Expression von Parathormone-related Peptide in koronaren Endothelzellen und der glatten Gefäßmuskulatur

Das Parathormon related peptide (PTHrP) wird im gesamten Gefäßsystem einschließlich der koronaren Endothelzellen exprimiert. Im Gegensatz zu den benachbarten Glattmuskelzellen, welche bislang hinsichtlich PTHrP eingehend untersucht wurden, hat man die koronaren Endothelzellen nur als Produktionsstätte des Ventrikels angesehen. Aus diesem Grund blieben bisher die PTHrP-Expression und seine biologische Rolle in diesen Zellen ungeklärt. Deshalb habe ich mich dafür interessiert, ob eine Stimulation a-adrenerger bzw. AT-Rezeptoren die Expression in den koronaren Endothelzellen steigert und ob das endogen exprimierte PTHrP einen sogenannten intrakrinen Effekt in diesem Zelltyp ausübt. Besonders interessant ist diese Fragestellung vor dem Hintergrund, daß diese Zellen keine klassischen Zielzellen des Peptidhormones darstellen, da sie nicht den korrespondierenden PTHrP-Rezeptor besitzen, wie das z. B. bei den Glattmuskelzellen der Fall ist. Was anhand der Ergebnisse gezeigt werden konnte ist die Tatsache, daß durch Stimulation der Zellen mit der a-adrenergen Substanz Phenylephrin im Vergleich zur Stimulation mit Angiotensin II, eine deutliche Steigerung in der Expression des PTHrP hervorgerufen werden konnte. Dieser Effekt konnte jedoch nicht im Stadium der Konfluenz reproduziert werden. Im Gegensatz dazu erhielten wir in den Glattmuskelzellen ein genau umgekehrtes Bild, da hier nur Angiotensin II eine signifikante Expressionssteigerung hervorrief. Deshalb wollte ich mehr über die Lokalisation des endogenen PTHrP in den verschiedenen Proliferationsstadien mit Hilfe der Immunfluoreszenz in Erfahrung bringen. Die Ergebnisse zeigten, daß der Anteil an PTHrP, welcher in den Kern transloziert wird, umso größer ist, je mehr sich die Zellen dem Konfluenzstadium nähern. Diese Tatsache war für mich Anlaß zu vermuten, daß das Peptidhormon einen intrakrinen Effekt auf die koronaren Endothelzellen ausübt. Mit Hilfe der Transfektion der Zellen mit Antisense-Oligonukleotiden gegen PTHrP und der daraus sich ergebenden Herabregulierung seiner Expression untersuchte ich seinen möglichen Einfluß auf die Proliferation bzw. Apoptose. Im Bezug auf die Proliferation ergaben sich keine Hinweise auf eine Beeinflussung. Bei der Betrachtung der Endothelzellen in den unterschiedlichen Konfluenzstadien konnte mit Hilfe einer UV-Bestrahlung in Zellen, in denen der Anteil des nukleären PTHrP normalerweise erhöht wäre, eine stärkere Apoptoserate als unter Basalbedingungen hervorgerufen werden. Im Gegensatz dazu wurde durch eine erhöhte Expression des PTHrP - welche durch Phenylephrin vermittelte wurde -der Anteil der apoptotischen Zellen sowohl gegenüber Basalbedingungen als auch unter UV-Induktion deutlich vermindert. Zusammenfassend läßt sich anhand der Ergebnisse der vorliegenden Studie sagen, daß in koronaren Endothelzellen die PTHrP-Expression durch eine a-Adrenozeptor-Stimulation in einer Zellzyklus-abhängigen und Zelltyp-spezifischen Art und Weise reguliert wird. Diese Studie zeigt eine neue biologische Rolle des PTHrP im Gefäßbett auf, da über den nachgewiesenen intrakrinen Effekt das Peptidhormon einen Anteil am Schutzmechanismus der Endothelschicht vor Apoptose besitzt.Parathyroid hormone related peptide (PTHrP) is expressed throughout the vascular system including coronary endothelial cells. The regulation of endothelial PTHrP expression and the role of PTHrP expression in endothelial cells is not clear. The present study investigates the question whether stimulation of a-adrenergic or angiotensin II receptors increases endothelial expression of PTHrP and whether endogenously expressed PTHrP exerts intracrine effects in coronary endothelial cells. It was found that stimulation of alpha1A-adrenoceptors, but not that of angiotensin II, increases cellular expression of PTHrP in growing, but not in growth arrested, coronary endothelial cells. Angiotensin II increases the expression of PTHrP in smooth muscle cells, but not in endothelial cells. PTHrP enters the nucleus of endothelial cells at the stadium of confluence. This suggests an intracrine effect of PTHrP. It was further investigated whether downregulation of endogenous PTHrP expression by transfection with antisense oligonucleotides alters cell proliferation or apoptosis resistance in growing or non-growing endothelial cells. Downregulation of PTHrP did not modify cell proliferation but increased the amount of UV-induced apoptosis. An increased expression of PTHrP in cells pre-treated with an a-adrenoceptor agonist reduced basal rate of apoptosis and improved resistance against UV-induced apoptosis. These results indicate a novel intracrine effect of PTHrP in coronary endothelial cells that improves cell survival. In endothelial cells, the expression of PTHrP is regulated by alpha-adrenoceptor stimulation in a cell-cycle dependent and cell-type specific manner

Positive allosteric modulation of the muscarinic M1 receptor improves efficacy of antipsychotics in mouse glutamatergic deficit models of behavior

Current antipsychotics are effective in treating the positive symptoms associated with schizophrenia, but they remain suboptimal in targeting cognitive dysfunction. Recent studies have suggested that positive allosteric modulation of the M1 muscarinic acetylcholine receptor (mAChR) may provide a novel means of improving cognition. However, very little is known about the potential of combination therapies in extending coverage across schizophrenic symptom domains. This study investigated the effect of the M1 mAChR positive allosteric modulator BQCA [1-(4-methoxybenzyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid], alone or in combination with haloperidol (a first-generation antipsychotic), clozapine (a second-generation atypical antipsychotic), or aripiprazole (a third-generation atypical antipsychotic), in reversing deficits in sensorimotor gating and spatial memory induced by the N-methyl-d-aspartate receptor antagonist, MK-801 [(5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine]. Sensorimotor gating and spatial memory induction are two models that represent aspects of schizophrenia modeled in rodents. In prepulse inhibition (an operational measure of sensorimotor gating), BQCA alone had minimal effects but exhibited different levels of efficacy in reversing MK-801–induced prepulse inhibition disruptions when combined with a subeffective dose of each of the three (currently prescribed) antipsychotics. Furthermore, the combined effect of BQCA and clozapine was absent in M1−/− mice. Interestingly, although BQCA alone had no effect in reversing MK-801–induced memory impairments in a Y-maze spatial test, we observed a reversal upon the combination of BQCA with atypical antipsychotics, but not with haloperidol. These findings provide proof of concept that a judicious combination of existing antipsychotics with a selective M1 mAChR positive allosteric modulator can extend antipsychotic efficacy in glutamatergic deficit models of behavior

Pseudomonas aeruginosa Adaptation to Lungs of Cystic Fibrosis Patients Leads to Lowered Resistance to Phage and Protist Enemies

Pathogenic life styles can lead to highly specialized interactions with host species, potentially resulting in fitness trade-offs in other ecological contexts. Here we studied how adaptation of the environmentally transmitted bacterial pathogen, Pseudomonas aeruginosa, to cystic fibrosis (CF) patients affects its survival in the presence of natural phage (14/1, ΦKZ, PNM and PT7) and protist (Tetrahymena thermophila and Acanthamoebae polyphaga) enemies. We found that most of the bacteria isolated from relatively recently intermittently colonised patients (1-25 months), were innately phage-resistant and highly toxic for protists. In contrast, bacteria isolated from long time chronically infected patients (2-23 years), were less efficient in both resisting phages and killing protists. Moreover, chronic isolates showed reduced killing of wax moth larvae (Galleria mellonella) probably due to weaker in vitro growth and protease expression. These results suggest that P. aeruginosa long-term adaptation to CF-lungs could trade off with its survival in aquatic environmental reservoirs in the presence of microbial enemies, while lowered virulence could reduce pathogen opportunities to infect insect vectors; factors that are both likely to result in poorer environmental transmission. From an applied perspective, phage therapy could be useful against chronic P. aeruginosa lung infections that are often characterized by multidrug resistance: chronic isolates were least resistant to phages and their poor growth will likely slow down the emergence of beneficial resistance mutations

Variations of training load, monotony, and strain and dose-response relationships with maximal aerobic speed, maximal oxygen uptake, and isokinetic strength in professional soccer players

This study aimed to identify variations in weekly training load, training monotony, and training strain across a 10-week period (during both, pre- and in-season phases); and to analyze the dose-response relationships between training markers and maximal aerobic speed (MAS), maximal oxygen uptake, and isokinetic strength. Twenty-seven professional soccer players (24.9±3.5 years old) were monitored across the 10-week period using global positioning system units. Players were also tested for maximal aerobic speed, maximal oxygen uptake, and isokinetic strength before and after 10 weeks of training. Large positive correlations were found between sum of training load and extension peak torque in the right lower limb (r = 0.57, 90%CI[0.15;0.82]) and the ratio agonist/antagonist in the right lower limb (r = 0.51, [0.06;0.78]). It was observed that loading measures fluctuated across the period of the study and that the load was meaningfully associated with changes in the fitness status of players. However, those magnitudes of correlations were small-to-large, suggesting that variations in fitness level cannot be exclusively explained by the accumulated load and loading profile

Photonic Doppler velocimetry probe used to measure grain boundaries of dynamic shocked materials

Author Institution: Mission Support and Test Services, LLC; Los Alamos National LaboratorySlides presented at the 2018 Photonic Doppler Velocimetry (PDV) Users Workshop, Drury Plaza Hotel, Santa Fe, New Mexico, May 16-18, 2018

Recommendations for selecting drug-drug interactions for clinical decision support

To recommend principles for including drug-drug interactions (DDIs) in clinical decision support

Using creative co-design to develop a decision support tool for people with malignant pleural effusion

Abstract: Background: Malignant pleural effusion (MPE) is a common, serious problem predominantly seen in metastatic lung and breast cancer and malignant pleural mesothelioma. Recurrence of malignant pleural effusion is common, and symptoms significantly impair people’s daily lives. Numerous treatment options exist, yet choosing the most suitable depends on many factors and making decisions can be challenging in pressured, time-sensitive clinical environments. Clinicians identified a need to develop a decision support tool. This paper reports the process of co-producing an initial prototype tool. Methods: Creative co-design methods were used. Three pleural teams from three disparate clinical sites in the UK were involved. To overcome the geographical distance between sites and the ill-health of service users, novel distributed methods of creative co-design were used. Local workshops were designed and structured, including video clips of activities. These were run on each site with clinicians, patients and carers. A joint national workshop was then conducted with representatives from all stakeholder groups to consider the findings and outputs from local meetings. The design team worked with participants to develop outputs, including patient timelines and personas. These were used as the basis to develop and test prototype ideas. Results: Key messages from the workshops informed prototype development. These messages were as follows. Understanding and managing the pleural effusion was the priority for patients, not their overall cancer journey. Preferred methods for receiving information were varied but visual and graphic approaches were favoured. The main influences on people’s decisions about their MPE treatment were personal aspects of their lives, for example, how active they are, what support they have at home. The findings informed the development of a first prototype/service visualisation (a video representing a web-based support tool) to help people identify personal priorities and to guide shared treatment decisions. Conclusion: The creative design methods and distributed model used in this project overcame many of the barriers to traditional co-production methods such as power, language and time. They allowed specialist pleural teams and service users to work together to create a patient-facing decision support tool owned by those who will use it and ready for implementation and evaluation

'Collective Making' as knowledge mobilisation: the contribution of participatory design in the co-creation of knowledge in healthcare

The discourse in healthcare Knowledge Mobilisation (KMb) literature has shifted from simple, linear models of research knowledge production and action to more iterative and complex models. These aim to blend multiple stakeholders’ knowledge with research knowledge to address the researchpractice gap. It has been suggested there is no 'magic bullet', but that a promising approach to take is knowledge co-creation in healthcare, particularly if a number of principles are applied. These include systems thinking, positioning research as a creative enterprise with human experience at its core, and paying attention to process within the partnership. This discussion paper builds on this proposition and extends it beyond knowledge co-creation to co-designing evidenced based interventions and implementing them. Within a co-design model, we offer a specific approach to share, mobilise and activate knowledge, that we have termed 'collective making'. We draw on KMb, design, wider literature, and our experiences to describe how this framework supports and extends the principles of co-creation offered by Geenhalgh et al[1] in the context of the state of the art of knowledge mobilisation. We describe how collective making creates the right ‘conditions’ for knowledge to be mobilised particularly addressing issues relating to stakeholder relationships, helps to discover, share and blend different forms of knowledge from different stakeholders, and puts this blended knowledge to practical use allowing stakeholders to learn about the practical implications of knowledge use and to collectively create actionable products. We suggest this collective making has three domains of influence: on the participants; on the knowledge discovered and shared; and on the mobilisation or activation of this knowledge

Qualitative study on the implementation of professional pharmacy services in Australian community pharmacies using framework analysis

Abbreviations: BCT, Behavioural change techniques taxonomy; BCW, Behavioural change wheel; CFIR, Consolidated framework for implementation research; EPOC, Cochrane effective practice and organisation of care; FISpH, Framework for the implementation of services in pharmacy; GIF, Generic implementation framework; KPI, Key performance indicator; TDF, Theoretical domains frameworkBackground: Multiple studies have explored the implementation process and influences, however it appears there is no study investigating these influences across the stages of implementation. Community pharmacy is attempting to implement professional services (pharmaceutical care and other health services). The use of implementation theory may assist the achievement of widespread provision, support and integration. The objective was to investigate professional service implementation in community pharmacy to contextualise and advance the concepts of a generic implementation framework previously published. Methods: Purposeful sampling was used to investigate implementation across a range of levels of implementation in community pharmacies in Australia. Twenty-five semi-structured interviews were conducted and analysed using a framework methodology. Data was charted using implementation stages as overarching themes and each stage was thematically analysed, to investigate the implementation process, the influences and their relationships. Secondary analyses were performed of the factors (barriers and facilitators) using an adapted version of the Consolidated Framework for Implementation Research (CFIR), and implementation strategies and interventions, using the Expert Recommendations for Implementing Change (ERIC) discrete implementation strategy compilation. Results: Six stages emerged, labelled as development or discovery, exploration, preparation, testing, operation and sustainability. Within the stages, a range of implementation activities/steps and five overarching influences (pharmacys' direction and impetus, internal communication, staffing, community fit and support) were identified. The stages and activities were not applied strictly in a linear fashion. There was a trend towards the greater the number of activities considered, the greater the apparent integration into the pharmacy organization. Implementation factors varied over the implementation stages, and additional factors were added to the CFIR list and definitions modified/contextualised for pharmacy. Implementation strategies employed by pharmacies varied widely. Evaluations were lacking. Conclusions: The process of implementation and five overarching influences of professional services implementation in community pharmacy have been outlined. Framework analysis revealed, outside of the five overarching influences, factors influencing implementation varied across the implementation stages. It is proposed at each stage, for each domain, the factors, strategies and evaluations should be considered. The Framework for the Implementation of Services in Pharmacy incorporates the contextualisation of implementation science for pharmacy.The study was funded as part of a University of Technology Sydney (UTS) Research Excellence Scholarship (RES), comprising of an Australian Postgraduate Award (APA) Scholarship funded by the Australian Government, plus a Top-up funded by the University of Technology Sydney, received from the primary author (JCM)