Ca(2+ )binding to complement-type repeat domains 5 and 6 from the low-density lipoprotein receptor-related protein

BACKGROUND: The binding of ligands to clusters of complement-type repeat (CR)-domains in proteins of the low-density lipoprotein receptor (LDLR) family is dependent on Ca(2+ )ions. One reason for this cation requirement was identified from the crystal structure data for a CR-domain from the prototypic LDLR, which showed the burial of a Ca(2+ )ion as a necessity for correct folding and stabilization of this protein module. Additional Ca(2+ )binding data to other CR-domains from both LDLR and the LDLR-related protein (LRP) have suggested the presence of a conserved Ca(2+ )cage within CR-domains from this family of receptors that function in endocytosis and signalling. RESULTS: We have previously described the binding of several ligands to a fragment comprising the fifth and the sixth CR-domain (CR56) from LRP, as well as qualitatively described the binding of Ca(2+ )ions to this CR-domain pair. In the present study we have applied the rate dialysis method to measure the affinity for Ca(2+), and show that CR56 binds 2 Ca(2+ )ions with an average affinity of K(D )= 10.6 microM, and there is no indication of additional Ca(2+ )binding sites within this receptor fragment. CONCLUSIONS: Both CR-domains of CR56 bind a single Ca(2+ )ion with an affinity of 10.6 microM within the range of affinities demonstrated for several other CR-domains

The slow professor: challenging the culture of speed in the academy, by Maggie Berg and Barbara K. Seeber

This paper explores the principles of the Slow Movement to counter work-stress among university and college teachers. We believe that a Slow approach to teaching and learning may be the most effective way to counter the erosion of humanistic education by the corporate ethos of consumerism, efficiency, accountability, and standardisation We explore the principles of Slow not only to counter the consumer model of education but also to foster better teachers and learners. It is well-documented that changes in academic work have created significant stress among academic teachers (Catano, Francis, Haines, Kirpalani, Shannon, Stringer, & Lozanksi, 2007; Miller, Buckholdt & Shaw, 2008), and students (Dabney, 1995; Brown & Ralph 1999; Rowbotham and Julian 2006), but what requires further attention is the link between the corporate reliance on efficiency and the problem of lack of time in learning and teaching. Corporatisation has sped up the clock. The Slow Movement—originating in the Slow Food Movement—has gained recognition as a way to resist both globalization and the frantic pace of contemporary life. While slowness has been lauded in architecture, business, urban life and interpersonal relations, among others, it has yet to be applied to academia. Yet, if there is one sector of society that should be cultivating deep thought in themselves and others it is academic teachers. The consumerism that has taken hold in higher education propels the belief that time is money, resulting in superficial learning (Coté & Allahar, 2011b; Readings, 1996). Perhaps the most damaging effect of corporatisation in the universities is that individual educators feel paralysed in the face of overwhelming odds. Our focus on individuals and their own professional practice is conceived as political resistance to corporatisation

Translanguaging and Public Service Encounters: Language Learning in the Library

This article explores the information desk of a city library as a site for language learning. Using a linguistic ethnographic approach, the interactions between a customer experience and information assistant and the many library users who approach her information desk were analysed. Findings are that, in addition to providing information about library resources, information desks are sites at which bits and pieces of different languages are taught and learned. Such language teaching and learning episodes created interactions of inclusion and welcome that went far beyond purely transactional information. Rather, language‐related episodes created moments of human contact and engagement, which were upheld through the translanguaging practices of interactants, the disposition and workplace competence of library staff, and the spatial ecology of the information desk. Furthermore, the article contributes to ongoing theoretical debates about translanguaging by noting that normativity and pressure toward uniformity are as much a part of languaging processes as creativity and flexibility. Our definition of translanguaging recognises the opposing pull of centrifugal and centripetal forces. The article ends by asking what schools, and language education, might learn from public libraries in creating arenas that maintain communitarianism, diversity of expression, and the development of civic skills

Study of the bivariate survival data using frailty models based on Lévy processes

Frailty models allow us to take into account the non-observable inhomogeneity of individual hazard functions. Although models with time-independent frailty have been intensively studied over the last decades and a wide range of applications in survival analysis have been found, the studies based on the models with time-dependent frailty are relatively rare. In this paper, we formulate and prove two propositions related to the identifiability of the bivariate survival models with frailty given by a nonnegative bivariate Lévy process. We discuss parametric and semiparametric procedures for estimating unknown parameters and baseline hazard functions. Numerical experiments with simulated and real data illustrate these procedures. The statements of the propositions can be easily extended to the multivariate case

Applying refinement to the use of mice and rats in rheumatoid arthritis research

Rheumatoid arthritis (RA) is a painful, chronic disorder and there is currently an unmet need for effective therapies that will benefit a wide range of patients. The research and development process for therapies and treatments currently involves in vivo studies, which have the potential to cause discomfort, pain or distress. This Working Group report focuses on identifying causes of suffering within commonly used mouse and rat ‘models’ of RA, describing practical refinements to help reduce suffering and improve welfare without compromising the scientific objectives. The report also discusses other, relevant topics including identifying and minimising sources of variation within in vivo RA studies, the potential to provide pain relief including analgesia, welfare assessment, humane endpoints, reporting standards and the potential to replace animals in RA research

Septic AKI in ICU patients. diagnosis, pathophysiology, and treatment type, dosing, and timing: a comprehensive review of recent and future developments

Evidence is accumulating showing that septic acute kidney injury (AKI) is different from non-septic AKI. Specifically, a large body of research points to apoptotic processes underlying septic AKI. Unravelling the complex and intertwined apoptotic and immuno-inflammatory pathways at the cellular level will undoubtedly create new and exciting perspectives for the future development (e.g., caspase inhibition) or refinement (specific vasopressor use) of therapeutic strategies. Shock complicating sepsis may cause more AKI but also will render treatment of this condition in an hemodynamically unstable patient more difficult. Expert opinion, along with the aggregated results of two recent large randomized trials, favors continuous renal replacement therapy (CRRT) as preferential treatment for septic AKI (hemodynamically unstable). It is suggested that this approach might decrease the need for subsequent chronic dialysis. Large-scale introduction of citrate as an anticoagulant most likely will change CRRT management in intensive care units (ICU), because it not only significantly increases filter lifespan but also better preserves filter porosity. A possible role of citrate in reducing mortality and morbidity, mainly in surgical ICU patients, remains to be proven. Also, citrate administration in the predilution mode appears to be safe and exempt of relevant side effects, yet still requires rigorous monitoring. Current consensus exists about using a CRRT dose of 25 ml/kg/h in non-septic AKI. However, because patients should not be undertreated, this implies that doses as high as 30 to 35 ml/kg/h must be prescribed to account for eventual treatment interruptions. Awaiting results from large, ongoing trials, 35 ml/kg/h should remain the standard dose in septic AKI, particularly when shock is present. To date, exact timing of CRRT is not well defined. A widely accepted composite definition of timing is needed before an appropriate study challenging this major issue can be launched

The EndoC-βH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates

Objective: To characterize the EndoC-βH1 cell line as a model for human beta cells and evaluate its beta cell functionality, focusing on insulin secretion, proliferation, apoptosis and ER stress, with the objective to assess its potential as a screening platform for identification of novel anti-diabetic drug candidates. Methods: EndoC-βH1 was transplanted into mice for validation of in vivo functionality. Insulin secretion was evaluated in cells cultured as monolayer and as pseudoislets, as well as in diabetic mice. Cytokine induced apoptosis, glucolipotoxicity, and ER stress responses were assessed. Beta cell relevant mRNA and protein expression were investigated by qPCR and antibody staining. Hundreds of proteins or peptides were tested for their effect on insulin secretion and proliferation. Results: Transplantation of EndoC-βH1 cells restored normoglycemia in streptozotocin induced diabetic mice. Both in vitro and in vivo, we observed a clear insulin response to glucose, and, in vitro, we found a significant increase in insulin secretion from EndoC-βH1 pseudoislets compared to monolayer cultures for both glucose and incretins.Apoptosis and ER stress were inducible in the cells and caspase 3/7 activity was elevated in response to cytokines, but not affected by the saturated fatty acid palmitate.By screening of various proteins and peptides, we found Bombesin (BB) receptor agonists and Pituitary Adenylate Cyclase-Activating Polypeptides (PACAP) to significantly induce insulin secretion and the proteins SerpinA6, STC1, and APOH to significantly stimulate proliferation.ER stress was readily induced by Tunicamycin and resulted in a reduction of insulin mRNA. Somatostatin (SST) was found to be expressed by 1% of the cells and manipulation of the SST receptors was found to significantly affect insulin secretion. Conclusions: Overall, the EndoC-βH1 cells strongly resemble human islet beta cells in terms of glucose and incretin stimulated insulin secretion capabilities. The cell line has an active cytokine induced caspase 3/7 apoptotic pathway and is responsive to ER stress initiation factors. The cells' ability to proliferate can be further increased by already known compounds as well as by novel peptides and proteins. Based on its robust performance during the functionality assessment assays, the EndoC-βH1 cell line was successfully used as a screening platform for identification of novel anti-diabetic drug candidates. Keywords: EndoC-βH1, Pseudoislets, Glucose stimulated insulin secretion, Somatostatin signaling, Proliferatio