Template switching between the leading and lagging strands at replication forks generates inverted copy number variants through hairpin-capped extrachromosomal DNA.

Inherited and germ-line de novo copy number variants (CNVs) are increasingly found to be correlated with human developmental and cancerous phenotypes. Several models for template switching during replication have been proposed to explain the generation of these gross chromosomal rearrangements. We proposed a model of template switching (ODIRA-origin dependent inverted repeat amplification) in which simultaneous ligation of the leading and lagging strands at diverging replication forks could generate segmental inverted triplications through an extrachromosomal inverted circular intermediate. Here, we created a genetic assay using split-ura3 cassettes to trap the proposed inverted intermediate. However, instead of recovering circular inverted intermediates, we found inverted linear chromosomal fragments ending in native telomeres-suggesting that a template switch had occurred at the centromere-proximal fork of a replication bubble. As telomeric inverted hairpin fragments can also be created through double strand breaks we tested whether replication errors or repair of double stranded DNA breaks were the most likely initiating event. The results from CRISPR/Cas9 cleavage experiments and growth in the replication inhibitor hydroxyurea indicate that it is a replication error, not a double stranded break that creates the inverted junctions. Since inverted amplicons of the SUL1 gene occur during long-term growth in sulfate-limited chemostats, we sequenced evolved populations to look for evidence of linear intermediates formed by an error in replication. All of the data are compatible with a two-step version of the ODIRA model in which sequential template switching at short inverted repeats between the leading and lagging strands at a replication fork, followed by integration via homologous recombination, generates inverted interstitial triplications.

Recommendations of the Blue Ribbon Committee II for the Optimization of Surgical Education and Training in the United States: The Surgical Trainee Perspective.

OBJECTIVE: This study aims to appraise recommendations from an expert panel of surgical educators on optimizing surgical education and training in the setting of contemporary challenges. BACKGROUND: The Blue Ribbon Committee (BRC II), a group of surgical educators, was convened to make recommendations to optimize surgical training considering the current changes in the landscape of surgical education. Surgical trainees were recruited to assess their impressions of the recommendations. METHODS: A mixed-methods study design was employed, with a survey, followed by focus group interviews. Participating residents and fellows were recruited through a purposeful sampling approach. Descriptive statistics were applied to analyze the survey data, and a thematic data analysis on interview transcripts was employed. RESULTS: The majority of trainee respondents (n=16) thought that all of the subcommittee recommendations should be included in the final BRC II recommendations and paper. According to the interviews, overall, the feedback from the trainees was positive, with particular excitement around work-life integration, education support and faculty development, and funding pitfalls. Some themes about concerns included a lack of clarity about the recommendations, concern about some recommendations being in conflict with one another, and a disconnect between the initial BRC II survey and the subsequent recommendations. CONCLUSIONS: The residents gathered for this focus group were encouraged by the thought, effort, and intention that gathered the surgical leaders across the country to make the recommendations. While there were areas the trainees wanted clarity on, the overall opinion was in agreement with the recommendations.

Infection in osteotomy around the knee: Incidence, management and outcomes in a high-volume case series.

PURPOSE: Current evidence around the management of osteotomy-related infection is insufficient to robustly underpin the expert statements formulated by a recent European consensus statement. We present a review of a large case series in a high-volume osteotomy practice to contribute to the understanding of the incidence, management and outcome of infection in this subspecialty area. METHODS: Analyses of two prospectively collected databases for all osteotomy around the knee and infections related to osteotomy were performed, along with a review of hospital readmission data to capture all osteotomy-related infections. Clinical notes were reviewed to assess patient demographics, incidence of infection, how infection was managed and clinical outcome. RESULTS: In a series of 822 osteotomies in 755 patients, there were 21 (2.8%) cases of suspected infection. Twelve (1.6%) were contemporaneously deemed 'superficial' and nine confirmed 'deep' infections (1.2%). Deep infections were all successfully managed with wound debridement, with or without plate removal, depending on union and time from initial surgery. One of these infections was noted during a revision procedure, but no revision was carried out as a direct result of infection, no external fixation was required and no infected nonunions were experienced. CONCLUSION: All of the cases in this series were managed successfully with debridement ± removal of the plate, without the need for revision or external fixation. Any potential signs of infection around an osteotomy, especially in the case of medial high tibial osteotomy, should raise awareness for deep infection and the need for further surgery due to the limited overlying soft tissue cover. This evidence supports the recent European Society of Sports Traumatology, Knee Surgery and Arthroscopy algorithm. LEVEL OF EVIDENCE: Level IV, case series.

Linking Cardiac Psychology and Cardiovascular Medicine via Self-Determination Theory and Shared Decision-Making.

Despite considerable progress in recent years, research in cardiac psychology is not widely translated into routine practice by clinical cardiologists or clinical health psychologists. Self-determination theory (SDT), which addresses how basic psychological needs of autonomy, competence, and relatedness contribute to the internalization of motivation, may help bridge this research-practice gap through its application to shared decision-making (SDM). This narrative review discusses the following: (a) brief background information on SDT and SDM, (b) the application of SDT to health behavior change and cardiology interventions, and (c) how SDT and SDM may be merged using a dissemination and implementation (D&I) framework. We address barriers to implementing SDM in cardiology, how SDM and SDT address the need for respect of patient autonomy, and how SDT can enhance D&I of SDM interventions through its focus on autonomy, competence, and relatedness and its consideration of other constructs that facilitate the internalization of motivation.

Lessons learnt from COVID-19 to reduce mortality and morbidity in the Global South: addressing global vaccine equity for future pandemics.

COVID-19, which killed more than 6 million people, will not be the last pandemic. Vaccines are key to preventing and ending pandemics. Therefore, it is critical to move now, before the next pandemic, towards global vaccine equity with shared goals, intermediate steps and long-term advocacy goals. Scientific integrity, ethical development, transparency, accountability and communication are critical. Countries can draw on lessons learnt from their response to the HIV pandemics, which has been at the vanguard of ensuring equitable access to rights-based services, to create shared goals and engage communities to increase access to and delivery of safe, quality vaccines. Access can be increased by: fostering the spread of mRNA intellectual property (IP) rights, with mRNA vaccine manufacturing on more continents; creating price transparency for vaccines; creating easily understandable, accessible and transparent data on vaccines; creating demand for a new international legal framework that allows IP rights to be waived quickly once a global pandemic is identified; and drawing on scientific expertise from around the world. Delivery can be improved by: creating strong public health systems that can deliver vaccines through the lifespan; creating or strengthening national regulatory agencies and independent national scientific advisory committees for vaccines; disseminating information from reliable, transparent national and subnational surveillance systems; improving global understanding that as more scientific data become available, this may result in changes to public health guidance; prioritising access to vaccines based on scientific criteria during an epidemic; and developing strategies to vaccinate those at highest risk with available vaccines.

Revision Total Ankle Arthroplasty Using a Novel Modular Fixed-Bearing Revision Ankle System.

INTRODUCTION: Large bone defects such as those encountered after failed total ankle replacement have previously been a relative contraindication to revision ankle replacement due to inadequate bone stock. We describe our experience and patient reported outcomes with a modular ankle replacement system with tibial and talar augments. METHODS: This is a retrospective case series analysis of patients who underwent a total ankle replacement using the INVISION system across 2 centers between 2016 and 2022. Patients completed the Manchester-Oxford Foot Questionnaire (MOXFQ), Ankle Osteoarthritis Scale (AOS), and EQ-5D pre-operatively and then post-operatively at 6 months, 1 year, 2 years, 3 years, and 5 years. Medical records were reviewed for complications and re-operations. X-rays were reviewed for lucencies and alignment. RESULTS: A total of 17 patients were included in the study; 14 men and 3 women with an average age at the time of surgery of 67.9 years (range 56-80 years). The average follow-up post-operatively was 40.5 months (range 7-78) at the time of this study. The indication for surgery was revision of failed total ankle replacement (TAR) in 16 and revision of failed ankle fusion in 1. An augmented tibia was used in 13, an augmented talus in 13, and both augmented tibia and talus in 9 cases. There were no early surgical complications. One patient required debridement and implant retention for late deep infection. No implants have been revised. The average MOXFQ score improved by 19.3 points at most recent follow-up. The average AOS score improved by 25.2 points. CONCLUSION: The early results of a modular augmented ankle arthroplasty system have shown satisfactory patient outcomes with a low complication and re-operation rate and present another option for patients with larger bone defects. This is a small series, and a larger series with long-term follow-up would be beneficial. LEVELS OF EVIDENCE: Level IV: Case series.

Using a quality improvement tool, Plan-Do-Study-Act cycle, to boost TB notification in India post-Covid-19 pandemic.

Quality improvement tools such as the Plan-Do-Study-Act (PDSA) cycle hold tremendous potential to improve the quality of healthcare in India. The electronic-PDSA tool was previously developed by CETI (Collaboration to Eliminate TB among Indians) and successfully piloted in small groups. In this study the e-PDSA was scaled up across the nation over a brief 10 week period to boost TB notification by training District Tuberculosis Officers (DTOs) virtually post-Covid-19 pandemic. Quality improvement counselors, who were interns from Masters in Public Health Institutions, were liaisons to "hand-hold" and assist the DTOs through the PDSA cycle. The course was voluntary and offered to all DTOs through Central TB Division and State TB Officers from May 2022 to July 2022. Of the 779 Districts in India and nearly equal number of DTOs, 546 (70%) DTOs enrolled in the course and of these 437 (80%) conducted a PDSA while 342 (43%) districts/DTOs did not enroll or did not complete a PDSA. With a baseline notification in February-March-April 2022 and intervention in May-June-July 2022; 55% of the districts in the PDSA group showed improvement in TB Notification compared to 45% in the non-PDSA group. When data was analyzed by population (not district) there was a trend in increase in notification post-pandemic in both PDSA and non-PDSA groups, and the PDSA group had a slightly higher 5.6 per 100,000 population improvement compared to 5.0 per 100,000 in the non-PDSA group. The cost of intervention was $40,000 or $92 per DTO for three months. The course was highly acceptable with DTOs rating 4.3 out of 5 in recommending the course to other DTOs. Our data shows that a large scale-up of the PDSA cycle is feasible, economical and effective with little additional resources. The focus was on increasing the efficiency of the existing processes well within the authority of the DTO. Repeat cycle of PDSA with notification and other measures such as presumptive sputum examination could significantly impact the program and help to achieve TB Free India.

Adipose tissue-selective ablation of ADAM10 results in divergent metabolic phenotypes following long-term dietary manipulation.

A Disintegrin And Metalloproteinase domain-containing protein 10 (ADAM10), is involved in several metabolic and inflammatory pathways. We speculated that ADAM10 plays a modulatory role in adipose tissue inflammation and metabolism. To this end, we studied adipose tissue-specific ADAM10 knock-out mice (aKO). While young, regular chow diet-fed aKO mice showed increased insulin sensitivity, following prolonged (33 weeks) high-fat diet (HFD) exposure, aKO mice developed obesity and insulin resistance. Compared to controls, aKO mice showed less inflammatory adipokine profile despite the significant increase in adiposity. In brown adipose tissue, aKO mice on HFD had changes in CD8+ T cell populations indicating a lesser inflammatory pattern. Following HFD, both aKO and control littermates demonstrated decreased adipose tissue pro-inflammatory macrophages, and increased anti-inflammatory accumulation, without differences between the genotypes. Collectively, our observations indicate that selective deletion of ADAM10 in adipocytes results in a mitigated inflammatory response, leading to increased insulin sensitivity in young mice fed with regular diet. This state of insulin sensitivity, following prolonged HFD, facilitates energy storage resulting in increased fat accumulation which ultimately leads to the development of a phenotype of obesity and insulin resistance. In conclusion, the data indicate that ADAM10 has a modulatory effect of inflammation and whole-body energy metabolism.

Enhancing research culture through PhD training: a systems approach to identifying leverage points for policy formation.

This article examines the role of PhD training programmes in identifying and implementing positive interventions in research culture in the biosciences. Using a data set consisting of transcripts from interviews and group discussions with 179 participants from 18 of the current 23 (78%) UK-based Wellcome-funded PhD programmes, we apply a systems theory methodology to the system of higher education and PhD training. Using system mapping as an investigative tool, this approach identifies points of leverage within the system where policy interventions might be best targeted to affect changes to research culture in the global higher education sector. The results of this investigation highlight the student-supervisor relationship as a nexus for these interventions and recommends the programme structure as a global policy for PhD training.

Association between Self-reported Sleep Quality and Single-task Gait in Young Adults: A Study Using Machine Learning.

Objective The objective of the present study was to find biomechanical correlates of single-task gait and self-reported sleep quality in a healthy, young population by replicating a recently published study. Materials and Methods Young adults ( n = 123) were recruited and were asked to complete the Pittsburgh Sleep Quality Inventory to assess sleep quality. Gait variables ( n = 53) were recorded using a wearable inertial measurement sensor system on an indoor track. The data were split into training and test sets and then different machine learning models were applied. A post-hoc analysis of covariance (ANCOVA) was used to find statistically significant differences in gait variables between good and poor sleepers. Results AdaBoost models reported the highest correlation coefficient (0.77), with Support-Vector classifiers reporting the highest accuracy (62%). The most important features associated with poor sleep quality related to pelvic tilt and gait initiation. This indicates that overall poor sleepers have decreased pelvic tilt angle changes, specifically when initiating gait coming out of turns (first step pelvic tilt angle) and demonstrate difficulty maintaining gait speed. Discussion The results of the present study indicate that when using traditional gait variables, single-task gait has poor accuracy prediction for subjective sleep quality in young adults. Although the associations in the study are not as strong as those previously reported, they do provide insight into how gait varies in individuals who report poor sleep hygiene. Future studies should use larger samples to determine whether single task-gait may help predict objective measures of sleep quality especially in a repeated measures or longitudinal or intervention framework.