LPS-induced Pellino3 degradation is mediated by p62-dependent autophagy

Background: In macrophages Toll-like receptor 4 (TLR4) is activated in response to lipopolysaccharide (LPS) and induces proinflammatory cytokine expression. Therefore, mechanisms terminating proinflammatory gene expression are important. Autophagy plays a central role in controlling innate immune responses by lysosomal degradation of signaling proteins, thus contributing to the resolution of inflammation. Autophagic proteins like p62 directly interact with molecules involved in the TLR4-signaling pathway, but a correlation with the IRAK E3 ligase and scaffold protein Pellino3 remains obscure. Hence, we are interested in elucidating the function of Pellino3 to prove our hypothesis that it is a key regulator in the TLR4-signaling cascade. Methods: We used the cecal ligation and puncture (CLP) mouse model causing polymicrobial sepsis to analyze Pellino3 protein and mRNA expression. Furthermore, we induced endotoxemia in RAW264.7 mouse macrophages by LPS treatment to verify in vivo experiments. Lentiviral Pellino3 knockdown in RAW264.7 macrophages was used for cytokine measurements at mRNA level. To analyze potential Pellino3 binding partners in TLR4-signaling by mass spectrometry (MS), we overexpressed FLAG-tagged Pellino3 in RAW264.7 macrophages, treated cells for 3, 6 and 24 hours with LPS and immunoprecipitated Pellino3 via its FLAG-tag. To consider Pellino3 degradation as a result of p62-mediated autophagy, we transiently knocked down p62 by siRNA in RAW264.7 macrophages and also pharmacologically blocked LPS-induced autophagy by Bafilomycin A1. Results: We demonstrated Pellino3 protein degradation in primary CD11b+ splenocytes after 24 hours following CLP operation and confirmed this in RAW264.7 macrophages after 24-hour LPS stimulation. Knockdown of Pellino3 attenuates proinflammatory cytokines, for example IL-6 mRNA, after 6 hours of LPS. Furthermore, we found by MS and verifying immunoprecipitation experiments that p62 is a Pellino3 binding partner, thus targeting Pellino3 for degradation. In line, both p62 knockdown and Bafilomycin A1 treatment prevent Pellino3 degradation, supporting an autophagic mechanism. Conclusion: Our observations highlight a regulatory role of Pellino3 on TLR4 signaling. Thus, antagonism of Pellino3 in the hyperinflammatory phase of sepsis may counteract the cytokine storm. Furthermore, stabilization of Pellino3 by inhibition of autophagy in the hypoinflammatory phase of sepsis may improve immunity. In consideration of these two conflictive sepsis phases, modulation of Pellino3 may provide a new strategy for the development of a therapy approach in sepsis

EFFECTS OF AN EXTENSIVE RUNNING BOUT IN NOVICE FEMALE RUNNERS

The purpose of this study was to characterize the spatiotemporal, force, and subjective effects that occur during an extensive run in novice female runners. Foot sole pressure, rate of fatigue, and speed were recorded during a 45-min flat ecological run. No significant effects were found in spatiotemporal, force-time, or rate of fatigue responses outside of the initiation phase of the running bout. When participants were grouped according to their pacing strategy, those with a decreasing speed over time exhibited significant decreases in rate of force development as time progressed. Participants tended to decrease mechanical loading variables, and increase their rate of fatigue throughout the run. Future studies should investigate a larger number of subjects to determine if these tendencies are characteristic of novice female runners

Vergleich von zwei Protokollen zur perioperativen Flüssigkeitssubstitution: Hydroxyethylstärke/Vollelektrolytlösung und Humanalbumin/Vollelektrolytlösung

Background: In the last years some large trials were conducted in critically ill patients with severe sepsis which demonstrated a higher risk for acute kidney failure and death after the use of HES. These assumptions were confirmed by some meta-analyses. Subsequently, based on these results, in October 2013 the PRAC recommended suspending marketing authorizations for infusion solutions containing HES. After Re-evaluation the PRAC changed the recommendation and the use of HES is currently allowed except for critically ill patient as well as those suffering from sepsis or burns. But the uncertainness remains especially for the administration for surgical patients due to the lack of large trials in the perioperative setting. Aim: This study compares two time periods, one with the use of 6% HES 130/0.4 and one without the use the 6% HES 130/0.4 during surgeries at the Clinic of Anaesthsiology, Ludwig-Maximilians-University. The aim was to investigate differences caused by the different administration of the infusions and to assess side effects especially acute kidney failures und the risk of death. Methods: This controlled observational study was performed in a single-Centre university Hospital and compared two time periods including 12.089 patients. In the first period (1. August - 31. October 2012) the use of HES for fluid resuscitation during surgeries was allowed. In the second period (1. August - 31. October 2013) there was no administration of HES. For the assessment two groups were generated, one group with overall patients and one subgroup just for the assessment of the RIFLE-criteria. The primary outcome was the incidence of postoperative acute kidney failure within 90 days after surgery due to requiring RRT. Secondary outcomes were in-hospital mortality, blood loss, blood transfusions, fluid requirements, hemodynamic stability and the need for postoperative intensive care unit treatment. Results: The administration of HES 130/0.4 for fluid resuscitation was not associated with an increased frequency of postoperative acute kidney failure. The in-hospital mortality and the hemodynamic stability did not differ significantly between the groups. There was a significantly higher blood loss in the HES-group, however the need for blood transfusions, the fluid requirements and the need for intensive care unit treatment were significantly higher in the Non-HES group. Conclusions: Except of the higher intraoperative blood loss, there were no associations between intraoperative HES therapy and postoperative complications. There was no higher incidence of postoperative acute kidney failure and no higher risk of death after intraoperative administration of HES. Due to the limitations of the study, the results of this investigation are not valid for general recommendation for the use of HES 130/0.4 in the perioperative setting. For the evidence of the safety of HES 130/0.4, there must be conduct large, randomized trials with appropriate patients, a suitable control-group/control-fluid, a high power, a proper observation period and proper Follow-Up time in the future

Vergleich von zwei Protokollen zur perioperativen Flüssigkeitssubstitution: Hydroxyethylstärke/Vollelektrolytlösung und Humanalbumin/Vollelektrolytlösung

Background: In the last years some large trials were conducted in critically ill patients with severe sepsis which demonstrated a higher risk for acute kidney failure and death after the use of HES. These assumptions were confirmed by some meta-analyses. Subsequently, based on these results, in October 2013 the PRAC recommended suspending marketing authorizations for infusion solutions containing HES. After Re-evaluation the PRAC changed the recommendation and the use of HES is currently allowed except for critically ill patient as well as those suffering from sepsis or burns. But the uncertainness remains especially for the administration for surgical patients due to the lack of large trials in the perioperative setting. Aim: This study compares two time periods, one with the use of 6% HES 130/0.4 and one without the use the 6% HES 130/0.4 during surgeries at the Clinic of Anaesthsiology, Ludwig-Maximilians-University. The aim was to investigate differences caused by the different administration of the infusions and to assess side effects especially acute kidney failures und the risk of death. Methods: This controlled observational study was performed in a single-Centre university Hospital and compared two time periods including 12.089 patients. In the first period (1. August - 31. October 2012) the use of HES for fluid resuscitation during surgeries was allowed. In the second period (1. August - 31. October 2013) there was no administration of HES. For the assessment two groups were generated, one group with overall patients and one subgroup just for the assessment of the RIFLE-criteria. The primary outcome was the incidence of postoperative acute kidney failure within 90 days after surgery due to requiring RRT. Secondary outcomes were in-hospital mortality, blood loss, blood transfusions, fluid requirements, hemodynamic stability and the need for postoperative intensive care unit treatment. Results: The administration of HES 130/0.4 for fluid resuscitation was not associated with an increased frequency of postoperative acute kidney failure. The in-hospital mortality and the hemodynamic stability did not differ significantly between the groups. There was a significantly higher blood loss in the HES-group, however the need for blood transfusions, the fluid requirements and the need for intensive care unit treatment were significantly higher in the Non-HES group. Conclusions: Except of the higher intraoperative blood loss, there were no associations between intraoperative HES therapy and postoperative complications. There was no higher incidence of postoperative acute kidney failure and no higher risk of death after intraoperative administration of HES. Due to the limitations of the study, the results of this investigation are not valid for general recommendation for the use of HES 130/0.4 in the perioperative setting. For the evidence of the safety of HES 130/0.4, there must be conduct large, randomized trials with appropriate patients, a suitable control-group/control-fluid, a high power, a proper observation period and proper Follow-Up time in the future

Community-based water markets and collective payment for ecosystem services : toward a theory of community-based environmental markets

Unidad de excelencia María de Maeztu CEX2019-000940-MIn the last few decades, the scope of governanceauthr solutions for environmental problems has increased substantially. The old trichotomy of governance by government, governance by markets, and governance by communities has been replaced by a new interest in hybrid solutions in the recognition that no single-governance mode possesses the capabilities to address the multiple facets, interdependencies, and scales of current environmental problems. This paper takes stock on experiences that combine community-based natural resource management and market-based solutions, or as we call them community-based environmental markets (CBEMs). Specifically, we draw lessons from the literature on community-based payment for ecosystem services in the forest context, and from water markets in the context of water user associations (WUA markets). Similarities across the two contexts include the role of communities to ensure participation, compliance, and distributional equity, and the importance of markets as a source of revenue for communities, among others. Differences across highlight the importance to pay attention to the authority held by the communities (stronger in the context of WUA markets) and the nature of the market (i.e. whether it is a service or a resource market). These commonalities and differences motivate the interest of generating new theory on CBEMs, that is, one that builds on but also transcends community-based natural resource management and environmental market theory and allows to compare experiences from different resource contexts

Zur Anschlussfähigkeit von Bevölkerungswachstum und Wohnungsmarktentwicklung

Der Beitrag unternimmt den Versuch, das Verständnis für die Ableitung des Wohnungsbedarfs aus der engen Beziehung zwischen Wanderungsgewinn, Einwohnerzuwachs und Wohnungsmarktentwicklung zu lösen. Die Differenzierung der Einwohnerentwicklung in die Stromgrößen des Wanderungsgeschehens sowie die demographischen Veränderungen, mit denen die Einwohnerentwicklung unterlegt ist, legen eine stärkere Berücksichtigung der (generativen) Veränderung der Haushaltestrukturen zur Beschreibung der Nachfrageentwicklung am Wohnungsmarkt nahe

Improvements of Sound Localization Abilities by the Facial Ruff of the Barn Owl (Tyto alba) as Demonstrated by Virtual Ruff Removal

BACKGROUND:When sound arrives at the eardrum it has already been filtered by the body, head, and outer ear. This process is mathematically described by the head-related transfer functions (HRTFs), which are characteristic for the spatial position of a sound source and for the individual ear. HRTFs in the barn owl (Tyto alba) are also shaped by the facial ruff, a specialization that alters interaural time differences (ITD), interaural intensity differences (ILD), and the frequency spectrum of the incoming sound to improve sound localization. Here we created novel stimuli to simulate the removal of the barn owl's ruff in a virtual acoustic environment, thus creating a situation similar to passive listening in other animals, and used these stimuli in behavioral tests. METHODOLOGY/PRINCIPAL FINDINGS:HRTFs were recorded from an owl before and after removal of the ruff feathers. Normal and ruff-removed conditions were created by filtering broadband noise with the HRTFs. Under normal virtual conditions, no differences in azimuthal head-turning behavior between individualized and non-individualized HRTFs were observed. The owls were able to respond differently to stimuli from the back than to stimuli from the front having the same ITD. By contrast, such a discrimination was not possible after the virtual removal of the ruff. Elevational head-turn angles were (slightly) smaller with non-individualized than with individualized HRTFs. The removal of the ruff resulted in a large decrease in elevational head-turning amplitudes. CONCLUSIONS/SIGNIFICANCE:The facial ruff a) improves azimuthal sound localization by increasing the ITD range and b) improves elevational sound localization in the frontal field by introducing a shift of iso-ILD lines out of the midsagittal plane, which causes ILDs to increase with increasing stimulus elevation. The changes at the behavioral level could be related to the changes in the binaural physical parameters that occurred after the virtual removal of the ruff. These data provide new insights into the function of external hearing structures and open up the possibility to apply the results on autonomous agents, creation of virtual auditory environments for humans, or in hearing aids