Ongoing Synthesis Research Articles

Human CCR4-NOT is a global regulator of gene expression and is a novel silencer of retrotransposon activation.

CCR4-NOT regulates multiple steps in gene regulation, including transcription, mRNA decay, protein ubiquitylation, and translation. It has been well studied in budding yeast; however, relatively less is known about its regulation and functions in mammals. To characterize the functions of the human CCR4-NOT complex, we developed a rapid auxin-induced degron system to deplete CNOT1 (the scaffold of the complex) and CNOT4 (E3 ubiquitin ligase) in cell culture. Transcriptome-wide measurements of gene-expression revealed that depleting CNOT1 changed several thousand transcripts, wherein most mRNAs were increased and resulted in a global decrease in mRNA decay rates. In contrast to what was observed in CNOT1-depleted cells, CNOT4 depletion only modestly changed RNA steady-state levels and, surprisingly, led to a global acceleration in mRNA decay. To further investigate the role of CCR4-NOT in transcription, we used transient transcriptome sequencing (TT-seq) to measure ongoing RNA synthesis. Depletion of either subunit resulted in increased RNA synthesis of several thousand genes. In contrast to most of the genome, a rapid reduction in the synthesis of KRAB-Zinc-Finger-proteins (KZNFs) genes, especially those clustered on chromosome 19, was observed. KZNFs are transcriptional repressors of retro-transposable elements (rTEs), and consistent with the decreased KZNFs expression, we observed a significant and rapid activation of rTEs, mainly Long interspersed Nuclear Elements (LINEs). Our data reveal that CCR4-NOT regulates gene expression and silences retrotransposons across the genome by maintaining KZNF expression. These data establish CCR4-NOT as a global regulator of gene expression, and we have identified a novel mammalian-specific function of the complex, the suppression of rTEs.

Abstract PO3-26-09: A novel class of RNR inhibitors that induces genomic instability and breast cancer cell death without inducing overt toxicities

Abstract Despite the development of screening techniques and therapies, nearly 30% of breast cancers progress to Stage IV metastatic disease. These advanced cancers are highly proliferative, engaging in ongoing DNA synthesis that requires ribonucleotide reductase (RNR), the rate-limiting enzyme for dNTP synthesis. RRM2, the small subunit of RNR, activates the enzyme, increases deoxynucleotide pools, and is overexpressed in many cancers, including breast cancer. Thus, RNR is an established therapeutic target in advanced breast cancer. Currently, gemcitabine is the only FDA-approved RNR inhibitor used to treat Stage IV disease. As a nucleoside analog, gemcitabine also causes severe dose-limiting toxicities (i.e. myelosuppression and hepatotoxicity) due to off-target DNA replication termination. We hypothesize that a novel, non-nucleoside, RNR inhibitor may overcome these limitations. To address this, we developed TMU27a, a derivative of naphthyl salicylic acyl hydrazone (NSAH), which was identified in an in silico screen for its high-affinity binding to the RNR catalytic site. To measure direct interactions between TMU27a and purified RNR protein, we used surface plasmon resonance, revealing on-target, tight binding (Kd = 150 nM). Unbiased docking and molecular dynamics simulations (using lowest energy calculations) predicted the binding sight of TMU27a to be the catalytic site of RNR, where it is stabilized through extensive interactions formed with conserved residues. Preliminary docking predictions also indicate multiple clash sites between TMU27a and the DNA polymerase lambda nucleotide pocket even in the lowest energy pose (unlike gemcitabine), suggesting that TMU27a inhibits RNR activity through a novel mechanism without significant off-target binding to other nucleotide binding proteins that contribute to toxicity. Demonstrating on-target inhibition, TMU27a reduces dATP levels and induces S and G2/M-phase stalling, leading to significant DNA damage and chromosomal instability. This excessive genomic damage causes growth repression, apoptosis, and senescence in several triple negative breast cancer (TNBC) cell lines. In xenograft mouse models of TNBC, TMU27a fully suppressed tumor growth without impacting mouse weight, suggesting strong drug efficacy in the absence of overt toxicity. Moreover, while short-term exposure to gemcitabine suppresses red and white blood cell populations, TMU27a does not. Together, these data indicate that TMU27a and its potential analogs are novel, non-toxic RNR inhibitors that overcome the limitations of gemcitabine for treating breast cancer. We are currently investigating how and where TMU27a binds human RNR using cryo-electron microscopy with the goal of generating a novel class of RNR inhibitors through structure-based drug design, improving binding affinity and selectivity. Citation Format: Taylor Baker, Kristen Weber-Bonk, Wei Huang, Derek Taylor, Ruth Keri. A novel class of RNR inhibitors that induces genomic instability and breast cancer cell death without inducing overt toxicities [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO3-26-09.

Use of multiple pharmacodynamic measures to deconstruct the Nix-TB regimen in a short-course murine model of tuberculosis.

A major challenge for tuberculosis (TB) drug development is to prioritize promising combination regimens from a large and growing number of possibilities. This includes demonstrating individual drug contributions to the activity of higher-order combinations. A BALB/c mouse TB infection model was used to evaluate the contributions of each drug and pairwise combination in the clinically relevant Nix-TB regimen [bedaquiline-pretomanid-linezolid (BPaL)] during the first 3 weeks of treatment at human equivalent doses. The rRNA synthesis (RS) ratio, an exploratory pharmacodynamic (PD) marker of ongoing Mycobacterium tuberculosis rRNA synthesis, together with solid culture CFU counts and liquid culture time to positivity (TTP) were used as PD markers of treatment response in lung tissue; and their time-course profiles were mathematically modeled using rate equations with pharmacologically interpretable parameters. Antimicrobial interactions were quantified using Bliss independence and Isserlis formulas. Subadditive (or antagonistic) and additive effects on bacillary load, assessed by CFU and TTP, were found for bedaquiline-pretomanid and linezolid-containing pairs, respectively. In contrast, subadditive and additive effects on rRNA synthesis were found for pretomanid-linezolid and bedaquiline-containing pairs, respectively. Additionally, accurate predictions of the response to BPaL for all three PD markers were made using only the single-drug and pairwise effects together with an assumption of negligible three-way drug interactions. The results represent an experimental and PD modeling approach aimed at reducing combinatorial complexity and improving the cost-effectiveness of in vivo systems for preclinical TB regimen development.

Advancing gender inclusivity for Two-Spirit, trans, nonbinary and other gender-diverse blood and plasma donors.

Two-Spirit, trans, nonbinary and other gender-diverse (2STGD) donors face challenges in donation. While many blood operators aim to address these challenges, to date, no empirical study with these donors has been conducted to guide their efforts. This paper reports 2STGD donors' views on a two-step approach asking donors their gender and sex assigned at birth (SAAB), and expanding gender options in donor registration. A qualitative community-based study was conducted with 2STGD donors (n = 85) in Canada. Semi-structured, in-depth interviews were conducted from July to October2022, audio-recorded and transcribed. Data were analysed using a thematic analytic framework. Participants were divided on their views of a two-step approach asking gender and SAAB. Themes underlying views in favour of this approach included the following: demonstrating validation and visibility, and treating 2STGD donors and cisgender donors alike. Themes underlying views not in favour or uncertain included potential for harm, compromising physical safety, and invalidation. All participants were in favour of expanding gender options if blood operators must know donors' gender. Results indicate that a two-step approach for all donors is not recommended unless the blood operator must know both a donor's gender and SAAB to ensure donor and/or recipient safety. Gender options should be expanded beyond binary options. Ongoing research and evidence synthesis are needed to determine how best to apply donor safety measures to nonbinary donors.

Enumeration, Nomenclature, and Stability Rules of Carbon Nanobelts.

With recent breakthroughs and advances in synthetic chemistry, carbon nanobelts (CNBs) have become an emerging hot topic in chemistry and materials science. Owing to their unique molecular structures, CNBs have intriguing properties with applications in synthetic materials, host-guest chemistry, optoelectronics, and so on. Although a considerable number of CNBs with diverse forms have been synthesized, no systematic nomenclature is available yet for this important family of macrocycles. Moreover, little is known about the detailed isomerism of CNBs, which, in fact, exhibits greater complexity than that of carbon nanotubes. The copious variety of CNB isomers, along with the underlying structure-property relationships, bears fundamental relevance to the ongoing design and synthesis of novel nanobelts. In this paper, we propose an elegant approach to systematically enumerate, classify, and name all possible isomers of CNBs. Besides the simplest, standard CNBs defined by chiral indices (n, m), the nonstandard CNBs (n, m, l) involve an additional winding index l. Based on extensive quantum chemical calculations, we present a comprehensive study of the relative isomer stability of CNBs containing up to 30 rings. A simple Hückel-based model with a high predictive power reveals that the relative stability of standard CNBs is governed by the π stabilization and the strain destabilization induced by the cylindrical carbon framework, and the former effect prevails over the latter. For nonstandard CNBs, a third stability factor, the H···H repulsion in the benzo[c]phenanthrene-like motifs, is also shown to be important and can be incorporated into the simple quantitative model. In general, lower-energy CNB isomers have a larger HOMO-LUMO gap, suggesting that their thermodynamic stability coincides with kinetic stability. The most stable CNB isomers determined can be considered the optimal targets for future synthesis. These results lay an initial foundation and provide a useful theoretical tool for further research on CNBs and related analogues.

Lung microenvironments harbor Mycobacterium tuberculosis phenotypes with distinct treatment responses.

Tuberculosis lung lesions are complex and harbor heterogeneous microenvironments that influence antibiotic effectiveness. Major strides have been made recently in understanding drug pharmacokinetics in pulmonary lesions, but the bacterial phenotypes that arise under these conditions and their contribution to drug tolerance are poorly understood. A pharmacodynamic marker called the RS ratio® quantifies ongoing rRNA synthesis based on the abundance of newly synthesized precursor rRNA relative to mature structural rRNA. Application of the RS ratio in the C3HeB/FeJ mouse model demonstrated that Mycobacterium tuberculosis populations residing in different tissue microenvironments are phenotypically distinct and respond differently to drug treatment with rifampin, isoniazid, or bedaquiline. This work provides a foundational basis required to address how anatomic and pathologic microenvironmental niches may contribute to long treatment duration and drug tolerance during the treatment of human tuberculosis.

Charting the Growth and Structure of Early ChatGPT-Education Research: A Bibliometric Study

Charting the Growth and Structure of Early ChatGPT-Education Research: A Bibliometric Study

CAN-SAR: A database of Canadian species at risk information

Threatened species lists describe the conservation status of species and are key tools used to inform decisions for biodiversity conservation. These lists are rich in information obtained during status assessment and recovery planning processes, ranging from biological attributes to actions that support recovery. Data compiled from species lists allow for analyses, including assessing trends in threats, prioritizing actions, and identifying barriers to achieving recovery objectives. For legally protected species at risk of extinction in Canada, such analyses are challenging owing to a lack of comprehensive and accessible data reflecting information compiled from listing and recovery documents. To encourage ongoing synthesis and minimise duplication of efforts, we initiated CAN-SAR: a database of Canadian Species at Risk information. This transparent, open-access, and searchable database contains information transcribed from listing documents, including listing date, and derived variables. Derived variables required interpretation for which we developed standardised criteria to record information, including classification of recovery actions. The CAN-SAR database is updateable, and will contribute towards improved recovery planning to safeguard species of conservation concern.

SUCCESSFUL DOUBLE LUNG TRANSPLANTATION FROM DONOR WITH PREVIOUS SARS-COV-2 INFECTION

TYPE: Case Report TOPIC: Transplantation INTRODUCTION: Continued community prevalence of SARS-CoV-2 is impacting center-level organ acceptance practices due to an anticipated increase in potential lung donors with exposure to or previous infection with SARS-CoV-2 causing concern for donor-derived viral transmission and impaired graft quality. We present the first report in the USA of a successful lung transplant (LTx) from a donor with prior COVID-19. CASE PRESENTATION: A 63-year-old man with pulmonary fibrosis received a donor offer in August 2020 from a 20-year-old donor with prior (2-3 months before death) COVID-19 with full recovery without medical care. Two nucleic acid tests for SARS-CoV-2 from nasopharyngeal and bronchial specimens, repeated 24 hours apart before procurement, were negative. SARS-CoV-2 serology demonstrated IgG. Bronchoscopy and imaging were unremarkable, and PaO2/FiO2 ratio was 417 mmHg. After multidisciplinary agreement, the donor was deemed acceptable. Our patient experienced an uneventful post-LTx course. His peak FEV1 and FVC were at 3.28 and 3.85 L; trimonthly bronchoscopies during the first year were normal; he repeatedly tested negative for SARS-CoV-2. He continues to do well at 1 year, with negative SARS-CoV-2 IgG and stable pulmonary function and chest imaging. DISCUSSION: An immunocompetent donor with resolved COVID-19 may carefully be considered for lung donation. The decision is driven by the candidates’ risk of remaining on the waitlist. Mild COVID-19 should be differentiated from ARDS/fibrosis. Our donor had mild COVID-19, excellent oxygenation capacity, and no lung damage. CONCLUSIONS: If recommendations are followed, donor-derived SARS-CoV-2 transmission is rare. Ongoing synthesis of data on long-term allograft quality is required. DISCLOSURE: Nothing to declare. KEYWORD: Lung transplantation

An unexpected epigraph: exploring the personal and philosophical relevance of Ralph Waldo Emerson to Carl Ransom Rogers

ABSTRACT This article takes as its starting point the Emerson epigraph chosen by Rogers for his seminal book Client-Centered Therapy, and discovers that Rogers also acknowledged the ‘deep influence’ that Emerson and the transcendental school had on his personal life and philosophy. It proposes evidence of this influence, noticing similarities in the writings and biographical details of the two men. It also extrapolates a question and proposes it for further research – the possibility that Rogers integrated Emersonian thinking into the development of the person-approach. The limited and interpretive reasoning for such an extrapolation is acknowledged. The article concludes that Emerson was one of many writers in literature, theology, philosophy, psychology, art and science with whom Rogers engaged over his lifetime, an eclectic contributed to Rogers’ fluid, ongoing, and ever-evolving creative synthesis. It also concludes that the question of whether Rogers’ use of the epigraph at the beginning of Client-Centered Therapy indicates a connection between Emerson and Rogers’ person-centered theory, is worthy of being asked.

Tobacco-specific nitrosamine 1-(N-methyl-N-nitrosamino)-1-(3-pyridinyl)-4-butanal (NNA) causes DNA damage and impaired replication/transcription in human lung cells.

Thirdhand smoke (THS) is a newly described health hazard composed of toxicants, mutagens and carcinogens, including nicotine-derived tobacco specific nitrosamines (TSNAs), one of which is 1-(N-methyl-N-nitrosamino)-1-(3-pyridinyl)-4-butanal (NNA). Although TSNAs are generally potent carcinogens, the risk of NNA, which is specific to THS, is poorly understood. We recently reported that THS exposure-induced adverse impact on DNA replication and transcription with implications in the development of cancer and other diseases. Here, we investigated the role of NNA in THS exposure-induced harmful effects on fundamental cellular processes. We exposed cultured human lung epithelial BEAS-2B cells to NNA. The formation of DNA base damages was assessed by Long Amplicon QPCR (LA-QPCR); DNA double-strand breaks (DSBs) and NNA effects on replication and transcription by immunofluorescence (IF); and genomic instability by micronuclei (MN) formation. We found increased accumulation of oxidative DNA damage and DSBs as well as activation of DNA damage response pathway, after exposure of cells to NNA. Impaired S phase progression was also evident. Consistent with these results, we found increased MN formation, a marker of genomic instability, in NNA-exposed cells. Furthermore, ongoing RNA synthesis was significantly reduced by NNA exposure, however, RNA synthesis resumed fully after a 24h recovery period only in wild-type cells but not in those deficient in transcription-coupled nucleotide excision repair (TC-NER). Importantly, these cellular effects are common with the THS-exposure induced effects. Our findings suggest that NNA in THS could be a contributing factor for THS exposure-induced adverse health effect.

Combination of Mycobacterium tuberculosis RS Ratio and CFU Improves the Ability of Murine Efficacy Experiments to Distinguish between Drug Treatments.

ABSTRACTMurine tuberculosis drug efficacy studies have historically monitored bacterial burden based on CFU of Mycobacterium tuberculosis in lung homogenate. In an alternative approach, a recently described molecular pharmacodynamic marker called the RS ratio quantifies drug effect on a fundamental cellular process, ongoing rRNA synthesis. Here, we evaluated the ability of different pharmacodynamic markers to distinguish between treatments in three BALB/c mouse experiments at two institutions. We confirmed that different pharmacodynamic markers measure distinct biological responses. We found that a combination of pharmacodynamic markers distinguishes between treatments better than any single marker. The combination of the RS ratio with CFU showed the greatest ability to recapitulate the rank order of regimen treatment-shortening activity, providing proof of concept that simultaneous assessment of pharmacodynamic markers measuring different properties will enhance insight gained from animal models and accelerate development of new combination regimens. These results suggest potential for a new era in which antimicrobial therapies are evaluated not only on culture-based measures of bacterial burden but also on molecular assays that indicate how drugs impact the physiological state of the pathogen.

Nursing surveillance: A concept analysis.

To provide a framework to assess and evaluate nursing surveillance of patients. The Nursing Interventions Classifications define surveillance as the purposeful and ongoing acquisition, interpretation, and synthesis of patient data for clinical decision making and is essential for improving patient safety. The existing literature was searched using CINAHL, OVID, EmCare, and 11 ScienceDirect databases. The Walker and Avant method was used to analyze the concept of surveillance. Technology that facilitates surveillance in the community is ubiquitous in acute care settings. Nurses caring for patients use a tremendous volume of patient data to inform their clinical decision-making. Five attributes are associated with nursing surveillance: systematic process, pattern recognition, coordinated communication, the anticipation of problems of instability, and decision making. Surveillance is dynamic and extends over time. Antecedents to nursing surveillance include sufficient nurse education, nurse expertise, nurse staffing, as well as an organizational culture that supports nursing surveillance. When nursing surveillance is present, patient safety is enhanced and adverse events that harm patients are reduced. The concept of nursing surveillance is complex and defies empirical measurement, though it is possible to measure the attributes and outcomes. Nursing surveillance is essential to the safe management of complex patient cases. Surveillance is more than monitoring or simple patient observation. Monitoring is an essential part of surveillance but incorporating the critical attributes of surveillance lead to improved patient outcomes.

Symptom clusters in patients receiving chemotherapy: A systematic review

Background and purposeSince 2001, symptom cluster research has grown considerably. However, because multiple methodological considerations remain, ongoing synthesis of the literature is needed to identify gaps in this area of...

Mapping levels of COVID-19 vaccine hesitancy worldwide: preliminary results of a systematic review

Abstract Background Several factors can determine the success of a vaccination campaign, regardless of the availability of an effective and safe vaccine. Among these, vaccine hesitancy (VH) has long represented one of the main barriers to achieving vaccination coverage goals in many countries. The aim of this systematic review was to summarize the global levels of COVID-19 VH and to identify their determinants. Methods A systematic review of the literature was carried out following the PRISMA guideline and searching on PubMed, Web of Science, Scopus and PsycINFO for observational studies published up to April 2021 in English or Italian language. Levels of hesitancy and data on determinants, according to the SAGE working group matrix of determinants, were extracted. The systematic review protocol was registered on Prospero (ID: CRD42021235328). Results Of the 5447 articles initially retrieved, 89 were included in the qualitative synthesis. The included studies investigated COVID-19 VH on 1.954.331 individuals from 68 different countries (North America 19; South America 23; Africa 3; Asia 18; Europe 55; Oceania 4) between March 2020 and January 2021. Rates of hesitancy, before to the worldwide availability of vaccinations (82 studies), ranged from 84.6% (Cameroon, May-August 2020) to 10.37% (China, March-June 2020). Studies carried out on January 2021 (N = 7) showed rates ranging from 42.8% (Turkey) to 5.3% (UK). A subgroup analysis of data referring to healthcare workers showed the highest hesitancy rates in Honk Kong (60%) and Turkey (26.6%), before and after vaccine availability, respectively. Conclusions Preliminary results confirm the wide variability of VH across the world. The ongoing quantitative synthesis on prevalence of VH and the analysis on determinants will allow to update intervention strategies to address VH, both in general population and healthcare workers, orienting public health policies. Key messages Vaccine Hesitancy continues to be one of the greatest threats to global health and, in the long term, one of the potential barriers to the success of the global COVID-19 vaccination campaign. Identifying and synthesizing the specific determinants of COVID-19 vaccine hesitancy is essential to guide intervention strategies to promote vaccine confidence and, finally, vaccine acceptance.

Exploring the co-involvement of disabled adolescents in participatory action research; protocol for a critical interpretative synthesis

Background: Participatory action research (PAR) provides an opportunity for academic researchers and adolescents to co-conduct research within an area of shared interest. Reciprocal learning occurs as co-researchers acquire research skills and knowledge, and academic researchers gain understanding of the issue being examined, from the perspective of those with lived experience. All members of the research team have a shared responsibility for the research and decision-making processes. PAR has predominantly involved adults as co-researchers. However, in recent years more effort has been made to co-conduct research with adolescents. The aim of this review is to interrogate the practices of academic researchers employing a PAR approach when working along-side disabled adolescents. Methods/design: A critical interpretive synthesis (CIS) will be conducted, allowing for a diverse range of evidence to be drawn from. A systematic search of nine databases, from 1990 onwards, will be conducted first. Reference checking will occur to elicit further relevant data. Following screening, further purposive sampling will be completed to facilitate the development of concepts and theory in line with the on-going analysis and synthesis of findings. Data analysis will involve interpretation of included papers in relation to the principles of PAR and a ‘best-practice’ framework will be developed. During analysis particular emphasis will be given to the identification of potential social barriers to the participation of disabled adolescents in PAR. Discussion: PAR is widely employed but little is known about its use when working with disabled adolescents. This current CIS will critically question the current practices of academic researchers employing PAR when working along-side disabled adolescents and future research through the best practice framework we will develop.

Strengthening nursing, midwifery and allied health professional leadership in the UK – a realist evaluation

PurposeThis paper aims to share the findings of a realist evaluation study that set out to identify how to strengthen nursing, midwifery and allied health professions (NMAHP) leadership across all health-care contexts in the UK conducted between 2018 and 2019. The collaborative research team were from the Universities of Bangor, Ulster, the University of the West of Scotland and Canterbury Christ Church University.Design/methodology/approachRealist evaluation and appreciative inquiry were used across three phases of the study. Phase 1 analysed the literature to generate tentative programme theories about what works, tested out in Phase 2 through a national social media Twitter chat and sense-making workshops to help refine the theories in Phase 3. Cross-cutting themes were synthesised into a leadership framework identifying the strategies that work for practitioners in a range of settings and professions based on the context, mechanism and output configuration of realist evaluation. Stakeholders contributed to the ongoing interrogation, analysis and synthesis of project outcomes.FindingsFive guiding lights of leadership, a metaphor for principles, were generated that enable and strengthen leadership across a range of contexts. – “The Light Between Us as interactions in our relationships”, “Seeing People’s Inner Light”, “Kindling the Spark of light and keeping it glowing”, “Lighting up the known and the yet to be known” and “Constellations of connected stars”.Research limitations/implicationsThis study has illuminated the a-theoretical nature of the relationships between contexts, mechanisms and outcomes in the existing leadership literature. There is more scope to develop the tentative programme theories developed in this study with NMAHP leaders in a variety of different contexts. The outcomes of leadership research mostly focussed on staff outcomes and intermediate outcomes that are then linked to ultimate outcomes in both staff and patients (supplemental). More consideration needs to be given to the impact of leadership on patients, carers and their families.Practical implicationsThe study has developed additional important resources to enable NMAHP leaders to demonstrate their leadership impact in a range of contexts through the leadership impact self-assessment framework which can be used for 360 feedback in the workplace using the appreciative assessment and reflection tool.Social implicationsWhilst policymakers note the increasing importance of leadership in facilitating the culture change needed to support health and care systems to adopt sustainable change at pace, there is still a prevailing focus on traditional approaches to individual leadership development as opposed to collective leadership across teams, services and systems. If this paper fails to understand how to transform leadership policy and education, then it will be impossible to support the workforce to adapt and flex to the increasingly complex contexts they are working in. This will serve to undermine system integration for health and social care if the capacity and capability for transformation are not attended to. Whilst there are ambitious global plans (WHO, 2015) to enable integrated services to be driven by citizen needs, there is still a considerable void in understanding how to authentically engage with people to ensure the transformation is driven by their needs as opposed to what the authors think they need. There is, therefore, a need for systems leaders with the full skillset required to enable integrated services across place-based systems, particularly clinicians who are able to break down barriers and silo working across boundaries through the credibility, leadership and facilitation expertise they provide.Originality/valueThe realist evaluation with additional synthesis from key stakeholders has provided new knowledge about the principles of effective NMAHP leadership in health and social care, presented in such a way that facilitates the use of the five guiding lights to inform further practice, education, research and policy development.

Exploring the co-involvement of disabled adolescents in participatory action research; protocol for a critical interpretative synthesis.

Background: Participatory action research (PAR) provides an opportunity for academic researchers and adolescents to co-conduct research within an area of shared interest. Reciprocal learning occurs as co-researchers acquire research skills and knowledge, and academic researchers gain understanding of the issue being examined, from the perspective of those with lived experience. All members of the research team have a shared responsibility for the research and decision-making processes. PAR has predominantly involved adults as co-researchers. However, in recent years more effort has been made to co-conduct research with adolescents. The aim of this review is to interrogate the practices of academic researchers employing a PAR approach when working along-side disabled adolescents. Methods/design: A critical interpretive synthesis (CIS) will be conducted, allowing for a diverse range of evidence to be drawn from. A systematic search of nine databases, from 1990 onwards, will be conducted first. Reference checking will occur to elicit further relevant data. Following screening, further purposive sampling will be completed to facilitate the development of concepts and theory in line with the on-going analysis and synthesis of findings. Data analysis will involve interpretation of included papers in relation to the principles of PAR and a 'best-practice' framework will be developed. During analysis particular emphasis will be given to the identification of potential social barriers to the participation of disabled adolescents in PAR. Discussion: PAR is widely employed but little is known about its use when working with disabled adolescents. This current CIS will critically question the current practices of academic researchers employing PAR when working along-side disabled adolescents and future research through the best practice framework we will develop.

The Development of Living, Rapid Practice Points: Summary of Methods From the Scientific Medical Policy Committee of the American College of Physicians.

In response to the COVID-19 pandemic, the Scientific Medical Policy Committee (SMPC) of the American College of Physicians (ACP) began developing "practice points" to provide clinical advice based on the best available evidence for the public, patients, clinicians, and public health professionals. As one of the first organizations in the United States to develop evidence-based clinical guidelines, ACP continues to lead and advance the science of evidence-based medicine by implementing new methods to rapidly publish practice points and maintain them as living advice that regularly assesses and incorporates new evidence. The overarching aim of practice points is to answer targeted key questions for which there is a timely need to synthesize evidence for decision making. The SMPC believes these methods can potentially be adapted to address various clinical and public health topics beyond the COVID-19 pandemic. This article presents an overview of the SMPC's living, rapid practice points development process, which includes a rapid systematic review, use of the GRADE (Grading of Recommendations Assessment, Development and Evaluation) method, use of stringent policies on the disclosure of interests and management of conflicts of interest, incorporating a public (nonclinician) perspective, and maintenance of the documents as living through ongoing surveillance and synthesis of new evidence as it emerges.

Reciprocal regulation of mTORC1 and ribosomal biosynthesis determines cell cycle progression in activated T cells

Abstract Anabolic metabolism of antigen-stimulated T cells is coordinated by mTOR which is activated by TCR and IL-2 signals. We show that mTORC1 regulates both the exit of quiescence and later divisions as Raptor-deficient T cells proceed 3x slower through each cell division in vivo. We noticed that following stimulation the amount of RNA per cell increases dramatically (10–40x) and is reduced to ~50% in Raptor-, but not Rictor-deficient T cells. RNAseq data indicate that this applies to all RNA polymerase I- and II-dependent biotypes. Using Coulter volume measurements, RNA-, FISH- and imaging-flow cytometry we determined that following stimulation the cell volume increases five-fold. It consists of the increase of nuclear volume and nucleolar rRNA synthesis on day 1 and a further increase of cytoplasmic volume on day 2. Raptor-deficiency delays and diminishes the growth of these cellular compartments, decreases ribosomal biosynthesis and thus the cell volume. Inhibitors of rRNA transcription block cell cycle progression at two checkpoints, of which only the G1/S transition is regulated by Raptor. We found that mTORC1 activity depends on ongoing rRNA synthesis in early activation and regulates the further synthesis rate. Altogether our data show that mTOR-adjusted cell physiology and the biosynthesis of ribosomal ‘hardware’ are reciprocally regulated during clonal T cell expansion.