bluuebunny/crossref_metadata_2025 huggingface dataset:
Quantitative imaging of cell dynamics in mouse embryos using light-sheet microscopy
Authors: Udan Ryan S. , Piazza Victor G. , Hsu Chih-wei , Hadjantonakis Anna-Katerina , Dickinson Mary E.
Published: October 2014

Single/selective-plane illumination, or light-sheet, systems offer several advantages over other fluorescence microscopy methods for live, 3D microscopy. These systems are valuable for studying embryonic development in several animal systems, such as Drosophila, C. elegans and zebrafish. The geometry of the light path in this form of microscopy requires the sample to be accessible from multiple sides and fixed in place so that it can be rotated around a single axis. Popular methods for mounting include hanging the specimen from a pin or embedding it in 1-2% agarose. These methods can be particularly problematic for certain samples, such as post-implantation mouse embryos, that expand significantly in size and are very delicate and sensitive to mounting. To overcome the current limitations and to establish a robust strategy for long-term (24 h) time-lapse imaging of E6.5-8.5 mouse embryos with light-sheet microscopy, we developed and tested a method using hollow agarose cylinders designed to accommodate for embryonic growth, yet provide boundaries to minimize tissue drift and enable imaging in multiple orientations. Here, we report the first 24-h time-lapse sequences of post-implantation mouse embryo development with light-sheet microscopy. We demonstrate that light-sheet imaging can provide both quantitative data for tracking changes in morphogenesis and reveal new insights into mouse embryogenesis. Although we have used this approach for imaging mouse embryos, it can be extended to imaging other types of embryos as well as tissue explants.
The polycomb component Ring1B regulates the timed termination of subcerebral projection neuron production during mouse neocortical development
Authors: Morimoto-Suzki Nao , Hirabayashi Yusuke , Tyssowski Kelsey , Shinga Jun , Vidal Miguel , Koseki Haruhiko , Gotoh Yukiko
Published: October 2014

In the developing neocortex, neural precursor cells (NPCs) sequentially generate various neuronal subtypes in a defined order. Although the precise timing of the NPC fate switches is essential for determining the number of neurons of each subtype and for precisely generating the cortical layer structure, the molecular mechanisms underlying these switches are largely unknown. Here, we show that epigenetic regulation through Ring1B, an essential component of polycomb group (PcG) complex proteins, plays a key role in terminating NPC-mediated production of subcerebral projection neurons (SCPNs). The level of histone H3 residue K27 trimethylation at and Ring1B binding to the promoter of Fezf2, a fate determinant of SCPNs, increased in NPCs as Fezf2 expression decreased. Moreover, deletion of Ring1B in NPCs, but not in postmitotic neurons, prolonged the expression of Fezf2 and the generation of SCPNs that were positive for CTIP2. These results indicate that Ring1B mediates the timed termination of Fezf2 expression and thereby regulates the number of SCPNs.
The zinc-finger protein basonuclin 2 is required for proper mitotic arrest, prevention of premature meiotic initiation and meiotic progression in mouse male germ cells
Authors: Vanhoutteghem Amandine , Messiaen Sébastien , Hervé Françoise , Delhomme Brigitte , Moison Delphine , Petit Jean-Maurice , Rouiller-Fabre Virginie , Livera Gabriel , Djian Philippe
Published: October 2014

Absence of mitosis and meiosis are distinguishing properties of male germ cells during late fetal and early neonatal periods. Repressors of male germ cell meiosis have been identified, but mitotic repressors are largely unknown, and no protein repressing both meiosis and mitosis is known. We demonstrate here that the zinc-finger protein BNC2 is present in male but not in female germ cells. In testis, BNC2 exists as several spliced isoforms and presumably binds to DNA. Within the male germ cell lineage, BNC2 is restricted to prospermatogonia and undifferentiated spermatogonia. Fetal prospermatogonia that lack BNC2 multiply excessively on embryonic day (E)14.5 and reenter the cell cycle prematurely. Mutant prospermatogonia also engage in abnormal meiosis; on E17.5, Bnc2−/− prospermatogonia start synthesizing the synaptonemal protein SYCP3, and by the time of birth, many Bnc2−/− prospermatogonia have accumulated large amounts of nonfilamentous SYCP3, thus appearing to be blocked at leptonema. Bnc2−/− prospermatogonia do not undergo proper male differentiation, as they lack almost all the mRNA for the male-specific methylation protein DNMT3L and have increased levels of mRNAs that encode meiotic proteins, including STRA8. Bnc2−/− prospermatogonia can produce spermatogonia, but these enter meiosis prematurely and undergo massive apoptotic death during meiotic prophase. This study identifies BNC2 as a major regulator of male germ stem cells, which is required for repression of meiosis and mitosis in prospermatogonia, and for meiosis progression during spermatogenesis. In view of the extreme evolutionary conservation of BNC2, the findings described here are likely to apply to many species.
Specification of neuronal subtypes by different levels of Hunchback
Authors: Moris-Sanz Marta , Estacio-Gómez Alicia , Álvarez-Rivero Javier , Díaz-Benjumea Fernando J.
Published: October 2014

During the development of the central nervous system, neural progenitors generate an enormous number of distinct types of neuron and glial cells by asymmetric division. Intrinsic genetic programs define the combinations of transcription factors that determine the fate of each cell, but the precise mechanisms by which all these factors are integrated at the level of individual cells are poorly understood. Here, we analyzed the specification of the neurons in the ventral nerve cord of Drosophila that express Crustacean cardioactive peptide (CCAP). There are two types of CCAP neurons: interneurons and efferent neurons. We found that both are specified during the Hunchback temporal window of neuroblast 3-5, but are not sibling cells. Further, this temporal window generates two ganglion mother cells that give rise to four neurons, which can be identified by the expression of empty spiracles. We show that the expression of Hunchback in the neuroblast increases over time and provide evidence that the absolute levels of Hunchback expression specify the two different CCAP neuronal fates.
Induction of diverse cardiac cell types by reprogramming fibroblasts with cardiac transcription factors
Authors: Nam Young-Jae , Lubczyk Christina , Bhakta Minoti , Zang Tong , Fernandez-Perez Antonio , McAnally John , Bassel-Duby Rhonda , Olson Eric N. , Munshi Nikhil V.
Published: October 2014

Various combinations of cardiogenic transcription factors, including Gata4 (G), Hand2 (H), Mef2c (M) and Tbx5 (T), can reprogram fibroblasts into induced cardiac-like myocytes (iCLMs) in vitro and in vivo. Given that optimal cardiac function relies on distinct yet functionally interconnected atrial, ventricular and pacemaker (PM) cardiomyocytes (CMs), it remains to be seen which subtypes are generated by direct reprogramming and whether this process can be harnessed to produce a specific CM of interest. Here, we employ a PM-specific Hcn4-GFP reporter mouse and a spectrum of CM subtype-specific markers to investigate the range of cellular phenotypes generated by reprogramming of primary fibroblasts. Unexpectedly, we find that a combination of four transcription factors (4F) optimized for Hcn4-GFP expression does not generate beating PM cells due to inadequate sarcomeric protein expression and organization. However, applying strict single-cell criteria to GHMT-reprogrammed cells, we observe induction of diverse cellular phenotypes, including those resembling immature forms of all three major cardiac subtypes (i.e. atrial, ventricular and pacemaker). In addition, we demonstrate that cells induced by GHMT are directly reprogrammed and do not arise from an Nxk2.5+ progenitor cell intermediate. Taken together, our results suggest a remarkable degree of plasticity inherent to GHMT reprogramming and provide a starting point for optimization of CM subtype-specific reprogramming protocols.
Clustered phosphatidylinositol 4,5 bisphosphate accumulation and ezrin phosphorylation in response to CLIC5A
Authors: Al-Momany Abass , Li Laiji , Alexander R. Todd , Ballermann Barbara
Published: October 2014

CLIC5A is a component of the ezrin-NHERF2-podocalyxin complex in renal glomerular podocyte foot processes. We explored the mechanism(s) by which CLIC5A regulates ezrin function. In COS-7 cells, CLIC5A augmented ezrin phosphorylation without changing ezrin abundance, increased the association of ezrin with the cytoskeletal fraction, enhanced actin polymerization and the formation of cell surface projections. CLIC5A caused the PI(4,5)P2 reporter RFP-PH-PLC to translocate from cytosol to discrete plasma membrane clusters at the cell surface, where it co-localized with CLIC5A. Transiently expressed HA-PIP5Kα co-localized with GFP-CLIC5A and was pulled from cell lysates by GST-CLIC5A, and silencing of endogenous PIP5Kα abrogated CLIC5A-dependent ERM phosphorylation. N- and C-terminal deletion mutants of CLIC5A, which failed to associate with the plasma membrane failed to co-localize with PIP5Kα, did not alter the abundance of PI(4,5)P2 plasma membrane clusters and failed to enhance ezrin phosphorylation. Relative to wild-type mice, in CLIC5 deficient mice glomerular ezrin phosphorylation was diminished and the cytoskeletal association of both ezrin and NHERF2 was reduced. Therefore, the mechanism of CLIC5A action involves clustered plasma membrane PI(4,5)P2 accumulation through an interaction of CLIC5A with PI(4,5)P2 generating kinases, in turn facilitating ezrin activation, and actin-dependent cell surface remodeling.
Podosome-regulating kinesin KIF1C translocates to the cell periphery in a CLASP-dependent manner
Authors: Efimova Nadia , Grimaldi Ashley , Bachmann Alice , Frye Keyada , Zhu Xiaodong , Feoktistov Alexander , Straube Anne , Kaverina Irina
Published: October 2014

The kinesin KIF1C is known to regulate podosomes, actin-rich adhesion structures, which remodel the extracellular matrix during physiological processes. Here we show that KIF1C is a player in the podosome-inducing signaling cascade. Upon induction of podosome formation by protein kinase C, KIF1C translocation to the cell periphery intensifies and KIF1C accumulates in the proximity of peripheral microtubules enriched with plus tip-associated proteins CLASPs and around podosomes. Importantly, without CLASPs, both KIF1C trafficking and podosome formation are suppressed. Moreover, chimeric mitochondria-targeted CLASP2 recruits KIF1C, suggesting a transient CLASP-KIF1C association. We propose that CLASP creates preferred microtubule tracks for KIF1C to promote podosome induction downstream of PKC.
CLIC4 regulates cell adhesion and β1 integrin trafficking
Authors: Argenzio Elisabetta , Margadant Coert , Leyton-Puig Daniela , Janssen Hans , Jalink Kees , Sonnenberg Arnoud , Moolenaar Wouter H.
Published: October 2014

Chloride intracellular channel (CLIC) protein CLIC4 exists in both soluble and membrane-associated forms, and is implicated in diverse cellular processes, ranging from ion channel formation to intracellular membrane remodeling. CLIC4 is rapidly recruited to the plasma membrane by lysophosphatidic acid (LPA) and serum, suggesting a possible role for CLIC4 in exocytic-endocytic trafficking. However, the function and subcellular target(s) of CLIC4 remain elusive. Here we show that in HeLa and MDA-MB-231 cells, CLIC4 knockdown decreases cell-matrix adhesion, cell spreading and integrin signalling, while increasing cell motility. LPA stimulates the recruitment of CLIC4 to β1 integrins at the plasma membrane and in Rab35-positive endosomes. CLIC4 is required for both the internalization and the serum/LPA-induced recycling of β1 integrins, but not for EGF receptor trafficking. Furthermore, we show that CLIC4 suppresses Rab35 activity and antagonizes Rab35-dependent regulation of β1-integrin trafficking. Our results define CLIC4 as a regulator of Rab35 activity and serum/LPA-dependent integrin trafficking.
Epithelial cell division – multiplying without losing touch
Authors: Le Bras Stéphanie , Le Borgne Roland
Published: October 2014

Epithelia are compact tissues comprising juxtaposed cells that function as mechanical and chemical barriers between the body and the environment. This barrier relies, in part, on adhesive contacts within adherens junctions, which are formed and stabilized by E-cadherin and catenin proteins linked to the actomyosin cytoskeleton. During development and throughout adult life, epithelia are continuously growing or regenerating, largely as a result of cell division. Although persistence of adherens junctions is needed for epithelial integrity, these junctions are continually remodelled during cell division. In this Commentary, we will focus on cytokinesis, the final step of mitosis, a multiparty phenomenon in which the adherens junction belt plays an essential role and during which a new cell–cell interface is generated between daughter cells. This new interface is the site of intense remodelling, where new adhesive contacts are assembled and cell polarity is transmitted from mother to daughter cells, ultimately becoming the site of cell signalling.
Synaptotagmin-7 links fusion-activated Ca2+ entry (FACE) and fusion pore dilation
Authors: Neuland Kathrin , Sharma Neeti , Frick Manfred
Published: October 2014

Ca2+-dependent regulation of fusion pore dilation/closure is a key mechanism determining the output of cellular secretion. We have recently described „fusion-activated“ Ca2+-entry (FACE) following exocytosis of lamellar bodies (LBs) in alveolar type II cells. FACE regulates fusion pore expansion and facilitates secretion. Yet, mechanisms linking this locally restricted Ca2+ signal and fusion pore expansion were still elusive. Here we demonstrate that synaptotagmin-7 (syt-7) is expressed on LBs and links FACE and fusion pore dilation. We directly assessed dynamic changes in fusion pore diameters analysing diffusion of fluorophores across fusion pores. Expressing wt or mutant syt-7 with impaired Ca2+-binding to the C2 domains revealed that binding of Ca2+ to the C2A domain facilitates FACE-induced pore dilation, likely inhibiting translocation of complexin-2 to fused vesicles. However, the C2A domain hampered Ca2+-dependent exocytosis of LBs. These findings support that syt-7 modulates fusion pore expansion in large secretory organelles and extend our picture that LBs contain the necessary molecular inventory to facilitate secretion during the exocytic post-fusion phase. Moreover, regulating syt-7 levels on LBs appears essential to not impede exocytosis during the pre-fusion phase.
Nuclear actin and Mrtfs control disuse muscle atrophy via Srf activity regulation
Authors: Collard Laura , Herledan Gaëlle , Pincini Alessandra , Randrianarison-Huetz Voahangy , Guerci Aline , Sotiropoulos Athanassia
Published: October 2014

Skeletal muscle atrophy is a debilitating process associated with a wide variety of conditions including inactivity, disease and aging. Here, we demonstrate that the actin/Mrtfs/Srf pathway is specifically downregulated in muscle atrophy induced by disuse in mice. We show in vivo that the abolition of mechanical signals leads to rapid accumulation of G-actin in myonuclei and export of the Srf coactivator Mrtf-A, resulting in Mrtfs/Srf-dependent transcription decrease that contributes to atrophy. We demonstrate that inhibition of the actin/Mrtfs/Srf axis by overexpression of nuclear non-polymerizable actin, pharmacological inhibition of Mrtfs/Srf and muscle-specific Srf deletion worsens denervation-induced atrophy. Conversely, maintenance of high Srf or Mrtfs activity in denervated muscle, through overexpression of constitutively active derivatives, counteracts atrophy. Altogether, our data provide new mechanistic insights into the control of muscle mass upon disuse atrophy by the actin/Mrtfs/Srf pathway, highlighting Srf as a key mediator of mechanotransduction in muscle.
Epiprofin orchestrates epidermal keratinocyte proliferation and differentiation
Authors: Nakamura Takashi , Yoshitomi Yasuo , Sakai Kiyoshi , Patel Vyomesh , Fukumoto Satoshi , Yamada Yoshihiko
Published: October 2014

The basal layer of the epidermis contains stem cells and transit amplifying (TA) cells that rapidly proliferate and differentiate further into the upper layers of the epidermis. A number of molecules have been identified as regulators for this process including p63 and Notch1. However, little is known about the mechanisms that regulate the transitions from stem cells to proliferating or differentiating TA cells. Here we demonstrate that Epiprofin (Epfn) plays critical distinct roles in these transition stages as a cell cycle regulator and a transcription factor. Epfn knockout mice have a thickened epidermis, in which p63-expressing basal cells formed multiple layers due to accumulation of premature TA cells with reduced proliferation, and a reduction in differentiating keratinocytes expressing Notch1. We found that low levels of Epfn expression increased proliferation of human immortalized keratinocyte (HaCaT) cells by increasing EGF-responsiveness and superphosphorylation of Rb. In contrast, high levels of Epfn expression promoted cell cycle exit and differentiation, by reducing E2F transactivation and inducing Notch1 expression. Our findings identify multiple novel functions of Epiprofin in epidermal development.
Don't break a leg: running birds from quail to ostrich prioritise leg safety and economy on uneven terrain
Authors: Birn-Jeffery Aleksandra V. , Hubicki Christian M. , Blum Yvonne , Renjewski Daniel , Hurst Jonathan W. , Daley Monica A.
Published: October 2014

Cursorial ground birds are paragons of bipedal running that span a 500-fold mass range from quail to ostrich. Here we investigate the task-level control priorities of cursorial birds by analysing how they negotiate single-step obstacles that create a conflict between body stability (attenuating deviations in body motion) and consistent leg force–length dynamics (for economy and leg safety). We also test the hypothesis that control priorities shift between body stability and leg safety with increasing body size, reflecting use of active control to overcome size-related challenges. Weight-support demands lead to a shift towards straighter legs and stiffer steady gait with increasing body size, but it remains unknown whether non-steady locomotor priorities diverge with size. We found that all measured species used a consistent obstacle negotiation strategy, involving unsteady body dynamics to minimise fluctuations in leg posture and loading across multiple steps, not directly prioritising body stability. Peak leg forces remained remarkably consistent across obstacle terrain, within 0.35 body weights of level running for obstacle heights from 0.1 to 0.5 times leg length. All species used similar stance leg actuation patterns, involving asymmetric force–length trajectories and posture-dependent actuation to add or remove energy depending on landing conditions. We present a simple stance leg model that explains key features of avian bipedal locomotion, and suggests economy as a key priority on both level and uneven terrain. We suggest that running ground birds target the closely coupled priorities of economy and leg safety as the direct imperatives of control, with adequate stability achieved through appropriately tuned intrinsic dynamics.
The role of antisense long noncoding RNA in small RNA-triggered gene activation
Authors: Zhang Xizhe , Li Haitang , Burnett John C. , Rossi John J.
Published: October 2014

Long noncoding RNAs (lncRNAs) are known to regulate neighboring protein-coding genes by directing chromatin remodeling complexes, imprinting, and X-chromosome inactivation. In this study, we explore the function of lncRNAs in small RNA-triggered transcriptional gene activation (TGA), a process in which microRNAs (miRNAs) or small interfering RNAs (siRNAs) associated with Argonaute (Ago) proteins induce chromatin remodeling and gene activation at promoters with sequence complementarity. We designed a model system with different lncRNA and chromatin environments to elucidate the molecular mechanisms required for mammalian TGA. Using RNA-fluorescence in situ hybridization (FISH) and rapid amplification of cDNA ends (RACE)-PCR, we demonstrated that small RNA-triggered TGA occurs at sites where antisense lncRNAs are transcribed through the reporter gene and promoter. Small RNA-induced TGA coincided with the enrichment of Ago2 at the promoter region, but Ago2-mediated cleavage of antisense lncRNAs was not observed. Moreover, we examined the allele-specific effects of lncRNAs through a Cre-induced inversion of a poly(A) sequence that was designed to block the transcription of antisense lncRNAs through the reporter gene region in an inducible and reversible manner. Termination of nascent antisense lncRNAs abrogated gene activation triggered by small RNAs, and only allele-specific cis-acting antisense lncRNAs, but not trans-acting lncRNAs, were capable of rescuing TGA. Hence, this model revealed that antisense lncRNAs can mediate TGA in cis and not in trans, serving as a molecular scaffold for a small RNA–Ago2 complex and chromatin remodeling.
Distinctive profiles of small RNA couple inverted repeat-induced post-transcriptional gene silencing with endogenous RNA silencing pathways in Arabidopsis
Authors: Wroblewski Tadeusz , Matvienko Marta , Piskurewicz Urszula , Xu Huaqin , Martineau Belinda , Wong Joan , Govindarajulu Manjula , Kozik Alexander , Michelmore Richard W.
Published: October 2014

The experimental induction of RNA silencing in plants often involves expression of transgenes encoding inverted repeat (IR) sequences to produce abundant dsRNAs that are processed into small RNAs (sRNAs). These sRNAs are key mediators of post-transcriptional gene silencing (PTGS) and determine its specificity. Despite its application in agriculture and broad utility in plant research, the mechanism of IR-PTGS is incompletely understood. We generated four sets of 60 Arabidopsis plants, each containing IR transgenes expressing different configurations of uidA and CHALCONE SYNTHASE (At-CHS) gene fragments. Levels of PTGS were found to depend on the orientation and position of the fragment in the IR construct. Deep sequencing and mapping of sRNAs to corresponding transgene-derived and endogenous transcripts identified distinctive patterns of differential sRNA accumulation that revealed similarities among sRNAs associated with IR-PTGS and endogenous sRNAs linked to uncapped mRNA decay. Detailed analyses of poly-A cleavage products from At-CHS mRNA confirmed this hypothesis. We also found unexpected associations between sRNA accumulation and the presence of predicted open reading frames in the trigger sequence. In addition, strong IR-PTGS affected the prevalence of endogenous sRNAs, which has implications for the use of PTGS for experimental or applied purposes.
Generation of a neuro-specific microarray reveals novel differentially expressed noncoding RNAs in mouse models for neurodegenerative diseases
Authors: Gstir Ronald , Schafferer Simon , Scheideler Marcel , Misslinger Matthias , Griehl Matthias , Daschil Nina , Humpel Christian , Obermair Gerald J. , Schmuckermair Claudia , Striessnig Joerg , Flucher Bernhard E. , Hüttenhofer Alexander
Published: October 2014

We have generated a novel, neuro-specific ncRNA microarray, covering 1472 ncRNA species, to investigate their expression in different mouse models for central nervous system diseases. Thereby, we analyzed ncRNA expression in two mouse models with impaired calcium channel activity, implicated in Epilepsy or Parkinson's disease, respectively, as well as in a mouse model mimicking pathophysiological aspects of Alzheimer's disease. We identified well over a hundred differentially expressed ncRNAs, either from known classes of ncRNAs, such as miRNAs or snoRNAs or which represented entirely novel ncRNA species. Several differentially expressed ncRNAs in the calcium channel mouse models were assigned as miRNAs and target genes involved in calcium signaling, thus suggesting feedback regulation of miRNAs by calcium signaling. In the Alzheimer mouse model, we identified two snoRNAs, whose expression was deregulated prior to amyloid plaque formation. Interestingly, the presence of snoRNAs could be detected in cerebral spine fluid samples in humans, thus potentially serving as early diagnostic markers for Alzheimer's disease. In addition to known ncRNAs species, we also identified 63 differentially expressed, entirely novel ncRNA candidates, located in intronic or intergenic regions of the mouse genome, genomic locations, which previously have been shown to harbor the majority of functional ncRNAs.
Undetected oesophageal perforation and feeding-tube misplacement
Authors: Taylor Stephen J , Ross Catherine , Hooper Timothy
Published: October 2014

This is a case report of an electromagnetically (EM)-guided Cortrak feeding tube that perforated the lower oesophagus and was not detected by the EM trace or by plain X-ray. Misplacement was diagnosed from computed tomography (CT) following injection of radio-contrast medium down the tube. Recommendations are offered for use of the EM trace in patients at high risk of oesophageal perforation.
Jordanian intensive care nurses' perspectives on evidence-based practice in nutritional care
Authors: Kalaldeh Mahmoud Al , Watson Roger , Hayter Mark
Published: October 2014

This study aimed to explore Jordanian nurses' perspectives on the implementation of evidence-based practice and team-working related to nutritional care in the critically ill. Poor adherence to evidence-based practice and poor collaboration may contribute to nutritional failure. Fourteen critical care nurses from three healthcare sectors in Jordan were selected purposively and interviewed using semi-structured interviews. The information obtained was subjected to a thematic analysis. Four themes emerged from the study: ‘undertaking nutritional responsibilities’, ‘approaching evidence-based practice’, ‘multidisciplinary team working’ and ‘consequences of enteral nutrition care deficits’. Although evidence-based practice was emphasised by nurses, lack of evidence-based resources, and ineffective aspiration reduction measures were found to impede adherence to evidence-based practice. Multidisciplinary team working was introduced as means to improve practice. However, ineffective nursing involvement and poor interaction were obstacles to effective sharing.
Implementing a new urinary catheter insertion and removal pack
Authors: Laws Alyson
Published: October 2014

This article describes the implementation of a community urinary catheter insertion and removal pack that provides equipment to maintain safe and effective practice at a cost-effective price within Newcastle upon Tyne Hospitals NHS Foundation Trust. This development was achieved through service integration as a result of two large trusts amalgamating, offering the opportunity for practice development resulting in the development of a joint catheter product formulary, the implementation of a patient catheter record booklet and the design of a community urinary catheterisation pack. The impact of service integration has provided scope for proactive working between acute and community continence services, resulting in innovative approaches generating better quality outcomes for patients who require urinary catheterisation.
The National Early Warning Score in practice: a reflection
Authors: Day Tina , Oxton Julie
Published: October 2014

In 2012, The Royal College of Physicians (RCP) developed a National Early Warning Score (NEWS) as a standardised approach to assessment and response to critical illness. This paper reports the authors' experiences whilst implementing NEWS across one large inner London NHS Trust. NEWS was introduced to all adult areas between November 2013 and January 2014. All healthcare staff completed the RCP's online e-learning module and received scenario-based teaching. One of the challenges was the Trust's geographical location over three sites. Comparisons across all sites will be made over time as data become available. Introducing NEWS has been a challenging but exciting initiative.
The art of good patient conversation in health care
Authors: Tingle John
Published: October 2014

John Tingle discusses two recent publications that consider, along with a number of other matters, issues relating to improving patient communication
New rules to improve food intake for NHS patients and staff
Authors: Glasper Alan
Published: October 2014

Professor Alan Glasper from the University of Southampton discusses the ramifications of the new Hospital Food Standards Panel rules to serve up better food for NHS patients and staff, which is recommending five legally binding food standards across the whole service
ECG or X-ray as the ‘gold standard’ for establishing PICC-tip location?
Authors: Oliver Gemma , Jones Matt
Published: October 2014

Recently there has been an increase in evidence that the tip position of a peripherally inserted central catheter (PICC) needs to be accurately placed in the lower third of the superior vena cava at the junction with the right atrium in order to minimise potential complications ( Royal College of Nursing, 2010 ; Infusion Nurses Society, 2011 ). The current ‘gold standard’ practice of performing a chest X-ray post-insertion of PICC can be fraught with complications with regard to accurately placing the PICC in this position. The purpose of this evaluation is to discuss how using an electrocardiogram-guided PICC placement system may be a preferable method with which to accurately measure the exact position of the PICC within the venous system.
Pig bladders and feather quills: a history of vascular access devices
Authors: Kelly Linda J
Published: October 2014

Vascular access is a requirement for many hospitalised patients. Over the years there have been many technological refinements and advances in the equipment used to provide this necessary procedure and subsequent treatments. This article will take a journey from the Renaissance period, the discovery of the circulatory system in the 15 th century, to the present day, and will give the reader a notion of the origins and subsequent development of intravenous therapy and devices used to deliver necessary treatments.
Peripheral intravenous cannulation: managing distress and anxiety
Authors: McGowan Donna
Published: October 2014

It is recognised that peripheral intravenous (IV) cannulation is an increasingly performed procedure within the hospital setting. This invasive, yet necessary, procedure often causes patients considerable anxiety and distress, especially those patients who have to endure multiple and possibly painful and difficult cannulations. Nurses can play a valuable role in minimising the associated physical discomfort and complications that patients may experience as a result of having a peripheral cannula inserted through maintaining their knowledge and skills in relation to patient preparation, assessment and the care and management of the cannula. However, nurses also play an equally valuable role in recognising the potential psychological complications that are associated with peripheral IV cannulation. Demonstrating a knowledge and understanding of the effective methods that are used to manage and minimise these complications can ensure patients' confidence is maintained and their experience of cannulation is positive.
Beacon Communities’ Public Health Initiatives: A Case Study Analysis
Authors: Massoudi Barbara L. , Marcial Laura H. , Haque Saira , Bailey Robert , Chester Kelley , Cunningham Shellery , Riley Amanda , Soper Paula
Published: October 2014

Introduction: The Beacon Communities for Public Health (BCPH) project was launched
in 2011 to gain a better understanding of the range of activities currently being
conducted in population- and public health by the Beacon Communities. The project
highlighted the successes and challenges of these efforts with the aim of sharing this
information broadly among the public health community.Background: The Beacon Community
Program, designed to showcase technology-enabled, community-based initiatives to improve
outcomes, focused on: building and strengthening health information technology (IT)
infrastructure and exchange capabilities; translating investments in health IT to
measurable improvements in cost, quality, and population health; and, developing
innovative approaches to performance measurement, technology, and care delivery.Methods:
Four multimethod case studies were conducted based on a modified sociotechnical
framework to learn more about public health initiative implementation and use in the
Beacon Communities. Our methodological approach included using document review and
semistructured key informant interviews. NACCHO Model Practice Program criteria were
used to select the public health initiatives included in the case studies.Findings:
Despite differences among the case studies, common barriers and facilitators were found
to be present in all areas of the sociotechnical framework application including
structure, people, technology, tasks, overarching considerations, and sustainability.
Overall, there were many more facilitators (range = 7–14) present for each Beacon
compared to barriers (range = 4–6).Discussion: Four influential promising practices were
identified through the work: forging strong and sustainable partnerships; ensuring a
good task-technology fit and a flexible and iterative design; fostering technology
acceptance; and, providing education and demonstrating value.Conclusions: A common
weakness was the lack of a framework or model for the Beacon Communities evaluation
work. Sharing a framework or approach to evaluation at the beginning of implementation
made the work more effective. Supporting evaluation to inform future implementations is
important.
Children's necessities: trends over time in perceptions and ownership
Authors: Main Gill , Bradshaw Jonathan
Published: October 2014

Child poverty remains high on the UK political agenda. This paper informs these debates by examining trends over time in perceptions of child necessities, rates of child poverty and parental behaviours. Results indicate high levels of stability between 1999 and 2012 in public perceptions of child necessities, stable or increasing rates of child poverty and similarities in the profile of child poverty vulnerability. In both 1999 and 2012, findings show that the majority of parents prioritise children's needs, posing a challenge for coalition rhetoric, and suggesting that a focus on structural rather than individual causes of poverty may be needed.
Interaction of Participant Characteristics and Type of AAC With Individuals With ASD: A Meta-Analysis
Authors: Ganz Jennifer B. , Mason Rose A. , Goodwyn Fara D. , Boles Margot B. , Heath Amy K. , Davis John L.
Published: October 2014

Individuals with autism spectrum disorders (ASD) and complex communication needs often rely on augmentative and alternative communication (AAC) as a means of functional communication. This meta-analysis investigated how individual characteristics moderate effectiveness of three types of aided AAC: the Picture Exchange Communication System (PECS), speech-generating devices (SGDs), and other picture-based AAC. Effectiveness was measured via the Improvement Rate Difference. Results indicated that AAC has small to moderate effects on speech outcomes, and that SGDs appear to be most effective when considering any outcome measure with individuals with ASD without comorbid intellectual/developmental disorders (IDD). PECS appears to be most effective when considering any outcome measure with individuals with ASD and IDD. SGDs and PECS were the most effective type of AAC for preschoolers, when aggregating across outcome measures. No difference was found between systems for elementary-aged and older individuals.
Acute and Sustained Effects of Methylphenidate on Cognition and Presynaptic Dopamine Metabolism: An [18F]FDOPA PET Study
Authors: Schabram Ina , Henkel Karsten , Shali Siamak Mohammadkhani , Dietrich Claudia , Schmaljohann Jörn , Winz Oliver , Prinz Susanne , Rademacher Lena , Neumaier Bernd , Felzen Marc , Kumakura Yoshitaka , Cumming Paul , Mottaghy Felix M. , Gründer Gerhard , Vernaleken Ingo
Published: October 2014

Methylphenidate (MPH) inhibits the reuptake of dopamine and noradrenaline. PET studies with MPH challenge show increased competition at postsynaptic D2/3-receptors, thus indirectly revealing presynaptic dopamine release. We used [18F]fluorodopamine ([18F]FDOPA)-PET in conjunction with the inlet–outlet model (IOM) of Kumakura et al. (2007) to investigate acute and long-term changes in dopamine synthesis capacity and turnover in nigrostriatal fibers of healthy subjects with MPH challenge. Twenty healthy human females underwent two dynamic [18F]FDOPA PET scans (124 min; slow bolus-injection; arterial blood sampling), with one scan in untreated baseline condition and the other after MPH administration (0.5 mg/kg, p.o.), in randomized order. Subjects underwent cognitive testing at each PET session. Time activity curves were obtained for ventral putamen and caudate and were analyzed according to the IOM to obtain the regional net-uptake of [18F]FDOPA (K; dopamine synthesis capacity) as well as the [18F]fluorodopamine washout rate (kloss, index of dopamine turnover). MPH substantially decreasedklossin putamen (−22%;p= 0.003). In the reversed treatment order group (MPH/no drug),Kwas increased by 18% at no drug follow-up. The magnitude ofKat the no drug baseline correlated with cognitive parameters. Furthermore, individualklosschanges correlated with altered cognitive performance under MPH. [18F]FDOPA PET in combination with the IOM detects an MPH-evoked decrease in striatal dopamine turnover, in accordance with the known acute pharmacodynamics of MPH. Furthermore, the scan-ordering effect onKsuggested that a single MPH challenge persistently increased striatal dopamine synthesis capacity. Attenuation of dopamine turnover by MPH is linked to enhanced cognitive performance in healthy females.
How stem cells get “turned on”
Authors: Wagers Amy J
Published: October 2014

Recent research began to link autophagic processes to the functional integrity of certain stem cells. A novel study published in this issue of The EMBO Journal reports on autophagic flux as crucial checkpoint to meet the energy demands during muscle stem cell activation.
MicroRNA‐15b promotes neurogenesis and inhibits neural progenitor proliferation by directly repressing TET3 during early neocortical development
Authors: Lv Xiaohui , Jiang Huihui , Liu Yanli , Lei Xuepei , Jiao Jianwei
Published: October 2014

MicroRNAs (miRNAs) are important regulators of mouse brain development. However, their precise roles in this context remain to be elucidated. Through screening of expression profiles from a miRNA microarray and experimental analysis, we show here that miR‐15b controls several aspects of cortical neurogenesis. miR‐15b inhibits cortical neural progenitor cell (NPC) proliferation and promotes cell‐cycle exit and neuronal differentiation. Additionally, miR‐15b expression decreases the number of apical progenitors and increases basal progenitors in the VZ/SVZ. We also show that miR‐15b binds to the 3′ UTR of TET3, which plays crucial roles during embryonic development by enhancing DNA demethylation. TET3 promotes cyclin D1 expression, and miR‐15b reduces TET3 expression and 5hmC levels. Notably, TET3 expression rescues miR‐15b‐induced impaired NPC proliferation and increased cell‐cycle exit in vivo. Our results not only reveal a link between miRNAs, TET, and DNA demethylation but also demonstrate critical roles for miR‐15b and TET3 in maintaining the NPC pool during early neocortical development.
Constant rate of p53 tetramerization in response to DNA damage controls the p53 response
Authors: Gaglia Giorgio , Lahav Galit
Published: October 2014

The dynamics of the tumor suppressor protein p53 have been previously investigated in single cells using fluorescently tagged p53. Such approach reports on the total abundance of p53 but does not provide a measure for functional p53. We used fluorescent protein‐fragment complementation assay (PCA) to quantify in single cells the dynamics of p53 tetramers, the functional units of p53. We found that while total p53 increases proportionally to the input strength, p53 tetramers are formed in cells at a constant rate. This breaks the linear input–output relation and dampens the p53 response. Disruption of the p53‐binding protein ARC led to a dose‐dependent rate of tetramers formation, resulting in enhanced tetramerization and induction of p53 target genes. Our work suggests that constraining the p53 response in face of variable inputs may protect cells from committing to terminal outcomes and highlights the importance of quantifying the active form of signaling molecules in single cells.
Determinants of Working Capital Management in Latin American Companies
Authors: Mongrut Samuel , Fuenzalida O’Shee Darcy , Cubillas Zavaleta Claudio , Cubillas Zavaleta Johan
Published: October 2014

The aim of this study is to determine the factors that affect working capital management in Latin American companies. Using an unbalanced panel data analysis for companies quoted in five Latin American capital markets it is shown that companies in Argentina, Brazil, Chile and Mexico are holding cash excesses, which could destroy firm value. Results show that the industry cash conversion cycle, the company market power, its future sales and country risk have an influence on the way Latin American companies manage their working capital with significant differences among countries in the region.
Relación entre la creación de valor y la inversión en I+D: una aproximación mediante redes neuronales artificiales
Authors: Samaniego Alcántar Ángel , Mongrut Samuel
Published: October 2014

Este documento proporciona nueva evidencia sobre la interacción de los efectos del tipo de interés a cinco años, el nivel de apalancamiento (la relación del pasivo y activo), las pérdidas históricas, el crecimiento del PIB, la intensidad del gasto en I+D y los ingresos con el rendimiento de la acción en el corto plazo. El modelo no lineal utilizado (combinación de modelos mediante redes neuronales artificiales) explica 31.59% de la varianza del rendimiento de las acciones en los primeros 5 meses del año, entre 2000-2006, en los mercados AMEX, NASDAQ y NYSE. Existe evidencia en la literatura que el 32% de explicación es alto, en comparación al estudio de la literatura realizado por Coad (2009) donde se utilizan modelos lineales que buscan explicar el crecimiento de la empresa. Se observa que los modelos lineales explican entre el 17% al 32% de la variabilidad de los datos. Al igual que en los trabajos de Bode (1998), Tsai (2005), Wang y Chien (2006) y Chien et al. (2010) se utilizan las redes neuronales artificiales para el estudio de la interacción entre la I+D y la empresa.
Sistemas locais de produção: uma análise dos fatores inibidores das relações de cooperação e interação
Authors: Iacono Antonio , Seido Nagano Marcelo , Escrivão Filho Edmundo
Published: October 2014

O presente artigo tem por objetivo apresentar os fatores organizacionais e ambientais que inibem as relações de cooperação e interação entre os diversos tipos de atores, conformados em um Sistema Local de Produção (SLP) de Pequenas e Médias Empresas (PMEs). Para a exploração desses fatores inibidores, realizou-se um estudo exploratório em um SLP do tipo embrionário, que atua no segmento de máquinas e implementos agrícolas, localizado na região oeste do estado do Paraná, Brasil. A partir da avaliação sobre as relações de cooperação e interação das empresas do SLP, foram identificados seus fatores inibidores, e estes foram relacionados às especificidades das empresas. As evidências empíricas demonstraram que os fatores inibidores identificados de maior impacto sobre as interações e ações conjuntas são os organizacionais, e que estes expressam uma forte relação com as características específicas ao porte das empresas.
Una aproximación holística a la determinación y gestión de precios en las compañías hoteleras
Authors: Martín Samper Rosario Del Carmen , Moreno Rojas José
Published: October 2014

En este trabajo se propone un doble objetivo: por una parte, identificar los factores que intervienen en la determinación de las tarifas hoteleras y, por otra, analizar los procedimientos utilizados para la gestión de los mismos. Para ello, se realiza un estudio de caso longitudinal en un establecimiento hotelero perteneciente a una prestigiosa cadena española con fuerte presencia a nivel internacional.Como resultado, los autores han identificado que los principales factores que influyen en las decisiones de precio en la empresa hotelera son el nivel de demanda, las percepciones de los clientes y las actuaciones de la competencia. El escaso coste marginal del servicio de alojamiento origina que el coste, ante niveles de demanda considerados normales, no incida de forma significativa, aunque pasa a ser un factor clave en la política de descuentos cuando la demanda se encuentra en recesión. En cuanto a los procedimientos de gestión empleados, se destacan los cuestionarios de calidad, los procesos de negociación con las cuentas claves y los sistemas Yield Management, cuyas potencialidades y limitaciones han sido descritas en el trabajo.
Determinantes do endividamento da administração local: estudo exploratório nos municípios do norte de Portugal
Authors: Baptista Ribeiro Nuno Adriano , Faustino Jorge Susana Margarida
Published: October 2014

A gestão do endividamento dos Estados, uma questão sempre pertinente no que concerne à análise da sua sustentabilidade financeira, está, no atual contexto de crise financeira à escala global, cada vez mais, no topo da agenda política. Como seria expectável, a Administração Local não poderá ser excluída desta análise, o que contribui para o esforço de controle que é exigido aos Estados no seu todo.Assim, o objetivo desta investigação é o de estudar, para os anos de 2006 e 2007, os fatores determinantes do endividamento dos municípios do norte de Portugal tentando identificar quais o influenciam e de que forma. Para além de uma análise bivariada, testa-se um modelo de regressão linear múltipla.Os resultados mostram que os principais fatores determinantes são o limite legal máximo de endividamento, as despesas totais, as receitas de capital e o saldo orçamental corrente. Adicionalmente permitem verificar que, tanto na análise bivariada quanto na multivariada, os fatores mais importantes são os mesmos para os dois anos em análise. Finalmente, consta-se que não existem diferenças estatisticamente significativas no endividamento líquido nos anos de 2006 e 2007, embora, neste último, ter sido aprovada uma nova Lei das Finanças Locais.