Since the discovery two decades ago that transgenes are efficiently integrated into the genome by homologous recombination in the moss Physcomitrella patens, it has been a premier model system to study evolutionary development (evo-devo) questions, stem cell reprogramming, as well as explore the biology of non-vascular plants. P. patens was the first non-seed plant to have its genome sequenced and with this level of genomic information together with increasing molecular genetic tools, a large number of reverse genetic studies have propelled the use of this model system. However, a number of technological advances have recently opened the doors to forward genetics as well as extremely efficient and precise genome editing. Additionally, careful phylogenetic studies with increased resolution suggest that P. patens emerged from within Physcomitrium Thus, rather than Physcomitrella patens the species should be named Physcomitrium patens Here we review these advances and describe the areas where P. patens has had the most impact on plant biology. © 2020 American Society of Plant Biologists. All rights reserved.BACKGROUND Despite the recent increase in the number of publications on diagnostic cerebral angiograms using transradial access (TRA), there have been relatively few regarding TRA for neurointerventional cases. Questions of feasibility and safety may still exist among physicians considering TRA for neurointerventional procedures. METHODS A systematic literature review was performed following PRISMA guidelines. Three online databases (MedLine via PubMed, Scopus and Embase) were searched for articles published between January 2000 and December 2019. Search terms included "Transradial access", "Radial Access", "Radial artery" AND "Neurointerventions". The reference lists of selected articles and pertinent available non-systematic analysis were reviewed for other potential citations. Primary outcomes measured were access site complications and crossover rates. RESULTS Twenty-one studies (n=1342 patients) were included in this review. Two of the studies were prospective while the remaining 19 were retrospective. Seuroendovascular devices, and the continued reports of its success in the literature, TRA is expected to become more widely used by neurointerventionalists. © Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.Mouse spermatogenesis is supported by spermatogenic stem cells (SSCs). https://www.selleckchem.com/products/ng25.html SSCs maintain their pool while migrating over an open (or facultative) niche microenvironment of testicular seminiferous tubules, where ligands that support self-renewal are likely distributed widely. This contrasts with the classic picture of closed (or definitive) niches in which stem cells are gathered and the ligands are highly localized. Some of the key properties observed in the dynamics of SSCs in the testicular niche in vivo, which show the flexible and stochastic (probabilistic) fate behaviors, are found to be generic for a wide range of, if not all, tissue stem cells. SSCs also show properties characteristic of an open niche-supported system, such as high motility. Motivated by the properties of SSCs, in this review, I will reconsider the potential unity and diversity of tissue stem cell systems, with an emphasis on the varying degrees of ligand distribution and stem cell motility. Copyright © 2020 Cold Spring Harbor Laboratory Press; all rights reserved.Microtubules dynamics is regulated by the plus end-tracking proteins (+TIPs) in cells. End binding protein 1 (EB1) acts as a master regulator in +TIPs networks by targeting microtubule growing ends and recruiting other factors. However, the molecular mechanism of how EB1 binds to microtubule ends with a high affinity remains to be an open question. Using single-molecule imaging, we show that the end-binding kinetics of EB1 changes along with the polymerizing and hydrolysis rate of tubulin dimers, confirming the binding of EB1 to GTP/GDP-Pi tubulin at microtubule growing ends. The affinity of wild-type EB1 to these sites is higher than monomeric EB1 mutants, suggesting that two CH domains in the dimer contribute to the end-binding. Introducing phosphomimicking mutations into the linker domain of EB1 weakens the end-binding affinity and confers a more curved conformation to EB1 dimer without compromising dimerization, suggesting that the overall architecture of EB1 is important for the end-binding affinity. Taken together, our results provide insights into understanding how the high-affinity end-binding of EB1 can be achieved and how this activity may be regulated in cells. © 2020. Published by The Company of Biologists Ltd.Podosomes are actin-based adhesion and invasion structures in a variety of cell types, with podosome-forming cells displaying up to several hundreds of these structures. Podosome number, distribution and composition can be affected by experimental treatments or during regular turnover, necessitating a tool that is able to detect even subtle differences in podosomal properties. Here, we present a Fiji-based macro code termed "Poji" ("podosome analysis by Fiji"), which serves as an easy-to-use tool to characterise a variety of cellular and podosomal parameters including area, fluorescence intensity, relative enrichment of associated proteins, and radial podosome intensity profiles. This tool should be useful to gain more detailed insight into regulation, architecture and functions of podosomes. Moreover, we show that Poji is easily adaptable for the analysis of invadopodia and associated extracellular matrix degradation, and likely also of other micron-size punctate structures. This article describes the work flow of the Poji macro, presents several examples of its applications, and also points out limitations, as well as respective solutions, and adaptable features to streamline the analysis. © 2020. Published by The Company of Biologists Ltd.The mechanisms that control intrinsic axon growth potential, and thus axon regeneration following injury, are not well understood. Developmental axon regrowth of Drosophila mushroom body γ neurons during neuronal remodeling offers a unique opportunity to study the molecular mechanisms controlling intrinsic growth potential. Motivated by the recently uncovered developmental expression atlas of γ neurons, we here focus on the role of the actin severing protein cofilin during axon regrowth. We show that Twinstar (Tsr), the fly cofilin, is a crucial regulator of both axon growth and branching during developmental remodeling of γ neurons. tsr mutant axons demonstrate growth defects both in vivo and in vitro and also exhibit actin rich filopodial-like structures at failed branch points in vivo Our data is inconsistent with Tsr being important for increasing G-actin availability. Furthermore, analysis of microtubule localization suggests that Tsr is required for microtubule infiltration into the axon tips and branch points. Taken together, we show that Tsr promotes axon growth and branching, likely by clearing F-actin to facilitate microtubules protrusion. © 2020. Published by The Company of Biologists Ltd.To gain a comprehensive view of the changes in host gene expression underlying Zika virus (ZIKV) pathogenesis, we performed whole-genome mRNAseq of ZIKV infected Drosophila adult flies. RNA-seq analysis revealed that ZIKV infection alters several and diverse biological processes including stress, locomotion, lipid metabolism, imaginal disc morphogenesis and regulation of JAK/STAT signaling, To explore the interaction between ZIKV infection and JAK/STAT signaling regulation, we generated genetic constructs overexpressing ZIKV-specific non-structural proteins NS2A, NS2B, NS4A and NS4B. We find that ectopic expression of non-structural proteins in the developing Drosophila eye significantly restricts growth of the larval and adult eye and correlates with a considerable repression of the in vivo JAK/STAT reporter, 10XStat92E-GFP At the cellular level, eye growth defects are associated with reduced rate of proliferation without affecting the overall rate of apoptosis. In addition, ZIKV NS4A genetically interacts with the JAK/STAT signaling components; co-expression of NS4A along with dominant negative form of domeless or StatRNAi results in aggravated reduction in eye size while co-expression of NS4A in HopTuml mutant background partially rescues the Hop-induced eye overgrowth phenotype. The function of ZIKV NS4A in regulating growth is maintained in the wing, where ZIKV NS4A overexpression in the pouch domain results in reduced growth linked with diminished expression of Notch targets, Wingless and Cut and the Notch reporter, NRE-GFP Thus, our study provides evidence that ZIKV infection in Drosophila results in restricted growth of the developing eye and wing, wherein eye phenotype is induced through regulation of JAK/STAT signaling while restricted wing growth is through regulation of Notch signaling. The interaction of ZIKV non-structural proteins with the conserved host signaling pathways further advance our understanding of ZIKV-induced pathogenesis. © 2020. Published by The Company of Biologists Ltd.The circadian clock provides a time-keeping mechanism for synchronizing various biological activities with the surrounding environment. Arabidopsis CIRCADIAN CLOCK ASSOCIATED1 (CCA1), which encodes a Myb-related transcription factor, is a key component of the core oscillator of Arabidopsis circadian clock with peaking expression in the morning. The molecular mechanisms regulating light induction and rhythmic expression of CCA1 still remain elusive. In this study, we show that two phytochrome signal transducers, FAR-RED ELONGATED HYPOCOTYL3 (FHY3) and its paralogue FAR-RED IMPAIRED RESPONSE1 (FAR1) are essential for light induction of CCA1 expression by direct binding to the promoter of CCA1 and activating its expression, whereas PHYTOCHROME INTERACTING FACTOR 5 (PIF5) directly binds to the promoter of CCA1 to repress its expression. Further, PIF5 and TIMING OF CAB EXPRESSION1 (TOC1) physically interact with FHY3 and FAR1 to repress the transcriptional activation activity of FHY3 and FAR1 on CCA1 expression. Our combined results demonstrate that the photosensory signaling pathway integrates with the circadian oscillators to orchestrate clock gene expression, which may form the molecular basis of light regulation of clock system in response to daily changes in the light environment, thus increasing plant fitness. © 2020 American Society of Plant Biologists. All rights reserved.BACKGROUND Data on discomfort and complications from research bronchoscopy in chronic obstructive pulmonary disease (COPD) and asthma is limited. We present complications and discomfort occurring within a week after bronchoscopy, and investigate personal and procedural risk factors. METHODS 239 subjects with COPD, asthma or without lung disease underwent research bronchoscopies as part of a microbiome study of the lower airways (the MicroCOPD study). Bronchoscopy was done in the supine position with oral scope insertion with the option of light conscious alfentanil sedation. Sampling consisted of protected specimen brushes, bronchoalveolar lavage, small volume lavage and for some, endobronchial biopsies. Bleeding, desaturation, cough, haemodynamic changes, dyspnoea and other events that required an unplanned intervention or early termination of bronchoscopy were prospectively recorded. Follow-up consisted of a telephone interview where subjects rated discomfort and answered questions about fever sensation and respiratory symptoms in the week following bronchoscopy.