mafba and mafbb differentially regulate lymphatic endothelial cell migration in topographically distinct manners.

Lymphangiogenesis, formation of lymphatic vessels from pre-existing vessels, is a dynamic process that requires cell migration. Regardless of location, migrating lymphatic endothelial cell (LEC) progenitors probe their surroundings to form the lymphatic network. Lymphatic-development regulation requires the transcription factor MAFB in different species. Zebrafish Mafba, expressed in LEC progenitors, is essential for their migration in the trunk. However, the transcriptional mechanism that orchestrates LEC migration in different lymphatic endothelial beds remains elusive. Here, we uncover topographically different requirements of the two paralogs, Mafba and Mafbb, for LEC migration. Both mafba and mafbb are necessary for facial lymphatic development, but mafbb is dispensable for trunk lymphatic development. On the molecular level, we demonstrate a regulatory network where Vegfc-Vegfd-SoxF-Mafba-Mafbb is essential in facial lymphangiogenesis. We identify that mafba and mafbb tune the directionality of LEC migration and vessel morphogenesis that is ultimately necessary for lymphatic function.

Using coaching and action learning to support staff leadership development.

During the coronavirus disease 2019 pandemic, nurse leaders and managers have been compelled to prioritise immediate issues in their clinical areas and put aside the professional development of staff. However, leadership development for individuals and teams is essential to ensure nurses feel valued and develop the skills required for team cohesion, problem-solving, decision-making and innovation. Simple and effective approaches to staff leadership development are needed. Two such approaches are coaching and action learning. This article provides an introduction to coaching and action learning as approaches nurse leaders and managers can use to promote leadership development among individual team members and within the team. It describes how coaching and action learning work and their potential benefits and challenges. It explains how the two approaches can be used to underpin effective problem-solving and goal setting, and support nurses in their professional development, the ultimate aim being to deliver safe and effective patient care.

The RNA helicase Ddx21 controls Vegfc-driven developmental lymphangiogenesis by balancing endothelial cell ribosome biogenesis and p53 function.

The development of a functional vasculature requires the coordinated control of cell fate, lineage differentiation and network growth. Cellular proliferation is spatiotemporally regulated in developing vessels, but how this is orchestrated in different lineages is unknown. Here, using a zebrafish genetic screen for lymphatic-deficient mutants, we uncover a mutant for the RNA helicase Ddx21. Ddx21 cell-autonomously regulates lymphatic vessel development. An established regulator of ribosomal RNA synthesis and ribosome biogenesis, Ddx21 is enriched in sprouting venous endothelial cells in response to Vegfc-Flt4 signalling. Ddx21 function is essential for Vegfc-Flt4-driven endothelial cell proliferation. In the absence of Ddx21, endothelial cells show reduced ribosome biogenesis, p53 and p21 upregulation and cell cycle arrest that blocks lymphangiogenesis. Thus, Ddx21 coordinates the lymphatic endothelial cell response to Vegfc-Flt4 signalling by balancing ribosome biogenesis and p53 function. This mechanism may be targetable in diseases of excessive lymphangiogenesis such as cancer metastasis or lymphatic malformation.

Co-developing and implementing a community nursing simulated learning resource for undergraduate nursing students.

Policy agendas across the developed world privilege systems transformation, notably shifting the balance of service provision from hospital to community settings. Primary and community health services have pivotal roles in the United Kingdom's longstanding policy ambition of healthcare services transformation, and it is imperative that undergraduate nursing students comprehend community settings as valuable learning environments, places of care and community nurses' roles therein. However, limited community placement learning opportunities means nursing students may be inadequately prepared to work in community settings at the point of initial registration. High fidelity simulated learning is well established within undergraduate nursing curricula. Currently, this learning approach predominantly focuses on acute and secondary care nursing and takes place in simulation centres. Within undergraduate nursing curricula there is limited evidence to support that community-based simulation is utilised with the result that students may struggle to recognise the value of learning opportunities in the community environment. This underpins the rationale for the development of this educational initiative which reflects current health policy agenda of care being provided closer to home. In addition, the new pre-registration 'Future nurse: Standards of proficiencies' (NMC, 2019) acknowledge that registered nurses need to be able to care for people in their own home and in the community setting. This paper describes the co-development and implementation of a pilot, community nursing focused, simulated learning resource within an undergraduate adult field nursing programme in Wales, UK. TWEETABLE ABSTRACT: How one university co-developed and implemented a community focused simulation resource for undergraduate nursing students.

[Surgery during pregnancy - a taboo?] / Operieren in der Schwangerschaft ­ ein Tabu?

When a pregnancy is announced, the normal working day is transformed for the urologists concerned. For fear of legal consequences, pregnant employees are still hardly allowed to operate. However, the new provisions of the Maternity Protection Act do not fundamentally exclude pregnant urologists from working and operating. Why do hospitals nevertheless allow such a deplorable state of affairs, which endangers the daily work routine and makes further training of pregnant urologists more difficult?

[Work and Training of Young Doctors in Germany - a Comprehensive Analysis of Survey Results from Six Disciplines]. / Die Arbeits- und Weiterbildungssituation junger Ärztinnen und Ärzte in Deutschland ­ eine zusammenfassende Analyse von Befragungsergebnissen aus sechs Fachrichtungen.

OBJECTIVE: The working and training conditions of young physicians in Germany have changed over the last few years, as a result of far-reaching changes in the healthcare system. Therefore, Germany-wide surveys among young physicians of several disciplines were evaluated in a pooled analysis, in order to obtain a current interdisciplinary impression of conflicts in their daily work. MATERIAL AND METHODS: Data from web-based surveys from residents training in six disciplines were analyzed together retrospectively. One focus was a gratification crises model for the assessment of psychosocial workload. RESULTS: Data on 4041 participants were evaluated. In day-to-day work, young physicians were burdened with a high proportion of tasks that were not directly medical. Instruments of good subspecialty training, such as training contracts, curricula and regular feedback were associated with a lower psychosocial workload, which was generally significant among the participants. An economic influence on medical-professional decisions was subjectively clearly present among the participants. CONCLUSION: Many young doctors find the current work and training conditions in the medical work environment unsatisfactory. This might have consequences not only for the doctors themselves, but also for the patients they care for. A healthy work environment with health professionals in good health is therefore crucial and should be in everyone's interest.

Improved PCR diagnostics using up-to-date in silico validation: An F-gene RT-qPCR assay for the detection of all four lineages of peste des petits ruminants virus.

Peste des petits ruminants (PPR) is a globally significant disease of small ruminants caused by the peste des petits ruminants virus (PPRV) that is considered for eradication by 2030 by the United Nations Food and Agriculture Organisation (FAO). Critical to the eradication of PPR are accurate diagnostic assays. RT-qPCR assays targeting the nucleocapsid gene of PPRV have been successfully used for the diagnosis of PPR. We describe the development of an RT-qPCR assay targeting an alternative region (the fusion (F) gene) based on the most up-to-date PPRV sequence data. In silico analysis of the F-gene RT-qPCR assay performed using PCRv software indicated 98% sensitivity and 100% specificity against all PPRV sequences published in Genbank. The assay indicated the greatest in silico sensitivity in comparison to other previously published and recommended PPRV RT-qPCR assays. We evaluated the assay using strains representative of all 4 lineages in addition to samples obtained from naturally and experimentally-infected animals. The F-gene RT-qPCR assay showed 100% diagnostic specificity and demonstrated a limit of detection of 10 PPRV genome copies per µl. This RT-qPCR assay can be used in isolation or in conjunction with other assays for confirmation of PPR and should support the global efforts for eradication.

A rapid RT-LAMP assay for the detection of all four lineages of Peste des Petits Ruminants Virus.

Peste des petits ruminants (PPR) is a viral disease of small ruminants that is caused by the PPR virus (PPRV) and is a significant burden on subsistence farmers across the developing world. Loop-mediated isothermal amplification (LAMP) provides cost-effective, rapid, specific and sensitive detection of nucleic acid and has been demonstrated to have field application for a range of viruses. We describe the development of a novel PPRV RT-LAMP assay utilising carefully-selected primers (targeting the N-gene) allowing for the detection of all known PPRV lineages in < 20 min. The assay was evaluated in comparison with a "gold standard" real-time RT-PCR assay using more than 200 samples, comprising samples from recent PPRV outbreaks, experimentally-infected goats, well-characterised cell culture isolates and samples collected from uninfected animals. The RT-LAMP assay demonstrated 100% diagnostic specificity and greater than 97% diagnostic sensitivity in comparison with the real-time RT-PCR assay. The limit of detection was between 0.3 and 0.8 log10 TCID50 ml-1 equating to a CT value of 31.52 to 33.48. In experimentally-infected animals, the RT-LAMP could detect PPRV as early as 4 days post infection (dpi) - before clinical signs were observed at 7 dpi. The RT-LAMP assay can support the global PPR eradication campaign.

Training, Research, and Working Conditions for Urology Residents in Germany: A Contemporary Survey.

BACKGROUND: Excellent uniform training of urology residents is crucial to secure both high-quality patient care and the future of our specialty. Residency training has come under scrutiny following the demands of subspecialized care, economical aspects, and working hour regulations. OBJECTIVE: To comprehensively assess the surgical training, research opportunities, and working conditions among urology residents in Germany. DESIGN, SETTING, AND PARTICIPANTS: We sent a 29-item online survey via email to 721 members of the German Society of Residents in Urology. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Descriptive analyses were conducted to describe the surveys' four domains: (1) baseline characteristics, (2) surgical training (cumulative completed case volume for all minor-, medium-, and major-complexity surgeries), (3) research opportunities, and (4) working conditions. RESULTS AND LIMITATIONS: Four hundred and seventy-two residents completed the online survey (response rate 65%). Surgical training: the median number of cumulative completed cases for postgraduate yr (PGY)-5 residents was 113 (interquartile range: 76-178). Minor surgeries comprised 57% of all surgeries and were performed by residents in all PGYs. Medium-complexity surgeries comprised 39% of all surgeries and were mostly performed by residents in PGYs 2-5. Major surgeries comprised 4% of all surgeries and were occasionally performed by residents in PGYs 3-5. Research opportunities: some 44% have attained a medical thesis (Dr. med.), and 39% are currently pursuing research. Working conditions: psychosocial work-related stress was high and for 82% of residents their effort exceeded their rewards. Some 44% were satisfied, 32% were undecided, and 24% were dissatisfied with their current working situation. Limitations include self-reported survey answers and a lack of validated assessment tools. CONCLUSIONS: Surgical exposure among German urology residents is low and comprises minor and medium-complex surgeries. Psychosocial work-related stress is high for the vast majority of residents indicating the need for structural improvements in German urology residency training. PATIENT SUMMARY: In this study, we evaluated the surgical training, research opportunities, and working conditions among urology residents in Germany. We found low surgical exposure and high rates for psychosocial work-related stress, indicating the need for structural improvements in German urology residency training.

Results of the TOP Study: Prospectively Randomized Multicenter Trial of an Ex Vivo Tacrolimus Rinse Before Transplantation in EDC Livers.

BACKGROUND: Organ shortage results in the transplantation of extended donor criteria (EDC) livers which is associated with increased ischemia-reperfusion injury (IRI). Experimental studies indicate that an organ rinse with the calcineurin inhibitor tacrolimus before implantation protects against IRI. The tacrolimus organ perfusion study was initiated to examine the effects of ex vivo tacrolimus perfusion on IRI in transplantation of EDC livers. METHODS: A prospective randomized multicenter trial comparing ex vivo perfusion of marginal liver grafts (≥2 EDC according to Eurotransplant manual) with tacrolimus (20 ng/mL) or histidine-tryptophane-ketoglutarate solution (control) was carried out at 5 German liver transplant centers (Munich Ludwig-Maximilians University, Berlin, Heidelberg, Mainz, Regensburg) between October 2011 and July 2013. Primary endpoint was the maximum alanine transaminase (ALT) level within 48 hours after transplantation. Secondary endpoints were aspartate transaminase (AST), prothrombine ratio, and graft-patient survival within an observation period of 1 week. After an interim analysis, the study was terminated by the scientific committee after the treatment of 24 patients (tacrolimus n = 11, Control n = 13). RESULTS: Tacrolimus rinse did not reduce postoperative ALT peaks compared with control (P = 0.207; tacrolimus: median, 812; range, 362-3403 vs control: median, 652; range, 147-2034). Moreover, ALT (P = 0.100), prothrombine ratio (P = 0.553), and bilirubin (P = 0.815) did not differ between the groups. AST was higher in patients treated with tacrolimus (P = 0.011). Survival was comparable in both groups (P > 0.05). CONCLUSIONS: Contrary to experimental findings, tacrolimus rinse failed to improve the primary endpoint of the study (ALT). Because 1 secondary endpoint (AST) was even higher in the intervention group, the study was terminated prematurely. Thus, tacrolimus rinse cannot be recommended in transplantation of EDC livers.

Combined use of toll-like receptor agonists and prostaglandin E(2) in the FastDC model: rapid generation of human monocyte-derived dendritic cells capable of migration and IL-12p70 production.

Phenotypical maturation, IL-12p70 production and migration upon chemokine receptor CCR7 ligation are currently proposed as requirements for the use of human monocyte-derived dendritic cells (DC) in antitumoral vaccination. We have previously described a short-term protocol for DC generation from monocytes including stimulation with TNF-alpha, IL-1beta and PGE(2) (FastDC). These "conventional" FastDC are mature, migrate in response to CCR7 ligation and effectively stimulate autologeous T cells in vitro, but are deficient in IL-12p70 production. Here, conventional FastDC were compared to FastDC activated with different TLR ligands. High levels of IL-12p70 were induced by combined activation of FastDC with TLR4 and TLR7/8 ligands. IL-12 secretion could be maximized by additional T cell-derived stimulation. However, TLR-stimulated FastDC failed to migrate upon CCR7 ligation, independent of additional activation with CD40 ligand and IFN-gamma. The presence of PGE(2) during TLR ligation fully restored migratory capacity of FastDC, but left IL-12p70 production and activation of tumor antigen-specific cytotoxic T cells unaffected, challenging previous findings obtained with standard 7-day monocyte-derived DC. The FastDC model thus not only represents an effective tool for antitumoral vaccination, but may also provide novel insights into human DC biology.