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O filme “Valente” (2012) foi o primeiro filme da Pixar com uma protagonista feminina, além de ser o primeiro filme do estúdio a ser dirigido por uma mulher. O filme é fruto da união entre Pixar e Disney, sendo que a Disney possui uma conhecida tradição em “filmes de princesa”. A partir de um conjunto de atributos comuns às primeiras princesas da Disney, constituindo-se assim a “Princesa Clássica”, verificamos a mudança da figura da princesa na personagem Merida, do filme “Valente”, que sugere uma quebra de padrões narrativos da Princesa Clássica ao se construir conforme uma imagem de mulher contemporânea. Também são identificadas novas funções dramáticas da protagonista Merida, em comparação às princesas antecedentes, bem como novas abordagens de relações entre personagens, como por exemplo a relação com a figura maternal e seu papel como antagonista nos filmes. “Valente” se apropria e ressignifica temas comuns aos filmes de princesa, tais como a bruxa, o casamento, o dever etc., e propõe uma conclusão que foge ao tradicional “felizes para sempre” da Princesa Clássica.

Receptor-interacting protein kinases 1 and 3 (RIPK1/3) have best been described for their role in mediating a regulated form of necrosis, referred to as necroptosis. During this process, RIPK3 phosphorylates mixed lineage kinase domain-like (MLKL) to cause plasma membrane rupture. RIPK3-deficient mice have recently been demonstrated to be protected in a series of disease models, but direct evidence for activation of necroptosis in vivo is still limited. Here, we sought to further examine the activation of necroptosis in kidney ischemia-reperfusion injury (IRI) and from TNFα-induced severe inflammatory response syndrome (SIRS), two models of RIPK3-dependent injury. In both models, MLKL-ko mice were significantly protected from injury to a degree that was slightly, but statistically significantly exceeding that of RIPK3-deficient mice. We also demonstrated, for the first time, accumulation of pMLKL in the necrotic tubules of human patients with acute kidney injury. However, our data also uncovered unexpected elevation of blood flow in MLKL-ko animals, which may be relevant to IRI and should be considered in the future. To further understand the mode of regulation of cell death by MLKL, we screened a panel of clinical plasma membrane channel blockers and we found phenytoin to inhibit necroptosis. However, we further found that phenytoin attenuated RIPK1 kinase activity in vitro, likely due to the hydantoin scaffold also present in necrostatin-1, and blocked upstream necrosome formation steps in the cells undergoing necroptosis. We further report that this clinically used anti-convulsant drug displayed protection from kidney IRI and TNFα-induces SIRS in vivo. Overall, our data reveal the relevance of RIPK3-pMLKL regulation for acute kidney injury and identifies an FDA-approved drug that may be useful for immediate clinical evaluation of inhibition of pro-death RIPK1/RIPK3 activities in human diseases.

When cells undergo necrotic cell death in either physiological or pathophysiological settings in vivo, they release highly immunogenic intracellular molecules and organelles into the interstitium and thereby represent the strongest known trigger of the immune system. With our increasing understanding of necrosis as a regulated and genetically determined process (RN, regulated necrosis), necrosis and necroinflammation can be pharmacologically prevented. This review discusses our current knowledge about signaling pathways of necrotic cell death as the origin of necroinflammation. Multiple pathways of RN such as necroptosis, ferroptosis, and pyroptosis have been evolutionary conserved most likely because of their differences in immunogenicity. As the consequence of necrosis, however, all necrotic cells release damage associated molecular patterns (DAMPs) that have been extensively investigated over the last two decades. Analysis of necroinflammation allows characterizing specific signatures for each particular pathway of cell death. While all RN-pathways share the release of DAMPs in general, most of them actively regulate the immune system by the additional expression and/or maturation of either pro- or anti-inflammatory cytokines/chemokines. In addition, DAMPs have been demonstrated to modulate the process of regeneration. For the purpose of better understanding of necroinflammation, we introduce a novel classification of DAMPs in this review to help detect the relative contribution of each RN-pathway to certain physiological and pathophysiological conditions.

Death of renal cells is central to the pathophysiology of acute tubular necrosis, autoimmunity, necrotizing glomerulonephritis, cystic kidney disease, urosepsis, delayed graft function and transplant rejection. By means of regulated necrosis, immunogenic damage-associated molecular patterns (DAMPs) and highly reactive organelles such as lysosomes, peroxisomes and mitochondria are released from the dying cells, thereby causing an overwhelming immunologic response. The rupture of the plasma membrane exhibits the “point of no return” for the immunogenicity of regulated cell death, explaining why apoptosis, a highly organized cell death subroutine with long-lasting plasma membrane integrity, elicits hardly any immune response. Ferroptosis, an iron-dependent necrotic type cell death, results in the release of DAMPs and large amounts of lipid peroxides. In contrast, anti-inflammatory cytokines are actively released from cells that die by necroptosis, limiting the DAMP-induced immune response to a surrounding microenvironment, whereas at the same time, inflammasome-associated caspases drive maturation of intracellularly expressed interleukin-1β (IL-1β). In a distinct setting, additionally interleukin-18 (IL-18) is expressed during pyroptosis, initiated by gasdermin-mediated plasma membrane rupture. As all of these pathways are druggable, we provide an overview of regulated necrosis in kidney diseases with a focus on immunogenicity and potential therapeutic interventions., Key Points Regulated necrosis is a genetically determined process that contributes to acute kidney injury and causes antibody-mediated rejection (ABMR).Necroptosis, ferroptosis and pyroptosis are the best-studied pathways of regulated necrosis in acute kidney injury and transplantation.Necrotic cell death results in the release of DAMPs and is often prone to elicit an immune response.Failure to efficiently remove necrotic debris by LC3-associated phagocytosis (LAP) results in autoimmunity.During the process of kidney transplantation, necrotic cells are capable of priming memory B cells that may drive ABMR years after transplantation.Necroptosis, ferroptosis and pyroptosis can be therapeutically interfered with by small molecules.

The kidney has been studied as an organ to investigate cell death in vivo for a number of reasons. The unique vasculature that does not contain collateral vessels favors the kidney over other organs for the investigation of ischemia-reperfusion injury. Unilateral uretic obstruction has become the most prominently studied model for fibrosis with impact far beyond postrenal kidney injury. In addition, the tubular elimination mechanisms render the kidney susceptible to toxicity models, such as cisplatin-induced acute kidney injury. During trauma of skeletal muscles, myoglobulin deposition causes tubular cell death in the model of rhabdomyolysis-induced acute kidney injury. Here, we introduce these clinically relevant in vivo models of acute kidney injury (AKI) and critically review the protocols we use to effectively induce them.

The glomerulus functions as the filtration unit of the kidney. The mesangial, endothelial, and podocyte cells of the glomerulus exhibit the three clinically most important cell types, which are involved in diverse pathologic processes. Cell death has hardly been investigated in these cells but may be of critical importance to the pathogenesis of nephrotic syndrome, nephritic syndrome, focal segmental glomerulosclerosis (FSGS), mesangial proliferation, and thrombonic microangiopathy (which involves dysfunction and death of glomerular endothelial cells). The complexity of the glomerulus is frequently affected in autoimmune disorders, which may elicit cell death in mesangial cells and glomerular endothelia. Artificial antisera are used to induce anti-mesangial cell serum-induced mesangiolysis and selective endothelial cell injury, respectively. Genetic variations result in loss of function of podocytes and nephrotic syndrome, which may encompass similar cell death mechanisms as the ones that are observed in the model of secondary focal segmental glomerulosclerosis (FSGS). The following protocols describe our current arsenal to target glomerular cells in vivo.

Hypoxia-inducible factor (HIF)-2–triggered erythropoietin production in renal interstitial fibroblast–like cells is the physiologically relevant source of erythropoietin for regulating erythropoiesis. During renal fibrosis, these cells transform into myofibroblasts and lose their ability to produce sufficient erythropoietin leading to anemia. To find if other cells for erythropoietin production might exist in the kidney we tested for the capability of nonepithelial glomerular cells to elaborate erythropoietin. Therefore, HIF transcription factors were stabilized by cell-specific deletion of the von Hippel–Lindau ( VHL ) gene. Inducible deletion of VHL in glomerular connexin40-expressing cells (endothelial, renin-expressing, and mesangial cells) markedly increased glomerular erythropoietin mRNA expression levels, plasma erythropoietin concentrations, and hematocrit values. These changes were mimicked by inducible cell-specific VHL deletion in renin-expressing and in mesangial cells but not in endothelial cells. The increases of erythropoietin production were absent, when VHL was co-deleted with HIF-2. The induction of glomerular erythropoietin expression was associated with the downregulation of juxtaglomerular renin expression, again in a HIF-2-dependent manner. Thus, VHL deletion in renin-expressing and in mesangial cells induces the capability to produce relevant amounts of erythropoietin and to suppress renin expression in the adult kidney if HIF-2 is stabilized.

Basierend auf neuen pathophysiologischen Erkenntnissen der vorausgehenden Jahre, erfolgte im Jahr 2010 die Einfuhrung einer neuen Klassifikation einer Glomerulonephritis mit dominanten oder kodominanten C3-Ablagerungen, deren wesentliche Untergruppe die C3-Glomerulopathie (C3G) darstellt. Wahrend sekundare Ursachen zu einer immunkomplexvermittelten membranoproliferativen Glomerulonephritis (MPGN) fuhren, liegen die Hauptursachen der C3-Glomerulopathie primar in Storungen des Komplementsystems, die dann haufig in Form einer MPGN, aber auch anderer lichtmikroskopischer Erscheinungsbilder apparent werden. Die Kenntnis der zugrunde liegenden Pathophysiologie ist fur die weiterfuhrende Diagnostik zur Abklarung sekundarer Ursachen wegweisend. Eine umfassende Komplementanalytik, begleitet von Antikorperscreening und humangenetischer Analyse, sollte konsequent erfolgen. Wenn auch bis heute durch Studien nicht systematisch validiert, bieten die vorliegenden Erkenntnisse doch eine robuste Grundlage fur die Anwendung verfugbarer Therapieansatze fur diese haufig rasch progredienten und nach Nierentransplantation oftmals wiederkehrenden Krankheitsbilder. In der vorliegenden Ubersicht werden die derzeitigen Erkenntnisse systematisch dargestellt und in Form eines (nationalen) Expertenkonsensus zur Diagnostik der C3-dominanten Glomerulonephritis und der MPGN zusammengefasst.

Background Severe forms of mono- and polygenetic hypercholesterolemia as well as elevated Lipoprotein (a) (LP(a)) with progressing cardiovascular (CV) disease are indication for lipoprotein apheresis (LA) in Germany. Many studies investigated pleiotropic effects of LA that might contribute to beneficial effects in advanced atherosclerosis. The present study aimed at investigating the potential role of Proangiogenic Cells (PAC) in patients with new onset or chronic LA using the heparin induced extracorporeal LDL-precipitation (H.E.L.P.) apheresis system. Methods Patients (n = 10) new to LA and HELP treatment were investigated immediately before, shortly after, 24 h later and 4 weeks following LA. Peripheral blood was used to count PAC in circulation via flow cytometry. In a second step, blood cells from patients were cultured in endothelial selective medium and further evaluated for adhesion in fibronectin coated chamber slides and migratory capacity (stromal cell-derived factor-1 (SDF-1) induced migration). Results Cells expressing typical EPC markers were rarely detected in blood samples. No differences occurred over time in CD34 + ; CD34 + CD133 + CD45 − ; CD34 + /KDR + and CXCR4 + /CD14 + positive PAC. We found no differences in cell adhesion at the different time points, while significantly more cells migrated into the SDF-1 medium following four weeks of continuing apheresis therapy. Conclusion Using well established systems, this study was not able to demonstrate relevant acute effects of LA on PAC in patients new to LA. The increased migratory capacity of PAC might be an indicator of chronic beneficial pleiotropic effects in patients undergoing H.E.L.P. apheresis.

Cardiovascular disease is the major cause of morbidity and mortality in the world. Diabetes and its complications, such as diabetic nephropathy, dramatically increase cardiovascular risk. Association studies suggest that asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthases, plays a role in the pathogenesis of diabetic nephropathy. The major pathway of ADMA catabolism is hydrolysis by dimethylarginine dimethylaminohydrolase 1 (DDAH1). The goal of this study was to test the hypothesis that lowering ADMA by overexpression of DDAH1 protects from development of diabetic nephropathy.Diabetes was induced with streptozotocin (STZ) in wild type and DDAH1 transgenic mice. Healthy mice served as controls. Mice were sacrificed after 20 weeks of diabetes. ADMA levels were assessed by isotope-dilution tandem mass spectrometry, creatinine by standard laboratory methods and albumin by ELISA. Kidney tissues were stained for markers of glomerular cells, cell matrix, inflammation and cell proliferation.STZ led to development of diabetes in all injected mice. Transgenic overexpression of DDAH1 led to a decrease in plasma ADMA levels in healthy animals. Diabetic state itself did not lead to elevation of plasma ADMA levels. Diabetic mice of both genotypes developed albuminuria (27 and 25 vs. 9 and 6 μg albumin/mg creatinine) (p 0.01). There were no changes in glomerular matrix expansion, podocyte injury, inflammatory or proliferative response.STZ-induced diabetes led to the development of early features of diabetic nephropathy. Overexpression of DDAH1 and lowering of systemic ADMA levels did not prevent these changes, indicating that ADMA is not the major mediator of the early diabetic changes reflected by this experimental model.

ABO incompatible (ABOi) organ transplantation requires pre-transplant reduction of the recipient's IgG and IgM isoagglutinin titer against the donor to prevent hyperacute rejection. Over the past four years we primarily used unspecific IgG immunoadsorption (IA) for this purpose and combined this selectively with membrane filtration (IAc) to reduce IgM isoagglutinines. In patients with an initial IgG titer against donor below 1:64, plasma exchange (PE) was initiated. In this retrospective analysis covering January 2012 to August 2015 we compared how efficiently IgG and IgM isoagglutinines in a total of 22 ABOi kidney transplant recipients were reduced by either IA (n = 75 sessions), IAc (n = 14 sessions) or PE (n = 40 sessions). Median pre-treatment IgG isoagglutinin titers were 32 (4-4096) while IgM titers were 16 (1-256) respectively. Mean IgG reduction by either treatment modality was 1.3 ± 0.9 (IA), 1.8 ± 1.0 (IAc) and 2.6 ± 1.3 (PE) titer steps per session (p 0.001 IA vs. PE; p 0.04 PE vs. IAc). Mean IgM reduction was 0.6 ± 0.6 (IA), 1.8 ± 0.8 (IAc) and 2.4 ± 1.9 (PE) titer steps (p 0.001 for both IA vs. PE and IA vs. IAc). Our data indicate that PE efficiently removed IgG- and IgM isoagglutinines. By processing only half the plasma volume per treatment PE was twice as effective as IA in terms of IgG-type isoagglutinin removal in our patient group. This is best explained by the presence of soluble AB0 antigens in the FFP used as plasma replacement. These advantages in efficacy have to be weighed against the potential hazards of PE. Combination of IA and plasma filtration effectively removes IgM-type and even enhances net IgG-type isoagglutinin elimination compared to IA alone. When trying to avoid PE, combined application of IA and IAc is a possible and effective way to reduce isoagglutinin titers before ABOi transplantation.

The paper analyzes the TV show Seinfeld and its breaking with the genre conventions historically constructed on american television. To this end, our analysis points out two dimensions of program aesthetics: the narrative and the stylistic ones. From the narrative aesthetics point of view, we indicated a break with the goal oriented dramatic model (or the “Aristotelian model”, hegemonic in the television of such period). From a stylistic point of view, we consider how the increasing adoption of unusual production practices led to the formation of a hybrid between the multi-camera sitcom and the single camera programs. El artículo analiza la serie Seinfeld y su ruptura con las convenciones de género históricamente construidas en las TV americana. Para ello, nuestro análisis observa dos dimensiones de la estética del programa: la narrativa y la estilística. Desde el punto de vista de la narrativa, señalamos la ruptura con el modelo dramático orientado hacia un objetivo (o un “modelo aristotélico”, hegemónico en la TV de este período). Desde el punto de vista estilístico, consideramos cómo la creciente adopción de prácticas de producción poco comunes llevó a la construcción de un Sitcom híbrido entre la multicámara y los programas de cámara única. O artigo analisa a série Seinfeld e sua ruptura com as convenções de gênero historicamente construídas na TV americana. Para tal, nossa análise observa duas dimensões da estética do programa: a narrativa e a estilística. Do ponto de vista da estética narrativa, indicamos a ruptura com o modelo dramático orientado a um objetivo (ou o “modelo aristotélico”, hegemônico na TV do período). Do ponto de vista estilístico, consideramos o quanto a crescente adoção de práticas de produção pouco comuns levou à constituição de um sitcom híbrido entre o multicâmera e os programas de câmera única.

We reported earlier that PPAR-gamma regulates renin transcription through a human-specific atypical binding sequence termed hRen-Pal3. Here we developed a mouse model to investigate the functional relevance of the hRen-Pal3 sequence in vivo since it might be responsible for the increased renin production in obesity and thus for the development of accompanying arterial hypertension. We used bacterial artificial chromosome construct and co-placement strategy to generate two transgenic mouse lines expressing the human renin gene from identical genomic locus without affecting the intrinsic mouse renin expression. One line carried a wild-type hRen-Pal3 in the transgene (Pal3wt strain) and the other a mutated non-functional Pal3 (Pal3mut strain). Human renin expression was correctly targeted to the renin-producing juxtaglomerular (JG) cells of kidney in both lines. However, Pal3mut mice had lower basal human renin expression. Since human renin does not recognize mouse angiotensinogen as substrate, the blood pressure was not different between the strains. Stimulation of renin production with the angiotensin-converting enzyme inhibitor enalapril equipotentially stimulated the human renin expression in Pal3wt and Pal3mut mice. High-fat diet for 10 weeks which is known to activate PPAR-gamma failed to increase human renin mRNA in kidneys of either strain. These findings showed that the human renin PPAR-gamma-binding sequence hRen-Pal3 is essential for basal renin expression but dispensable for the cell-specific and high-fat diet regulated renin expression in the kidney.

Transplant international (2017). doi:10.1111/tri.13110, Published by Wiley-Blackwell, Oxford [u.a.]

There is an art of which every human being should be a master, the art of reflection. Very pertinent is the question which the English poet Samuel Taylor Coleridge puts, – “If you are not a thinking man, to what purpose are you a man at all?” (Coleridge, 2015). Daniel Kahneman differentiates between “Thinking, Fast and Slow”.

Faced with the ruins of the house of risk management, we now recognize that the first way to managing extreme and systemic risks effectively (Part I) turned out to be not promising or ineffective (Propositions 6 and 8) and that the same holds for the second way in this Part II, which dealt with not quantitative, but qualitative approaches (Conjecture 9). Yet, criticizing something is always considerably easier than proposing a constructive alternative. The call for a Third Way, for alternative risk models and measures, better: an alternative class of risk models for complex financial systems thus becomes unmistakable.

The comet may have taken an infinite number of other trajectories from point to point, the finitely many points observed on its previous path do not say how the actual trajectory continues in the future or at the other points, respectively. In a similar way, a finite number of statistical data may not be safely extrapolated into the future. Nonetheless, we may rely on the “habits of nature”, which, as experience according to Leibniz shows, let her causes recur in a similar manner throughout time – put in the image of the comet: it may be safely assumed a path with no wild or radical changes.

The moral of the story of the preceding discussion in Part I could well be that quantitative approaches to assessing and managing (extreme and systemic) risk ought to be abandoned, which goes beyond the conclusions reached in Part I. One might argue in favor of reinventing finance (Shiller, 2012: xii) and that uncertainties, as contrasted with clearly measurable risks which are not to be expected (see Chapter 7, where “our [supposedly] best tools [might have] prove[d] to be our worst instruments”; Blyth, 2010: 453), require a broader, more forward-looking and strategy-oriented approach to risk management.

In this inquiry, we envision the behavior of LBR by taking up the case of LTCM from Chapter 5.2. The LTCM case is a predestined application example of LBR since its fall counts as one of the most impressive instances of massive losses in derivative markets. This chapter is structured as follows. We start by reconstructing its dynamic trading and so-called ‘fixed income’ strategy with an example trade (16.1.), before elaborating on how the portfolio is described in a concrete LBR model (16.2.). The out-come of our exemplary risk model for the portfolio will be discussed at the end, in the remaining two sections, 16.3. based on the three steps of visual data mining (overview – zoom/filter – details); and 16.4. as an overall bottom line.

The purpose of this chapter is to argue for the relevance of systemic risks for (private) banks (as opposed to regulators) (5.1.) and to provide empirical evidence for systemic risk events (5.2.) by drawing on an illustrative case which will reappear in the course of this work. This chapter is essential, not only for contextualizing RQ1 as this study is interested in banks’, and primarily in-house, risk management approaches, but also for clarifying the centrality of RQ1 for all participants in financial markets.

The distinctions between qualitative (see this Part II) and quantitative (see Part I above) approaches have been sharply etched and, as underscored by Forrester (1975: 48) or Rapoport (1953), somewhat exaggerated. Yet, “the proper impression emerges – the ‘art’ of the former is still better able to deal with decisions of great consequence than the ‘science’ of the latter” (Forrester, 1975: 48). The anthropomorphic desiderata of good and effective (risk management) decisions, resilience, and flexibility (Brose et al., 2014a: 369; Helbing, 2013; Detken & Nymand-Andersen, 2013; Sheffi, 2005) become more important than preciseness, efficiency and optimization under conditions of volatility, uncertainty, complexity, and ambiguity that characterize our world, a so-named “VUCA world” (Bennett & Lemoine, 2014; Kleindorfer, 2010: 185; see Chapter 6 for a deeper analysis of complex systems).

Another treatise on systemic risk is a risky venture for any author. There are simply so many of them. However, despite the amount of research undertaken in the area of systemic risk in the financial sector (e.g., Battiston et al., 2012; Billio et al., 2012; Blyth, 2010; Sornette, 2009; Gorton & Metrick, 2009; Huang et al., 2009a; Hellwig, 2008; Schwarcz, 2008; Taleb, 2007a; etc.), a holistic and, especially, a complexity/systems theories driven and logic-based perspective can be very fruitful for contributing to how to be better equipped for assessing and managing systemic risks more effectively in the future (Klimek et al., 2015; Thurner et al., 2010); a critical perspective on systemic and extreme risks in financial systems and quantitative risk models that lie within banks’ scope. To start with, it ought to be acknowledged that systems theoretic notions are beginning to provide new insights on risk and resilience of banks (e.g., Battiston et al., 2016; Bonabeau, 2007: 64).

“The financial markets, seen by many as the most efficient and farsighted, should rapidly evolve to near-optimality due to the huge stakes and enormous talent brought to bear. Yet even the highly sophisticated hedge funds [like LTCM, C.H.] bear the scars of self-inflicted wounds from open-loop thinking [see 5.1.; because their risk models have missed to consider that other actors might pursue the same or a similar strategy as an input to the models; C.H.].” (Sterman, 2000: 697). At best, the result is inefficiency; at worst, it is ineffectivity which hurts the organization, the people in it, and not too rarely third parties or even the larger society.

The fact that simple mathematical structures can simulate complex, dynamic systems is well-known and exemplified by, for instance, the “Game of Life” (Berlekamp et al., 2001). Our symbolic models of risk are designed in the tradition of such devices and embedded in a novel LBR approach which consists of three components.

The Central Argument has shaken the house of modern risk management literature and practice with their standard extreme risk modeling approaches in banking to its very foundations; it further serves the purpose of shaking risk management professionals out of a ‘dogmatic slumber’ (with regard to Part III). In a broader sense, it also illustrates the confusion (folly and nonsense) that can occur when a concept that belongs properly to one realm, namely that of disorganized complexity and of gambling, wagering, betting and games of chance, in particular (Hacking, 2006), is uncritically and improperly applied to another (risk management in banking): the borrowing of any method on the sole ground that is has been successful somewhere else is inadmissible (Schumpeter, 1954/1994: 17; von Hayek, 1943).

The purpose of this chapter is to outline and discuss different concepts of risk (4.1.), systemic risk (4.2.), and to define a proper meaning of the two terms in the realm of banking. This is essential as the central research question RQ1 revolves around quantitative risk assessment and management approaches and it should be clear what the object of interest, of assessment and management endeavors is.

The presence, if not prevalence, of crises is not unusual in many systems. Current risk management and financial systems are not an exception. It does not follow, however, that they must decline and face doom.

The renal disorder C3 glomerulopathy with dense deposit disease (C3G-DDD) pattern results from complement dysfunction and primarily affects children and young adults. There is no effective treatment, and patients often progress to end-stage renal failure. A small fraction of C3G-DDD cases linked to factor H or C3 gene mutations as well as autoantibodies have been reported. Here, we examined an index family with 2 patients with C3G-DDD and identified a chromosomal deletion in the complement factor H-related (CFHR) gene cluster. This deletion resulted in expression of a hybrid CFHR2-CFHR5 plasma protein. The recombinant hybrid protein stabilized the C3 convertase and reduced factor H-mediated convertase decay. One patient was refractory to plasma replacement and exchange therapy, as evidenced by the hybrid protein quickly returning to pretreatment plasma levels. Subsequently, complement inhibitors were tested on serum from the patient for their ability to block activity of CFHR2-CFHR5. Soluble CR1 restored defective C3 convertase regulation; however, neither eculizumab nor tagged compstatin had any effect. Our findings provide insight into the importance of CFHR proteins for C3 convertase regulation and identify a genetic variation in the CFHR gene cluster that promotes C3G-DDD. Monitoring copy number and sequence variations in the CFHR gene cluster in C3G-DDD and kidney patients with C3G-DDD variations will help guide treatment strategies.

Background Impaired cytomegalovirus (CMV)-specific cell-mediated immunity (CMV-CMI) is a major cause of uncontrolled CMV reactivation and associated complications in both solid-organ transplantation (SOT) and hematopoietic stem cell transplantation (HSCT). Reliably assessing CMV-CMI is desirable to individually adjust antiviral and immunosuppressive therapy. We demonstrate here the suitability of a novel IFN-γ ELISpot assay (T-Track® CMV), based on the stimulation of PBMC with pp65 and IE-1 CMV proteins, to monitor CMV-CMI in SOT and HSCT patients. Methods Two independent prospective, longitudinal, observational, multicenter studies were conducted: in 86 intermediate-risk (D−/R+, D+/R+) renal transplant recipients (completed), and in 175 intermediate- or high-risk (D+/R+, D+/R−, D−/R+) HSCT recipients (ongoing). In both studies, patients underwent pre-emptive antiviral therapy. CMV-CMI, CMV load and clinical complications were monitored over ~6 months post-transplantation. Results In the kidney transplantation setting, 95% and 88–92% of IFN-γ ELISpot test results were positive pre- and post-transplantation, respectively. CMV-specific response was reduced following immunosuppressive therapy and increased in patients with graft rejection, indicating the ability of the assay to monitor the patients’ immunosuppressive state. Interestingly, median pp65-specific response was 9-fold higher in patients with self-clearing viral load compared with antivirally-treated patients prior to first detection of CMV (MWU; P < 0.001), suggesting that reactivity to pp65 is a potential immunocompetence marker. In HSCT patients, interim data analysis indicates that pp65-specific CMI measured after resolution of a primary CMV reactivation (requiring antiviral treatment) is a fair predictor of occurrence of recurrent CMV reactivation. Out of 71 patients (25 D+/R+, 3 D+/R−, 43 D−/R+) who experienced a primary CMV reactivation, 27 encountered a recurrent CMV reactivation. Interestingly, 39/44 (89%) patients free of recurrent reactivation had a positive pp65-specific test result following primary CMV reactivation. Conclusion Altogether, this novel IFN-γ ELISpot assay is a highly sensitive immune-monitoring tool with a potential use for the risk assessment of CMV-related clinical complications after SOT and HSCT. Disclosures All authors, Lophius Biosciences: Investigator, Research support.

Immunofluorescent staining is a widespread tool in basic science to understand organ morphology and (patho-) physiology. The analysis of imaging data is often performed manually, limiting throughput and introducing human bias. Quantitative analysis is particularly challenging for organs with complex structure such as the kidney. In this study we present an approach for automatic quantification of fluorescent markers and histochemical stainings in whole organ sections using open source software. We validate our novel method in multiple typical challenges of basic kidney research and demonstrate its general relevance and applicability to other complex solid organs for a variety of different markers and stainings. Our newly developed software tool “AQUISTO”, applied as a standard in primary data analysis, facilitates efficient large scale evaluation of cellular populations in various types of histological samples. Thereby it contributes to the characterization and understanding of (patho-) physiological processes.

The role of bone marrow marrow-derived cells after kidney endothelial injury is controversial. In this study, we investigated if and to what extent extrarenal cells incorporate into kidney endothelium after acute as well as during chronic endothelial injury. Fischer F-344 wt (wild type) rat kidney grafts were transplanted into R26- hPAP (human placental alkaline phosphatase) transgenic Fischer F-344 recipient rats to allow identification of extrarenal cells by specific antibody staining. A severe model of renal thrombotic microangiopathy was induced via graft perfusion with antiglomerular endothelial cell (GEN) antibody and resulted in eradication of 85% of the glomerular and 69% of the peritubular endothelium (GEN group). At week 4 after injury, renal endothelial healing as well as recovery of the kidney function was seen. Endothelial chimerism was evaluated by double staining for hPAP and endothelial markers RECA-1 or JG-12. Just 0.25% of the glomerular and 0.1% of the peritubular endothelium was recipient derived. In a second experiment, chronic endothelial injury was induced by combination of kidney transplantation with 5/6 nephrectomy (5/6 Nx group). After 14 wk, only 0.86% of the peritubular and 0.05% of the glomerular endothelium was of recipient origin. In summary, despite demonstration of extensive damage and loss as well as excellent regeneration, just a minority of extrarenal cells were incorporated into kidney endothelium in rat models of acute and chronic renal endothelial cell injury. Our results highlight that kidney endothelial regeneration after specific and severe injury is almost exclusively of renal origin.

Nierentransplantierte Patienten haben im Vergleich zur Gesamtbevolkerung ein 2- bis 4-fach erhohtes Tumorrisiko, wobei das Risiko besonders fur Hauttumoren, Lymphome und Nierenzellkarzinome erhoht ist. Ursachlich fur die erhohte Tumorinzidenz ist die durch die immunsuppressive Therapie bedingte eingeschrankte Uberwachungskapazitat des Immunsystems in Bezug auf maligne Veranderungen sowie die erhohte Anfalligkeit fur sog. Onkoviren. Hervorzuheben ist hier die Bedeutung der Primarinfektion mit Epstein-Barr-Virus (EBV) bei der Entstehung von Posttransplantationslymphomen. Allerdings ist auch eine chronische Nierenerkrankung selbst mit einem erhohten Risiko, insbesondere fur Nieren- und Blasentumoren, vergesellschaftet. Bezuglich der Art der immunsuppressiven Medikation bestehen Unterschiede fur das Tumorrisiko. Das gehaufte Auftreten bestimmter Tumoren und ihre Risikofaktoren haben Implikationen fur die Pravention, Fruherkennung und Behandlung von Tumoren nach Nierentransplantation.

Over the past decade, researchers have made substantial progress in characterizing a network of mononuclear phagocytes in the kidney, which variously have been referred to as resident macrophages or dendritic cells. Two recent studies, published in Cell and Kidney International, have identified these resident macrophages/dendritic cells as local immune monitors of peritubular capillaries for circulating antigens and immune complexes. These cells appear to represent an early line of defense against circulating infectious particles and immune complexes, but the resulting inflammatory response may also contribute to interstitial inflammation and kidney disease progression.

Este artigo tem por objetivo empreender uma análise hermenêutica dos textos críticos veiculados nos principais jornais, revistas e sites sobre o filme O Som ao Redor, do cineasta Kleber Mendonça Filho. Foram selecionados 16 textos críticos, os quais forneceram subsídios para a observação, entre outros elementos, da recorrência de três pressupostos estéticos: o aspecto sociológico do filme, pelo qual é possível compreendê-lo como uma metáfora ou um discurso sobre o Brasil; a estética da mistura de gêneros, a partir, principalmente, dos recursos sonoros empregados no longa-metragem e, por fim, o caráter inovador, critério moderno que valoriza o potencial de novidade contido numa obra. El artículo tiene como fin emprender un análisis hermenéutico de los textos críticos en circulación en los principales periódicos, revistas y sites acerca de la película O Som ao Redor, del cineasta Kleber Mendonça Filho. Se seleccionaron 16 textos críticos, los que brindaron subsidios para la observación, entre otros elementos, de la recurrencia de tres presupuestos estéticos: el aspecto sociológico de la película, mediante el cual es posible comprenderla como una metáfora o un discurso sobre Brasil; la estética de la mezcla de géneros, desde, sobre todo, los recursos sonoros utilizados en el largometraje, y, por fin, el carácter innovador, criterio moderno que valora el potencial de novedoso contenido en una obra. The article undertakes a hermeneutical analysis ofa series ofcritical reviews appearing in major newspapers, magazines and media sites on O Som ao Redor, by the filmmaker Kleber Mendonça Filho. Sixteen reviews were selected. They provide support for observing, among other elements, the recurrence of three aesthetic proposals: the sociological aspect, whereby it is possible to understand the film as a metaphor or a discourse on Brazil; the aesthetics of the mixture of genres, particularly the sound resources used in this feature film, and, lastly, its innovative nature or modern criterion that values the potential of novel content in a work of art.

DNA binding protein A (DbpA) is a member of the human cold shock domain-containing protein superfamily, with known functions in cell proliferation, differentiation, and stress responses. DbpA mediates tight junction-associated activities in tubular epithelial cells, but the function of DbpA in mesangial cells is unknown. Here, we found DbpA protein expression restricted to vascular smooth muscle cells in healthy human kidney tissue but profound induction of DbpA protein expression within the glomerular mesangial compartment in mesangioproliferative nephritis. In vitro, depletion or overexpression of DbpA using lentiviral constructs led to inhibition or promotion, respectively, of mesangial cell proliferation. Because platelet-derived growth factor B (PDGF-B) signaling has a pivotal role in mesangial cell proliferation, we examined the regulatory effect of PDGF-B on DbpA. In vitro studies of human and rat mesangial cells confirmed a stimulatory effect of PDGF-B on DbpA transcript numbers and protein levels. Additional in vivo investigations showed DbpA upregulation in experimental rat anti-Thy1.1 nephritis and murine mesangioproliferative nephritis models. To interfere with PDGF-B signaling, we injected nephritic rats with PDGF-B neutralizing aptamers or the MEK/ERK inhibitor U0126. Both interventions markedly decreased DbpA protein expression. Conversely, continuous PDGF-B infusion in healthy rats induced DbpA expression predominantly within the mesangial compartment. Taken together, these results indicate that DbpA is a novel target of PDGF-B signaling and a key mediator of mesangial cell proliferation.

The Global Sequence Protocol (GSP) is an operational model for replicated data stores, in which updates propagate asynchronously. We introduce the GSP-calculus as a formal model for GSP. We give a formal account for its proposed implementation, which addresses communication failures and compact representation of data, and use simulation to prove that the implementation is correct. Then, we use the GSP-calculus to reason about execution histories and prove ordering guarantees, such as read my writes, monotonic reads, causality and consistent prefix. We also prove that GSP extended with synchronous updates provides strong consistency guarantees. Fil: Melgratti, Hernan Claudio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Computación; Argentina Fil: Roldán, Christian Hugo. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Computación; Argentina

Background: Elevation of the endogenous inhibitor of nitric oxide synthase asymmetric dimethylarginine (ADMA) has been shown to be associated with increased risk of cardiovascular diseases. There are two major pathways of ADMA catabolism: hydrolysis to citrulline by dimethylarginine dimethylaminohydrolases (DDAH) and transamination by alanine-glyoxylate aminotransferase 2 (AGXT2) with formation of asymmetric dimethylguanidino valeric acid (ADGV). The second pathway is poorly characterized. The goal of the current study was to test the hypothesis that transgenic overexpression of AGXT2 leads to lowering of systemic levels of ADMA and improvement of vasomotor function. Methods and Results: We generated transgenic mice (TG) with ubiquitous overexpression of AGXT2 under control of the chicken beta actin (CAG) promoter. qPCR and Western Blot were used to confirm the ubiquitous expression of the transgene. There were no developmental or phenotypic changes in the TG animals. Biochemical data were generated using HPLC-MS/MS. ADMA plasma levels were decreased by 15% (p Conclusion: In the current study we demonstrated that upregulation of AGXT2 leads to lowering of ADMA levels and improvement of endothelium-dependent relaxation in vivo. AGXT2 thereby may be a potential drug target for long-term reduction of systemic ADMA levels in cardiovascular pathologies. This is especially important, because all the efforts to develop pharmacological ADMA-lowering interventions by means of upregulation of DDAH have not been successful so far.