspiratory phase) followed by a forced expiratory effort against a closed glottis (compressive phase) followed by opening of the glottis and rapid expiratory airflow (expulsive phase). The expiration reflex differs from the cough reflex in that it lacks the preparatory inspiration  acute or chronic (micro-) aspiration of refluxed material into the airways or (b) by the activation of the vagal afferent nerves in the esophagus that may directly trigger cough or, more likely, may lead to cough reflex up-regulation at central (brainstem) level. Animal studies have identified and characterized the neural pathways for two putative mechanisms, reviewed in a recent paper  result in hyperventilation, whereas apnea and a series of expiration reflexes may cause hypoventilation (Korpas and Tomori, Chronic cough is frequent in childhood and should be treated based on etiology. The true prevalence of cough is difficult to evaluate given the methodological difficulties (Chang et al., The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Deception is a central component of the personality 'Dark Triad' . However, whether individuals exhibiting high scores on Dark Triad measures have a heightened deceptive ability has received little experimental attention. The present study tested whether the ability to lie effectively, and to detect lies told by others, was related to Dark Triad, Lie Acceptability, or Self-Deceptive measures of personality using an interactive group-based deception task. At a group level, lie detection accuracy was correlated with the ability to deceive others—replicating previous work. No evidence was found to suggest that Dark Triad traits confer any advantage either to deceive others, or to detect deception in others. Participants who considered lying to be more acceptable were more skilled at lying, while self-deceptive individuals were generally less credible and less confident when lying. Results are interpreted within a framework in which repeated practice results in enhanced deceptive ability. Many studies have shown humans to be relatively poor lie detectors, performing little better than chance at discriminating truthful from deceptive statements ,2. DespiDeceptive Interaction Task ‘DeceIT’ )A potential explanation for the inconsistent findings with regard to an association between Machiavellianism and the ability to produce successful lies may be found in the significant correlation between the degree to which participants rated lying as acceptable and their ability to produce successful lies. Machiavellianism and Lie Acceptability were significantly correlated . This correlation was only of moderate strength however (r = .384) and so sampling bias in terms of Lie Acceptability within a high Machiavellian sample may determine the extent to which the high Machiavellian sample outperform a low Machiavellian sample via the mediating influence of Lie Acceptability. More generally, the association between Lie Acceptability and success in producing lies merits further investigation. At least two mechanisms may explain why those for whom lying is more acceptable are better able to lie. First, the endorsement of lying as an interpersonal strategy may permit an individual to lie more, and thus garner greater opportunities to practice deceiving others. Second, those who consider lying more acceptable might experience less of the guilt and anxiety brought about by lying  and therCSender and the confidence with which people lie (relative to their confidence when telling the truth). CSender indexes an individual’s general credibility (or Demeanour Bias)—irrespective of whether they are telling the truth or lying. Individuals with higher credibility reported higher relative confidence when lying than those with lower credibility. Sender demeanour has been shown to have a strong influence on the outcome of deceptive encounters [An experiential account is also likely to explain the association between counters , and thedʹSender—and the relationship between self-deception and general credibility was significantly negative. Thus, individuals high in self-deception were seen as generally less credible than those low in self-deception. This finding is in contrast to the hypothesis that self-deception contributes to deceptive success .Self-deception was not associated with an increased ability to lie effectively in these data: no relationship was observed between the ability to deceive as indexed by ss  use response latency as a cue to deception, and that good liars can control the extent to which they exhibit this cue. The use of response latency as a cue to deceit by those judging deception is valid; response latencies for deceptive statements were significantly longer than those for true statements. Whether the use / control of response latency by Receivers / Senders is deliberate or implicit is an interesting avenue for future investigation.As in any experimental study of deception one must question the validity of the deceptive behaviour elicited by the experimental context. It could be argued that in the DeceIT paradigm the experimenter sanctions lies, and therefore the guilt associated with deception is reduced. We have discussed this issue, and the wider issue of laboratory-based deception research previously , and intIn summary, despite the way in which its component traits are conceptualised, the Dark Triad appears unrelated to deceptive ability, either in the role of lie detector or liar. Although the current experimental sample (n = 75) is modest in comparison with individual differences questionnaire based research samples, sufficient variability was observed in Dark Triad traits to suggest any meaningful relationship with deceptive performance might have emerged. Indeed, power analysis suggests even the strongest relationship between Dark Triad measures and deceptive ability would require a sample of 191 participants in order to reach significance and would suggest poorer deception detection associated with Dark Triad traits rather than any deceptive skill.In contrast, lie acceptability, the extent to which one endorses deceit and manipulative behaviour, relates positively to deceptive success. Furthermore, contrary to the hypothesis that self-deception is evolutionarily selected to promote the effective deception of others, these results suggest that high self-deceivers are less credible overall, and less confident when lying, than those low in self-deception. This study replicated the key findings of Wright et al. : the abi


Human embryonic stem cells (hESCs) can be differentiated into structurally and electrically functional myocardial tissue and have the potential to regenerate large regions of infarcted myocardium. One of the key challenges that needs to be addressed towards full-scale clinical application of hESCs is enhancing survival of the transplanted cells within ischaemic or scarred, avascular host tissue. Shortly after transplantation, most hESCs are lost as a result of multiple mechanical, cellular and host factors, and a large proportion of the remaining cells undergo apoptosis or necrosis shortly thereafter, as a result of loss of adhesion-related signals, ischaemia, inflammation or immunological rejection. Blocking the apoptotic signalling pathways of the cells, using pro-survival cocktails, conditioning hESCs prior to transplant, promoting angiogenesis, immunosuppressing the host and using of bioengineered matrices are among the emerging techniques that have been shown to optimize cell survival. This review presents an overview of the current strategies for optimizing cell and host tissue to improve the survival and efficacy of cardiac cells derived from pluripotent stem cells. IntroductionCell differentiation, selection and protection– Impact of cell differentiation and selection– Improving cell survival: conditioning the cellsOptimizing host myocardium– Modifying Inflammation and Immunogenicity– Angiogenesis– Extracellular and bioengineering solutionsConclusions and future challenges of functional integrationOver the past decade, significant advances have been made in generating viable cardiac cells from pluripotent stem cells. Several protocols have been developed to guide the differentiation of human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSCs) into functioning cardiomyocytes Following transplant, a high rate of stem cell loss is observed, as a result of multiple mechanical, cellular and host factors. Several studies have demonstrated that only 10–30% of stem cells remain within a few days of transplantation, with that number decreasing to 5–15% over 10–12 weeks Several investigators have described successful production of definitive, beating cardiomyocytes from human embryonic stem cells Once cardiomyocytes have been derived from hESCs, two principal strategies have been explored to improve their survival in infarcted myocardium: pre-treating cells to induce endogenous cellular survival mechanisms and using chemical or biologic inhibitors of major cell death pathways e.g. 43°C for 30 min.) induces a highly conserved transcriptional pathway that results in synthesis of cytoprotective proteins, including heat shock proteins such as Hsp60, Hsp70 and Hsp90, as well as antioxidants. Heat-shocked rat and hESC-derived cardiomyocytes have increased survival following exposure to death stimuli in vitro , cyclosporine  and the caspase inhibitor ZVAD-fmk, we observed greatly improved engraftment and survival 4 weeks post-transplant compared with cells transplanted with Matrigel only or cells delivered in serum-free medium  or Bcl2 , treatment with the pro-survival cytokine IGF-1, blocking ‘danger signals’ from nucleic acid breakdown with allopurinol and uricase, natural killer cell depletion and broad-spectrum immunosuppression. None of these individual interventions prevented graft cell death. However, when we combined heat shock with a ‘pro-survival cocktail’ made up of Matrigel, IGF-1, a Bcl XMyocardium that has been acutely or chronically infarcted presents an unwelcoming environment for the survival of transplanted cardiomyocytes. The large amount of cell necrosis in acutely infarcted tissue is associated with significant quantities of inflammatory cells, reactive oxygen species and cytokines that are toxic to the grafted cell, while the scar tissue of an old infarction can be poorly receptive to the engraftment and integration of new cells. In both situations, the ischaemia associated with the non-vascularized cell grafts presents an immediate challenge to cell survival, and the positive outcome of blocking cell death and augmenting cell survival pathways speaks to the need to overcome the inhospitable ischaemic and inflammatory environment.The inflammatory cytokine interleukin (IL)-1 is elevated in acutely infarcted myocardium, and mediates cardiac myocyte apoptosis and adverse remodelling e.g. through haematopoietic chimerism) In contrast to autotransplant studies with adult stem cells or circulating progenitor cells, allograft transplant of hESCs elicits an adaptive immunological response and compounds the problem of cells trying to survive in this inflammatory milieu Improving blood flow to infarcted myocardium has also been explored as a means to reduce the persistent ischaemia of the host tissue that threatens the survival of grafts. Pre-treating the host myocardium with adenovirus encoding VEGF 3 weeks prior to transplanting foetal cardiomyocytes led to increased capillary density in the infarct and higher rate of survival of the transplanted cells While direct intramyocardial injection of cells results in a greater retention of cells than intracoronary or intravenous methods, as much as 90% of the cells can be immediately lost from the injection site because of mechanical extrusion from the injection track and washout of cells into the circulation In vitro, hESCs cultured and transplanted in collagen have reduced apoptosis The most widely used hydrogels have properties resembling extracellular matrix that can provide adhesive peptides to maintain survival signalling of transplanted hESCs In vitro, cells survived, proliferated and aggregated, and following injection into the infarct, the chitosan formed a temporary scaffold that improved cell retention and survival, resulting in a significantly larger graft compared with direct cell injections alone. A study performed with Rat H9c2 neonatal heart cells injected into infarcted myocardium with a mixture of collagen gel and Matrigel had a threefold increase in survival compared with direct cell injections or either vehicle alone (Fig. Fibrin glue is generated by mixing fibrinogen and thrombin and has been successfully used in the injection track to prevent extrusion of skeletal myoblasts transplanted into a myocardial infarction, resulting in a greater number of cells in the target site, smaller scar and increased arteriole density The hydrogels themselves may have ameliorative effects on ischaemic myocardium by providing structural support to the heart and promoting angiogenesis. Cell-free fibrin glue injected into infarcted myocardium prevented scar expansion and wall thinning compared with control injections Anti-apoptotic, angiogenic and anti-inflammatory factors can be added to the hydrogels, providing a depot of pro-survival factors for controlled release. The use of VEGF- and FGF-loaded hydrogels promotes increased capillary ingrowth and angiogenesis when added to stem cell transplants Injection of a foreign substance, particularly a synthetic material, however, may not always be beneficial. Hyaluronic acid has also been shown to impair IGF-1 signalling Other novel methods for improving cell retention have been described. One such technique uses superparamagnetic microspheres to magnetize cardiac-derived cells that are then localized and retained with a magnet superimposed over target tissue The past decade has provided promising techniques for improving the survival of hESC-derived cardiomyocyte grafts. One of the future challenges for cardiac regeneration in the coming years will be learning how to integrate graft cell conditioning, angiogenesis, anti-apoptosis, immunosuppression, use of bioengineered matrices and other emerging strategies to optimize cell survival. In addition, the complex interaction of transplanted cardiomyocytes and host tissue must also be co-ordinated in the proper temporal sequence. Formation of a rich vascular bed, modulation of inflammation and up-regulation of anti-apoptotic factors and homing cytokines need to be timed to coincide with cell transplant, when these cells are most likely to respond to these signals Tissue-engineered cardiac cell sheets have been successfully constructed from pluripotent stem cells and may provide a solution to the problem of cell death as a result of loss of matrix/cell attachments et al. demonstrated that the hESC cardiomyocytes can form structural and electromechanical connections with host tissue Beyond simply achieving survival of the hESC-derived cardiomyocytes, attention needs to be given to ensuring the optimal electromechanical function of the graft and integration with the host tissue. Kehat While great strides have been made towards the goal of regenerating cardiac tissue with pluripotent stem cells, the road to carrying out clinical human studies requires further optimization of these pro-survival techniques. We will need to achieve large-scale remuscularization of an infarct in larger animals with grafts that are integrated electrically and mechanically into host tissue, before the ultimate goal of regenerating the human heart can be realized.


The online AFCC course performed strongly in its first years, yielding both high participant satisfaction and substantial improvement in workplace asthma management activity. This training holds promise for introducing and improving multidimensional asthma management strategies at childcare facilities nationwide.In 2011, the Florida Asthma Coalition (FAC) began offering its Asthma-Friendly Childcare Center (AFCC) training online. This course teaches childcare center employees the fundamentals of effective asthma management. It covers basic asthma physiology, ways to recognize asthma attacks, techniques to help children experiencing attacks, and strategies to create healthy environments for asthmatics. A team of health services researchers evaluated both years of the online training. Evaluators used a quasi-experimental design with pretest, posttest, and follow-up assessment. Questions measured knowledge gain and retention, user satisfaction, and implementation of management strategies. Over 650 people from nearly all 67 Florida counties took AFCC training online between 2011 and 2013. Test scores improved by a minimum of 11% points in all program years evaluated. Gains in both knowledge and confidence were substantial and highly significant across years. While individual trainees did forget some content on follow-up, they seemed to retain the specific messages most relevant for their own workplaces. Most trainees also planned to implement multiple management strategies recommended by the training. A large majority of participants rated the training as excellent on all quality metrics, including relevance of content and time efficiency of the online format. Nearly all respondents perceived the training as useful for both providing improved care and fulfilling licensure or certification requirements. Many participants also indicated that their centers would pursue formal certification as AFCCs  In 2011, the Florida Asthma Coalition (FAC) began offering its Asthma-Friendly Childcare Center (AFCC) training online. This single-session course was developed in partnership with St. Petersburg College, which also served as the data steward for pilot sessions of the training. The 1-h training covers basic concepts of asthma physiology, ways to recognize that a child is having an asthma attack, techniques to help a child who is having an asthma attack, and strategies to help create healthier environments for children with asthma.Participants in the course take a pretest to capture their knowledge at baseline. After completing the course, participants take a posttest in which they respond to the same knowledge questions, as well as a supplemental survey with questions about the quality of the course itself. Starting in spring 2012, participants have also been sent a follow-up survey roughly 2 months following completion of the course. This follow-up survey tracks knowledge retention from taking the course, as well as changes in asthma management behavior implemented by course participants.The AFCC program has been active from summer 2011 through the present. This article focuses on results from the preliminary evaluation of AFCC. Initial evaluation efforts used data from the first two program years: 2011–2012 and 2012–2013. A subsequent evaluation was conducted with the same instruments for the online version of the training using a modified content management platform in the 2013–2014 program year.The prevalence of childhood asthma in the United States has increased steadily over time . Asthma Zahran et al.  stress tvia telephone. The 6-month training course yielded significant positive changes in asthma management competencies and behaviors.Educational interventions with families were some of the earliest to demonstrate success. Multiple teams have developed asthma self-management curricula for urban low-income families, both in general  and specSeveral research teams have assessed the effectiveness and promise of computer-based training for asthma management. Early efforts to use computer technology to help children develop good self-management skills yielded positive results . ParticiBuilding on the increasing level of success observed with newer versions of traditional multimedia trainings using curricula on CD-ROM, more recent efforts have extended the successes of these trainings to online and hybridized formats. Early data suggest a possible advantage for online training formats that is likely to persist due to progressively higher levels of connectivity in younger populations. Web-based education has yielded excellent results in domains such as consistent use of controller medications, reduced use of reliever medications, and fewer urgent physician visits . Cicuttovia mutually supportive committees for quality, safety, and service. Araújo-Martins et al.  in collaboration with St. Petersburg College incorporates these four principles in its integrative approach to asthma management. To date, the AFCC training is the first of its kind – an online intervention that not only assesses but also builds capacity for asthma management in childcare centers. However, an instrument for evaluating readiness to work with young children with asthma in day care settings has recently been released.The Preparing for Asthma in Child Care (PACC) questionnaire uses national guidelines and standards along with evidence from scientific literature, onsite case studies, and key informant interviews to assess how prepared childcare centers are to work with young asthmatics . The PACHow many people are participating in the AFCC training?How much knowledge are participants gaining from the AFCC training?How satisfied are participants with the AFCC training?How much have participants changed their behavior/taken action since participating in the AFCC training?The overall goal of the evaluation was to see how well the online version of the AFCC training builds capacity to create asthma-friendly environments for young children in day care settings. Evaluation of the AFCC training addressed the following specific questions:The FAC will share evaluation results with key stakeholders including training participants, program partners, FAC Childcare Workgroup members, local asthma coalitions, and FAP staff at the Centers for Disease Control and Prevention (CDC). Findings will be used to improve course content and evaluation tools in future years.T-tests to check for significant differences in scores for each pretest and posttest question, as well as overall scores. Likewise, paired T-tests were conducted to assess knowledge gain or loss between posttest and follow-up.Scoring reports for pretest and posttest data were shared with evaluators by FAP staff. These scoring reports consisted of Excel spreadsheets with one row for each participant. Pretest and posttest responses were recorded by question, and demographic information about participants was also captured. Information about asthma management in each content domain was captured using matrix questions with Likert response scales, indicating increasing levels of agreement . These data were recoded in Excel to reflect correct and incorrect responses (for knowledge questions) or level of agreement . Follow-up survey data were downloaded from SurveyMonkey into Excel, where they were likewise recoded into dummy or polytomous categorical variables as appropriate. Data on perceived course quality were analyzed by computing frequencies of spreadsheet cell values. Knowledge gain from pretest to posttest was assessed using paired A total of 296 people took the online AFCC training between September 1, 2011 and June 30, 2012; pretest and posttest data are available for all 296 of these participants. Of those individuals, 96 also completed the 2-month follow-up survey. Only people who participated through June 30 were included in the 2012 analysis due to timing constraints for outcomes reporting during Grant Year 3.A total of 365 people took the online AFCC training between September 1, 2012 and August 31, 2013. Of these individuals, 94 reported working for Head Start Programs. Pretest and posttest data are available for all 365 of these participants. Of the 365, 113 also completed the 2-month follow-up survey, which concluded in early November 2013 for the August cohort. Participants were distributed across multiple Florida counties, as shown in Table T-tests to detect significant differences in average percentage of respondents answering each question correctly between pretest and posttest, as well as between posttest and follow-up for knowledge questions that were repeated on the follow-up survey. Results from statistical testing are displayed in Table Responses from knowledge questions on the pretest, posttest, and follow-up survey were compared to assess knowledge gain and retention. These data were compared using paired In both years, trainees showed significant knowledge gain from pretest to posttest on all questions except two. No consistent change was observed across program years for knowledge of management strategies or triggers. In addition, participants’ total scores increased markedly from pretest to posttest, with a minimum of 11% improvement in both program years. In 2011–2012, average posttest scores increased to 95% from a baseline average score of 82% on pretest. In 2012–2013, similar increases were observed, from an average baseline of 84% on pretest to an average posttest score of 95%.Results were also compared for knowledge questions that matched on the posttest and follow-up survey. Follow-up assessment of knowledge yielded poor outcomes, with trainees showing slightly decreased knowledge on two out of three questions. Given the smaller battery of questions (3 versus 11) as well as the smaller sample of people (96 versus 296 in 2011–2012 and 113 versus 365 in 2012–2013) who completed the online follow-up survey compared to the pre- and posttests, caution should be exercised in interpreting these results.Trainees were asked to indicate their level of confidence about specific asthma management activities on the follow-up survey. Differences in scores are expressed as points in a five-point attitudinal scale. Table Gains in confidence about asthma management activities after taking the training were highly significant across all seven questions in both program years. Taken together with findings from knowledge retention questions, these results suggest that while individual trainees may not remember every specific piece of information they learned in the training, they are likely retaining the specific training messages that are most relevant for implementation in their own work settings.Participants were asked to rate the quality of specific elements of the training. Distribution of ratings across quality categories is shown in Table On the posttest, trainees were asked to indicate how useful the training was for their professional development and activities. Nearly all respondents indicated that the training was useful for both providing improved care and fulfilling licensure or certification requirements. The 2-month follow-up survey also asked questions about the utility of the training; these questions were mapped to specific asthma management activities. In both program years, a large majority of respondents found the training either extremely influential or very influential in helping them build asthma management skills.The 2-month follow-up survey asked participants to indicate whether or not they had implemented specific asthma management strategies since taking the training. Although we had fewer people taking the follow-up survey than completing the course in all evaluated years, we did not observe significant differences in demographics between people who took the follow-up assessment and people who did not. Results are shown in Table Smaller numbers of trainees indicated that these activities were already in place at their facilities before they took the training. Likewise, several participants were not sure of which activities were already in place or currently being implemented. In 2011–2012, about the same amount of trainees answered “yes” and “no” for each question. The following program year, however, “yes” became the most common response to each question.Finally, trainees were asked about their use of the guidance resources provided by the FAC, as well as their intent to pursue the AFCC certification for their centers. While many participants were already using the resource guide, fewer were certain of their center’s intent to pursue AFCC recognition. However, a great many more were not sure whether or not their center would pursue the designation. Comparatively few respondents said that their center had no intention of pursuing the AFCC recognition opportunity. In 2012–2013, five respondents also said that they had already secured AFCC recognition for their centers prior to taking the training.Overall, findings from this evaluation suggest that the online AFCC training offers valuable information in an accessible format that makes learning easy. The quality of the training may explain why it attracted substantial numbers of people from almost all of Florida’s 67 counties in its first 2 years of availability. Likewise, the diversity of professions represented among people who took the course suggests that it is useful not just for people who work in traditional childcare centers but also for anyone else who interacts with children affected by asthma.Preliminary results for knowledge retention from posttest to follow-up are weak. However, the small number of follow-up questions may not capture the types of information from the course that have actually proven most useful to trainees. This is especially likely since the specific questions included in the follow-up to measure knowledge address behaviors and strategies rather than asthma pathology or possible triggers. Likewise, results from questions about which management strategies trainees have implemented suggest that knowledge retention and the ability to translate this knowledge into action may be stronger than indicated by the follow-up questions on asthma basics.Given the strength of results on the posttest, we were unsurprised to see a small slip in scores on follow-up due to the time lag. However, we also note that assessment of knowledge change from posttest to follow-up was not as thorough as assessment of knowledge gain from taking the course, due to the much smaller sample size and smaller number of questions. We might have seen overall results more similar to those obtained on posttest if we had repeated the full battery of knowledge questions instead of choosing three. In either case, participants retained a high level of knowledge on follow-up despite not scoring quite as highly as they did on posttest.Participants corroborated the quality of the training almost unanimously, rating it highly on a variety of different quality assessments and consistently stating that it enhanced their professional lives. Likewise, trainees gave overwhelmingly positive reports of how their confidence about asthma management changed after taking the course. These increases in confidence are solidly borne out by available data on behavior change – despite the comparatively small sample sizes of only 96 and 113 people for the follow-up surveys, implementation of recommended management strategies appeared to be widespread. No behavior change was reported by fewer than 17% of participants. More typical rates of behavior change were well above 50%, with a maximum of 82%.p-values below 0.01 and often close to 0. Likewise, data collected on time required to complete the course suggest that the online AFCC training is an efficient means of delivering this content and provides busy professionals with the flexibility required to fit continuing education activities into their work schedules.Increases in participant’s knowledge from pretest to posttest also suggest that the training is highly effective at developing multidimensional competencies in asthma management. In both program years, all but two of the knowledge gain questions showed highly significant improvement in scores upon posttest, with Many people who have taken the course have reviewed the supplementary materials  and are using them at work. A substantial number of participants also indicated that their centers had already decided to pursue the AFCC recognition, and almost twice as many indicated that pursuing the recognition was a possibility by choosing “not sure.” The high number of “not sure” responses on this question may owe in part to phrasing – some respondents may not have decision-making authority about whether or not their centers pursue the certification, despite independent interest in taking advantage of this opportunity.Overall, data quality was a leading strength of the AFCC training and its component evaluation activities. The high level of data quality achieved across multiple years of AFCC implementation appeared to stem largely from the online delivery system developed by St. Petersburg College and linkage with SurveyMonkey follow-up assessment managed by FAP. Completeness of data was also a substantial strength of the training and our evaluation thereof for pretest and posttest, and a modest strength for follow-up assessment as well. Given adequate sample sizes for records with three timepoints’ worth of data, we could make reasonable inferences about the long-term potential for positive impact on asthma management behaviors and policies without imputing any values for participants who were missing data on the follow-up assessment. However, we do note that our projections about long-term impact could be much stronger given more timepoints of follow-up data collection.The AFCC training performed extremely well in its first years and holds tremendous promise for changing the landscape of asthma management at childcare facilities nationwide. To strengthen the training further, evaluators offered recommendations based on results from preliminary evaluation. Researchers suggested four action items for FL DOH program staff and St. Petersburg College partners. Recommendations are outlined below, along with implementation strategies for each.The slight knowledge loss observed in this evaluation did not present a major concern due to unmatched sample sizes between posttest and follow-up. Evaluators projected that analyzing complete data for all cohorts in future program years would afford better understanding of why participants may lose some knowledge over time. However, evaluators also noted that some score attrition might continue to occur as a function of regression to the mean, given extremely high scores observed on posttesting for the preliminary evaluation.Wording of the original evaluation question made causal inference impossible regarding why people felt unsure, because it only assessed organizational strategy as opposed to participant interest. Evaluators and FL DOH program staff changed this question language slightly to reflect participants’ actual interest in the certification.Strong results for behavior change across the board did not indicate taking in-depth action to address discrepancies between rates of behavior change on specific measures. However, analysis of additional cohorts in future program years may reveal systemic trends in which behavior changes might not be sufficiently emphasized in the training or easily achievable in specific workplaces. The latter point may become especially salient if AFCC is offered in other states.Evaluation results from the online course’s first year suggested that AFCC offers an efficient, effective, engaging, impactful, and empowering opportunity for childcare professionals to increase their skill in managing children’s asthma. Given the ease of offering the training and the quality of evaluation data the online delivery method provides, evaluators recommended making every effort to increase exposure and enrollment for this training in years to come.via online public health listservs. As AFCCs reach continues to expand in Florida, other states may wish to explore the utility of adapting this training for use in their own childcare facilities.Enrollment data for more recent program years show a consistent upward trend in the popularity of AFCC. Program managers and evaluators have also worked actively to disseminate information about the AFCC training to public health professionals from other states. Successes and lessons learned from the program are shared regularly at national conferences, with chronic disease prevention and management coalitions, and AN completed all data management and analysis, wrote the first drafts of most manuscript sections, and edited the whole manuscript. HC assisted with all data management and analysis and contributed edits during the manuscript writing process. NP wrote the literature review and contributed edits during the manuscript writing process. JD designed the intervention, supplied data for analysis, and contributed edits during the manuscript writing process. JF assisted with designing the intervention, supplied data for analysis, and contributed edits during the manuscript writing process.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Eutropis longicaudata) living on Orchid Island (Taiwan). This species typically does not provide protection to its offspring. Using a common garden experiment, we show that maternal care is genetically determined in this population. Through field manipulations, we demonstrate that care provides a significant increase in egg survival on Orchid Island by reducing predation from egg-eating snakes (Oligodon formosanus); this predator is not abundant in other populations of the lizard, which do not display parental care. Finally, using extensive field surveys, we show that the seasonal availability of green sea turtle (Chelonia mydas) nests is the cause for the high abundance of snake predators on Orchid Island, with the snakes consuming lizard eggs when green turtle eggs are not available. Together, these lines of evidence provide the first full demonstration of how predation can trigger the evolution of parental care in a species derived from a non-caring ancestor.Animals display a great diversity of parental care tactics that ultimately enhance offspring survival, but how such behaviors evolve remains unknown for most systems. Here, we studied the evolution of maternal care, in the form of nest guarding, in a single population of long-tailed sun skink ( Parental care, defined as any form of behavior that increases the fitness of offspring by providing protection or food, is an important feature of most endotherms  and a number of ectotherms . Across many taxa, a wide diversity of parental care strategies have evolved, ranging from relatively primitive nest-guarding behaviors to more complex interdependency between parents and offspring, such as active food provisioning to offspring123436789101213141516et al. 20101For many taxa, the driving force behind the evolution of parental care is thought to be the protection of young or eggs from predation19221Eutropis longicaudata). Mothers from a single population on Orchid Island, off the coast of Taiwan, attend their egg clutches after laying, aggressively warding off egg predators, whereas lizards from other populations always abandon egg clutches immediately after layingOligodon formosanus is the most likely cause for the evolution of care on Orchid Island, as the abundance of this predator is unusually high at that locationChelonia mydas) might be an important food source for this snakeOne species that does exhibit intraspecific variation in parental care is the long-tailed sun skink (Chelonia mydas) nesting, egg-eating snake population dynamics and density, and predation rates on long-tailed sun skink nests on Orchid Island, to understand the role of this trophic link between terrestrial and marine habitats in the evolution of lizard maternal care.Here, we show that parental care on Orchid Island is genetically determined, and quantify the role of predation in favouring this behaviour on Orchid Island, as well as investigating why predation pressure is unusually high at this location. We do so by first performing a common garden experiment to study the expression of anti-snake behavior in captive lizards from Orchid Island as well as from two non-caring populations (Green Island and mainland Taiwan) which show low levels of genetic differentiation from one anotherOur common garden maternal attendance experiment used 10 captive-raised females with eggs from Orchid Island, 14 from Green Island, and 12 from the mainland Taiwan site, none of which had prior experience nesting or encountering snakes. When confronted with egg-eating snakes, all 10 females from Orchid island violently attacked snakes, thereby demonstrating egg-guarding behaviour, but none of the females from the other two sites did so, indicating that the guarding behaviour has a genetic basis despite the low level of overall genetic differentiation among these populations22 = 106.74, P < 0.0001; F2,415 = 1.57, P = 0.21) and relative humidity , which therefore do not explain differences in hatching success among populations.Hatching success of lizard eggs in the absence of maternal care was lowest on Orchid Island (18%), and much higher in the two populations lacking maternal care , with snakes preying upon most eggs on Orchid Island, but ants and fungal infections causing most mortality at the other two sites .The causes of lizard egg mortality differed significantly among populations  was over ten times higher at Orchid Island (Mean ± SE = 12.1 ± 0.95) relative to the other two sites , and within each year density peaked in the months (May-October) when sea turtles nest . By contrast, there was no relationship between annual sea turtle and lizard nest abundances , nor between snake and lizard nest abundance ; thus the relationship between sea turtle nesting and snake abundance is driven by processes that are different from those driving lizard nesting.Orchid Island is a major nesting site for green turtles in Taiwan, and provides a potential seasonal source of food for egg-eating snakes. To determine whether this food source is an important driver of the snake density, and thus of the predation pressure experienced by lizards, we surveyed the abundance of egg-sneaking snakes at Orchid Island, running monthly surveys and distinguishing between snakes captured in inland habitat where lizards nest, and beach habitat where sea turtles nest. Over the whole island (pooling estimates from inland and beach sites), we found snake density to be highest in years of greater sea turtle nesting effort  on Orchid Island is a response to egg predation by snakes. Guarding is highly effective, since lizards are large enough to aggressively attack snakes, but the snakes are obligate egg predators and thus poses no physical threat to the adult lizardsOncorhynchus spp.) from the North Pacific Ocean into freshwater streams and lakes, where they spawn and die, is possibly the best-studied example of this phenomenon. Adult salmon deliver nutrients to freshwater and terrestrial ecosystems and support the populations of many animal species (e.g. grizzly bear)33There is evidence that marine animals, such as sea turtles, can sometimes provide seasonal influxes of nutrients to terrestrial communities that can exert significant influence upon evolutionary dynamics on the structure and dynamics of terrestrial communities, even if they are extrinsic to those communities for most of their life histories33We cannot determine when egg guarding may have evolved in skinks nesting on Orchid Island. The concrete barriers used as nesting sites by most lizards on Orchid Island have only been present since 1995. Even though the resulting concentration of nests on this artificial substratum might have made the nests more attractive to egg-eating snakes, it seems unlikely that the trait has become fixed in the population within such a short period of time. It seems more likely that the seasonal nature of turtle nesting has imposed a great evolutionary pressure on the lizards to start defending nests, and this might have been exacerbated since the construction of the concrete barriers. The low level of genetic differentiation between Orchid Island and neighbouring populations makes this species a prime target to look for the genetic mutations that might underlie guarding behaviorEutropis longicaudata) that may be unique among vertebrates in that only a single population is known to display parental care. This species typically nests in natural grassland/lowland rainforest ecotones where eggs are buried beneath rocks and abandoned immediately after nestingOligodon formosanus), which only eat eggs, and not lizards3832We studied a widely distributed tropical lizard  is an obligate egg-eater that primarily consumes soft-shelled reptile eggsChelonia mydas)Eutropis longicaudata) (283737The Taiwan kukri snake (ata) 2837. There aata) 2837 than enta) 283737 and sea a) 28373740. Consua) 283737.Eutropis longicaudata eggs from populations on each of the Taiwan mainland, Green Island, and Orchid Island, and raised them in captivity to determine if egg-guarding behavior has a strong genetic basis (Oligodon formosanus) into the cage, and documented any egg-guarding behavior of the mother lizards collected from the wild. Because anti-snake behaviors were displayed dichotomously  is a 110 m long green sea turtle nesting beach with a long-tailed sun skink nesting site located 600 m inland . The sea32During our study, sea turtle eggs were available for snakes to consume from 1997–2008. After 2008, most sea turtle nests at Orchid Island were relocated for conservation purposes (due to high levels of snake predation) and thus were no longer available for consumption by snakes during July and August, the peak nesting months.We surveyed sea turtle nesting beaches six times nightly from May-October 1997–2008 and recorded all sea turtle nests laid and captured each snake observed. Snakes were individually marked with a PIT tag inserted between the trunk muscles and skin. We surveyed the inland lizard nesting site every six hours (4 times daily) from May-October 1997–2008 to monitor snake and lizard numbers, and snake depredation rates of lizard nestsAcross all sites, the main causes of lizard egg failure are predation .How to cite this article: Pike, D. A. et al. Surf and turf: predation by egg-eating snakes has led to the evolution of parental care in a terrestrial lizard. Sci. Rep.6, 22207; doi: 10.1038/srep22207 (2016).


High-level analysis of the results reveals expected connections between tissue-specific developmental processes and diseases linked to those tissues, and widespread connections to developmental disorders and cancers. Yet interesting new hypotheses may be derived from examining the unexpected connections. We highlight and discuss the implications of three such connections, linking dementia with bone development, polycystic ovary syndrome with cardiovascular development, and retinopathy of prematurity with lung development. Our results provide additional evidence that VEGF pathway and downstream NFKB signaling may explain the complex relationship between bronchopulmonary dysplasia and retinopathy of prematurity, and may form a bridge between two currently-competing hypotheses about the molecular origins of bronchopulmonary dysplasia. Further data exploration and similar queries about other gene sets may generate a variety of new information about the molecular relationships between additional diseases.Identifying molecular connections between developmental processes and disease can lead to new hypotheses about health risks at all stages of life. Here we introduce a new approach to identifying significant connections between gene sets and disease genes, and apply it to several gene sets related to human development. To overcome the limits of incomplete and imperfect information linking genes to disease, we pool genes within disease subtrees in the MeSH taxonomy, and we demonstrate that such pooling improves the power and accuracy of our approach. Significance is assessed through permutation. We created a web-based visualization tool to facilitate multi-scale exploration of this large collection of significant connections ( Understanding the roles that genes involved in normal human development can play in disease processes is an important part of predicting disease risk and designing novel treatment approaches. In this study, we have identified classes of disease that are associated with a surprisingly large number of genes involved in any of several tissue-specific developmental processes. To do so, we developed a novel approach whose strength comes from pooling genetic information across related diseases, overcoming problems ordinarily posed by limited information about individual gene-disease relationships. We demonstrate the method's efficacy both by examining its ability to highlight connections between gene sets and disease classes that are known to be related, and by demonstrating that the approach recovers expected broad classes of connections, such as those between heart development and cardiovascular disorders. However, by examining unexpected connections in this data set, we are able to develop new understanding of some surprising disease relationships, such as the one between dementia and osteoporosis. Such connections may lead to a better overall understanding of the role of development in lifelong health, as well as to the design of new methods to treat a range of diseases. The study of the health implications of developmental processes has now entered the genomic era. The recent sequencing of an entire fetal genome Despite considerable progress, our understanding of the molecular etiology of most complex diseases is still limited. Yet by combining weak signals from multiple genes, we may identify patterns that provide clinically significant insights into disease processes. We hypothesized that by examining the relationships between sets of genes related to specific developmental processes and reported disease genes, we could develop novel insights into developmental impacts on health. To test this hypothesis, we created a novel approach and tool to assess the overrepresentation of various developmental gene sets among groups of genes linked to specific diseases. Our approach derives its strength from combining signals of sets of genes and from pooling disease-gene links across disease subtypes using a hierarchical taxonomy of disease. We demonstrate that this pooling approach improves accuracy over a comparable enrichment-detection method without pooling. Our approach has the advantage of potentially generalizing incomplete disease gene data and overcoming variation in how genes are associated with specific disease terms, improving our ability to detect novel and interesting connections.We note that a similar principle - that of pooling many weak signals to provide a stronger one - has led to the creation of many highly effective “gene-set analysis” methods for expression data The choice of a disease taxonomy for this analysis is vitally important, yet most existing hierarchies lack the molecular focus inherent in the proposed analysis We derive our disease-gene links from two sources: OMIM, a curated collection of genes linked to human disease query gene sets.For this study, we focus on identifying connections to genes involved in developmental processes. The gene sets chosen were based on Biological Process terms from the Gene Ontology (GO), a hierarchically-organized collection of controlled-vocabulary functional annotation of genes and gene products Additional related work assesses significant enrichment of GO functional annotation terms in query gene sets using the directed-acyclic graph structure of the Gene Ontology. Such approaches adjust enrichment calculations by accounting for relationships between the genes at a given annotation node and those at the parent or child To identify significant connections between gene sets and disease, we used a novel method of assessing overlaps between disease genes and the designated query gene sets. We first created a computational representation of the MeSH disease taxonomy in which each node represents a MeSH disease concept. We extracted and combined gene-disease links from the HuGE Genopedia database and from OMIM, and mapped the resulting 119,400 gene-disease links to the MeSH forest see . Taking For this study we considered nine DFLAT gene sets, broadly representing development in brain, bone, heart, kidney, liver, lung, nerve, blood vessels, and skin. We identified the overlaps between each of these gene sets and the disease genes at each node of our MeSH tree by counting the number of genes in both.  to assess significance of linkage using exactly the same random permutations of gene labels, but counting only the genes directly linked to disease node D (rather than those linked to the node or any of its descendants).The intuition behind this experiment is that disease classes that are We note that the traditional method used here is really just a randomized approximation to the classical hypergeometric calculation, but one that maintains the correlation structure of genes between different diseases. We have separately computed the hypergeometric probabilities (data not shown), and found them to give very similar overall results to those derived using permutation. Accordingly, we present just the permutation-based method, which is the most direct control for our pooling approach, in the comparison below.We withheld 100 randomly chosen links, each connecting a gene in the query gene set to a specific associated disease. We recomputed enrichment at each disease node without the withheld links, using both the pooling method and the traditional one. Counting then allows us to estimate the probability We repeated this experiment with a different set of 100 withheld links 100 times for each of the 9 developmental gene sets. While it is relatively easy to provide a list, for each developmental gene set, of MeSH terms whose gene set enrichment p-value is below some cutoff, interpreting those lists is complex. Because enrichment calculations are based on subtrees, there is important information available at different scales, ranging from high-level overviews of the MeSH disease forest to specific enriched gene-disease links, their significance scores, and the genes involved. For these results to lead to new discoveries, we must select from this large collection of significant links a few that are surprising yet plausible. Doing this requires a considerable amount of domain knowledge in molecular medicine.http://gda.cs.tufts.edu/development). For a high-level overview, we visualize each disjoint hierarchy of disease terms  in a simplified triangular form .The root node of MeSH category C4, “Neoplasms,” is significantly associated . This is to be expected in a subset of genes involved in bone development. Yet we also saw enrichment of retinoic acid receptor proteins  and heparin-binding proteins . These links are of interest because both heparin derivatives and retinoic acid are candidates for new Alzheimers therapies One surprising link is a significant overlap  are among those with the GO Biological Process annotation “response to steroid hormone stimulus,” a finding whose adjusted false discovery rate (as computed in DAVID via the Benjamini-Hochberg method) is below 0.015.Evidence supporting this connection has recently been proposed in empirical observations of an association between lower bone mineral density and dementia in postmenopausal women MDK, a still relatively unstudied, retinoic acid-responsive, heparin-binding protein appears to be involved in both neuron and bone growth This analysis also suggests a possible connection between dementia and bone density through additional signaling pathways. For example, the growth factor The link between heart development and polycystic ovary syndrome  has a p-value below 0.0001. PCOS is an endocrine disorder that causes hormonal changes, ovarian “cysts” , and decreased female fertility. It has been associated with an increased risk of diabetes, dyslipidemia, and cardiovascular disease TGFB/SMAD complexes are known to mediate the DNA damage response It has long been known that cardiovascular symptoms are associated with PCOS, but the molecular etiology of this connection is not clear. One study proposed that oxidative stress caused by insulin resistance may lead to cardiovascular injury in nonobese PCOS patients, but did not implicate specific molecular pathways A role for upregulation. However, there are four genes in the KEGG TGFb pathway that are known to inhibit LTBP1, DCN, Lefty, and Activin. For all of these genes there are mouse mutant strains (in a variety of backgrounds) that disrupt the homologous proteins' expression, thus potentially upregulating DCN and Activin) have knockout mutations that cause reduced female fertility We looked for further corroborating evidence in mouse models, but could not identify an existing, well-characterized mutant that is a good model of The lung development gene set was linked, with a p-value below 0.0001, to retinopathy of prematurity (MeSH term C16.614.521.731). Retinopathy of prematurity (ROP) is a complication that occurs primarily in infants delivered before approximately 28 weeks' gestational age, before the infants' visual system has been fully formed IGF1, NOS3, EPAS1, KDR, and VEGFA. These genes are all related to blood vessel or tube development. Excessive but disordered VEGF-mediated vascularization of the retina is known as the cause of ROP The significant connection we observed was based on five genes linked to both ROP and lung development: We therefore hypothesized that there might be similar molecular enrichment of the ROP genes in bronchopulmonary dysplasia (BPD), another complication of prematurity characterized by extended need for supplemental oxygen and, in extreme cases, long-term respiratory insufficiency. Like ROP, BPD also affects some, but not all, extremely premature infants. Its exact cause is unknown. Current hypotheses include one in which inflammation plays a major role VEGF pathway , using the Unified Medical Language System (UMLS) We assembled a combined set of disease-gene links for 11,831 genes using 116,117 human gene-disease associations from the Genopedia compendium in the HuGE database of Human Genetic Epidemiology We estimate the distribution of the expected number of shared genes between the query gene set and the genes associated with a disease under the null hypothesis that there is no meaningful relationship between the query gene set and the disease class. We do so by randomly choosing gene sets of the query-set size from among all the genes in our MeSH tree. This is equivalent to randomly permuting the labels of the genes in the data to determine whether or not they are in the query set. Such permutation leaves the gene-disease connections intact and maintains the complex correlation structure of genes between related diseases. Assuming that Density of enrichment was computed between the 9 query gene sets and the 26 top-level MeSH disease categories, each represented by its own tree. Because many diseases are represented multiple times at different places in each tree, we first created a listing of all the unique MeSH disease terms in each tree. If different instances of the same disease in the same tree had different p-values, they were averaged. We then compared the p-values to the chosen significance cutoff of 0.005. The fraction of unique terms in the tree with lower significance was computed. This fraction represents the “density” of significant enrichment of the query gene set in the chosen MeSH category.To create the heatmap, we z-score normalized the densities across each row (query gene set). To identify expected enrichment, we manually selected the 9 top-level MeSH disease categories thought to be most relevant to the 9 query gene sets .We performed the following experiment to compare the accuracy of our proposed pooling approach to a comparable enrichment analysis using only the genes directly associated with a given disease term. To describe the experiment, we first introduce new terminology:Assume that we are discussing only a single, fixed query gene set. Let Then we will repeatedly randomly withhold some links from supported by gene-disease link We say a node Table S1List of 26 top-level categories in the MeSH disease (C) forest.(PDF)Click here for additional data file.Table S2The 9 query gene sets and their sizes.(XLSX)Click here for additional data file.


Centrocercus urophasianus) across Wyoming. We found that pairwise resistance derived from the distribution of low-quality nesting and winter, but not summer, seasonal habitat had the strongest correlation with genetic differentiation. Although the patterns were not as strong as with habitat distribution, multivariate models with sagebrush cover and landscape ruggedness or forest cover and ruggedness similarly had a much stronger fit with genetic differentiation than an undifferentiated landscape. In most cases, landscape resistance surfaces transformed with 17.33-km-diameter moving windows were preferred, suggesting small-scale differences in habitat were unimportant at this large spatial extent. Despite the emergence of these overall patterns, there were differences in the selection of top models depending on the model selection criteria, suggesting research into the most appropriate criteria for landscape genetics is required. Overall, our results highlight the importance of differences in seasonal habitat preferences to patterns of gene flow and suggest the combination of habitat suitability modeling and linear mixed models with our resistance parameterization is a powerful approach to discerning the effects of landscape on gene flow.Given the significance of animal dispersal to population dynamics and geographic variability, understanding how dispersal is impacted by landscape patterns has major ecological and conservation importance. Speaking to the importance of dispersal, the use of linear mixed models to compare genetic differentiation with pairwise resistance derived from landscape resistance surfaces has presented new opportunities to disentangle the menagerie of factors behind effective dispersal across a given landscape. Here, we combine these approaches with novel resistance surface parameterization to determine how the distribution of high- and low-quality seasonal habitat and individual landscape components shape patterns of gene flow for the greater sage-grouse ( Animal dispersal has important and well-documented effects on population dynamics and persistence  to high  dispersal resistance. Thus, instead of investigating multiple landscape features individually, we transform one biologically meaningful surface and then compare with the genetic data. Generally, resistance values derived from suitability indices have improved the fit over geographic distance, suggesting the distribution and quality of habitat is important in driving gene flow  is distributed across western North America, with a range largely consonant with the distribution of sagebrush (Artemisia spp.). With population reductions occurring across the range of sage-grouse  What moving window size and resistance parameterization best characterize functional connectivity for sage-grouse? (2) Is effective dispersal in sage-grouse driven by the distribution of habitat preferences in a particular season? and (3) What is the added value of using habitat suitability indices over individual landscape components in landscape genetics? Lastly, the use of linear mixed models and model selection is a relatively new venture in landscape genetics  represented all unique individuals that amplified at seven or more loci.The Wyoming Game and Fish Department collected feather and blood samples from sage-grouse between the years of 2007 and 2010 across Wyoming and provided these samples for this study. Most of these samples were feathers collected noninvasively and well distributed across lek sites within the state Fig.. Our finThe majority of the feathers used in the genetic analysis were collected from leks, and thus, we used these as the basis for defining our groupings (hereafter “lek group”) for pairwise analysis. We do not suggest that our groupings represent biologically unique populations or management units , and agricultural fields , as well as from a terrain ruggedness index (RUGG) and a road decay function from primary and secondary paved roads (ROAD). All of the landscape components were consistent with those used in Fedy et al.  represent known regions of influence for habitat selection and movement (Holloran and Anderson Ri) with low values and place an emphasis on differences in high-resistance habitat,α being a scale parameter controlling the steepness of the exponential function. Higher values of α resulted in greater separation between low and high-resistance values (See Rmax) ensured consistency in the transformation between variables with different ranges of resistance values in the original layers , we reversed the resistance values  and used the resulting values in eq. 1. Subsequently, we reversed these values and added 0.1 to ensure no zero values. This equation results in the converse of the previous transformation and essentially emphasizes variation in the favorable habitats  and homogenizes high-resistance habitats  for each transformation and thus produced five resistance surfaces for each original surface )). We assessed convergence of the MCMC models by comparing results across multiple runs. Estimates from lmer and MCMCglmm were nearly identical, and thus, we present coefficients from lmer, but confidence intervals from MCMCglmm. Because we were interested in determining the relative influence of each landscape metric, variables were standardized and centered around their mean  and those that do not (coded as 0)  are generally regarded as not appropriate for models estimated with REML  to models with random and fixed effects. Unlike traditional R2, marginal R2 can decrease with the addition of variables  by ranking the candidate models separately for each criteria and then determining the mean rank and standard deviation across models. This provided a consensus value (mean) and an estimate of agreement (standard deviation) between the different model selection criteria.In total, we had 15 resistance surfaces for each landscape variable  resulting in a single combined resistance surface. We validated this approach by calculating pairwise resistances from the combined surfaces and compared the fit of the resulting pairwise resistances with genetic data. An increased fit of the combined resistance surfaces compared to models with the individual components alone would suggest some value to this approach.Hexp ranging from 0.70 to 0.82  to 0.84  was the poorest ranking model. This model also had a low standard deviation in model rank, suggesting it performed poorly for all model selection criteria Fig.; Table4.dels Fig.. The besdels Fig.; Table4.dels Fig..Examining relative model coefficients, values for WINTER and NEST were similarly higher than all other coefficients Fig.. FOR andEliminating the anthropogenic models, which had a much lower fit, one of the top three landscape component models was within each of the remaining model groupings, with a large gap in rank for the next highest ranking model in the group Fig.. Thus, tThrough model selection and parameter estimation, we were able to discern the major ecological factors driving functional connectivity for a mobile terrestrial vertebrate. Overall, the broad-scale  distribution of high-resistance  nesting and winter seasonal habitat appeared most important in driving patterns of effective dispersal for sage-grouse across this spatial extent. Comparing the results of models containing habitat suitability indices and individual landscape components suggested some convergence between dispersal and daily-use habitat, and a benefit  to using habitat suitability modeling in landscape genetics.The spatial scale with which organisms respond to landscape structure is largely determined through their inherent dispersal ability and sensitivity to changes in a given landscape feature  that likely limit sage-grouse dispersal. This is evidenced by the fact that all of the resistance values that had the best fit with genetic data were derived from surfaces that placed high-resistance values to these regions. Given the extent of these mountain ranges, these large-scale barriers may have a disproportionate influence and mask the effect of some of the other landscape features , which may have more subtle impacts. Future studies conducted at smaller spatial extents could test for regional variation in the patterns observed here and make valuable contributions to our understanding of the influence of varying spatial extents on our conclusions. Further, the importance of individual landscape variables can vary with their availability . Overall, these results indicate that there needs to be more research on the most appropriate model selection criteria for different circumstances. The selection criteria may vary in their accuracy when attempting to explain different processes and sampling variation. In the absence of a clearly preferred criterion, we recommend the assessment of agreement across multiple selection criteria as presented here.Mixed models are an important tool to account for dependence in pairwise datasets in landscape genetics (Pavlacky et al. A potential disadvantage of using model selection approaches is that although the approach will establish a model describing functional connectivity, it will not lead to an overall resistance surface if the top model is multivariate. We attempted to circumvent this problem by using standardized regression coefficients that combine individual resistance surfaces into a combined surface. In general, pairwise resistances derived from the combined surfaces provided a better fit than values derived from a single landscape component, suggesting that this may be a promising approach. Despite this improvement, the level of fit with genetic data was not as great as the multivariate models or the HSI models, and thus, more research is required into the best methods for translating multivariate models into individual resistance surfaces. In the meantime, our results combined with other studies linking HSIs and genetic differentiation (Laiolo and Tella Across their range, sage-grouse have experienced population declines and range contractions (Schroeder et al.


Malarial retinopathy allows detailed study of central nervous system vascular pathology in living patients with severe malaria. An adult with cerebral malaria is described who had prominent retinal whitening with corresponding retinal microvascular obstruction, vessel dilatation, increased vascular tortuosity, and blood retinal barrier leakage with decreased visual acuity, all of which resolved on recovery. Additional study of these features and their potential role in elucidating the pathogenesis of cerebral malaria is warranted. The pathogenesis of coma in falciparum malaria and its rapid reversibility are potential targets for adjunctive therapies, but they are not well-understood. Microvascular obstruction is probably an important contributor. The brain microvasculature is relatively inaccessible; it can be studied in detail only at post-mortem. Similarly, microvascular obstruction in the retina is thought to be a major contributor to the unique retinopathy of severe falciparum malaria, and, because it is easily visualized in living subjects, in-depth study is providing new and valuable insights. We describe an adult patient with cerebral malaria who had prominent retinal changes with some previously unrecognized features that resolved on recovery.Plasmodium falciparum malaria (parasitemia = 0.3%) with coma, generalized convulsions, hyperlactatemia, renal failure, and black urine. He had no prior medical history. Retinal photography showed bilateral patchy macular whitening with corresponding capillary non-perfusion and leakage of fluorescein caused by blood retinal barrier breakdown on fluorescein angiography , which are distributed particularly around the optic disk and typically more opaque. Malarial retinopathy is considered reversible,The angiogram in this patient showed that the whitening corresponds closely to capillary non-perfusion. This finding has not been described previously in adults but is common in Malawian children with cerebral malaria.This patient had mildly increased tortuosity of retinal blood vessels that decreased on recovery. Although increased vascular tortuosity has not been well-described in malaria, it is a recognized feature of other vascular occlusive diseases of the retina. Vessel tortuosity is caused by a combination of vessel dilation from radial stretching and the vessel taking a more serpentine path because of longitudinal stretching.1Increased vascular tortuosity has not been well-described previously in severe falciparum malaria, possibly because the normal appearance of retinal vessels varies significantly between individuals and subtle changes are difficult to identify. Ophthalmoscopy revealed engorgement and tortuosity of retinal veins in 26% of children with cerebral malaria in Ghana, which mostly resolved by 1 week.The angiogram in this patient showed focal leakage of fluorescein across the blood–retinal barrier (BRB) in areas of non-perfusion, suggesting a common etiology. The BRB is analogous to the blood–brain barrier, which is also mildly disrupted in cerebral malaria. Leakage from larger retinal vessels crossing ischemic areas is a well-known phenomenon in retinal ischemia. The significance of this finding as a contributor to the pathogenesis of malarial coma is not known. More angiographic studies are needed.This patient had decreased visual acuity, which had resolved at follow-up. Although it is not possible to give a cause, it is the first report of an association between macular retinal whitening and decreased visual acuity with subsequent recovery.Additional studies of malarial retinopathy have great potential to enhance our understanding of vascular changes in severe malaria. To maximize their impact, studies should use retinal photography, where possible, to allow detailed examination of the full range of fundus signs by multiple blinded observers. This examination should be done both acutely and at follow-up. Fluorescein angiography provides a highly detailed map of CNS retinal perfusion. There is a need for additional detailed studies to include assessment of vascular tortuosity to investigate its role as a potential early and sensitive marker in studies of severe malaria.The rate of reversibility of malarial retinopathy has potential as an end point in intervention studies of severe malaria, particularly for adjunctive therapies that directly target the pathogenesis. Additional information on the speed of reversibility of the various components of malarial retinopathy is needed, and studies are underway to investigate this.


A growing number of treatment options exist to treat metastatic castrate-resistant prostate cancer (mCRPC), and with these newer options, many questions about optimising treatment remain unanswered. One recommendation that may potentially be overlooked by practitioners is that androgen deprivation therapy (ADT) should be maintained when CRPC develops and when treatment with any of the newer agents is initiated.However, to emphasise this recommendation, it is valuable to interrogate the evidence for maintaining ADT in different clinical situations.This statement, reflecting the views of the authors, provides a discussion of this evidence and the rationale behind the recommendation that ADT should be continued in CRPC. The European Association of Urology (EAU) guideline clearly states that when castrate-resistant prostate cancer (CRPC) develops, androgen deprivation therapy (ADT) should be continued indefinitely; this recommendation applies to metastatic CRPC (mCRPC) and non-metastatic CRPC (nmCRPC) . Other gHowever, there appears to be a risk that the need to continue ADT may be overlooked by some practitioners. This aspect of the guidelines is sometimes not mentioned in subsequent discussions. Furthermore, although randomised controlled trials of the many newer agents for mCRPC, including abiraterone, enzalutamide, sipuleucel-T, radium 223 and cabazitaxel –10, all Lowering testosterone levels is associated with improved survival in prostate cancer , and thin = 85) and using chemotherapy regimens that are now outdated [Likewise, when chemotherapy is initiated in CRPC patients, the EAU recommends continuing the backbone ADT . This guoutdated . TherefoHowever, it is becoming clear that within the prostate and prostate tumour microenvironment androgen activity continues even when serum testosterone levels are suppressed by ADT , and intIn December 2013, a group of experts (the authors of this paper) met to explore the evidence and rationale for continuing ADT in CRPC when other treatments are initiated. The aim is to provide clear statements on this issue in this manuscript. Before the meeting, participants were assigned to specific topics and conducted PUBMED searches on the recent literature on these topics. Assigned participants presented on these topics during the meeting, and all participants then developed the recommendations and contents of this paper based on all the reports presented at the meeting. As such, the contents of this paper represent the conclusions of the authors only. References in the text have been assessed according to their level of scientific evidence  and thus inhibits androgen biosynthesis . In CRPCCrucially, experimental evidence suggests that the testosterone suppression achieved by abiraterone monotherapy is not sustained in non-castrated men and is overcome by a subsequent twofold–threefold surge in luteinising hormone (LH) levels  level o, 23. AltThis rationale has been used in phase III trials of abiraterone. The efficacy of abiraterone (plus prednisolone) was demonstrated in two pivotal trials in patients with mCRPC; in one study, abiraterone was used before chemotherapy, and in one study, it was used after chemotherapy , 5  monotherapy with abiraterone plus ADT to confirm the need for continued ADT when initiating abiraterone therapy in patients with mCRPC. The planned German multicentre trial SPARE will investigate the impact of continuing ADT when initiating abiraterone. This study  will investigate abiraterone monotherapy (plus prednisolone) versus abiraterone plus ADT (plus prednisolone) in 70 men with chemotherapy-naïve mCRPC .Interestingly, there are no published data assessing abiraterone (with or without ADT) in patients with non-metastatic CRPC (nmCRPC). More data on the use of newer agents for the treatment of nmCRPC are needed.Androgen receptor signalling persists during castration, and several mechanisms, even in individual patients , may explain this persistence . AdditioThe androgen receptor blocker bicalutamide has been used for many years in combination with ADT to achieve complete androgen blockade. However, in advanced metastatic prostate cancer, monotherapy with bicalutamide was inferior to ADT in prolonging overall survival (OS)  http://www.nih.gov/news/health/dec2013/nci-05.htm). Conversely, an open-label phase III trial showed no benefit of adding chemotherapy to ADT as first-line treatment compared with ADT alone in hormone-sensitive men with metastatic prostate cancer [Recent pivotal trials of chemotherapy in prostate cancer have stipulated that ADT should be continued when chemotherapy was initiated , 41  are investigating the possible advantage of maintaining ADT during chemotherapy. One of these studies (NCT01487902) that is comparing docetaxel plus prednisolone with docetaxel plus prednisolone and leuprolide in approximately 90 men with CRPC was due for completion in October 2013, and results may therefore be available soon. Until these results are available ADT should be continued when chemotherapy is initiated in mCRPC in daily practice.All clinical trials of newer agents  in mCRPC include patients who maintain castrate levels of testosterone, and so clinical practice should adhere to this principle of continuing ADT when initiating abiraterone, enzalutamide or chemotherapy. Likewise, experimental agents that are in advanced stages of clinical development for treatment of CRPC are all being assessed in patients who maintain their castration status with ADT, and so this recommendation is likely to apply to other agents that may be registered in the next few years. However, not only are more prospective data needed to assess the importance of backbone ADT in CRPC, but also reliable prognostic and predictive biomarkers are urgently needed to individualise treatment with newer agents, their combination with ADT, and the optimum treatment sequences.Another consideration is that ADT as the standard of care may be moving earlier in the disease continuum, and the availability of newer agents means that optimum treatment sequences that include ADT need to be ascertained in CRPC and in less advanced prostate cancer.http://www.medscape.com/viewarticle/781457). Whereas, the 147 trial suggested that denosumab may delay the onset of skeletal-related events (which included bone metastases) in patients with CRPC, but the adverse event profile was not favourable [Finally, more trial data are urgently needed on the management of patients with nmCRPC; EAU definition of CRPC does not require the presence of metastases but it is possible that all patients with CRPC have metastatic disease and those classified as having nmCRPC have metastases that cannot be detected with current techniques. Irrespective of this, very few studies have included appreciable numbers of patients that would currently be considered nmCRPC. In practice, the lack of data means that maintaining ADT in these nmCRPC patients is the recommended treatment option—it is unclear if bone-targeted treatment is also useful in this setting. For example, in the Zometa European Study (ZEUS), zoledronic acid did not prevent bone metastases or improve OS in M0 patients .As all randomised prospective data for abiraterone use in CRPC include the continuation of backbone ADT, and based upon the findings of O’Donnell et al.  that sugAll phase III trial data of enzalutamide in CRPC include the continuation of backbone ADT. This combined with the potential adverse event of gynaecomastia with enzalutamide monotherapy suggest that ADT should be continued when initiating enzalutamide in men with CRPC (Grade B recommendation).When initiating chemotherapy in CRPC, data on the benefits of continuing ADT are conflicting. However, in the absence of any clear evidence of harm and because prospective phase III trial data maintain ADT, it is recommended that backbone ADT is maintained when chemotherapy is initiated in CRPC (Grade B recommendation).


Blocking or evading apoptosis is one of the hallmarks of cancer . Recent Chemotherapy remains the mainstay of treatment for most cancers. Paradoxically, the intrinsic or acquired resistance often displayed by cancer cells of different origin to chemotherapeutic agents that damage cells in disparate ways, coupled with drug-related toxicities, reduce drug efficacy and pose major barriers to effective treatment . TreatmeOur recent study published in Cancer Research  detailedWe now show that the tyrosine kinase BMX, acting in concert with PTPN21, phosphorylates BAK on Y108 to suppress BAK activation to favor survival of cells exposed to cytotoxic agents (5; Figure With increasing interest in targeting apoptotic proteins to re-activate apoptosis in cells with either intrinsic or acquired resistance to existing chemotherapeutic agents, targeting BMX represents a novel approach to directly modulate the level of apoptotic priming in the cell. The ability to make a cell more ‘primed to die’ has enormous clinical relevance, not only to increase the efficacy of current and emerging treatments, but also potentially to decrease side effects by lowering the dose of cytotoxic drugs patients require. Furthermore, BMX may also be an important biomarker for survival dependency in specific cancer types.


A receptor function. From the pioneering work of Hans Selye, who described the sedative properties of steroids, synthetic compounds resembling the progesterone metabolite AP have been developed. If some of them have been used as anesthetics, it seems difficult to propose them as a therapeutic option for pain since they display several adverse side effects such as sedation, amnesia and functional tolerance. An alternative strategy, chosen by few laboratories around the world, is aimed at stimulating the local production of 3-alpha reduced neurosteroids in order to limit these well-known side effects. This pharmacological approach has the advantage of targeting specific structures, fully equipped with the necessary biosynthetic enzymatic machinery, where neurosteroids already act as endogenous pain modulators. The various pharmacological trials which attempted to treat pain symptoms by stimulating the production of 3-alpha reduced neurosteroids are reviewed here, as well as novel neurotransmitter systems possibly regulating their endogenous production.A growing number of studies indicate that 3-alpha reduced neurosteroids are remarkable analgesics in various pain states. This is the case for allopregnanolone (AP), one of the most potent endogenous positive allosteric modulators of GABA A receptors (GABAARs), expressed either at extrasynaptic , but also by neural cells in the nervous system. This discovery led Baulieu and collaborators to propose the term “neurosteroids” for neuroactive steroids produced by neural cells independently of the endocrine steroidogenic glands  the recent attempts to use TSPO agonists for the production of steroid analgesics; and (ii) the identification of endogenous signaling pathways which may regulate neurosteroidogenesis and could be targeted in the near future for therapeutic purposes.ARs in the spinal cord of rats displaying mechanical or thermal pain symptoms . Interestingly, glycinergic synapses during proestrus are also converted into mixed glycine/GABAergic synapses. Together, this led to an increased inhibitory control of layer II neurons processing pain informations and in a reduction in the intensity of pain symptoms, as previously demonstrated  or directly into rodent brain structures. These observations also raised critical issues linked to the well-known sex-specific differences in pain responses  is prescribed in several countries as a non-benzodiazepine anxiolytic  to animals exhibiting generalized neuropathic pain symptoms after chemotherapy with vincristine (VCR) sulphate, oxaliplatin or paclitaxel  in the spinal cord of rats, exhibiting neuropathic and osteoarthritic pain symptoms (Miller et al., 3H] PK11195 binding in the spinal cord seems to occur in microglial cells. TSPO is strongly expressed in activated microglia (Benavides et al., Indeed, TSPO protein levels are upregulated in the ipsilateral spinal cord in rats displaying inflammatory pain symptoms after knee injection of complete Freund’s adjuvant (Hernstadt et al., AR binding sites (Costa and Guidotti, Many endogenous ligands of TSPO have been identified and are described in detail in a recent review (Rupprecht et al., Beside TSPO ligands, we showed that oxytocin, a neuropeptide released by hypothalamic neurons projecting onto spinal nociceptive neurons, is exerting its long-lasting analgesic effect by stimulating the production of AP (Juif et al., In conclusion, there are now several convergent experimental evidences demonstrating that the local production of AP-like steroids in the nociceptive system constitutes a diffuse analgesic system. The development of numerous TSPO ligands capable of stimulating this system is on the way, but little is known regarding other possible physiological stimulators. In line with this idea, neuropeptides could play a key role.Pierrick Poisbeau is currently collaborating with Biocodex laboratories to decipher etifoxine action.


Heat shock response and homeostatic plasticity are mechanisms that afford functional stability to cells in the face of stress. Each mechanism has been investigated independently, but the link between the two has not been extensively explored. We explore this link. The heat shock response enables cells to adapt to stresses such as high temperature, metabolic stress and reduced oxygen levels. This mechanism results from the production of heat shock proteins (HSPs) which maintain normal cellular functions by counteracting the misfolding of cellular proteins. Homeostatic plasticity enables neurons and their target cells to maintain their activity levels around their respective set points in the face of stress or disturbances. This mechanism results from the recruitment of adaptations at synaptic inputs, or at voltage-gated ion channels. In this perspective, we argue that heat shock triggers homeostatic plasticity through the production of HSPs. We also suggest that homeostatic plasticity is a form of neuroprotection. The nervous system is particularly vulnerable to heat damage  that control neuronal excitability, or both , synaptic strengths were attenuated above room temperature (22°C)  which heat shock protects to afford synaptic homeostasis? At the Drosophila larval NMJs, intracellular calcium-handling mechanisms that contribute toward sustaining neurotransmitter release at elevated temperatures were protected  (Barclay and Robertson, + currents increased action potential duration to prevent action potential failure at elevated temperatures (Wu et al., + channels at elevated temperatures (Money et al., + and K+ channels are altered to enable action potential firing at elevated temperatures.In motor neurons that control locust flight muscles, heat shock increased the upper temperature limit at which action potentials could fire, preventing action potential failure at elevated temperatures (Wu et al., v2.3 calcium channel (Krieger et al., v7.4 voltage-gated potassium channels are found on the outer hair cell membranes of the inner ear and are involved in the conduction of sound (Gao et al., v7.4 channels on HEK cells (Gao et al., HSP interactions with voltage-gated ion channels that aid action potential firing are poorly understood. In previous work, HSPs were found to interact with VGCCs (Krieger et al., + that depolarizes the membrane, preventing the generation of action potentials. Heat shock prevented this build up by causing the clearance of extracellular K+ through insertion of more Na+/K+-ATPase pumps into the cell membrane. These pumps are designed to move K+ into the cell and this movement probably contributed to the hyperpolarization (Money et al., Drosophila larval motor neurons, changes in Na+/K+-ATPase pump function can lead to long-lasting changes in membrane excitability (Pulver and Griffith, +, or Ca2+ or another non-selective cation channel is thought to produce these afterdepolarizations. Alternatively, a reduction in K+ conductance could be another means for generating afterdepolarizations (Money et al., Studies have shown that there are additional benefits afforded by heat shock in the reliable generation of action potentials at elevated temperatures (Money et al., +/K+-ATPase pumps to the cytoskeletal network located just beneath the neuronal membrane (Riordan et al., + at elevated temperatures could be reduced to promote membrane hyperpolarization and action potential firing. In another example, hyperthermia caused the production of HSP27 and HSP32 in perisynaptic glial cells which surround synapses. These HSPs were then transferred into the synapses to potentially afford them protection (Bechtold and Brown, Early work from the Tytell laboratory conducted on the squid giant axon showed that heat shock caused HSP70 to be rapidly synthesized and transferred from the surrounding glia into the axon (Tytell et al., Drosophila and locusts suggest that heat shock affords functional stability to neural circuits under stress by recruiting homeostatic plasticity. Motor patterns are rhythmic bursts of electrical activity that are generated by motor circuits to drive movements (Barclay et al., Drosophila and locusts (Robertson et al., Drosophila, previous work has shown that, by overexpressing HSP70 in motor neurons, synaptic performance is improved at elevated temperatures, protecting larval locomotion (Xiao et al., It is thought that if homeostatic plasticity preserves the normal activity of individual neural circuit elements, the circuit itself should be afforded functional stability (Turrigiano, Heat shock response is a mechanism that preserves cellular functions under stress through the upregulation of HSPs. Homeostatic plasticity is a mechanism that preserves the activities of neurons and their target cells around their respective set points in the face of stress. Therefore we propose that the heat shock response may afford homeostatic plasticity. In this review, we have sought to provide examples from literature suggesting that the heat shock response affords homeostatic plasticity. Currently, investigations of the link between the two mechanisms are scant. By providing such examples, we hope to stimulate the interest of readers to further explore the interface between these two mechanisms.Certain disorders, such as seizures, sleep disturbances and cognitive disorders are thought to result, in part, from disruptions of homeostatic plasticity (Burke and Barnes, The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.