An Intron-Retaining Splice Variant of Human Cyclin A2, Expressed in Adult Differentiated Tissues, Induces a G1/S Cell Cycle Arrest In Vitro

BACKGROUND: Human cyclin A2 is a key regulator of S phase progression and entry into mitosis. Alternative splice variants of the G1 and mitotic cyclins have been shown to interfere with full-length cyclin functions to modulate cell cycle progression and are therefore likely to play a role in differentiation or oncogenesis. The alternative splicing of human cyclin A2 has not yet been studied. METHODOLOGY/PRINCIPAL FINDINGS: Sequence-specific primers were designed to amplify various exon-intron regions of cyclin A2 mRNA in cell lines and human tissues. Intron retaining PCR products were cloned and sequenced and then overexpressed in HeLa cells. The subcellular localization of the splice variants was studied using confocal and time-lapse microscopy, and their impact on the cell cycle by flow cytometry, immunoblotting and histone H1 kinase activity. We found a splice variant of cyclin A2 mRNA called A2V6 that partly retains Intron 6. The gene expression pattern of A2V6 mRNA in human tissues was noticeably different from that of wild-type cyclin A2 (A2WT) mRNA. It was lower in proliferating fetal tissues and stronger in some differentiated adult tissues, especially, heart. In transfected HeLa cells, A2V6 localized exclusively in the cytoplasm whereas A2WT accumulated in the nucleus. We show that A2V6 induced a clear G1/S cell cycle arrest associated with a p21 and p27 upregulation and an inhibition of retinoblastoma protein phosphorylation. Like A2WT, A2V6 bound CDK2, but the A2V6/CDK2 complex did not phosphorylate histone H1. CONCLUSION/SIGNIFICANCE: This study has revealed that some highly differentiated human tissues express an intron-retaining cyclin A2 mRNA that induced a G1/S block in vitro. Contrary to full-length cyclin A2, which regulates cell proliferation, the A2V6 splice variant might play a role in regulating nondividing cell states such as terminal differentiation or senescence

Centrality evolution of the charged-particle pseudorapidity density over a broad pseudorapidity range in Pb-Pb collisions at root s(NN)=2.76TeV

Peer reviewe

Duration of androgen deprivation therapy with postoperative radiotherapy for prostate cancer: a comparison of long-course versus short-course androgen deprivation therapy in the RADICALS-HD randomised trial

Background Previous evidence supports androgen deprivation therapy (ADT) with primary radiotherapy as initial treatment for intermediate-risk and high-risk localised prostate cancer. However, the use and optimal duration of ADT with postoperative radiotherapy after radical prostatectomy remains uncertain. Methods RADICALS-HD was a randomised controlled trial of ADT duration within the RADICALS protocol. Here, we report on the comparison of short-course versus long-course ADT. Key eligibility criteria were indication for radiotherapy after previous radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to add 6 months of ADT (short-course ADT) or 24 months of ADT (long-course ADT) to radiotherapy, using subcutaneous gonadotrophin-releasing hormone analogue (monthly in the short-course ADT group and 3-monthly in the long-course ADT group), daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as metastasis arising from prostate cancer or death from any cause. The comparison had more than 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 75% to 81% (hazard ratio [HR] 0·72). Standard time-to-event analyses were used. Analyses followed intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov , NCT00541047 . Findings Between Jan 30, 2008, and July 7, 2015, 1523 patients (median age 65 years, IQR 60–69) were randomly assigned to receive short-course ADT (n=761) or long-course ADT (n=762) in addition to postoperative radiotherapy at 138 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 8·9 years (7·0–10·0), 313 metastasis-free survival events were reported overall (174 in the short-course ADT group and 139 in the long-course ADT group; HR 0·773 [95% CI 0·612–0·975]; p=0·029). 10-year metastasis-free survival was 71·9% (95% CI 67·6–75·7) in the short-course ADT group and 78·1% (74·2–81·5) in the long-course ADT group. Toxicity of grade 3 or higher was reported for 105 (14%) of 753 participants in the short-course ADT group and 142 (19%) of 757 participants in the long-course ADT group (p=0·025), with no treatment-related deaths. Interpretation Compared with adding 6 months of ADT, adding 24 months of ADT improved metastasis-free survival in people receiving postoperative radiotherapy. For individuals who can accept the additional duration of adverse effects, long-course ADT should be offered with postoperative radiotherapy. Funding Cancer Research UK, UK Research and Innovation (formerly Medical Research Council), and Canadian Cancer Society

Adding 6 months of androgen deprivation therapy to postoperative radiotherapy for prostate cancer: a comparison of short-course versus no androgen deprivation therapy in the RADICALS-HD randomised controlled trial

Background Previous evidence indicates that adjuvant, short-course androgen deprivation therapy (ADT) improves metastasis-free survival when given with primary radiotherapy for intermediate-risk and high-risk localised prostate cancer. However, the value of ADT with postoperative radiotherapy after radical prostatectomy is unclear. Methods RADICALS-HD was an international randomised controlled trial to test the efficacy of ADT used in combination with postoperative radiotherapy for prostate cancer. Key eligibility criteria were indication for radiotherapy after radical prostatectomy for prostate cancer, prostate-specific antigen less than 5 ng/mL, absence of metastatic disease, and written consent. Participants were randomly assigned (1:1) to radiotherapy alone (no ADT) or radiotherapy with 6 months of ADT (short-course ADT), using monthly subcutaneous gonadotropin-releasing hormone analogue injections, daily oral bicalutamide monotherapy 150 mg, or monthly subcutaneous degarelix. Randomisation was done centrally through minimisation with a random element, stratified by Gleason score, positive margins, radiotherapy timing, planned radiotherapy schedule, and planned type of ADT, in a computerised system. The allocated treatment was not masked. The primary outcome measure was metastasis-free survival, defined as distant metastasis arising from prostate cancer or death from any cause. Standard survival analysis methods were used, accounting for randomisation stratification factors. The trial had 80% power with two-sided α of 5% to detect an absolute increase in 10-year metastasis-free survival from 80% to 86% (hazard ratio [HR] 0·67). Analyses followed the intention-to-treat principle. The trial is registered with the ISRCTN registry, ISRCTN40814031, and ClinicalTrials.gov, NCT00541047. Findings Between Nov 22, 2007, and June 29, 2015, 1480 patients (median age 66 years [IQR 61–69]) were randomly assigned to receive no ADT (n=737) or short-course ADT (n=743) in addition to postoperative radiotherapy at 121 centres in Canada, Denmark, Ireland, and the UK. With a median follow-up of 9·0 years (IQR 7·1–10·1), metastasis-free survival events were reported for 268 participants (142 in the no ADT group and 126 in the short-course ADT group; HR 0·886 [95% CI 0·688–1·140], p=0·35). 10-year metastasis-free survival was 79·2% (95% CI 75·4–82·5) in the no ADT group and 80·4% (76·6–83·6) in the short-course ADT group. Toxicity of grade 3 or higher was reported for 121 (17%) of 737 participants in the no ADT group and 100 (14%) of 743 in the short-course ADT group (p=0·15), with no treatment-related deaths. Interpretation Metastatic disease is uncommon following postoperative bed radiotherapy after radical prostatectomy. Adding 6 months of ADT to this radiotherapy did not improve metastasis-free survival compared with no ADT. These findings do not support the use of short-course ADT with postoperative radiotherapy in this patient population

The Elements to Enhance the Successful Start and Completion of Program and Policy Evaluations: The Injury & Violence Prevention (IVP) Program & Policy Evaluation Institute

Background: Public health practitioners, including injury and violence prevention (IVP) professionals, are responsible for implementing evaluations, but often lack formal evaluation training. Impacts of many practitioner-focused evaluation trainings—particularly their ability to help participants successfully start and complete evaluations—are unknown. Objectives: We assessed the impact of the Injury and Violence Prevention (IVP) Program & Policy Evaluation Institute (“Evaluation Institute”), a team-based, multidisciplinary, and practitioner-focused evaluation training designed to teach state IVP practitioners and their cross-sector partners how to evaluate program and policy interventions. Design: Semi-structured interviews were conducted with members of 13 evaluation teams across eight states at least one year after training participation (24 participants in total). Document reviews were conducted to triangulate, supplement, and contextualize reported improvements to policies, programs, and practices. Intervention: Teams of practitioners applied for and participated in the Evaluation Institute, a five-month evaluation training initiative that included a set of online training modules, an in-person workshop, and technical support from evaluation consultants. Main Outcome Measure(s): The successful start and/or completion of a program or policy evaluation focused on an IVP intervention. Results: Of the 13 teams studied, a total of 12 teams (92%) reported starting or completing an evaluation. Four teams (31%) reported fully completing their evaluations; eight teams (61%) reported partially completing their evaluations. Teams identified common facilitators and barriers that impacted their ability to start and complete their evaluations. Nearly half of the 13 teams (46%) – whether or not they completed their evaluation – reported at least one common improvement made to a program or policy as a result of engaging in an evaluative process. Conclusion: Practitioner-focused evaluation trainings are essential to build critical evaluation skills among public health professionals and their multidisciplinary partners. The process of evaluating an intervention—even if the evaluation is not completed—has substantial value and can drive improvements to public health interventions. The Evaluation Institute can serve as a model for training public health practitioners and their partners to successfully plan, start, complete, and utilize evaluations to improve programs and policies. Keywords: Evaluation; injury; multidisciplinary partnerships; practitioner-focused evaluation training; professional development; program and policy evaluation; public health; technical assistance; violenc

The Elements to Enhance the Successful Start and Completion of Program and Policy Evaluations: The Injury & Violence Prevention (IVP) Program & Policy Evaluation Institute: The Injury & Violence Prevention (IVP) Program & Policy Evaluation Institute

Background: Public health practitioners, including injury and violence prevention (IVP) professionals, are responsible for implementing evaluations, but often lack formal evaluation training. Impacts of many practitioner-focused evaluation trainings—particularly their ability to help participants successfully start and complete evaluations—are unknown. Objectives: We assessed the impact of the Injury and Violence Prevention (IVP) Program & Policy Evaluation Institute (“Evaluation Institute”), a team-based, multidisciplinary, and practitioner-focused evaluation training designed to teach state IVP practitioners and their cross-sector partners how to evaluate program and policy interventions. Design: Semi-structured interviews were conducted with members of 13 evaluation teams across eight states at least one year after training participation (24 participants in total). Document reviews were conducted to triangulate, supplement, and contextualize reported improvements to policies, programs, and practices. Intervention: Teams of practitioners applied for and participated in the Evaluation Institute, a five-month evaluation training initiative that included a set of online training modules, an in-person workshop, and technical support from evaluation consultants. Main Outcome Measure(s): The successful start and/or completion of a program or policy evaluation focused on an IVP intervention. Results: Of the 13 teams studied, a total of 12 teams (92%) reported starting or completing an evaluation. Four teams (31%) reported fully completing their evaluations; eight teams (61%) reported partially completing their evaluations. Teams identified common facilitators and barriers that impacted their ability to start and complete their evaluations. Nearly half of the 13 teams (46%) – whether or not they completed their evaluation – reported at least one common improvement made to a program or policy as a result of engaging in an evaluative process. Conclusion: Practitioner-focused evaluation trainings are essential to build critical evaluation skills among public health professionals and their multidisciplinary partners. The process of evaluating an intervention—even if the evaluation is not completed—has substantial value and can drive improvements to public health interventions. The Evaluation Institute can serve as a model for training public health practitioners and their partners to successfully plan, start, complete, and utilize evaluations to improve programs and policies. Keywords: Evaluation; injury; multidisciplinary partnerships; practitioner-focused evaluation training; professional development; program and policy evaluation; public health; technical assistance; violenc

Awareness of and Confidence to Address Equity-Related Concepts Across the US Governmental Public Health Workforce

OBJECTIVE: To assess the governmental public health (GPH) workforce\u27s awareness of and confidence to address health equity, social determinants of health (SDoH), and social determinants of equity (SDoE) in their work. DESIGN, SETTING, AND PARTICIPANTS: A nationally representative population of US local and state GPH employees (n = 41 890) were surveyed through the 2021 Public Health Workforce Interests and Needs Survey (PH WINS 2021). MAIN OUTCOME MEASURES: Self-reported awareness and confidence were explored by self-identified racial and ethnic group identity, public health degree attainment, and supervisory status. RESULTS: GPH employees reported higher levels of awareness across concepts (health equity-71%, 95% confidence interval [CI]: 70.5-71.6; SDoH-62%, 95% CI: 62.3-63.5; SDoE-48%, 95% CI: 47.2-48.4) than confidence (health equity-48%, 95% CI: 47.8-49.0; SDoH-46%, 95% CI: 45.4-46.7; SDoE-34%, 95% CI: 33.4-34.6). Self-identified Black or African American employees reported higher confidence across all concepts (health equity-56%, 95% CI: 54.3-57.6; SDoH-52%, 95% CI: 50.8-54.1; SDoE-43%, 95% CI: 41.3-44.6) compared to other self-identified racial groups. Employees with a PH degree reported higher confidence across all concepts (health equity-65%, 95% CI: 63.8-68.8; SDoH-73%, 95% CI: 71.3-74.1; SDoE-39%, 95% CI: 36.9-40.1) compared with employees without a PH degree (health equity-45%, 95% CI: 44.8-46.1; SDoH-41%, 95% CI: 40.6-41.9; SDoE-33%, 95% CI: 32.6-33.8). We found an inverse relationship between supervisory status and confidence to address SDoE: Nonsupervisors reported higher confidence (35%, 95% CI: 29.2-31.9) than supervisors (31%, 95% CI: 29.2-31.9), managers (31%, 95% CI: 28.8-32.6), and executives (32%, 95% CI: 27.5-34.4). CONCLUSION: PH WINS 2021 reveals that GPH employees are aware of equity-related concepts but lack confidence to address them. Public health agencies should build employees\u27 confidence by prioritizing and operationalizing equity internally and externally in collaboration with communities and partners

Complete streets state laws & provisions: An analysis of legislative content and the state policy landscape, 1972–2018

Across the U.S., states have adopted Complete Streets legislative statutes—state laws that direct transportation agencies to routinely design and operate roadways to provide safe access for all users, including pedestrians, bicyclists, motorists, and public transit users. To date, there has not been a systematic and comprehensive analysis of the content and provisions of these laws. In this study, Complete Streets state statutes were identified using legal research databases. Using established legal mapping methods, a qualitative analysis was conducted of state laws that were effective through December 2018. A codebook and open-source data set were developed to support the public use of the data. Eighteen states and Washington, DC, have adopted Complete Streets legislative statutes. A total of 21 have been adopted, with 76% (n=16) of laws adopted since 2007. While the laws vary in content, detail, and specificity, several common provisions were identified across statutes. Complete Streets legislative statutes may be essential to ensure that road networks throughout states are safe, connected, and accessible for all users. This study provides key insights into the legislative landscape of Complete Streets state laws and makes available a new data set that can support future evaluations of these laws

Direct observation of the dead-cone effect in quantum chromodynamics

The direct measurement of the QCD dead cone in charm quark fragmentation is reported, using iterative declustering of jets tagged with a fully reconstructed charmed hadron