Search

This study examined co-teaching -two teachers collaboratively leading one course -in undergraduate applied psychology courses. Previous research has established potential benefits of co-teaching on student success; however, none have examined students’ perspectives on co-teaching in applied psychology courses. The present study hypothesized that students would report more benefits than drawbacks to co-teaching. For this purpose, 50 college students aged 18 to 25 completed an online questionnaire after completing one of two co-taught applied psychology courses in the United States. Results revealed that students perceived more benefits to co-teaching than drawbacks, and this difference was statistically significant. Additional findings suggest that students believe co-teaching to be beneficial, and most report a preference for it. Consensual qualitative coding further revealed that ‘diversity of instructor perspectives’ was the most commonly reported benefit, as related to course content, and the most commonly reported drawback, as related to course management. Study limitations and implications for future research and teaching were also considered.

Early intervention (EI) services are provided for families and children at risk for or with developmental delays. Early intervention includes services that are provided in the natural environment as mandated by the Individuals with Disabilities Education Act (IDEA; 2004). The natural environment is

Using adenine base editors, we aim to treat sickle cell disease by generating single nucleotide polymorphisms in human CD34+ hematopoietic stem and progenitor cells (HSPCs) at specific target sites by mediating A-T to G-C base conversions. While ex vivo gene editing approaches show great therapeutic promise, access is limited due to the requirement of an autologous hematopoietic stem cell (HSC) transplant to deliver the ex vivo edited cells. To further increase the number of patients eligible for base editing therapy, we are developing an alternative approach to directly deliver base editors to HSCs in vivo through non-viral delivery methods. Lipid Nanoparticles (LNPs) are a clinically validated, non-viral approach that enables the delivery of nucleic acid payloads, which may avoid the challenges associated with ex vivo approaches including the transplantation of edited CD34+ HSPCs. Here we describe the development and characterization of LNPs for the delivery of messenger RNA (mRNA) to HSPCs in vivo in both mice and cynomolgus macaques. By screening >1,000 chemically distinct LNPs in vivo utilizing a DNA barcoding technology, we identified several hit LNPs capable of biodistribution to HSPCs. Upon individual validation of these hit LNPs by delivery of Cre recombinase mRNA in a Cre-reporter mouse model (Ai14), which expresses the fluorescent protein tdTomato under a constitutive CAG promoter following Cre-meditate gene editing, we confirmed that several LNPs efficiently delivered Cre recombinase mRNA to mouse Lin-Sca-1+c-Kit+ (LSK) HSPCs. We next confirmed the most potent hit LNP (LNP-HSC1) identified from the in vivo screen to transfect LSK HSPCs in a dose-dependent manner between 0.1 and 1.0 mg/kg Cre recombinase mRNA, transfecting over 40% of LSK HSPCs in Ai14 mice at 1.0mg/kg. In a transfection durability study using Ai14 mice, we observed maintenance of tdTomato+ LSK HSPCs levels in the bone marrow at 10 weeks post-LNP delivery. As LNP-HSC1 had been identified and validated in mice of a C57BL6/j background, we next confirmed its ability to transfect a reporter mRNA into HSPCs in Balb/c mice and in 5 cynomolgus macaques. LNP-HSC1 efficiently transfected LSK HSPCs in Balb/c mice at doses ranging from 0.3 to 1.0 mg/kg. In 5 cynomolgus macaques (n=5 across two experiments), we observed a dose-dependent increase in reporter mRNA delivery with an average of 19% of bone marrow-derived CD34+ HSPCs (n=3) expressing the reporter protein at the highest dose tested. Taken together, these data demonstrate the value of our in vivo high-throughput LNP screening approach to identify novel LNPs capable of delivering to HSPCs, providing a promising delivery platform for an in vivo HSC gene editing approach for the treatment of hemoglobinopathies. Disclosures Sago: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Campbell: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Lutz: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Patwardhan: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Hamilton: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Wong: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Lee: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Keating: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Murray: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Singh: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company. Ciaramella: Beam Therapeutics: Current Employment, Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company, Membership on an entity's Board of Directors or advisory committees.

Using available cryo-EM and x-ray crystal structures of the nonstructural proteins that are responsible for SARS-CoV-2 viral RNA replication and transcription, we have constructed an atomistic model of how the proteins assemble into a functioning superstructure. Our principal finding is that the complex is hexameric, centered around nsp15. The nsp15 hexamer is capped on two faces by trimers of nsp14/nsp16/(nsp10)2, where nsp14 is seen to undergo a large conformational change between its two domains. This conformational change facilitates binding of six nsp12/nsp7/(nsp8)2 polymerase subunits to the complex. To this, six subunits of nsp13 are arranged around the superstructure, but not evenly distributed. Two of the six polymerase subunits are each proposed to carry dimers of nsp13, while two others are proposed to carry monomers. The polymerase subunits that coordinate nsp13 dimers also bind the nucleocapsid, which positions the 5’-UTR TRS-L RNA over the polymerase active site, a state distinguishing transcription from replication. Analyzing the path of the viral RNA indicates the dsRNA that exits the polymerase passes over the nsp14 exonuclease and nsp15 endonuclease sites before being unwound by a convergence of zinc fingers from nsp10 and nsp14. The template strand is then directed away from the complex, while the nascent strand is directed to the sites responsible for mRNA capping (the nsp12 NiRAN and the nsp14 and nsp16 methyltransferases). The model presents a cohesive picture of the multiple functions of the coronavirus replication-transcription complex and addresses fundamental questions related to proofreading, template switching, mRNA capping and the role of the endonuclease. It provides a platform to guide biochemical and structural research to address the stoichiometric and spatial configuration of the replication-transcription complex.Author SummaryThe replication of the coronavirus genome and the synthesis of subgenomic mRNA is a complex process involving multiple viral proteins. Despite a fairly complete structural picture of the individual proteins that are believed to coalesce into a larger replication-transcription complex, there is no clear model of how these proteins interact. Here we present the first detailed atomistic model of a complete replication-transcription complex for SARS-CoV-2, made up of the non-structural proteins nsp7-nsp16, as well as the nucleocapsid. Forming a large, hexameric superstructure centered around nsp15, the model provides new perspective on the function of its individual components, including the exonuclease, the endonuclease, the NiRAN site, the helicase, the multiple zinc fingers, and the nucleocapsid. It offers a cohesive view of replication, proofreading, template switching and mRNA capping, which should serve as a guide for future experimental exploration.

Research has found that perceived health locus of control is related to increased health-enhancing behaviors. However, studies have not examined if perceived health locus of control similarly impacts psychological health and have largely ignored women’s health. These studies have also overlooked contexts where individuals have little control over their behaviors. Therefore, in the current study, we examine how perceptions of health locus of control relate to psychological health in a sample of women with clinical suspicion of having gynecological cancer. Women (n = 301), ranging from 40 to 80 years-old and undergoing inpatient gynecological surgery at Michigan Medicine University of Michigan, USA, were included in our study. Data, including psychological measures of control and health perceptions, were collected preoperatively. Our findings indicate that increased perceptions of health locus of control were related to a significant increase in preoperative psychological health through a reduction in negative affect, controlling for the invasiveness of the planned procedure (more invasive Laparotomy or Laparoscopy) and the subsequent formal postoperative diagnosis (benign or malignant). These findings have implications for enhancing women’s preoperative health. We provide insight for promoting positive interactions between healthcare providers and patients, and we discuss postoperative implications.

For Christian college students, the journey toward vocation can be hindered by faith-based anxiety. The relationship between students and mentors in Christian higher education offers a unique, optimum space to explore these beliefs. Christians in academia have invaluable personal experience in integrating their faith and career, and have much to teach the next generation in courageously engaging both mind and heart when making vocational choices. Within the professor–student relationship, intentional curiosity and conversation are tools for inquiry and resolution. We have included four common anxieties, with Scripture-based affirmations and challenges to those viewpoints, to model investigating these beliefs with students.

Most studies examining the association of the food environment and health outcomes have focused on urban settings. However, rural adults and children have high rates of obesity, which may be related to their food environments. This study examines the food environment in 7 rural counties in Maryland. A cross-sectional study was conducted at 244 retail food stores from 2012 to 2013. Data on store-level characteristics were collected and a healthy food availability index (HFAI) ranging from 0 to 31 points was calculated for each store. Convenience stores (18.9%) and other nontraditional stores (16.8%) were the most common store types, and box stores were the least (4.9%). Supermarkets had the highest mean HFAI (24.8), whereas gas stations (8.7) had the lowest; convenience stores were also low (11.1). In rural Maryland, the most common food source types have low healthy food availability, and nontraditional food sources are an important part of the food environment.

Summary This chapter primarily addresses nutrition-related labeling policies in the United States of America (USA) and provides a limited overview of international policies. The policies covered in this chapter include mandatory and optional nutrition label information in specified formats and optional nutrition-related label claims. Nutrition labeling is required in the USA for most food products, and most of the policy detail provided in the chapter is for conventional foods in the USA. Dietary supplements are discussed in Volume I, Chapter 35. Since in the USA they are a regulatory subset of foods, this chapter also briefly addresses US nutrition labeling requirements for dietary supplements.

Recent advances in single particle cryogenic electron microscopy (cryo-EM) have enabled the structural determination of numerous protein assemblies at high resolution, yielding unprecedented insights into their function. However, despite its extraordinary capabilities, cryo-EM remains time-consuming and resource-intensive. It is therefore beneficial to have a means for rapidly assessing and optimizing the quality of samples prior to lengthy cryo-EM analyses. To do this, we have developed a native mass spectrometry (nMS) platform that provides rapid feedback on sample quality and highly streamlined biochemical screening. Because nMS enables accurate mass analysis of protein complexes, it is well-suited for routine evaluation of the composition, integrity, and homogeneity of samples prior to their plunge-freezing on EM grids. We demonstrate the utility of our nMS-based platform for facilitating cryo-EM studies using structural characterizations of exemplar bacterial transcription complexes as well as the replication-transcription assembly from the SARS-CoV-2 virus that is responsible for the COVID-19 pandemic. Funding: This work is supported by the Pels Foundation to The Rockefeller University, NIH grants P41 GM109824 and P41 GM103314 to BTC, R35 GM118130 to SAD, and R01 GM114450 to EAC. Access to the cryo-EM microscopes and support was through The Rockefeller University Evelyn Gruss Lipper Cryo-EM Resource Center and at The Simons Electron Microscopy Center (SEMC), National Resource for Automated Molecular Microscopy (NRAMM), and National Center for CryoEM Access and Training (NCCAT) at the NYSBC, supported by NIH NIGMS (P41 GM103310), NYSTAR, the Simons Foundation (SF349247), the NIH Common Fund Transformative High Resolution CryoElectron Microscopy program (U24 GM129539) and NY State Assembly Majority. Conflict of Interest: The authors declare there are no competing interests.

TraR and its homolog DksA are bacterial proteins that regulate transcription initiation by binding directly to RNA polymerase (RNAP) rather than to promoter DNA. Effects of TraR mimic the combined effects of DksA and its cofactor ppGpp, but the structural basis for regulation by these factors remains unclear. Here, we use cryo-electron microscopy to determine structures of Escherichia coli RNAP, with or without TraR, and of an RNAP-promoter complex. TraR binding induced RNAP conformational changes not seen in previous crystallographic analyses, and a quantitative analysis revealed TraR-induced changes in RNAP conformational heterogeneity. These changes involve mobile regions of RNAP affecting promoter DNA interactions, including the βlobe, the clamp, the bridge helix, and several lineage-specific insertions. Using mutational approaches, we show that these structural changes, as well as effects on σ70 region 1.1, are critical for transcription activation or inhibition, depending on the kinetic features of regulated promoters., eLife digest Cells need to make proteins in order to survive. To make a protein, a cell first needs to take the information from a gene coded in its DNA and copy it into a template made of a different molecule called RNA via a process called transcription. Not all genes are transcribed or expressed at the same time or the same rate, because a cell needs different proteins depending on its environment. The molecules that regulate when each gene is transcribed are called transcription factors. When the bacterium Escherichia coli is starved for the building blocks it needs to make proteins, it changes the expression of almost one quarter of its genes within 5 minutes. This broad response requires two transcription factors, ppGpp and DksA. Unlike most transcription factors, these two molecules bind directly to the enzyme responsible for transcribing DNA into RNA, the RNA polymerase, but they do not bind to DNA. Another transcription factor called TraR can mimic the combined effects of ppGpp and DksA on transcription, changing the conformation of RNA polymerase in the same way. Now, Chen et al. have used a high-resolution imaging technique called cryo-electron microscopy to reveal the details of how the structure of RNA polymerase changes in response to TraR binding, and by analogy, in response to ppGpp and DksA. The experiments showed that TraR interacts with regions of RNA polymerase that move when the protein binds DNA. These interactions can either help or hinder the start of transcription depending on which DNA sequence the RNA polymerase binds. Using the structures obtained via cryo-electron microscopy as guides, Chen et al. mutated each of the mobile parts of RNA polymerase individually to determine which interactions with TraR were necessary to change gene expression. These results shed light on a fundamental process in all living cells, the initiation of transcription, and how it changes in response to the cell's nutritional environment. This may help explain how different cells regulate gene expression and may also lead to the development of new antibiotics.

TraR and its homolog DksA are bacterial proteins that regulate transcription initiation by binding directly to RNA polymerase (RNAP) rather than to promoter DNA. Effects of TraR mimic the combined effects of DksA and its cofactor ppGpp. How TraR and its homologs regulate transcription is unclear. Here, we use cryo-electron microscopy to determine structures ofEscherichia coliRNAP, with or without TraR, and of an RNAP-promoter complex. TraR binding induced RNAP conformational changes not seen in previous crystallographic analyses, and a quantitative analysis of RNAP conformational heterogeneity revealed TraR-induced changes in RNAP dynamics. These changes involve mobile regions of RNAP affecting promoter DNA interactions, including the βlobe, the clamp, the bridge helix, and several lineage-specific insertions. Using mutational approaches, we show that these structural changes, as well as effects on σ70region 1.1, are critical for transcription activation or inhibition, depending on the kinetic features of regulated promoters.

In bacteria, a primary σ-factor associates with the core RNA polymerase (RNAP) to control most transcription initiation, while alternative σ-factors are used to coordinate expression of additional regulons in response to environmental conditions. Many alternative σ-factors are negatively regulated by anti–σ-factors. In Escherichia coli, Salmonella enterica, and many other γ-proteobacteria, the transcription factor Crl positively regulates the alternative σ(S)-regulon by promoting the association of σ(S) with RNAP without interacting with promoter DNA. The molecular mechanism for Crl activity is unknown. Here, we determined a single-particle cryo-electron microscopy structure of Crl-σ(S)-RNAP in an open promoter complex with a σ(S)-regulon promoter. In addition to previously predicted interactions between Crl and domain 2 of σ(S) (σ(S)(2)), the structure, along with p-benzoylphenylalanine cross-linking, reveals that Crl interacts with a structural element of the RNAP β′-subunit that we call the β′-clamp-toe (β′CT). Deletion of the β′CT decreases activation by Crl without affecting basal transcription, highlighting the functional importance of the Crl-β′CT interaction. We conclude that Crl activates σ(S)-dependent transcription in part through stabilizing σ(S)-RNAP by tethering σ(S)(2) and the β′CT. We propose that Crl, and other transcription activators that may use similar mechanisms, be designated σ-activators.

Bead reagents are used in a large number of assays in bioscience and biotechnology to collect and purify antibodies by immobilization. Bead-based immunoassays offer high-throughput analysis of multiple antibodies in a single sample. Although a variety of antibody-binding moieties on the collection beads have been studied, the physical and material properties of collection beads have not been optimized to isolate specific antibodies over a broad range of concentrations from complex environments containing cells. We present a study of how to optimally use microparticles coated with protein G to collect low concentrations of IgG antibodies from complex solutions. We study the impact of bead material, bead size, incubation time, and protein G density to more efficiently collect antibodies and detect specific antibodies via fluorescent antigen labeling. The minimum detectable limit and the minimum incubation time for antibody collection are used as metrics to evaluate the collection parameters. We found that larger silica beads can capture more antibodies from a low concentration of sample, with a minimum incubation time of 60 min to equilibrium binding, resulting in a minimum detectable concentration of antibodies of 26 nM. We show that simple biophysical optimization of antibody collection reagents can be used to improve the collection of low concentrations of antibodies in complex environments. We demonstrate that the technology may be useful for monitoring antibody secretions from hybridoma cultures.

In bacteria, a primary σ factor associates with the core RNA polymerase (RNAP) to control most transcription initiation, while alternative σ factors are used to coordinate expression of additional regulons in response to environmental conditions. Many alternative σ factors are negatively regulated by anti-σ factors. In Escherichia coli, Salmonella enterica, and many other γ-proteobacteria, the transcription factor Crl positively regulates the alternative σS regulon by promoting the association of σS with RNAP without interacting with promoter DNA. The molecular mechanism for Crl activity is unknown. Here, we determined a single-particle cryo-electron microscopy structure of Crl-σS-RNAP in an open promoter complex with a σS regulon promoter. In addition to previously predicted interactions between Crl and domain 2 of σS (σS), the structure, along with p-benzoylphenylalanine crosslinking, reveals that Crl interacts with a structural element of the RNAP β’ subunit we call the β’-clamp-toe (β’CT). Deletion of the β’CT decreases activation by Crl without affecting basal transcription, highlighting the functional importance of the Crl-β’CT interaction. We conclude that Crl activates σS-dependent transcription in part through stabilizing σS-RNAP by tethering σS and the β’CT. We propose that Crl, and other transcription activators that may use similar mechanisms, be designated σ-activators.Significance StatementIn bacteria, multiple σ factors can bind to a common core RNA polymerase (RNAP) to alter global transcriptional programs in response to environmental stresses. Many γ-proteobacteria, including the pathogens Yersinia pestis, Vibrio cholera, Escherichia coli, and Salmonella typhimurium, encode Crl, a transcription factor that activates σS-dependent genes. Many of these genes are involved in processes important for infection, such as biofilm formation. We determined a high-resolution cryo-electron microscopy structure of a Crl-σS-RNAP transcription initiation complex. The structure, combined with biochemical experiments, shows that Crl stabilizes σS-RNAP by tethering σS directly to the RNAP.

Background The transcription factor hypoxia inducible factor (HIF) -1 drives tumor growth and metastasis and is associated with poor prognosis in breast cancer. Ascorbate can moderate HIF-1 activity in vitro and is associated with HIF pathway activation in a number of cancer types, but whether tissue ascorbate levels influence the HIF pathway in breast cancer is unknown. In this study we investigated the association between tumor ascorbate levels and HIF-1 activation and patient survival in human breast cancer. Methods In a retrospective analysis of human breast cancer tissue, we analysed primary tumor and adjacent uninvolved tissue from 52 women with invasive ductal carcinoma. We measured HIF-1α, HIF-1 gene targets CAIX, BNIP-3 and VEGF, and ascorbate content. Patient clinical outcomes were evaluated against these parameters. Results HIF-1 pathway proteins were upregulated in tumor tissue and increased HIF-1 activation was associated with higher tumor grade and stage, with increased vascular invasion and necrosis, and with decreased disease-free and disease-specific survival. Grade 1 tumors had higher ascorbate levels than did grade 2 or 3 tumors. Higher ascorbate levels were associated with less tumor necrosis, with lower HIF-1 pathway activity and with increased disease-free and disease-specific survival. Conclusions Our findings indicate that there is a direct correlation between intracellular ascorbate levels, activation of the HIF-1 pathway and patient survival in breast cancer. This is consistent with the known capacity of ascorbate to stimulate the activity of the regulatory HIF hydroxylases and suggests that optimisation of tumor ascorbate could have clinical benefit via modulation of the hypoxic response.

Tasks are formative to what every working individual does, yet there is increasing ambiguity amid the disjointed state of the task literature which demands a much-needed state of clarification and ...

LifeRAFT, a helping skills training model for undergraduate paraprofessionals, addresses training needs for applied psychology skills for undergraduate psychology majors. LifeRAFT draws from three empirically supported psychotherapy treatmentsto introduce counselling theory and encourage helping skill progression. Trainees learn practical helping strategies appropriate for paraprofessional use that are drawn from cognitive, behavioural, and emotion–focused therapies and are taught to work interactively with thoughts, actions, and feelings to promote client exploration and transformation. In a study of LifeRAFT’s training effectiveness, 18 undergraduate students received LifeRAFT training. Training effectiveness was explored pre– and post–training using client, counsellor, and observer rating forms. Resultssuggest that LifeRAFT is an effective method for training undergraduates in helping skills

Forest insect outbreaks are influenced by ecological processes operating at multiple spatial scales, including host-insect interactions within stands and across landscapes that are modified by regional-scale variations in climate. These drivers of outbreak dynamics are not well understood for the western spruce budworm, a defoliator that is native to forests of western North America. Our aim was to assess how processes across multiple spatial scales drive western spruce budworm outbreak dynamics. Our objective was to assess the relative importance and influence of a set of factors covering the stand, landscape, and regional scales for explaining spatiotemporal outbreak patterns in British Columbia, Canada. We used generalized linear mixed effect models within a multi-model interference framework to relate annual budworm infestation mapped from Landsat time series (1996–2012) to sets of stand-, landscape-, and regional-scale factors derived from forest inventory data, GIS analyses, and climate models. Outbreak patterns were explained well by our model (R 2 = 93%). The most important predictors of infestation probability were the proximity to infestations in the previous year, landscape-scale host abundance, and dry autumn conditions. While stand characteristics were overall less important predictors, we did find infestations were more likely amongst pure Douglas-fir stands with low site indices and high crown closure. Our findings add to growing empirical evidence that insect outbreak dynamics are driven by multi-scaled processes. Forest management planning to mitigate the impacts of budworm outbreaks should thus consider landscape- and regional-scale factors in addition to stand-scale factors.

Western spruce budworm is a defoliator of coniferous forests of North America. Past research has shown that climate is a principal driver of budworm outbreaks, though the underlying relationships are not well understood. We utilized Landsat time series to investigate the relationships between spatiotemporal patterns of budworm outbreaks and weather variability. Landsat-based maps of budworm infestations from (1995–2013) were produced and used to describe patterns of the most recent budworm outbreak in British Columbia. Superposed epoch and regression analysis were used to explore relationships between outbreak patterns and annual temperature and precipitation anomalies. We found that western spruce budworm outbreaks were preceded by autumn precipitation deficits that were 12% lower than the long-term average, and co-occurred with summer precipitation up to 20% below the long-term average. Spring temperatures shortly before outbreaks were 0.7°C lower than normal, whereas winter temperatures durin...

Background Despite the importance of a collaborative leadership approach for successful interprofessional practice, little is known about the leadership capabilities and attitudes of health professions students at the beginning of their educational program. Purpose Evaluate the association of self-perceived servant leadership (SL) and leadership self-efficacy (LSE) ability with degree program and demographics of first-semester health professions students. Method A survey measuring SL and LSE was administered to 1014 students from 17 academic programs enrolled in the first semester of an interprofessional health curriculum. Discussion 453 students (45%) responded to the survey. No individual academic program differences for either SL or LSE were identified on post-hoc testing. SL and LSE scores were correlated with additional previous leadership roles and previous leadership trainings. Lower socioeconomic status was associated with higher total SL score (p Conclusions This single-site, cross-sectional evaluation of leadership self-perception identified an exposure-dependent increase in SL and LSE scores with additional leadership experience (both leadership training and leadership roles) along with higher SL scores in students from low socioeconomic status.

Hiring managers make inferences from signals to determine if job candidates are capable and committed to performing the job to which they are applying to work. Building on how prior research has sp...

A key regulated step of transcription is promoter melting by RNA polymerase (RNAP) to form the open promoter complex1–3. To generate the open complex, the conserved catalytic core of the RNAP combines with initiation factors to locate promoter DNA, unwind 12–14 base pairs of the DNA duplex and load the template-strand DNA into the RNAP active site. Formation of the open complex is a multi-step process during which transient intermediates of unknown structure are formed4–6. Here we present cryo-electron microscopy structures of bacterial RNAP–promoter DNA complexes, including structures of partially melted intermediates. The structures show that late steps of promoter melting occur within the RNAP cleft, delineate key roles for fork-loop 2 and switch 2—universal structural features of RNAP—in restricting access of DNA to the RNAP active site, and explain why clamp opening is required to allow entry of single-stranded template DNA into the active site. The key roles of fork-loop 2 and switch 2 suggest a common mechanism for late steps in promoter DNA opening to enable gene expression across all domains of life. Cryo-electron microscopy structures of bacterial RNAP–promoter DNA complexes, including structures of partially melted intermediates, suggest a universally conserved common mechanism for promoter DNA opening prior to gene expression.

Wetlands are important globally for supplying clean water and unique habitat, and for storing vast amounts of carbon and nutrients. The geographic extent and state of wetlands vary over time and represent a dynamic land condition rather than a permanent land cover state. Herein, we combined a time series of land cover maps derived from Landsat data at 30-m resolution to inform on spatial and temporal changes to non-treed and treed wetland extents over Canada’s forested ecosystems (>650 million ha) from 1984 to 2016. Overall, for the period, 1984 to 2016, we found the extent of wetlands (non-treed and treed combined) in Canada’s forested ecosystems to be stable, with some regional variability, often resulting from offsetting decreases and increases within a given ecozone. Notwithstanding difficulties in using optical satellite data for mapping a land condition, by accumulating wetland evidence via earth observations consistently through multiple decades, our results capture the trends in wetland cover over a previously unmapped, national extent at a level of spatial detail and temporal reach suitable for further focused interpretations of wetlands and drivers and projections of wetland dynamics.

Atmospheric CO

A mountain pine beetle (Dendroctonus ponderosae Hopkins) outbreak has recently spread into boreal forests, with unknown consequences for this ecosystem. We intensively sampled 12 stands affected by the current outbreak in northern British Columbia to determine the potential of western boreal forests to recover from this novel disturbance. We sampled the species composition, size structure, and spatial distribution (using 5 m × 5 m subplots, 40 per stand) of live and dead trees and used a variety of analyses, including ordinations, to assess potential developmental trajectories of stands. Advance regeneration (stems < 10 m tall) varied greatly in abundance among stands (50–18 280 stems·ha−1). However, most subplots contained at least one individual; only three stands had many empty subplots. We conclude that most stands have enough advance regeneration and residual canopy trees to form a nearly continuous new canopy. Ordinations indicate that species composition will shift substantially and become more divergent among stands. Species of high economic value will remain common, though, and active management will not be necessary in most stands to maintain productive forests. However, this novel disturbance will have very different effects on these forests than the typical fire-disturbance regime and is likely to deflect these forests into new successional trajectories.

Background Despite the significance of the role of caregivers among children with Obstructive Adenotonsillar hypertrophy (OAT), their caregiving experience and emotional well-being are given poor attention in research, policy and service design. This study represents a novel effort sought to investigate the burden experienced by caregivers of children with OAT and the impact of emotional distress on their caregiving burden. Methodology A total of one hundred consecutive caregivers of children with OAT were interviewed with designed socio-demographic questionnaire. This was subsequently followed by administration of General Health Questionnaire-12 (GHQ-12) to ascertain emotional distress using cut-off score ≥3 and Zarit Caregivers Burden of care scale was used to characterize the pattern of burden experienced by these caregivers. Results In this study, the mean ages of children with OAT and their caregivers were 36.7(±21.8) months and 34.4(±5.4) years, respectively. The affected children were mainly males (68%), while their caregivers were predominantly females (84%). Fifty-seven percent of children with OAT were schooling but 13% of them had academic delay. Majority of the caregivers (66%) had a tertiary level of education. Of the total participants, up to 43% reported various degrees of burden of care, with majority (34%) of them reporting mild to moderate burden of care. In the same vein, 48% were emotionally distressed. Following regression analyses, emotional distress was independently associated with burden of care among participants (Odds ratio [OR] = 0.108; 95% Confidence Interval [95% CI] = 0.043–0.272; p Conclusion Caregivers of children with OAT reported the experience of various degrees of burden, and worse among caregivers with emotional distress. Overall, the level of distress and burden observed in this study were many-fold what has been reported among caregivers of other populations of children. The results of this study support the proposal of proactive measures to address the psychosocial needs of caregivers as integral to the care of children with OAT. Further research on the well-being of caregivers is also justified.

To investigate the profile of functional impairment and the influence of psychopathology with other determinants among individuals with schizophrenia in a Nigerian tertiary health facility. The study participants consisted of one hundred adults with schizophrenia. Eligible participants were recruited using systematic random sampling technique. All participants were interviewed with questionnaire to elicit socio-demographic and treatment related information. This was followed by interview with brief psychiatric rating scale (BPRS) and the global assessment of functioning (GAF) to assess for psychopathology and impairment in functioning respectively. Analyses of data were done using SPSS-16. The mean age of the participants was 33.87(±11.6) years and a little above half (52 %) of the participants were female. Sixty (60 %) subjects had active psychopathology using BPRS cut-off score of >10; while participants with duration of illness of more than 10 years (X2 = 5.59, p = 0.018) and previous relapse (X2 = 6.34, p = 0.012) were more likely to have psychopathology. Overall, a little over two-thirds (69 %) had impairment following administration of GAF. Mild impairment was observed in 23 % of the participants, moderate impairment in 19 and up to 18 % of participants had severe impairment due to psychopathology. Subjects with lesser education, duration of illness of more than 10 years, extrapyramidal side effects (EPSE), previous relapse and psychopathology had significant levels of impairment (p

This paper is a response to requests from Christian psychologists for subject-specific class activities that integrate Christianity and psychology. Although Christian/ psychology integration is highly valued by many as an overarching pedagogical goal, few examples are available of how to achieve such integration in specific class activities. This paper describes an example of a subject-specific, Christian/psychology integration class activity by utilizing the serenity prayer to teach psychology students about stress management. The activity is described for the purpose of justification and replication. Conclusions and future directions are also discussed. ********** The value of teaching Christian integration in undergraduate psychology courses has been well documented (e.g., Entwistle, 2010; Watson & Eveleigh, 2014). Christian psychology instructors, particularly those teaching at Christian universities, widely express commitment to integration at a broad curricular level (Koch & Doughty, 1998). Despite the high value placed on teaching Christian/psychology integration, recent scholars have pointed out that specific methods are lacking for how to implement such integration in specific class activities (e.g., Garzon & Hall, 2012; Garzon, Hall, & Ripley, 2014). Without examples of integration methods, it is challenging for instructors to fulfill curricular integration goals to build a repertoire of class activities. The current lack of models can be particularly problematic for new instructors and those transferring from secular to Christian education. As Garzon and Hall (2012) warned, a lack of clear models for activities could "easily encourage individualistic, new age, or eastern mystical faith instead of a deepened relationship with Jesus Christ" (p. 158). Accordingly, scholars have requested descriptions of classroom activities that exemplify valuable Christian/psychology integration. For example, Garzon and Hall (2012) reviewed research on teaching Christian/ psychology integration and their recommendations included a need for "increasing applied integration resources in subject-specific areas" (p. 158). They placed particular emphasis on resources that incorporated, "active involvement and interaction with peers as key elements" (Garzon & Hall, 2012, p. 157). Thus, their conclusions suggest a need for Christian instructors to share examples of subject-specific class activities targeted at integrating Christianity and psychology. Subject-Specific Christian/Psychology Integration Activities Subject-specific class activities aimed at integrating Christianity and psychology serve multiple purposes. Class activities serve as concrete models that counteract messages students may have previously received about the incompatibility of faith and learning and introduce the idea that meaningful interaction can take place between the two fields (Garzon et al., 2014). In fact, class activities such as the one presented in this article can demonstrate to students how Christian/ psychology integration can actually strengthen their understanding and incorporation of faith-based principles into their daily lives. Providing modern psychological explanations and empirical support for biblical proclamations can help students understand and connect with their faith in new and powerful ways and can encourage a desire to learn more about both psychology and religion. The field of psychology, in particular, can benefit from explicit examples of faith integration since the field is often criticized for being fundamentally incompatible with Christianity, despite countless examples to the contrary (e.g., Adams, McMinn, & Thurston, 2014; Butman & Yarhouse, 2014; Cook & Leonard, 2014; Ripley & Dwiwardani, 2014). It follows that an introductory psychology course would benefit most from subject-specific Christian/psychology integration activities, since this is typically the first and most widely attended psychology course among college students and can foster students' expectations for integration in subsequent courses. …

Fidaxomicin (Fdx) is an antimicrobial RNA polymerase (RNAP) inhibitor highly effective againstMycobacterium tuberculosisRNAPin vitro, but clinical use of Fdx is limited to treatingClostridium difficileintestinal infections due to poor absorption. To enable structure-guided optimization of Fdx to treat tuberculosis, we report the 3.4 Å cryo-electron microscopy structure of a completeM. tuberculosisRNAP holoenzyme in complex with Fdx. We find that the actinobacteria general transcription factor RbpA contacts fidaxomycin and explains its strong effect onM. tuberculosis. We present additional structures that define conformational states ofM. tuberculosisRNAP between the free apo-holenzyme and the promoter-engaged open complex ready for transcription. The results establish that Fdx acts like a doorstop to jam the enzyme in an open state, preventing the motions necessary to secure promoter DNA in the active site. Our results provide a structural platform to guide development of anti-tuberculosis antimicrobials based on Fdx.

The purpose of this study was to explore the occurrence of anticipated work–family conflict (AWFC) in young adults and examine the assumption that AWFC and anticipated work and family barriers were interchangeable constructs. Previous research sought to examine AWFC but due to theoretical and measurement factors, an understanding of it remains contradictory and unclear. Clarification is needed for subsequent research on work and family planning. For this purpose, 120 college students between the ages of 18 to 25 completed six different self-report questionnaires measuring AWFC and barriers. Results were examined to establish construct validity and prevalence rates for AWFC. A multi-trait, multi-method matrix revealed construct validity for some, but not all, AWFC and barriers measures. Low to moderate AWFC prevalence rates were reported overall, and Friedman’s analysis of variance revealed that rates varied by different types of measurement instruments. Thus, methodology emerged as a critical factor for continued AWFC research. Additional implications and future directions for AWFC and barriers are discussed.

Endstage renal disease (ESRD) is increasingly being recognized as a major public health issue globally. Planning of intervention measures is preferably hinged on what is known about outcome parameters.This study investigated the influence of anxiety with depression and psychosocial- and treatment-related correlates on quality of life (QOL) in ESRD.Overall, 100 eligible individuals with ESRD were recruited by systematic random sampling technique. They were initially interviewed using the sociodemographic/clinical profile questionnaire, followed by assessment with the Hospital Anxiety and Depression Scale. Subsequently, subjective QOL of participants was assessed using the World Health Organization QOL-BREF.The mean age of participants was 41.9 ± 10.9 years, and males (55.0%) were preponderant. A total of 29 (29.0%) participants had diagnosable anxiety with depression psychopathology based on Hospital Anxiety and Depression Scale scores ≥ 8. Different degrees of impairment across domains of QOL were observed. Factors like being employed, married status, younger age, and spending less on treatment correlated positively with good QOL across specific domains, whereas comorbid anxiety with depression, history of dialysis, monthly income less than ₦50,000 ($300), and having up to 50 dialysis sessions correlated negatively with good outcome in specific domains of QOL (p0.05). However, only age, anxiety/depression, employment, and history of dialysis were independently related to QOL following logistic regression analyses.The care of ESRD should be matched with need-based mental health services, and psychosocial support across important illness trajectories is indicated for best outcome. Further research among people with ESRD is also warranted.

Triclosan (TCS), a broad-spectrum antimicrobial, is used in commercial toothpastes with reported dental benefits. Our studies on 22 popular manual toothbrushes in the U.S. showed that common toothbrush head components can accumulate substantial amounts of TCS after brushing with TCS-formulated toothpastes (TCS-TPs). After simulated 3-month brushing with a commercial best-selling TCS-TP, over one third of the adults' toothbrushes showed a cumulative TCS uptake of 21-37.5 mg, equivalent to 7-12.5 doses of the TCS used per brushing. Similar results were observed on children's toothbrushes with small pea-size heads. Elastomer components were found to be the main contributor while both nylon bristles and elastomers could act as absorptive sinks for TCS during brushing. Studies on six different TCS-TPs containing 0.3 wt% TCS showed similar profiles of TCS accumulation. The absorbed TCS was gradually released into toothpaste slurries after switching to TCS-free alternatives. Release of TCS, which typically measured at a fraction (75%) of the standard dose using the TCS-TPs, continued for over 2 weeks and occurred most rapidly in peroxide-containing "whitening" toothpastes, followed by alkaline and surfactant-rich toothpastes. The accumulating effect was not exclusive to TCS but was commonly observed on several chemicals identified in TCS-TPs and a range of regular toothpastes.

Summary Noncoding RNAs (ncRNAs) regulate gene expression in all organisms. Bacterial 6S RNAs globally regulate transcription by binding RNA polymerase (RNAP) holoenzyme and competing with promoter DNA. Escherichia coli ( Eco ) 6S RNA interacts specifically with the housekeeping σ 70 -holoenzyme (Eσ 70 ) and plays a key role in the transcriptional reprogramming upon shifts between exponential and stationary phase. Inhibition is relieved upon 6S RNA-templated RNA synthesis. We report here the 3.8 A resolution structure of a complex between 6S RNA and Eσ 70 determined by single-particle cryo-electron microscopy and validation of the structure using footprinting and crosslinking approaches. Duplex RNA segments have A-form C3′ endo sugar puckers but widened major groove widths, giving the RNA an overall architecture that mimics B-form promoter DNA. Our results help explain the specificity of Eco 6S RNA for Eσ 70 and show how an ncRNA can mimic B-form DNA to directly regulate transcription by the DNA-dependent RNAP.