Mice carrying ubiquitin-specific protease 2 (Usp2) gene inactivation maintain normal sodium balance and blood pressure

Ubiquitylation plays an important role in the control of Na⁺ homeostasis by the kidney. It is well established that the epithelial Na⁺ channel ENaC is regulated by the ubiquitin-protein ligase NEDD4-2, limiting ENaC cell surface expression and activity. Ubiquitylation can be reversed by the action of deubiquitylating enzymes (DUBs). One such DUB, USP2-45, was identified previously as an aldosterone-induced protein in the kidney and is also a circadian output gene. In heterologous expression systems, USP2-45 binds to ENaC, deubiquitylates it, and enhances channel density and activity at the cell surface. Because the role of USP2-45 in renal Na⁺ transport had not been studied in vivo, we investigated here the effect of Usp2 gene inactivation in this process. We demonstrate first that USP2-45 protein has a rhythmic expression with a peak at ZT12. Usp2-KO mice did not show any differences from wild-type littermates with respect to the diurnal control of Na⁺ or K⁺ urinary excretion and plasma levels either on a standard diet or after acute and chronic changes to low- and high-Na⁺ diets, respectively. Moreover, they had similar aldosterone levels on either a low- or high-Na⁺ diet. Blood pressure measurements using telemetry did not reveal variations compared with control mice. Usp2-KO mice did not display alterations in expression of genes involved in sodium homeostasis or the ubiquitin system, as evidenced by transcriptome analysis in the kidney. Our data suggest that USP2 does not play a primary role in the control of Na⁺ balance or blood pressure.

ICOMF: Towards a Multi-cloud Ecosystem for Dynamic Resource Composition and Scaling

Modern cloud-based applications and infrastructures may include resources and services (components) from multiple cloud providers, are heterogeneous by nature and require adjustment, composition and integration. The specific application requirements can be met with difficulty by the current static predefined cloud integration architectures and models. In this paper, we propose the Intercloud Operations and Management Framework (ICOMF) as part of the more general Intercloud Architecture Framework (ICAF) that provides a basis for building and operating a dynamically manageable multi-provider cloud ecosystem. The proposed ICOMF enables dynamic resource composition and decomposition, with a main focus on translating business models and objectives to cloud services ensembles. Our model is user-centric and focuses on the specific application execution requirements, by leveraging incubating virtualization techniques. From a cloud provider perspective, the ecosystem provides more insight into how to best customize the offerings of virtualized resources.

Assessment of a robust opportunistic routing for video transmission in dynamic topologies

Mobile multimedia ad hoc services run on dynamic topologies due to node mobility or failures and wireless channel impairments. A robust routing service must adapt to topology changes with the aim of recovering or maintaining the video quality level and reducing the impact of the user's experience. In those scenarios, beacon-less Opportunistic Routing (OR) increases the robustness by supporting routing decisions in a completely distributed manner based on protocol-specific characteristics. However, the existing beacon-less OR approaches do not efficiently combine multiple metrics for forwarding selection, which cause higher packet loss rate, and consequently reduce the video quality level. In this paper, we assess the robustness and reliability of our recently developed OR protocol under node failures, called cross-layer Link quality and Geographical-aware OR protocol (LinGO). Simulation results show that LinGO achieves multimedia dissemination with QoE support and robustness in scenarios with dynamic topologies.

Illuminating somatostatin analog action at neuroendocrine tumor receptors

Somatostatin analogs for the diagnosis and therapy of neuroendocrine tumors (NETs) have been used in clinical applications for more than two decades. Five somatostatin receptor subtypes have been identified and molecular mechanisms of somatostatin receptor signaling and regulation have been elucidated. These advances increased understanding of the biological role of each somatostatin receptor subtype, their distribution in NETs, as well as agonist-specific regulation of receptor signaling, internalization, and phosphorylation, particularly for the sst2 receptor subtype, which is the primary target of current somatostatin analog therapy for NETs. Various hypotheses exist to explain differences in patient responsiveness to somatostatin analog inhibition of tumor secretion and growth as well as differences in the development of tumor resistance to therapy. In addition, we now have a better understanding of the action of both first generation (octreotide, lanreotide, Octreoscan) and second generation (pasireotide) FDA-approved somatostatin analogs, including the biased agonistic character of some agonists. The increased understanding of somatostatin receptor pharmacology provides new opportunities to design more sophisticated assays to aid the future development of somatostatin analogs with increased efficacy.

Establishing optimal quantitative-polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F-associated myeloproliferative neoplasms: a joint European LeukemiaNet/MPN&MPNr-EuroNet (COST action BM0902) study.

Reliable detection of JAK2-V617F is critical for accurate diagnosis of myeloproliferative neoplasms (MPNs); in addition, sensitive mutation-specific assays can be applied to monitor disease response. However, there has been no consistent approach to JAK2-V617F detection, with assays varying markedly in performance, affecting clinical utility. Therefore, we established a network of 12 laboratories from seven countries to systematically evaluate nine different DNA-based quantitative PCR (qPCR) assays, including those in widespread clinical use. Seven quality control rounds involving over 21,500 qPCR reactions were undertaken using centrally distributed cell line dilutions and plasmid controls. The two best-performing assays were tested on normal blood samples (n=100) to evaluate assay specificity, followed by analysis of serial samples from 28 patients transplanted for JAK2-V617F-positive disease. The most sensitive assay, which performed consistently across a range of qPCR platforms, predicted outcome following transplant, with the mutant allele detected a median of 22 weeks (range 6-85 weeks) before relapse. Four of seven patients achieved molecular remission following donor lymphocyte infusion, indicative of a graft vs MPN effect. This study has established a robust, reliable assay for sensitive JAK2-V617F detection, suitable for assessing response in clinical trials, predicting outcome and guiding management of patients undergoing allogeneic transplant.

Site- and species-specific responses of forest growth to climate across the European continent

Aim To evaluate the climate sensitivity of model-based forest productivity estimates using a continental-scale tree-ring network. Location Europe and North Africa (30–70° N, 10° W–40° E). Methods We compiled close to 1000 annually resolved records of radial tree growth for all major European tree species and quantified changes in growth as a function of historical climatic variation. Sites were grouped using a neural network clustering technique to isolate spatiotemporal and species-specific climate response patterns. The resulting empirical climate sensitivities were compared with the sensitivities of net primary production (NPP) estimates derived from the ORCHIDEE-FM and LPJ-wsl dynamic global vegetation models (DGVMs). Results We found coherent biogeographic patterns in climate response that depend upon (1) phylogenetic controls and (2) ambient environmental conditions delineated by latitudinal/elevational location. Temperature controls dominate forest productivity in high-elevation and high-latitude areas whereas moisture sensitive sites are widespread at low elevation in central and southern Europe. DGVM simulations broadly reproduce the empirical patterns, but show less temperature sensitivity in the boreal zone and stronger precipitation sensitivity towards the mid-latitudes. Main conclusions Large-scale forest productivity is driven by monthly to seasonal climate controls, but our results emphasize species-specific growth patterns under comparable environmental conditions. Furthermore, we demonstrate that carry-over effects from the previous growing season can significantly influence tree growth, particularly in areas with harsh climatic conditions – an element not considered in most current-state DGVMs. Model–data discrepancies suggest that the simulated climate sensitivity of NPP will need refinement before carbon-cycle climate feedbacks can be accurately quantified.

The Nature of Sustainable Consumption and How to Achieve it. Results from the Focal Topic "From Knowledge to Action – New Paths towards Sustainable Consumption"

Is the online trade with second-hand products changing individual consumer behaviour? What is the sustainability potential of this activity? How can daily energy-consuming routines at the workplace be changed? Do major changes in the course of people's lives represent opportunities to modify their consumer behaviour towards greater sustainability? These are only some of the research questions studied in the focal topic "From Knowledge to Action - New Paths towards Sustainable Consumption" which is funded by the German Federal Ministry of Education and Research (BMBF) as part of the "Social-ecological Research Programme" (SÖF). This book gives an insight into the research results of the ten project groups. Their diversity highlights that there is much more to "sustainable consumption" than the simple purchase of organic or fair trade products.In addition, overarching conceptual and normative issues were treated across the project groups of the focal topic. Developed collaboratively and moderated by the accompanying research project, the results of the synthesis process are also presented here, as for example how the sustainability of individual consumer behaviour can be evaluated,or which theories of action are particularly useful for specific consumer behaviour phenomena.

Does Social Support Really Help to Eat a Low-Fat Diet? Main Effects and Gender Differences of Received Social Support within the Health Action Process Approach

BACKGROUND: Most theories of health-behavior change focus exclusively on individual self-regulation without taking social factors, such as social support, into account. This study's first aim was to systematically test the added value of received instrumental and emotional social support within the Health Action Process Approach (HAPA) in the context of dietary change. In the social support literature, gender effects emerge with regard to the effectiveness of social support. Thus, a second aim was the examination of gender differences in the association of social support with dietary behavior. METHODS: Participants were 252 overweight and obese individuals. At baseline and 12 months later, participants completed questionnaires on HAPA variables; diet-specific received social support and low-fat diet. RESULTS: For the prediction of intentions 12 months later, instrumental support was more beneficial for men than for women over and above individual self-regulation. In terms of dietary behavior at T2, a moderate main effect of instrumental support emerged. Moreover, received emotional social support was beneficial for men, but not for women in terms of a low-fat diet 12 months later. CONCLUSIONS: Effects of received instrumental social support found in this study provide new evidence for the added value of integrating social support into the HAPA.

Explaining Emergence and Consequences of Specific Formal Controls in IS Outsourcing – A Process-View

IS outsourcing projects often fail to achieve project goals. To inhibit this failure, managers need to design formal controls that are tailored to the specific contextual demands. However, the dynamic and uncertain nature of IS outsourcing projects makes the design of such specific formal controls at the outset of a project challenging. Hence, the process of translating high-level project goals into specific formal controls becomes crucial for success or failure of IS outsourcing projects. Based on a comparative case study of four IS outsourcing projects, our study enhances current understanding of such translation processes and their consequences by developing a process model that explains the success or failure to achieve high-level project goals as an outcome of two unique translation patterns. This novel process-based explanation for how and why IS outsourcing projects succeed or fail has important implications for control theory and IS project escalation literature.

From Framework to Action: The DESIRE Approach to Combat Desertification

It has become increasingly clear that desertification can only be tackled through a multi-disciplinary approach that not only involves scientists but also stakeholders. In the DESIRE project such an approach was taken. As a first step, a conceptual framework was developed in which the factors and processes that may lead to land degradation and desertification were described. Many of these factors do not work independently, but can reinforce or weaken one another, and to illustrate these relationships sustainable management and policy feedback loops were included. This conceptual framework can be applied globally, but can also be made site-specific to take into account that each study site has a unique combination of bio-physical, socio-economic and political conditions. Once the conceptual framework was defined, a methodological framework was developed in which the methodological steps taken in the DESIRE approach were listed and their logic and sequence were explained. The last step was to develop a concrete working plan to put the project into action, involving stakeholders throughout the process. This series of steps, in full or in part, offers explicit guidance for other organizations or projects that aim to reduce land degradation and desertification.

Cardiac-specific ablation of synapse-associated protein SAP97 in mice decreases potassium currents but not sodium current

BACKGROUND Membrane-associated guanylate kinase (MAGUK) proteins are important determinants of ion channel organization in the plasma membrane. In the heart, the MAGUK protein SAP97, encoded by the DLG1 gene, interacts with several ion channels via their PDZ domain-binding motif and regulates their function and localization. OBJECTIVE The purpose of this study was to assess in vivo the role of SAP97 in the heart by generating a genetically modified mouse model in which SAP97 is suppressed exclusively in cardiomyocytes. METHODS SAP97(fl/fl) mice were generated by inserting loxP sequences flanking exons 1-3 of the SAP97 gene. SAP97(fl/fl) mice were crossed with αMHC-Cre mice to generate αMHC-Cre/SAP97(fl/fl) mice, thus resulting in a cardiomyocyte-specific deletion of SAP97. Quantitative reverse transcriptase-polymerase chain reaction, western blots, and immunostaining were performed to measure mRNA and protein expression levels, and ion channel localization. The patch-clamp technique was used to record ion currents and action potentials. Echocardiography and surface ECGs were performed on anesthetized mice. RESULTS Action potential duration was greatly prolonged in αMHC-Cre/SAP97(fl/fl) cardiomyocytes compared to SAP97(fl/fl) controls, but maximal upstroke velocity was unchanged. This was consistent with the decreases observed in IK1, Ito, and IKur potassium currents and the absence of effect on the sodium current INa. Surface ECG revealed an increased corrected QT interval in αMHC-Cre/SAP97(fl/fl) mice. CONCLUSION These data suggest that ablation of SAP97 in the mouse heart mainly alters potassium channel function. Based on the important role of SAP97 in regulating the QT interval, DLG1 may be a susceptibility gene to be investigated in patients with congenital long QT syndrome.

Ubiquitin-specific protease USP2-45 acts as a molecular switch to promote α2δ-1-induced downregulation of Ca v1.2 channels

Availability of voltage-gated calcium channels (Cav) at the plasma membrane is paramount to maintaining the calcium homeostasis of the cell. It is proposed that the ubiquitylation/de-ubiquitylation balance regulates the density of ion channels at the cell surface. Voltage-gated calcium channels Cav1.2 have been found to be ubiquitylated under basal conditions both in vitro and in vivo. In a previous study, we have shown that Cav1.2 channels are ubiquitylated by neuronal precursor cell-expressed developmentally downregulated 4 (Nedd4-1) ubiquitin ligases, but the identity of the counterpart de-ubiquitylating enzyme remained to be elucidated. Regarding sodium and potassium channels, it has been reported that the action of the related isoform Nedd4-2 is counteracted by the ubiquitin-specific protease (USP) 2-45. In this study, we show that USP 2-45 also de-ubiquitylates Cav channels. We co-expressed USPs and Cav1.2 channels together with the accessory subunits β2 and α2δ-1, in tsA-201 and HEK-293 mammalian cell lines. Using whole-cell current recordings and surface biotinylation assays, we show that USP2-45 specifically decreases both the amplitude of Cav currents and the amount of Cav1.2 subunits inserted at the plasma membrane. Importantly, co-expression of the α2δ-1 accessory subunit is necessary to support the effect of USP2-45. We further show that USP2-45 promotes the de-ubiquitylation of both Cav1.2 and α2δ-1 subunits. Remarkably, α2δ-1, but not Cav1.2 nor β2, co-precipitated with USP2-45. These results suggest that USP2-45 binding to α2δ-1 promotes the de-ubiquitylation of both Cav1.2 and α2δ-1 subunits, in order to regulate the expression of Cav1.2 channels at the plasma membrane.

Modeling corrective experiences as motivational mountain pass

Aim: The landscape metaphor allows viewing corrective experiences (CE) as pathway to a state with relatively lower 'tension' (local minimum). However, such local minima are not easily accessible but obstructed by states with relatively high tension (local maxima) according to the landscape metaphor (Caspar & Berger, 2012). For example, an individual with spider phobia has to transiently tolerate high levels of tension during an exposure therapy to access the local minimum of habituation. To allow for more specific therapeutic guidelines and empirically testable hypotheses, we advance the landscape metaphor to a scientific model which bases on motivational processes. Specifically, we conceptualize CEs as available but unusual trajectories (=pathways) through a motivational space. The dimensions of the motivational state are set up by basic motives such as need for agency or attachment. Methods: Dynamic system theory is used to model motivational states and trajectories using mathematical equations. Fortunately, these equations have easy-to-comprehend and intuitive visual representations similar to the landscape metaphor. Thus, trajectories that represent CEs are informative and action guiding for both therapists and patients without knowledge on dynamic systems. However, the mathematical underpinnings of the model allow researchers to deduct hypotheses for empirical testing. Results: First, the results of simulations of CEs during exposure therapy in anxiety disorders are presented and compared to empirical findings. Second, hypothetical CEs in an autonomy-attachment conflict are reported from a simulation study. Discussion: Preliminary clinical implications for the evocation of CEs are drawn after a critical discussion of the proposed model.

Intentionality in Reference and Action

This essay asks whether there is a relation between action-serving and meaning-serving intentions. The idea that the intentions involved in meaning and action are nominally designated alike as intentionalities does not guarantee any special logical or conceptual connections between the intentionality of referential thoughts and thought-expressive speech acts with the intentionality of doing. The latter category is typified by overt physical actions in order to communicate by engaging in speech acts, but also includes at the origin of all artistic and symbolic expression such cerebral and linguistic doings as thinking propositional thoughts. There are exactly four possibilities by which meaning and action intentionalities might be related to be systematically investigated. Meaning-serving and action-serving intentionalities, topologically speaking, might exclude one another, partially overlap with one another, or subsume one in the other or the other in the one. The theoretical separation of the two ostensible categories of intendings is criticized, as is their partial overlap, in light of the proposal that thinking and artistic and symbolic expression are activities that favor the inclusion of paradigm meaning-serving intentions as among a larger domain of action-serving intentions. The only remaining alternative is then developed, of including action-serving intentions reductively in meaning-serving intentions, and is defended as offering in an unexpected way the most cogent universal reductive ontology in which the intentionality of doing generally relates to the specific intentionality of referring in thought to the objects of predications, and of its artistic and symbolic expression.

Evaluation of liver parenchyma and perfusion using dynamic contrast-enhanced computed tomography and contrast-enhanced ultrasonography in captive green iguanas (Iguana iguana) under general anesthesia.

BACKGROUND Contrast-enhanced diagnostic imaging techniques are considered useful in veterinary and human medicine to evaluate liver perfusion and focal hepatic lesions. Although hepatic diseases are a common occurrence in reptile medicine, there is no reference to the use of contrast-enhanced ultrasound (CEUS) and contrast-enhanced computed tomography (CECT) to evaluate the liver in lizards. Therefore, the aim of this study was to evaluate the pattern of change in echogenicity and attenuation of the liver in green iguanas (Iguana iguana) after administration of specific contrast media. RESULTS An increase in liver echogenicity and density was evident during CEUS and CECT, respectively. In CEUS, the mean ± SD (median; range) peak enhancement was 19.9% ± 7.5 (18.3; 11.7-34.6). Time to peak enhancement was 134.0 ± 125.1 (68.4; 59.6-364.5) seconds. During CECT, first visualization of the contrast medium was at 3.6 ± 0.5 (4; 3-4) seconds in the aorta, 10.7 ± 2.2 (10.5; 7-14) seconds in the hepatic arteries, and 15 ± 4.5 (14.5; 10-24) seconds in the liver parenchyma. Time to peak was 14.1 ± 3.4 (13; 11-21) and 31 ± 9.6 (29; 23-45) seconds in the aorta and the liver parenchyma, respectively. CONCLUSION CEUS and dynamic CECT are practical means to determine liver hemodynamics in green iguanas. Distribution of contrast medium in iguana differed from mammals. Specific reference ranges of hepatic perfusion for diagnostic evaluation of the liver in iguanas are necessary since the use of mammalian references may lead the clinician to formulate incorrect diagnostic suspicions.