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Translational neuroscience bridges insights from specific mechanisms in rodents to complex functions in humans and is key to advance our general understanding of central nervous function. A prime example of translational research is the study of cross-species mechanisms that underlie responding to learned threats, by employing Pavlovian fear conditioning protocols in rodents and humans. Hitherto, evidence for (and critique of) these cross-species comparisons in fear conditioning research was based on theoretical viewpoints. Here, we provide a perspective to substantiate these theoretical concepts with empirical considerations of cross-species methodology. This meta-research perspective is expected to foster cross-species comparability and reproducibility to ultimately facilitate successful transfer of results from basic science into clinical applications.

Laboratory experiments revealed the stress hormone cortisol to decrease memory retrieval of emotional material, but a translation to real-life settings is missing so far. In this study, 51 students encoded a list of neutral, positive, and negative words as well as two neutral, biographical notes one day before attendance at a seminar at the university. In the stress condition, students gave a graded oral presentation, whereas they just attended the same seminar in the control condition immediately before retrieval took place. Measures of state anxiety, salivary cortisol and alpha-amylase confirmed the oral presentation to constitute a potent stressor. Importantly, stress significantly impaired retrieval of negative words, but not retrieval of the biographical notes. These results indicate that a real-life stressor decreases memory retrieval for negative items. In contrast, delayed memory retrieval of neutral information and interrelated details of biographical notes seems to be less prone to stress effects. These results have critical implications for educational settings.

There has been an explosion of research on fear extinction in humans in the past 2 decades. This has not only generated major insights, but also brought a new goal into focus: how to maintain extinction memory over time (i.e., extinction retention). We argue that there are still important conceptual and procedural challenges in human fear extinction research that hamper advancement in the field. We use extinction retention and the extinction retention index to exemplarily illustrate these challenges. Our systematic literature search identified 16 different operationalizations of the extinction retention index. Correlation coefficients among these different operationalizations as well as among measures of fear/anxiety show a wide range of variability in four independent datasets, with similar findings across datasets. Our results suggest that there is an urgent need for standardization in the field. We discuss the conceptual and empirical implications of these results and provide specific recommendations for future work.

Stress hormones influence the processing of fear, anxiety, and related memory mechanisms. For example, they modulate consolidation and retrieval processes associated with emotional episodic memory, fear and extinction learning. In this review, we summarize recent laboratory findings on the timing-dependent effects of stress on extinction learning and extinction retrieval. Furthermore, we relate these experiments to clinical intervention approaches relying on extinction processes such as exposure therapy, for which beneficial effects of the administration of the stress hormone cortisol have been observed. The modulation of extinction-based interventions differs from findings obtained with reconsolidation manipulation procedures utilizing the restabilization of retrieved (or reactivated) memories. In this case, blockade of adrenergic beta-receptors during reconsolidation of the fearful stimulus might represent a promising intervention. The substantial progress made in the understanding of the interaction of stress hormones with memory processes associated with fear and anxiety has the potential to enhance therapeutic success and prevent relapse in the long run.

Individuals have a tendency to show enhanced vigilance to groups of which they themselves are not a member. Stress can up-regulate hypervigilance towards threatening stimuli and was shown to promote the reinstatement of out-group related biases in a previous study conducted in women only. The current study examines how exposure to acute stress affects the retrieval of out-group related extinction biases in male participants. Results showed that men exerted a specific out-group related bias at the beginning of extinction training indexed by higher skin conductance responses (SCRs) towards out-group faces, while stress led to a return of this extinguished out-group bias. Specifically, the stress group showed higher SCRs towards out-group faces during retrieval compared to the control group and the bias index was negatively related to post-stress cardiovascular recovery. These results indicate the important interaction between stress and intergroup bias in fear conditioning, along with a potential modulation of sex.

Continued improvements on existing reconstruction methods are vital to the success of high-energy physics experiments, such as the IceCube Neutrino Observatory. In IceCube, further challenges arise as the detector is situated at the geographic South Pole where computational resources are limited. However, to perform real-time analyses and to issue alerts to telescopes around the world, powerful and fast reconstruction methods are desired. Deep neural networks can be extremely powerful, and their usage is computationally inexpensive once the networks are trained. These characteristics make a deep learning-based approach an excellent candidate for the application in IceCube. A reconstruction method based on convolutional architectures and hexagonally shaped kernels is presented. The presented method is robust towards systematic uncertainties in the simulation and has been tested on experimental data. In comparison to standard reconstruction methods in IceCube, it can improve upon the reconstruction accuracy, while reducing the time necessary to run the reconstruction by two to three orders of magnitude., Comment: 39 pages, 15 figures, submitted to Journal of Instrumentation; added references

IntroductionNo evaluated therapeutic approaches, that can efficiently be established in routine mental healthcare, are currently available for traumatised adolescent refugees in Germany. This study evaluates the efficacy of the Stress-Traumasymptoms-Arousal-Regulation-Treatment (START) programme to reduce trauma-related symptoms and psychological distress in traumatised adolescent refugees based in Germany.Methods and analysisThis randomised, waiting-list-controlled, multicentre trial with a 12-week follow-up will include 174 refugee minors with partial or full post-traumatic stress disorder who are fluent in either Arabic, Dari, English, German or Somali. Eligible refugee minors will be randomised to the START or waiting-list control groups. The manualised 8-week START programme is based on techniques of dialectical behaviour therapy (DBT), fosters adaptive coping with emotional distress and traumatic symptoms and comprises eight therapy modules and a booster session. Study assessments are planned at baseline, post-treatment (ie, after programme participation or waiting time), booster session at week 12 or 12-week waiting time, and at the 12-week follow-up. Primary and coprimary outcomes are changes in psychological distress and traumatic symptoms at post-treatment and will be analysed as response variables in linear mixed regression models. Secondary outcomes are changes in further trauma-related and other psychopathological symptoms, emotion regulation and intermediate effects of the programme at follow-up. We will also assess effects of the programme with ecological momentary assessments and on neuroendocrine stress parameters using hair cortisol.Ethics and disseminationThis study has been approved by the lead ethics committee of Rhineland-Palatinate and the ethics committees of participating sites. The study results will be disseminated through peer-reviewed publications and scientific conferences.Trial registration numberDRKS00020771.

Research has shown that fear acquisition, in reaction to potentially harmful stimuli or situations, is characterized by pronounced interindividual differences. It is likely that such differences are evoked by variability in the macro- and microstructural properties of brain regions involved in the processing of threat or safety signals from the environment. Indeed, previous studies have shown that the strength of conditioned fear reactions is associated with the cortical thickness or volume of various brain regions. However, respective studies were exclusively targeted at single brain regions instead of whole brain networks. Here, we tested 60 young and healthy individuals in a differential fear conditioning paradigm while they underwent fMRI scanning. In addition, we acquired T1-weighted and multi-shell diffusion-weighted images prior to testing. We used task-based fMRI data to define global brain networks which exhibited increased BOLD responses towards CS+ or CS- presentations, respectively. From these networks, we obtained mean values of gray matter density, neurite density, and neurite orientation dispersion. We found that mean gray matter density averaged across the CS+ network was significantly correlated with the strength of conditioned fear reactions quantified via skin conductance response. Measures of neurite architecture were not associated with conditioned fear reaction in any of the two networks. Our results extend previous findings on the relationship between brain morphometry and fear learning. Most importantly, our study is the first to introduce neurite imaging to fear learning research and discusses how its implementation can be improved in future research.

Physical review letters 125(14), 141801 (1-11) (2020). doi:10.1103/PhysRevLett.125.141801, The results of a 3+1 sterile neutrino search using eight years of data from the IceCube Neutrino Observatory are presented. A total of 305 735 muon neutrino events are analyzed in reconstructed energy-zenith space to test for signatures of a matter-enhanced oscillation that would occur given a sterile neutrino state with a mass-squared differences between 0.01 and 100 eV$^2$. The best-fit point is found to be at sin$^2$($2θ_{24}$)=0.10 and $Δm_{41}^2$=4.5 eV$^2$, which is consistent with the no sterile neutrino hypothesis with a $p$ value of 8.0%., Published by APS, College Park, Md.

The observation of electromagnetic radiation from radio to $\gamma$-ray wavelengths has provided a wealth of information about the universe. However, at PeV (10$^{15}$ eV) energies and above, most of the universe is impenetrable to photons. New messengers, namely cosmic neutrinos, are needed to explore the most extreme environments of the universe where black holes, neutron stars, and stellar explosions transform gravitational energy into non-thermal cosmic rays. The discovery of cosmic neutrinos with IceCube has opened this new window on the universe. In this white paper, we present an overview of a next-generation instrument, IceCube-Gen2, which will sharpen our understanding of the processes and environments that govern the universe at the highest energies. IceCube-Gen2 is designed to: 1) Resolve the high-energy neutrino sky from TeV to EeV energies; 2) Investigate cosmic particle acceleration through multi-messenger observations; 3) Reveal the sources and propagation of the highest energy particles in the universe; 4) Probe fundamental physics with high-energy neutrinos. IceCube-Gen2 will increase the annual rate of observed cosmic neutrinos by a factor of ten compared to IceCube, and will be able to detect sources five times fainter than its predecessor. Furthermore, through the addition of a radio array, IceCube-Gen2 will extend the energy range by several orders of magnitude compared to IceCube. Construction will take 8 years and cost about \$350M. The goal is to have IceCube-Gen2 fully operational by 2033. IceCube-Gen2 will play an essential role in shaping the new era of multi-messenger astronomy, fundamentally advancing our knowledge of the high-energy universe. This challenging mission can be fully addressed only in concert with the new survey instruments across the electromagnetic spectrum and gravitational wave detectors which will be available in the coming years., Comment: 56 pages, 29 figures

Neuroscientific research has identified specific brain networks involved in the acquisition of fear memories. Using fMRI to assess changes in resting-state functional connectivity (RSFC) induced by fear acquisition, single brain regions from these networks have also been linked to fear memory consolidation. However, previous studies only examined RSFC changes within restricted sets of brain regions or without a proper control group, leaving our knowledge about fear consolidation outside of traditional fear networks incomplete. Here, we tested a group of 84 healthy participants in a differential fear conditioning paradigm and quantified RSFC changes between 358 cortical and 16 subcortical brain areas. Subsequent to fear learning, 21 functional connections exhibited significant RSFC changes. Importantly, these connections were not restricted to the traditional fear networks but also comprised various frontal and visual areas. Our findings indicate that fear memory consolidation is a complex process that integrates relevant information across the entire brain.

Cortisol administration prior to treatment can promote the efficacy of exposure-based treatments in specific phobia: cortisol has been proposed to reduce fear retrieval at the beginning of exposure and to enhance the acquisition and consolidation of corrective information learned during exposure. Whether cortisol exerts a beneficial therapeutic effect when given after exposure, e.g., by targeting the consolidation of new corrective information, has not been addressed so far to date. Here, we examined whether post-exposure cortisol administration promotes fear reduction and reduces return of fear following contextual change in specific phobia. Furthermore, the effect of cortisol on return of fear following contextual change (i.e., contextual renewal) was assessed. Patients with spider phobia (N = 43) were treated with a single session of in-vivo exposure, followed by cortisol administration (20 mg hydrocortisone) in a double-blind, placebo-controlled study design. Return of fear was assessed with behavioral approach tests (BATs) in the familiar therapy context (versus a novel unfamiliar context) at one-month and seven-month follow-up assessment. Exposure was effective in reducing fear from pre-treatment to post-treatment (i.e., 24 h after exposure) on fear-related behavioral (approach behavior during the BAT), psychophysiological (heart rate during the BAT) and subjective (fear during the BAT, spider-fear related questionnaires) measures of therapeutic outcome, with no add-on benefit of cortisol administration. Cortisol had no effect on contextual renewal at one-month follow-up. However, in a subsample (N = 21) that returned to the seven-month follow-up, an adverse effect of cortisol on fear renewal was found, with cortisol-treated patients showing an increase in subjective fear at the final approach distance of the BAT from post-treatment to seven-month follow-up. These and previous findings underline the importance of considering the exact timing of cortisol application when used as an add-on treatment for extinction-based psychotherapy: post-exposure cortisol administration does not seem to be effective, but might promote fear renewal at the subjective level.

Inhibition of the amygdala slows down acquisition of conditioned eyeblink responses (CRs). Based on the two-stage or two-factor theory of aversive conditioning, amygdala-dependent conditioned fear is a necessary prerequisite to acquire eyeblink CRs but is no longer needed after eyeblink CRs are attained. According to the sensory gating hypothesis of the amygdala, on the other hand, the amygdala modulates the salience of unconditioned stimuli (USs) and conditioned stimuli (CSs) in eyeblink conditioning. We tested these two opposing assumptions in five groups of 20 young and healthy men. On day 1, three groups underwent fear acquisition training followed by acquisition of eyeblink CRs. On the next day (day 2), extinction was tested. In group 1, fear and eyeblink extinction trials overlapped; in group 2, fear and eyeblink extinction trials alternated; and in group 3, fear extinction trials were followed by eyeblink extinction trials. Groups 4 and 5 were control conditions testing fear and eyeblink conditioning only. Preceding fear acquisition training facilitated acquisition of conditioned eyeblinks. Concomitant fear extinction impeded extinction of eyeblink CRs, which was accompanied by increased autonomic responses. Fear extinction, however, was not significantly altered by concomitant eyeblink extinction. Recall of fear CRs on day 2 was facilitated in group 1, suggesting additive response summation. Findings are difficult to explain with the two-stage theory of aversive conditioning, which predicts the suppression of conditioned fear once conditioned eyeblinks are acquired. Facilitated acquisition and impeded extinction of eyeblink CRs, however, are in accordance with the sensory-gating hypothesis of the amygdala.

Fear conditioning represents an experimental paradigm ideally suited to investigate aversive learning and memory mechanisms that are fundamental to the development, maintenance and treatment of mental disorders. Men and women seem to differ in their capability to learn and retrieve fear and extinction memories. This review outlines how sex may influence human fear conditioning, with an emphasis on the sex hormones and oral contraceptives. Available evidence suggests women with high estrogen levels to acquire fear more readily, but also to extinguish fear more easily, leading to an enhanced extinction memory trace. By contrast, women with low estrogens (e.g. due to oral contraceptives) seem to show deficits in extinction recall. These findings are highly relevant for future basic and applied studies alike.

While generalization of fear seems to be naturally acquired as frequently observed in fear-related disorders, extinction learning appears to be stimulus-specific. Thus, treatments aiming to generalize extinction learning comprise the chance to overcome stimulus-specificity and consequently reduce relapse. One suggested candidate is the timing-dependent administration of the stress hormone cortisol. In the present pre-registered, three-day fear conditioning study, we aimed to create a generalized extinction memory trace in 60 healthy men and women using multiple sizes of one conditioned stimulus (CS+G; generalized) during extinction training, whereas the other CS (CS+N; non-generalized) and the CS- were solely presented in their original sizes. Extinction training took place either after pharmacological administration of 20 mg cortisol or placebo. Following successful fear acquisition on day one, generalization effects during extinction training and retrieval were investigated in the comparison of CS+G and CS+N. Insula and dorsal anterior cingulate cortex (dACC) activation for CS+G as compared to CS+N extending to the second half of extinction training indicated prolonged fear processing during extinction training for the CS+G on day two. During retrieval on day three, an activation of the anterior hippocampus occurred for CS+N minus CS+G in the cortisol but not in the placebo group. Additionally, a more posterior hippocampal activation (compared to the other hippocampal activation) was observed for the contrast CS+G minus CS+N. In accordance with our hypotheses, amygdala and dACC responding during reinstatement test was reduced for the CS+G as compared to CS+N. However, cortisol did not modulate amygdala responding, but abolished the CS+G/CS+N differentiation in the dACC relative to placebo. Generalization and cortisol effects were not mirrored in skin conductance responses. In conclusion, extinction generalization processes appear to rely on prolonged fear processing still present in the second half of extinction training that in turn leads to reduced fear-related processing after reinstatement. Cortisol administration prior to extinction training, however, selectively reduced fear-related activation for standard extinction but did not further reduce fear-related activation for extinction generalization.

We present a high-energy neutrino event generator, called LeptonInjector, alongside an event weighter, called LeptonWeighter. Both are designed for large-volume Cherenkov neutrino telescopes such as IceCube. The neutrino event generator allows for quick and flexible simulation of neutrino events within and around the detector volume, and implements the leading Standard Model neutrino interaction processes relevant for neutrino observatories: neutrino-nucleon deep-inelastic scattering and neutrino-electron annihilation. In this paper, we discuss the event generation algorithm, the weighting algorithm, and the main functions of the publicly available code, with examples., Comment: 28 pages, 10 figures, 3 tables

Controversy has long surrounded the question of whether spontaneous lateral demixing of membranes into coexisting liquid phases can organize proteins and lipids on micron scales within unperturbed, living cells. A clear answer hinges on observation of hallmarks of a reversible phase transition. Here, by directly imaging micron-scale membrane domains of yeast vacuoles both in vivo and cell free, we demonstrate that the domains arise through a phase separation mechanism. The domains are large, have smooth boundaries, and can merge quickly, consistent with fluid phases. Moreover, the domains disappear above a distinct miscibility transition temperature (Tmix) and reappear below Tmix, over multiple heating and cooling cycles. Hence, large-scale membrane organization in living cells under physiologically relevant conditions can be controlled by tuning a single thermodynamic parameter.

Dense-core vesicles (DCVs) are secretory organelles that store and release modulatory neurotransmitters from neurons and endocrine cells. Recently, the conserved coiled-coil protein CCCP-1 was identified as a component of the DCV biogenesis pathway in the nematode Caenorhabditis elegans. CCCP-1 binds the small GTPase RAB-2 and colocalizes with it at the trans-Golgi. Here, we report a structure-function analysis of CCCP-1 to identify domains of the protein important for its localization, binding to RAB-2, and function in DCV biogenesis. We find that the CCCP-1 C-terminal domain (CC3) has multiple activities. CC3 is necessary and sufficient for CCCP-1 localization and for binding to RAB-2, and is required for the function of CCCP-1 in DCV biogenesis. In addition, CCCP-1 binds membranes directly through its CC3 domain, indicating that CC3 may comprise a previously uncharacterized lipid-binding motif. We conclude that CCCP-1 is a coiled-coil protein that binds an activated Rab and localizes to the Golgi via its C-terminus, properties similar to members of the golgin family of proteins. CCCP-1 also shares biophysical features with golgins; it has an elongated shape and forms oligomers.

Why do only some individuals develop pathological anxiety following adverse events? Fear acquisition, extinction and return of fear paradigms serve as experimental learning models for the development, treatment and relapse of anxiety. Individual differences in experimental performance were however mostly regarded as 'noise' by researchers interested in basic associative learning principles. Our work for the first time presents a comprehensive literature overview and methodological discussion on inter-individual differences in fear acquisition, extinction and return of fear. We tell a story from noise that steadily develops into a meaningful tune and converges to a model of mechanisms contributing to individual risk/resilience with respect to fear and anxiety-related behavior. Furthermore, in light of the present 'replicability crisis' we identify methodological pitfalls and provide suggestions for study design and analyses tailored to individual difference research in fear conditioning. Ultimately, synergistic transdisciplinary and collaborative efforts hold promise to not only improve our mechanistic understanding but can also be expected to contribute to the development of specifically tailored ('individualized') intervention and targeted prevention programs in the future.

Distinct stages of the menstrual cycle and the intake of oral contraceptives (OC) affect sex hormone levels, stress responses, and memory processes critically involved in the pathogenesis of mental disorders. To characterize the interaction of sex and stress hormones on memory encoding, 30 men, 30 women in the early follicular phase of the menstrual cycle (FO), 30 women in the luteal phase (LU), and 30 OC women were exposed to either a stress (socially evaluated cold-pressor test) or a control condition prior to memory encoding and immediate recall of neutral, positive, and negative words. On the next day, delayed free and cued recall was tested. Sex hormone levels verified distinct estradiol, progesterone, and testosterone levels between groups. Stress increased blood pressure, cortisol concentrations, and ratings of stress appraisal in all four groups as well as cued recall performance of negative words in men. Stress exposure in OC women led to a blunted cortisol response and rather enhanced cued recall of neutral words. Thus, pre-encoding stress facilitated emotional cued recall performance in men only, but not women with different sex hormone statuses pointing to the pivotal role of circulating sex hormones in modulation of learning and memory processes.

Background: Extracellular adenosine triphosphate (ATP) binds as a danger signal to purinergic receptor P2X 7 and promotes inflammasome assembly and interleukin-1β expression. We hypothesized a functional role of the signal axis ATP–P2X 7 in inflammasome activation and the chronic inflammation driving atherosclerosis. Methods: P2X 7 -competent and P2X 7 -deficient macrophages were isolated and stimulated with lipopolysaccharide, ATP, or both. To assess whether P2X 7 may have a role in atherosclerosis, P2X 7 expression was analyzed in aortic arches from low density lipoprotein receptor -/- mice consuming a high-cholesterol or chow diet. P2X 7 +/+ and P2X 7 −/− low density lipoprotein receptor −/− mice were fed a high-cholesterol diet to investigate the functional role of P2X 7 knockout in atherosclerosis. Human plaques were derived from carotid endarterectomy and stained against P2X 7 . Results: Lipopolysaccharide or ATP stimulation alone did not activate caspase 1 in isolated macrophages. However, priming with lipopolysaccharide, followed by stimulation with ATP, led to an activation of caspase 1 and interleukin-1β in P2X 7 -competent macrophages. In contrast, P2X 7 -deficient macrophages showed no activation of caspase 1 after sequential stimulation while still expressing a basal amount of interleukin-1β. P2X 7 receptor was higher expressed in murine atherosclerotic lesions, particularly by lesional macrophages. After 16 weeks of a high-cholesterol diet, P2X 7 -deficient mice showed smaller atherosclerotic lesions than P2X 7 -competent mice (0.162 cm 2 ±0.023 [n=9], P2X 7 −/− low density lipoprotein receptor −/− : 0.084 cm 2 ±0.01 [n=11], P =0.004) with a reduced amount of lesional macrophages. In accord with our in vitro findings, lesional caspase 1 activity was abolished in P2X 7 −/− mice. In addition, intravital microscopy revealed reduced leukocyte rolling and adhesion in P2X 7 -deficient mice. Last, we observe increased P2X 7 expression in human atherosclerotic lesions, suggesting that our findings in mice are relevant for human disease. Conclusions: P2X 7 deficiency resolved plaque inflammation by inhibition of lesional inflammasome activation and reduced experimental atherosclerosis. Therefore, P2X 7 represents an interesting potential new target to combat atherosclerosis.

Background: Extinction of conditioned fear represents an important mechanism in the treatment of anxiety disorders. Return of fear after successful extinction or exposure therapy in patients with anxiety disorders might be linked to poor temporal or contextual generalization of extinction due to individual differences in brain structural connectivity. The goal of this magnetic resonance imaging study was therefore to investigate the association of context-dependent extinction recall with brain structural connectivity. Methods: Diffusion-tensor imaging was used in order to determine the fractional anisotropy as a measure of white matter structural integrity of fiber tracts connecting central brain regions of the fear and extinction circuit (uncinate fasciculus, cingulum). Forty-five healthy men participated in a two-day fear conditioning experiment with fear acquisition in context A and extinction learning in context B on the first day. Extinction recall in the extinction context as well as renewal in the acquisition context and a novel context C took place one day later. Results: Renewal of conditioned fear (skin conductance responses) in the acquisition context was associated with higher structural integrity of the hippocampal part of the cingulum. Conclusions: Enhanced structural integrity of the cingulum might be related to stronger hippocampal modulation of the dorsal anterior cingulate cortex, a region important for modulating conditioned fear output by excitatory projections to the amygdala. This finding underpins the crucial role of individual differences in the structural integrity of relevant fiber tracts for context-dependent extinction recall and return of fear after exposure therapy in anxiety disorders. This article is protected by copyright. All rights reserved.

When using in-group and out-group faces as conditional stimuli (CS) in fear conditioning designs, extinction learning is selectively impaired for out-group faces. Additionally, stress seems to inhibit extinction retrieval leading to a higher return of fear, which might be especially the case for out-group faces. To test this hypothesis, 51 healthy women underwent fear acquisition training, consisting of repeated presentations of two in-group and two out-group faces. One of each (CS+) was paired with an electrical stimulation (unconditional stimulus, UCS), whereas the other was not coupled with the UCS (CS−). During immediate extinction training, all CS were presented again. On the next day, a retrieval and reinstatement test took place after a stress or a control procedure. Confirming previous research, impaired extinction learning occurred for out-group relative to in-group faces. During the reinstatement test, stress specifically increased responding towards the out-group CS−, thus reducing its safety signaling properties. So, stress seems to reduce the ability to adequately distinguish threat and safety cues after aversive experiences mimicked by reinstatement shocks.

SUMMARYSNARE chaperones of the Sec1/mammalian Unc-18 (SM) family have critical roles in SNARE-mediated membrane fusion. Using SNARE and Sly1 mutants, and a newin vitroassay of fusion, we separate and assess proposed mechanisms through which Sly1 augments fusion: (i) opening the closed conformation of the Qa-SNARE Sed5; (ii) close-range tethering of vesicles to target organelles, mediated by the Sly1-specific regulatory loop; and (iii) preferential nucleation of productivetrans-SNARE complexes. We show that all three mechanisms are important and operate in parallel, and we present evidence that close-range tethering is particularly important fortrans-complex assembly whencis-SNARE assembly is a competing process. In addition, the autoinhibitory N-terminal Habc domain of Sed5 has at least two positive activities: the Habc domain is needed for correct Sed5 localization, and it directly promotes Sly1-dependent fusion. Remarkably, “split Sed5,” with the Habc domain present only as a soluble fragment, is functional bothin vitroandin vivo.

The essential Golgi protein Sly1 is a member of the SM (Sec1/mammalian Unc-18) family of SNARE chaperones. Sly1 was originally identified through gain-of-function alleles that bypass requirements for diverse vesicle tethering factors. Employing genetic analyses and chemically defined reconstitutions of ER-Golgi fusion, we discovered that a loop conserved among Sly1 family members is not only autoinhibitory, but also acts as a positive effector. An amphipathic helix within the loop directly binds high-curvature membranes; membrane binding is required for relief of Sly1 autoinhibition and allows Sly1 to directly tether incoming vesicles to the Qa-SNARE on the target organelle. TheSLY1-20allele bypasses requirements for diverse tethering factors but loses this functionality if Sly1 membrane binding is impaired. We propose that long-range tethers, including Golgins and multisubunit tethering complexes, hand off vesicles to Sly1, which then tethers at close range to activate SNARE assembly and fusion in the early secretory pathway.

High-energy neutrinos are unique messengers of the high-energy universe, tracing the processes of cosmic ray acceleration. This paper presents analyses focusing on time-dependent neutrino point-source searches. A scan of the whole sky, making no prior assumption about source candidates, is performed, looking for a space and time clustering of high-energy neutrinos in data collected by the IceCube Neutrino Observatory between 2012 and 2017. No statistically significant evidence for a time-dependent neutrino signal is found with this search during this period, as all results are consistent with the background expectation. Within this study period, the blazar 3C 279, showed strong variability, inducing a very prominent gamma-ray flare observed in 2015 June. This event motivated a dedicated study of the blazar, which consists of searching for a time-dependent neutrino signal correlated with the gamma-ray emission. No evidence for a time-dependent signal is found. Hence, an upper limit on the neutrino fluence is derived, allowing us to constrain a hadronic emission model.

Introduction Cardiovascular diseases are the main cause of death worldwide. Acute ischemia results in cell death of cardiomyocytes accompanied with the release of so called damage associated molecular patterns (DAMPs) such as nucleotides. High concentration of ATP in the extracellular fluid leads to the opening of the ionotropic purinergic receptor P2X7. The following transmembranous ion flux triggers the assembly of the NLRP-3 inflammasome and proteolytic caspase-1 activation which cleaves pro-IL-1β and provokes the release of fully active pro-inflammatory IL-1β. Myocardial scar formation can be divided into an early remodelling IL-1β-dependent and a late scar forming TGFβ phase. Both phases negatively regulate each other. The current stuy aimed to investigate the role of this master regulator of NLRP3 inflammasome assembly P2Y7 in myocardial remodeling during prolonged ischemic conditions. Methods 10 weeks old P2X7 knock-out and C57Bl6/J mice received a full ligation of the left anterior descending (LAD) artery. Mice three and seven days post-infarction underwent echocardiography. Myocardial scar formation was assessed by histological staining and flow cytometry. Furthermore caspase-1 activity was measured using FLICA in histology. Gene expression was assessed using TaqMan realtime PCR. Results Macrophages in the myocardial infarct area showed high P2X7 expression by Co-staining with fluorescent antibodies against F4/80, CD68 and P2X7. Intriguingyl P2X7 deficient animals showed a significantly worse survival rate in a Kaplan-Meier survival analysis compared to wt littermated with LAD ligation (Mortality after 21 days P2X7+/+ 50%; P2X7−/− 100%, p Conclusion The knockout of the NLRP3 inflammasome activating P2X7 receptor impairs the outcome after myocardial infarction by reduced monocyte infiltration and deranged scar formation. Disruption of the fine-tuned IL-1β/TGFβ sequence with an early block of IL-1β signaling and premature TGFβ activation could explain the missing clean-up of necrotic debris and impaired scar formation. Taken together these findings highlight the importance of an early inflammasome phase during myocardial scar formation.

Background Metabolic syndrome clusters the main risk factors for cardiovascular diseases and endocrine dysfunction. Novel studies show that the important underlying mechanism is a long term inflammation of adipose tissue with a gradual shift from the anti-inflammatory M2 towards the pro-inflammatory M1 macrophages (pathognomonic). Massive hypertrophy induces adipocyte death, which releases DAMPs like nucleotides recognized by purinergic receptors orchestrating ongoing inflammatory process. Interestingly, we found Gi coupled P2Y13R only expressed on M1 macrophages, not in M0 (unstimulated) and M2 macrophages. Nevertheless, the role of P2Y13R in the immune system and especially macrophages is currently unknown. Given its pivotal role in central metabolic processes (P2Y13R has been described in insulin secretory signalling) together with its unique expression in its pathognomonic inflammatory macrophage subtype makes it an interesting candidate to investigate its role in metabolic syndrome. Purpose Due to the unique expression of P2Y13R on M1 macrophages we hypothesise an improved outcome in a high-fat diet induced metabolic syndrome by interfering with the P2Y13 signalling cascade. Methods BMDM differentiation to macrophages using M-CSF and subsequent stimulation with medium (M0), LPS and IFNγ (M1) or IL4 (M2); Expression of P2Rs quantified using Taqman qPCR. Male C57Bl6/J wild-type (WT) and P2Y13-deficient (KO) mice were fed with a high-fat diet (HFD) for 20 weeks; On week 15 we performed the ITT, on week 16 the GTT. Metabolic performance was monitored by metabolic caging. Results We observed a unique expression of P2Y13R on M1 macrophages. Adult P2Y13-deficient mice showed a higher O2 consumption compared to adult C57Bl6/J wild-type mice (AUC of O2 consumption 2nd day= KO: 61620±2261mL/kg vs WT: 53830±916.1mL/kg, p=0.0331). Although both P2Y13−/− mice and WT littermates consumed comparable amount of food (daily food intake per mouse → KO: 3.97±0.25g vs WT: 3.76±0.18g), P2Y13 deficient animals showed significantly decelerated weight gain (e.g. on week 15 → KO: 142±2% (n=10) vs WT: 198±5% (n=10), p Conclusion Global P2Y13 deficiency leads to an improved outcome in metabolic syndrome with an increased protection against developing an insulin resistance as shown through an improved glucose tolerance and basal glucose levels, a decelerated weight gain despite comparable food consumption and a better metabolic turnover. Observing these beneficial metabolic improvements, we hypothesise that antagonization of P2Y13R could be a promising therapeutic target in the field of metabolic syndrome.

Introduction Cardiomyopathies due to pathological remodelling are the most common myocardial disorders and leading indication for heart transplant in young adults. Cardiomyocytes rely on sessile macrophage phagocytosis for cell homeostasis. Purinergic receptor Y6, selectively binding to UDP, promotes phagocytosis and is involved in several tissue-remodelling processes. We hypothesize a critical role of P2Y6 in the maintenance of cardiac tissue homeostasis in vivo. Methods Echocardiography was performed on male C57Bl6 wild-type (WT) and P2Y6-deficient (KO) mice at different ages up to 20 weeks. In addition, WT and KO mice underwent bone marrow transplantation. For histological analysis, hearts from WT and KO mice were stained with HE, Masson's trichrome, wheat germ agglutinin, CD11b, LC3B and TUNEL. Cardiac ultrastructure of WT and KO hearts was investigated by electron microscopic imaging. Furthermore, uptake of fluorescent pHrodo bioparticles by WT and KO bone marrow derived macrophages (BMDMs), either in presence or absence of specific P2Y6 agonist UDP or full antagonist MRS2578, was measured in vitro. Results KO mice were significantly impaired in their LV function by reduced ejection fraction (WT: 57.37±1.27%, n=13, KO: 44.13±1.09%, n=16, p Conclusion Global P2Y6 deficiency leads to a progressive cardiomyopathy in mice, mainly characterized by an impaired left ventricular function and ultrastructural irregularities. Its exacerbation seems to be prevented by reconstitution with WT bone marrow. For the underlying mechanism, we conclude a deranged cardiomyocyte homeostasis in KO animals due to defective phagocytic activity of resident macrophages. Potential induction of cardiac clearance via P2Y6 signalling could be a promising therapeutic target in the field of cardiomyopathies.

Fear acquisition manifests in the development of conditioned fear responses (CRs), whereas a decrement in CRs as a consequence of unconditioned stimulus (UCS) omission is referred to as extinction learning. Time windows for CR scoring in physiological readouts are subject to discussion, especially regarding the subdivision of skin conductance responses (SCRs) into first- (FIR) and second-interval responses (SIR). However, distinct temporal CR trajectories within or across measures may reflect specific characteristics of the underlying associative processes. In this study, 41 participants underwent fear acquisition and extinction, while SCRs and pupillary responses were recorded and separated into different time bins to explore the temporal dynamics of CRs across both learning phases. For SCRs, we observed a shift from early (FIR) to late (SIR) time intervals during fear acquisition most likely reflecting subsequent learning processes, in which CS-UCS associations and their relative timing are formed. During extinction, only the FIR exhibited a CR decline and was thus able to track the learning progress. These results indicate that conditioned SCRs follow a dynamic temporal pattern that may be related to different learning dimensions. By contrast, pupillary CRs were generally better captured by a late pupillary response component, suggesting a rather stable temporal CR pattern for the pupil in both learning phases. Our findings underscore the importance of specifying CR quantification for different physiological readouts when evaluating learning performance in the context of fear acquisition and extinction and may motivate further investigation of time-specific CR patterns and their relation to specific associative dimensions.

Background Inflammation drives atherosclerosis and its complications. Anti-inflammatory therapy with interleukin 1 beta (IL-1β) antibody reduces cardiovascular events in patients with elevated high-sensitive C-reactive protein (hsCRP). This study aims to identify the share of patients with coronary heart disease (CHD) and residual inflammation who may benefit from anti-inflammatory therapy. Methods hsCRP and low-density lipoprotein (LDL) levels were determined in 2741 all-comers admitted to the cardiological ward of our tertiary referral hospital between June 2016 and June 2018. Patients without CHD, with acute coronary syndrome, chronic or recurrent systemic infection, use of immunosuppressant or anti-inflammatory agents, chronic inflammatory diseases, chemotherapy, terminal organ failure, traumatic injury and pregnancy were excluded. Results 856 patients with stable CHD were included. 42.7% of those had elevated hsCRP ≥ 2 mg/l. Within the group of patients with LDL-cholesterol

Emotion regulation is essential for adaptive behavior and mental health. Strategies applied to alter emotions are known to differ in their impact on psychological and physiological aspects of the emotional response. However, emotion regulation outcome has primarily been assessed via self-report, and studies comparing regulation strategies with regard to their peripheral physiological mechanisms are limited in number. In the present study, we therefore aimed to investigate the effects of different emotion regulation strategies on pupil dilation, skin conductance responses, and subjective emotional responses. Thirty healthy females were presented with negative and neutral pictures and asked to maintain or up- and downregulate their upcoming emotional responses through reappraisal or distraction. Pupil dilation and skin conductance responses were significantly enhanced when viewing negative relative to neutral pictures. For the pupil, this emotional arousal effect manifested specifically late during the pupillary response. In accordance with subjective ratings, increasing negative emotions through reappraisal led to the most prominent pupil size enlargements, whereas no consistent effect for downregulation was found. In contrast, early peak dilations were enhanced in all emotion regulation conditions independent of strategy. Skin conductance responses were not further modulated by emotion regulation. These results indicate that pupil diameter is modulated by emotional arousal, but is initially related to the extent of mental effort required to regulate automatic emotional responses. Our data thus provide first evidence that the pupillary response might comprise two distinct temporal components reflecting cognitive emotion regulation effort on the one hand and emotion regulation success on the other hand.

Stress influences emotional learning and memory processes. These effects are thought to underlie stress-associated mental disorders. Sex differences in stress reactivity and in central nervous system stress sensitivity illustrate the important modulatory role of sex hormones. This Review outlines how stress hormones influence different stages of the fear conditioning process, such as fear acquisition, extinction, and retrieval. Results will be compared with findings on the impact of stress on episodic memory. The focus is on the available human data on sex differences and the impact sex hormones have on the stress effects on emotional learning and memory. It will become apparent that the menstrual cycle but also the intake of hormonal contraceptives modulates the impact of stress on brain and behavior. Additional basic research is needed for a deeper insight regarding the interplay between stress and sex hormones in emotion and cognition. In addition, new treatment options might be derived to optimize existing strategies such as exposure therapy, which relies on the principles of fear conditioning. © 2016 Wiley Periodicals, Inc.

The renewal effect describes the recovery of extinguished responses that may occur after a change in context and indicates that extinction memory retrieval is sometimes prone to failure. Stress hormones have been implicated to modulate extinction processes, with mostly impairing effects on extinction retrieval. However, the neurobiological mechanisms mediating stress effects on extinction memory remain elusive. In this functional magnetic resonance imaging study, we investigated the effects of cortisol administration on the neural correlates of extinction memory retrieval in a predictive learning task. In this task, participants were required to predict whether certain food stimuli were associated with stomach trouble when presented in two different contexts. A two-day renewal paradigm was applied in which an association was acquired in context A and subsequently extinguished in context B. On the following day, participants received either cortisol or placebo 40 min before extinction memory retrieval was tested in both contexts. Behaviorally, cortisol impaired the retrieval of extinguished associations when presented in the extinction context. On the neural level, this effect was characterized by a reduced context differentiation for the extinguished stimulus in the ventromedial prefrontal cortex, but only in men. In the placebo group, ventromedial prefrontal cortex was functionally connected to the left cerebellum, the anterior cingulate and the right anterior parahippocampal gyrus to express extinction memory. This functional crosstalk was reduced under cortisol. These findings illustrate that the stress hormone cortisol disrupts ventromedial prefrontal cortex functioning and its communication with other brain regions implicated in extinction memory.

Previous work from our group has shown that cortisol enhances fear reconsolidation in men. Whether similar effects can be observed in women remains an open question. The effects of cortisol on the reconsolidation of fear memories were investigated in women. Based on results in men, we expected a specific enhancing effect of cortisol administration on the reactivated fear memory. In addition, possible interactions with oral contraceptive use were tested. We incorporated a differential fear conditioning paradigm in a 3-day reconsolidation design. A fear memory, which was created on the first day, was reactivated on the second day following cortisol administration in the target group. One control group was given cortisol without reactivation, and the other participated in the reactivation session following placebo intake. On the third day, the return of fear for all stimuli following reinstatement was tested. Skin conductance response served as measure of conditioned response. In contrast to the hypothesis, cortisol in combination with reactivation did not enhance fear reconsolidation. No differences between the three experimental groups were apparent. In addition, hormonal contraceptive use had no effect on any of the learning phases and did not interact with the cortisol manipulation. The lack of an effect in women might be the result of alternating concentrations of sex hormones during different phases of the menstrual cycle or following oral contraceptive use. Considering the higher vulnerability of women to stress-related mental disorders, further investigations in women are of great importance for both theory and treatment.

Return of fear is a serious problem in exposure-based treatments of anxiety disorders. Renewal of the fear response may occur when re-encountering the conditioned stimulus within a novel context. Findings in rodents underpin the hippocampus' role in conditioned fear renewal in novel contexts, but it has yet to be investigated in humans. Forty-six healthy men took part in a 2-day, context-dependent, cued fear conditioning paradigm with fear acquisition, extinction learning (day 1) and extinction recall in the acquisition, extinction and a novel context one day later. Conditioned evaluative, skin conductance responses (SCRs) and blood-oxygen-level-dependent responses served as dependent variables. Context-dependent fear renewal was reflected in stronger conditioned SCRs. In the acquisition context, individuals with a higher renewal of conditioned SCRs showed stronger activation of the fear circuit. Hippocampal activation distinguished conditioned responding in the novel compared with the extinction context. Individuals with a stronger renewal of conditioned SCRs in the novel context showed increased effective connectivity of hippocampal activation foci with structures in the fear and extinction network. These results outline the pivotal role of the hippocampus and its connectivity in conditioned fear renewal in a novel context in humans and might have important implications for exposure therapy in anxiety disorders.

Sterile inflammation of visceral fat, provoked by dying adipocytes, links the metabolic syndrome to cardiovascular disease. Danger-associated molecular patterns, such as adenosine triphosphate (ATP), are released by activated or dying cells and orchestrate leukocyte infiltration and inflammation via the purinergic receptor P2Y2. The gene expression of ATP receptor P2Y2 did not change in several tissues in the course of obesity, but was increased within epididymal fat. Adipose tissue from P2Y 2 −/− mice consuming high-fat diet (HFD) contained less crown-like structures with a reduced frequency of adipose tissue macrophages (ATMs). This was likely due to decreased leukocyte migration because of missing VCAM-1 exposition on P2Y2 deficient hypertrophic adipose tissue endothelial cells. Accordingly, P2Y 2 −/− mice showed blunted traits of the metabolic syndrome: they gained less weight compared to P2Y 2 +/+ controls, while intake of food and movement behaviour remained unchanged. Liver and adipose tissue were smaller in P2Y 2 −/− animals. Insulin tolerance testing (ITT) performed in obese P2Y 2 −/− mice revealed a better insulin sensitivity as well as lower plasma C-peptide and cholesterol levels. We demonstrate that interfering with somatic P2Y2 signalling prevents excessive immune cell deposition in diet-induced obesity (DIO), both attenuating adipose tissue inflammation and ameliorating the metabolic phenotype. Thus, blocking the P2Y2 cascade may be a promising strategy to limit metabolic disease and its sequelae.

In recent years, various research groups aimed to augment extinction learning (the most important underlying mechanism of exposure therapy) using glucocorticoids (GCs), in particular the stress hormone cortisol. In this review, we introduce the STaR (Stress Timing affects Relapse) model, a theoretical model of the timing-dependent effects of stress/GCs treatment on extinction and relapse. In particular, we show that (1) pre-extinction stress/GCs promote memory consolidation in a context-independent manner, making extinction memory more resistant to relapse following context change. (2) Post-extinction stress also enhances extinction consolidation, but in a context-bound manner. These differences may result from the timing-dependent effects of cortisol on emotional memory contextualization. At the neural level, extinction facilitation is reflected in alterations in the amygdala-hippocampal-prefrontal cortex network. (3) Stress/GCs before a retrieval test impair extinction retrieval and promote relapse. This may result from strengthening amygdala signaling or disruption of the inhibitory functioning of the prefrontal cortex. The STaR model can contribute to the understanding and prevention of relapse processes.