A Versatile Two-Step CRISPR- and RMCE-Based Strategy for Efficient Genome Engineering in Drosophila

The development of clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated (Cas) technologies promises a quantum leap in genome engineering of model organisms. However, CRISPR-mediated gene targeting reports in Drosophila melanogaster are still restricted to a few genes, use variable experimental conditions, and vary in efficiency, questioning the universal applicability of the method. Here, we developed an efficient two-step strategy to flexibly engineer the fly genome by combining CRISPR with recombinase-mediated cassette exchange (RMCE). In the first step, two sgRNAs, whose activity had been tested in cell culture, were co-injected together with a donor plasmid into transgenic Act5C-Cas9, Ligase4 mutant embryos and the homologous integration events were identified by eye fluorescence. In the second step, the eye marker was replaced with DNA sequences of choice using RMCE enabling flexible gene modification. We applied this strategy to engineer four different locations in the genome, including a gene on the fourth chromosome, at comparably high efficiencies. Our data suggest that any fly laboratory can engineer their favorite gene for a broad range of applications within approximately 3 months

Enhancing a sustainable healthy working life:design of a clustered randomized controlled trial

Background: To improve a sustainable healthy working life, we have developed the intervention 'Staying healthy at work', which endeavours to enhance work participation of employees aged 45 years and older by increasing their problem-solving capacity and stimulating their awareness of their role and responsibility towards a healthy working life. This research study aims to evaluate the process and the effectiveness of the intervention compared with care as usual.Methods/design: The study is a cluster-randomized controlled trial design (randomized at the supervisor level), with a 1-year follow-up. Workers aged 45 years and older have been enrolled in the study. Workers in the intervention group are receiving the intervention 'Staying healthy at work'. The main focus of the intervention is to promote a healthy working life of ageing workers by: (1) changing workers awareness and behaviour, by emphasizing their own decisive role in attaining goals; (2) improving the supervisors' ability to support workers in taking the necessary action, by means of enhancing knowledge and competence; and (3) enhancing the use of the human resource professionals and the occupational health tools available within the organization. The supervisors in the intervention group have been trained how to present themselves as a source of support for the worker. Workers in the control group are receiving care as usual; supervisors in the control group have not participated in the training. Measurements have been taken at baseline and will be followed up at 3, 6 and 12 months. The primary outcome measures are vitality, work ability and productivity. The secondary outcomes measures include fatigue, job strain, work attitude, self-efficacy and work engagement. A process evaluation will be conducted at both the supervisor and the worker levels, and satisfaction with the content of the intervention will be assessed.Discussion: The intervention 'Staying healthy at work' has the potential to provide evidence-based knowledge of an innovative method to promote a sustainable healthy working life in the older working population. The results of the study will be relevant for workers, employers, occupational health professionals and human resource professionals.</p

Wonen in Houwerzijl:vind je heil in Houwerzijl!

Deze managementsamenvatting omschrijft kort en bondig het onderzoek naar de woonomgeving van Houwerzijl. Deze samenvatting is bedoeld om de hoofdlijnen van het onderzoek weer te geven, voor het totale onderzoek wordt u doorverwezen naar het onderzoeksrapport ‘Wonen in Houwerzijl’. De doelstelling is het advies geven voor mogelijke aanpassingen ten aanzien van de leefomgeving. De probleemstelling en tevens de onderzoeksvraag van dit onderzoek luidt: ‘Wat vinden de inwoners van Houwerzijl van de kwaliteit van hun leefomgeving en hoe zien zij dit in 2020?’ Deze onderzoeksvraag is verder verdeeld in twee deelvragen, die de woonmotieven en de kwaliteit van de woningen onderzoeken. Studentonderzoek in het kader van het thema Werklandschappen

No association between preoperative physical activity level and time to return to work in patients after total hip or knee arthroplasty:A prospective cohort study

PURPOSE: It is important for patients of working age to resume work after total hip or knee arthroplasty (THA/TKA). A higher preoperative level of physical activity is presumed to lead to a better or faster recovery. Aim is to examine the association between preoperative physical activity (PA) level (total and leisure-time) and time to return-to-work (RTW). METHODS: A prospective multicenter survey study. Time to RTW was defined as the length of time (days) from surgery to RTW. PA level was assessed with the SQUASH questionnaire. Questionnaires were filled in before surgery and 6 weeks and 3, 6 and 12 months post-surgery. Multiple regression analyses were conducted separately for THA and TKA patients. RESULTS: 243 patients were enrolled. Median age was 56 years; 58% had undergone a THA. Median time to RTW was 85 (THA) and 93 (TKA) days. In the multiple regression analysis, neither preoperative total PA level nor leisure-time PA level were significantly associated with time to RTW. CONCLUSIONS: Preoperative physical activity level is not associated with a shorter time to RTW in either THA or TKA patients. Neither preoperative total PA level nor leisure-time PA level showed an association with time to RTW, even after adjusting for covariates. TRIAL REGISTRY: Dutch Trial Register: NTR3497

Burrowing Behavior of a Deposit Feeding Bivalve Predicts Change in Intertidal Ecosystem State

Behavior has a predictive power that is often underutilized as a tool for signaling ecological change. The burrowing behavior of the deposit feeding bivalve Macoma balthica reflects a typical food-safety trade-off. The choice to live close to the sediment surface comes at a risk of predation and is a decision made when predation danger, food intake rates or future fitness prospects are low. In parts of the Dutch Wadden Sea, Macoma populations declined by 90% in the late 1990s, in parallel with large-scale mechanical cockle-dredging activities. During this decline, the burrowing depth of Macoma became shallow and was correlated with the population decline in the following year, indicating that it forecasted population change. Recently, there has been a series of large recruitment events in Macoma. According to the food-safety trade-off, we expected that Macoma should now live deeper, and have a higher body condition. Indeed, we observed that Macoma now lives deeper and that living depth in a given year forecasted population growth in the next year, especially in individuals larger than 14 mm. As living depth and body condition were strongly correlated in individuals larger than 14 mm, larger Macoma could be living deeper to protect their reproductive assets. Our results confirmed that burrowing depth signals impending population change and, together with body condition, can provide an early warning signal of ecological change. We suggest that population recovery is being driven by improved intertidal habitat quality in the Dutch Wadden Sea, rather than by the proposed climate-change related effects. This shift in ecosystem state is suggested to include the recovery of diatom habitat in the top layer of the sediment after cockle-dredging ended

Excitons in a Photosynthetic Light-Harvesting System: A Combined Molecular Dynamics/Quantum Chemistry and Polaron Model Study

The dynamics of pigment-pigment and pigment-protein interactions in light-harvesting complexes is studied with a novel approach which combines molecular dynamics (MD) simulations with quantum chemistry (QC) calculations. The MD simulations of an LH-II complex, solvated and embedded in a lipid bilayer at physiological conditions (with total system size of 87,055 atoms) revealed a pathway of a water molecule into the B800 binding site, as well as increased dimerization within the B850 BChl ring, as compared to the dimerization found for the crystal structure. The fluctuations of pigment (B850 BChl) excitation energies, as a function of time, were determined via ab initio QC calculations based on the geometries that emerged from the MD simulations. From the results of these calculations we constructed a time-dependent Hamiltonian of the B850 exciton system from which we determined the linear absorption spectrum. Finally, a polaron model is introduced to describe quantum mechanically both the excitonic and vibrational (phonon) degrees of freedom. The exciton-phonon coupling that enters into the polaron model, and the corresponding phonon spectral function are derived from the MD/QC simulations. It is demonstrated that, in the framework of the polaron model, the absorption spectrum of the B850 excitons can be calculated from the autocorrelation function of the excitation energies of individual BChls, which is readily available from the combined MD/QC simulations. The obtained result is in good agreement with the experimentally measured absorption spectrum.Comment: REVTeX3.1, 23 pages, 13 (EPS) figures included. A high quality PDF file of the paper is available at http://www.ks.uiuc.edu/Publications/Papers/PDF/DAMJ2001/DAMJ2001.pd

Critical Dimensions in Architectural Photography: Contributions to Architectural Knowledge

This paper illustrates and explores three critical dimensions of photography in architecture, each of which informs the production of images, texts, and other artifacts which establish what might be called a building’s media footprint. The paper’s broad goal is to question the extent to which these critical dimensions are relevant to architectural decision-making processes. Acknowledging that such dimensions as the ones examined here rarely predict an architect’s specific design decisions in a transparent manner, the paper discusses not only the decisions made by architects during the process of designing buildings, but the decisions made by critics, visitors, and members of the general public as they engage in activities such as visiting buildings, writing about them and, particularly, photographing them. First, the text discusses the potential of buildings to operate as mechanisms for producing images, in the sense originated by Beatriz Colomina. The question is developed through the analysis of the space of photography – mapping of points of view, directions of view, and fields of view of defined photographic collections. Secondly, it considers photography’s complicity in the canonization of buildings, and specifically, the extent to which photography is responsible for distinguishing between major and minor architectural works. Finally, the essay examines the erosion over time of photography’s historical power to frame when confronted with contemporary technologies of virtual reality and photo realistically rendered digital models. Each of these critical dimensions, or concepts, develops a specific aspect of how photographic information about buildings is organized, structured, and disseminated, and is thus only part of the larger project of architectural epistemology, which inquires into this wider field. This will be done through an examination of the Mies van der Rohe-designed Commons Building at ITT in Chicago and the evolution of its relationship with architectural photography and photographic representation – both on its own terms and through the prism of the Rem Koolhaas-designed McCormick Tribune Student Center, which adds to and incorporates the Commons Building. Until the end of the twentieth century, the Commons Building on the campus of the Illinois Institute of Technology was generally considered one of Mies van der Rohe’s lesser works. Reportedly neglected by its own architect during the design process, and frequently marginalized in academic discussions of the campus, when mentioned at all the building was often cited as an unrefined prototype of Crown Hall. This discourse took a new direction when in 1998, Rem Koolhaas/OMA won a design competition for a student center on the IIT campus: uniquely among the competition entries, Koolhaas’s design incorporated the Commons Building within a new context – what ultimately became the McCormick Tribune Campus Center (MTCC). When critics concluded that the incorporation of the Commons Building into the larger whole could compromise its integrity as an exemplar of Mies’s work, the building became the object of renewed interest and controversy. The two projects considered here show a clear evolution in architecture’s relationship with the photographic image. Specifically, the history of the Commons Building can be traced through photographs: during and shortly following its construction, the building was photographed as part of Mies’s own attention to publicity; it was documented as part of historical analyses; and over time it was visited and photographed by casual and amateur photographers. Following the competition results, photographs of the Commons Building were strategically deployed by both proponents and critics of Koolhaas’s design. Contemporary photographs of the building appear in architectural and campus guidebooks and on websites such as Flickr.com. Examining the ways in which photographs of the Commons Building appear in these various contexts allows discussion of the critical dimensions identified above and permits us to trace the evolution of the mutually reinforcing relationship between architecture and photography

Behavioural stress responses predict environmental perception in European sea bass (Dicentrarchus labrax)

Individual variation in the response to environmental challenges depends partly on innate reaction norms, partly on experience-based cognitive/emotional evaluations that individuals make of the situation. The goal of this study was to investigate whether pre-existing differences in behaviour predict the outcome of such assessment of environmental cues, using a conditioned place preference/avoidance (CPP/CPA) paradigm. A comparative vertebrate model (European sea bass, Dicentrarchus labrax) was used, and ninety juvenile individuals were initially screened for behavioural reactivity using a net restraining test. Thereafter each individual was tested in a choice tank using net chasing as aversive stimulus or exposure to familiar conspecifics as appetitive stimulus in the preferred or non preferred side respectively (called hereafter stimulation side). Locomotor behaviour (i.e. time spent, distance travelled and swimming speed in each tank side) of each individual was recorded and analysed with video software. The results showed that fish which were previously exposed to appetitive stimulus increased significantly the time spent on the stimulation side, while aversive stimulus led to a strong decrease in time spent on the stimulation side. Moreover, this study showed clearly that proactive fish were characterised by a stronger preference for the social stimulus and when placed in a putative aversive environment showed a lower physiological stress responses than reactive fish. In conclusion, this study showed for the first time in sea bass, that the CPP/CPA paradigm can be used to assess the valence (positive vs. negative) that fish attribute to different stimuli and that individual behavioural traits is predictive of how stimuli are perceived and thus of the magnitude of preference or avoidance behaviour.European Commission [265957]; Portuguese Fundacao para a Ciencia e Tecnologia (FCT) [FRH/BPD/72952/2010]; FCT [SFRH/BD/80029/2011