Human red blood cells at work: identification and visualization of erythrocytic eNOS activity in health and disease

A nitric oxide synthase (NOS)-like activity has been demonstrated in human red blood cells (RBCs), but doubts about its functional significance, isoform identity and disease relevance remain. Using flow cytometry in combination with the NO-imaging probe DAF-FM we find that all blood cells form NO intracellularly, with a rank order of monocytes > neutrophils > lymphocytes > RBCs > platelets. The observation of a NO-related fluorescence within RBCs was unexpected given the abundance of the NO-scavenger oxyhemoglobin. Constitutive normoxic NO formation was abolished by NOS inhibition and intracellular NO scavenging, confirmed by laser-scanning microscopy and unequivocally validated by detection of the DAF-FM reaction product with NO using HPLC and LC-MS/MS. Employing immunoprecipitation, ESI-MS/MS-based peptide sequencing and enzymatic assay we further demonstrate that human RBCs contain an endothelial NOS (eNOS) that converts L-3H-Arginine to L-3H-Citrulline in a Ca2+/Calmodulin-dependent fashion. Moreover, in patients with coronary artery disease, red cell eNOS expression and activity are both lower than in age-matched healthy individuals and correlate with the degree of endothelial dysfunction. Thus, human RBCs constitutively produce NO under normoxic conditions via an active eNOS isoform the activity of which is compromised in patients with coronary artery disease.

Endothelin-converting enzyme-1 degrades internalized somatostatin-14

Agonist-induced internalization of somatostatin receptors (ssts) determines subsequent cellular responsiveness to peptide agonists and influences sst receptor scintigraphy. To investigate sst2A trafficking, rat sst2A tagged with epitope was expressed in human embryonic kidney cells and tracked by antibody labeling. Confocal microscopical analysis revealed that stimulation with sst and octreotide induced internalization of sst2A. Internalized sst2A remained sequestrated within early endosomes, and 60 min after stimulation, internalized sst2A still colocalized with beta-arrestin1-enhanced green fluorescence protein (EGFP), endothelin-converting enzyme-1 (ECE-1), and rab5a. Internalized (125)I-Tyr(11)-SST-14 was rapidly hydrolyzed by endosomal endopeptidases, with radioactive metabolites being released from the cell. Internalized (125)I-Tyr(1)-octreotide accumulated as an intact peptide and was released from the cell as an intact peptide ligand. We have identified ECE-1 as one of the endopeptidases responsible for inactivation of internalized SST-14. ECE-1-mediated cleavage of SST-14 was inhibited by the specific ECE-1 inhibitor, SM-19712, and by preventing acidification of endosomes using bafilomycin A(1). ECE-1 cleaved SST-14 but not octreotide in an acidic environment. The metallopeptidases angiotensin-1 converting enzyme and ECE-2 did not hydrolyze SST-14 or octreotide. Our results show for the first time that stimulation with SST-14 and octreotide induced sequestration of sst2A into early endosomes and that endocytosed SST-14 is degraded by endopeptidases located in early endosomes. Furthermore, octreotide was not degraded by endosomal peptidases and was released as an intact peptide. This mechanism may explain functional differences between octreotide and SST-14 after sst2A stimulation. Moreover, further investigation of endopeptidase-regulated trafficking of neuropeptides may result in novel concepts of neuropeptide receptor inactivation in cancer diagnosis.

The application of Regional Climate Model output for the simulation of high-mountain permafrost scenarios

This study investigates the possibilities and limitations of using Regional Climate Model (RCM) output for the simulation of alpine permafrost scenarios. It focuses on the general problem of scale mismatch between RCMs and impact models and, in particular, the special challenges that arise when driving an impact model in topographically complex high-mountain environments with the output of an RCM. Two approaches are introduced that take into account the special difficulties in such areas, and thus enable the use of RCM for alpine permafrost scenario modelling. Intended as an initial example, they are applied at the area of Corvatsch (Upper Engadine, Switzerland) in order to demonstrate and discuss the application of the two approaches, rather than to provide an assessment of future changes in permafrost occurrence. There are still many uncertainties and inaccuracies inherent in climate and impact models, which increase when driving one model with the output of the other. Nevertheless, our study shows that the use of RCMs offers new and promising perspectives for the simulation of high-mountain permafrost scenarios

Bridging the gap between research and practice in organizational network analysis: a conversation between Rob Cross and Martin Kilduff

The article features a conversation between Rob Cross and Martin Kilduff about organizational network analysis in research and practice. It demonstrates the value of using social network perspectives in HRM. Drawing on the discussion about managing personal networks; managing the networks of others; the impact of social networking sites on perceptions of relationships; and ethical issues in organizational network analysis, we propose specific suggestions to bring social network perspectives closer to HRM researchers and practitioners and rebalance our attention to people and to their relationships.

Building from scratch a marketing services giant by acquisition: case study and interview with Sir Martin Sorrell, Chief Executive of WPP

Purpose – Mergers and acquisitions are among the most intensely used strategic decisions. Yet research by both academics and consulting groups suggests that many mergers and acquisitions fail to add value. On the other hand there are many companies that successfully use mergers and acquisition to grow and add shareholder value. One such company is WPP. The aim of this paper is to explore why WPP has been successful in its acquisition strategy while so many other companies fail. Design/methodology/approach – The paper draws on documentary evidence and a semi-structured interview with Sir Martin Sorrell – Chief Executive and founder of WPP. Research limitations/implications – The case study offers a unique insight into thinking of a successful acquirer and sheds light on how mergers and acquisitions are managed by WPP. However, because of its design the findings are not generalisable. Originality/value – This case study sheds light on how mergers and acquisitions can be used to create a £9 billion company from a standing start. Furthermore, very few case studies offer insight into the thinking of entrepreneurial Chief Executives who established the business, grew it to become the largest and most profitable marketing services company in the world and engineered close to 300 acquisitions.

Stephen Martin Leake: a Victorian's view of the Common Law

This article considers the life and work of Stephen Martin Leake and seeks to locate his work within the wider context of the procedural and substantive transformation of the mid-to-late Victorian legal world. In particular, the article attempts to rescue Leake from obscurity and emphasise his importance in this process. It is argued that Leake’s work began the process whereby common lawyers conceived of their law as organised in a principled rather than procedural manner. Later common law jurists built upon this work. Consideration is also given to the philosophical and jurisprudential sources upon which Leake drew in constructing his treatises.

Plague of diagrams

Group exhibition curated by Plastique Fantastique. Organised by David Burrows and Dean Kenning in collaboration with Ami Clarke, Andrew Conio, John Cussans and David Osbaldeston. Contributors: David Burrows, Rachel Cattle & Jenna Collins, Neil Chapman & Gillian Wylde, Ami Clarke, Richard Cochrane, Andrew Conio, John Cussans, David Burrows, Benedict Drew, English Heretic, Nikolaus Gansterer, Joey Holder, Dean Kenning, Christoph Lueder, Stine Llungdalh, Adelheid Mers, Mike Nelson, Paul O'Kane, David Osbaldeston, Plastique Fantastique, Patricia Reed, John Russell, Erica Scourti, Andy Sharp, Kamini Vellodi, Martin Westwood and Carey Young.

Maktens ideologier och språk i svenska skolors värdegrunder : - Från Martin Luther till lokala likabehandlingsplaner

Syftet med studien Maktens ideologier och språk i svenska skolors värdegrunder är att systematiskt analysera innehållet i skriftliga dokument som kan sägas utgöra eller har utgjort värdegrunder för allmänna svenska skolor. Resultaten visar att det finns en relativt tydlig koppling mellan den rådande maktordningen i samhället och den värdegrund som råder i allmänna svenska skolor. Detta tycks gälla oavsett historisk epok och oavsett vilka ideologier - religiösa eller politiska - de dominerande maktgrupperna förespråkar. Allmänna skolor kan således i hög grad ses som redskap för att reproducera, återskapa, makten hos för tillfället dominerande samhällsgrupper och för att styra eleverna, samhällsmedborgarna, i önskvärd riktning.

Die bibelübersetzung Martin Luthers : eine soziolinguistische analyse der absicht, der methode und der auswirkung

Brundin (2004, S. 63) sagt, dass sich die Reformation „um einen Kampf handelte, der Auswirkungen auf die ganze gesellschaftliche Struktur hatte.“ Das Ziel dieser Arbeit ist die Absichten hinter, die linguistischen Methoden und die sozialen Auswirkungen der Bibelübersetzung Luthers festzustellen, und dadurch die Aussage Brundins zu bestätigen bzw. widerlegen. Es wurde gefunden, dass Martin Luther die Bibelübersetzung und die Reformation in enger Zusammenarbeit mit seinen Kollegen an der Leucorea Universität und unter Führung des sächsischen Kurfürsten, Friedrich III., durchgeführt hat. Dabei haben die verwendeten linguistischen Methoden eine Schlüsselrolle gespielt, und viele heute bekannten wissenschaftlichen Theorien sind praktisch umgesetzt worden. Dazu gehören die Sapir-Whorf-Hypothese, die Defizit- bzw. die Differenzhypothese und die Diskurstheorie. Die Reformation hat eine gewaltige Machtverschiebung zur Folge, wo der Klerus dem Adel viele Rechte abgeben müsste, und die neu erzeugte Sprache der Lutherbibel hat zu einer deutschen Einheitssprache und die Erstehung eines deutschen Nationalstaates geführt. Als Schlussergebnis kann die Aussage Brundins klar bestätigt werden.

In Their Footsteps, In Their Words: Special Section, 1813-1863

In Their Footsteps and In Their Words: Colby explores the first 50 years, from Jeremiah Chaplin to the Civil War.

A sonoridade do violão na execução musical : um estudo sobre os seus aspectos formadores e análise de duas gravações das Quatro Peças Breves de Frank Martin

A sonoridade é um aspecto importante para a execução violonística. Esse trabalho tem por objetivo estudar a sonoridade com um elemento da execução musical, entender os seus aspectos formadores dentro do âmbito técnico, textual e perceptivo, utilizando-se como exemplo duas gravações das Quatro Peças Breves de Frank Martin. Os resultado são discutidos à luz do embasamento teórico, fazendo-se considerações a respeito das possibilidades de execução musical e como a sonoridade do violão atua nesse contexto.