Cigarette craving in virtual reality cue exposure in abstainers and relapsed smokers.

Cue exposure therapy (CET) in substance-use disorders aims to reduce craving and ultimately relapse rates. Applying CET in virtual reality (VR) was proposed to increase its efficacy, as VR enables the presentation of social and environmental cues along with substance-related stimuli. However, limited success has been reported so far when applying VR-CET for smoking cessation. Understanding if effects of VR-CET differ between future abstainers and relapsing smokers may help to improve VR-CET. Data from 102 participants allocated to the intervention arm (VR-CET) of a recent RCT comparing VR-CET to relaxation in the context of smoking cessation was analyzed with respect to tolerability, presence, and craving during VR-CET. Cue exposure was conducted in four VR contexts (Loneliness/Rumination, Party, Stress, Café), each presented twice. Relapsed smokers compared to abstainers experienced higher craving during VR-CET and stronger craving responses especially during the Stress scenario. Furthermore, lower mean craving during VR-CET positively predicted abstinence at 6-month follow-up. Attempts to improve smoking cessation outcomes of VR-CET should aim to identify smokers who are more at risk of relapse based on high craving levels during VR-CET. Specifically measuring craving responses during social stress seems to be well suited to mark relapse. We propose to investigate individualized treatment approaches accordingly.

Designed Ankyrin Repeat Proteins as Actin Labels of Distinct Cytoskeletal Structures in Living Cells.

The orchestrated assembly of actin and actin-binding proteins into cytoskeletal structures coordinates cell morphology changes during migration, cytokinesis, and adaptation to external stimuli. The accurate and unbiased visualization of the diverse actin assemblies within cells is an ongoing challenge. We describe here the identification and use of designed ankyrin repeat proteins (DARPins) as synthetic actin binders. Actin-binding DARPins were identified through ribosome display and validated biochemically. When introduced or expressed inside living cells, fluorescently labeled DARPins accumulated at actin filaments, validated through phalloidin colocalization on fixed cells. Nevertheless, different DARPins displayed different actin labeling patterns: some DARPins labeled efficiently dynamic structures, such as filopodia, lamellipodia, and blebs, while others accumulated primarily in stress fibers. This differential intracellular distribution correlated with DARPin-actin binding kinetics, as measured by fluorescence recovery after photobleaching experiments. Moreover, the rapid arrest of actin dynamics induced by pharmacological treatment led to the fast relocalization of DARPins. Our data support the hypothesis that the localization of actin probes depends on the inherent dynamic movement of the actin cytoskeleton. Compared to the widely used LifeAct probe, one DARPin exhibited enhanced signal-to-background ratio while retaining a similar ability to label stress fibers. In summary, we propose DARPins as promising actin-binding proteins for labeling or manipulation in living cells.

Multiple Genes Core to ERAD, Macroautophagy and Lysosomal Degradation Pathways Participate in the Proteostasis Response in α1-Antitrypsin Deficiency.

BACKGROUND & AIMS: In the classic form of α1-antitrypsin deficiency (ATD), the misfolded α1-antitrypsin Z (ATZ) variant accumulates in the endoplasmic reticulum (ER) of liver cells. A gain-of-function proteotoxic mechanism is responsible for chronic liver disease in a subgroup of homozygotes. Proteostatic response pathways, including conventional endoplasmic reticulum-associated degradation and autophagy, have been proposed as the mechanisms that allow cellular adaptation and presumably protection from the liver disease phenotype. Recent studies have concluded that a distinct lysosomal pathway called endoplasmic reticulum-to-lysosome completely supplants the role of the conventional macroautophagy pathway in degradation of ATZ. Here, we used several state-of-the-art approaches to characterize the proteostatic responses more fully in cellular systems that model ATD. METHODS: We used clustered regularly interspaced short palindromic repeats (CRISPR)-mediated genome editing coupled to a cell selection step by fluorescence-activated cell sorter to perform screening for proteostasis genes that regulate ATZ accumulation and combined that with selective genome editing in 2 other model systems. RESULTS: Endoplasmic reticulum-associated degradation genes are key early regulators and multiple autophagy genes, from classic as well as from ER-to-lysosome and other newly described ER-phagy pathways, participate in degradation of ATZ in a manner that is temporally regulated and evolves as ATZ accumulation persists. Time-dependent changes in gene expression are accompanied by specific ultrastructural changes including dilation of the ER, formation of globular inclusions, budding of autophagic vesicles, and alterations in the overall shape and component parts of mitochondria. CONCLUSIONS: Macroautophagy is a critical component of the proteostasis response to cellular ATZ accumulation and it becomes more important over time as ATZ synthesis continues unabated. Multiple subtypes of macroautophagy and nonautophagic lysosomal degradative pathways are needed to respond to the high concentrations of misfolded protein that characterizes ATD and these pathways are attractive candidates for genetic variants that predispose to the hepatic phenotype.

Causal evidence for a coordinated temporal interplay within the language network.

Recent neurobiological models on language suggest that auditory sentence comprehension is supported by a coordinated temporal interplay within a left-dominant brain network, including the posterior inferior frontal gyrus (pIFG), posterior superior temporal gyrus and sulcus (pSTG/STS), and angular gyrus (AG). Here, we probed the timing and causal relevance of the interplay between these regions by means of concurrent transcranial magnetic stimulation and electroencephalography (TMS-EEG). Our TMS-EEG experiments reveal region- and time-specific causal evidence for a bidirectional information flow from left pSTG/STS to left pIFG and back during auditory sentence processing. Adapting a condition-and-perturb approach, our findings further suggest that the left pSTG/STS can be supported by the left AG in a state-dependent manner.

Public speaking training in front of a supportive audience in Virtual Reality improves performance in real-life.

Public speaking is a challenging task that requires practice. Virtual Reality allows to present realistic public speaking scenarios in this regard, however, the role of the virtual audience during practice remains unknown. In the present study, 73 participants completed a Virtual Reality practice session while audience was manipulated to be supportive or unsupportive or presentations were practiced without audience. Importantly, following the virtual practice, participants held the presentation during a real university course via Zoom. We measured emotional experience, self-efficacy, and the subjective evaluation of performance at baseline, after VR practice, and after the real presentation. Additionally, participants' performance in the real presentation was evaluated by instructors (blinded to condition). Supportive in contrast to unsupportive audiences led to more positive believes about one's own performance, while there were no changes in beliefs in the group without audience. Importantly, practice in front of a supportive compared to unsupportive audience resulted in a more positive evaluation of speaker confidence in real-life public speaking as rated by the instructors. These results demonstrate an impact of virtual social feedback during public speaking on subsequent subjective performance evaluation. This may increase self-confidence resulting in actual improved public speaking performance in real-life.

Identification of Brain-Penetrant ATP-Competitive mTOR Inhibitors for CNS Syndromes.

The allosteric inhibitor of the mechanistic target of rapamycin (mTOR) everolimus reduces seizures in tuberous sclerosis complex (TSC) patients through partial inhibition of mTOR functions. Due to its limited brain permeability, we sought to develop a catalytic mTOR inhibitor optimized for central nervous system (CNS) indications. We recently reported an mTOR inhibitor (1) that is able to block mTOR functions in the mouse brain and extend the survival of mice with neuronal-specific ablation of the Tsc1 gene. However, 1 showed the risk of genotoxicity in vitro. Through structure-activity relationship (SAR) optimization, we identified compounds 9 and 11 without genotoxicity risk. In neuronal cell-based models of mTOR hyperactivity, both corrected aberrant mTOR activity and significantly improved the survival rate of mice in the Tsc1 gene knockout model. Unfortunately, 9 and 11 showed limited oral exposures in higher species and dose-limiting toxicities in cynomolgus macaque, respectively. However, they remain optimal tools to explore mTOR hyperactivity in CNS disease models.

Autophagy in the commercial arena: a conversation with Dr. Leon Murphy, Chief Scientific Officer at Casma therapeutics.

As a maturing field that continues to provide fundamental insights into cell physiology, autophagy is also beginning to attract considerable interest from the biotechnology/pharmaceutical sector. For this Editor's corner, I thought it would be both useful and interesting to talk with somebody who has spent a lot of time in the commercial sphere, working on autophagy and related processes. I was fortunate that Dr. Leon Murphy, Chief Scientific Officer at Casma therapeutics, was willing and able to answer my questions. In addition to his insights on the commercial interest for autophagy, Dr. Murphy also shared his personal experience on the scientific life working in large and small pharmaceutical companies.

A terpene nucleoside from M. tuberculosis induces lysosomal lipid storage in foamy macrophages.

Induction of lipid-laden foamy macrophages is a cellular hallmark of tuberculosis (TB) disease, which involves the transformation of infected phagolysosomes from a site of killing into a nutrient-rich replicative niche. Here, we show that a terpenyl nucleoside shed from Mycobacterium tuberculosis, 1-tuberculosinyladenosine (1-TbAd), caused lysosomal maturation arrest and autophagy blockade, leading to lipid storage in M1 macrophages. Pure 1-TbAd, or infection with terpenyl nucleoside-producing M. tuberculosis, caused intralysosomal and peribacillary lipid storage patterns that matched both the molecules and subcellular locations known in foamy macrophages. Lipidomics showed that 1-TbAd induced storage of triacylglycerides and cholesterylesters and that 1-TbAd increased M. tuberculosis growth under conditions of restricted lipid access in macrophages. Furthermore, lipidomics identified 1-TbAd-induced lipid substrates that define Gaucher's disease, Wolman's disease, and other inborn lysosomal storage diseases. These data identify genetic and molecular causes of M. tuberculosis-induced lysosomal failure, leading to successful testing of an agonist of TRPML1 calcium channels that reverses lipid storage in cells. These data establish the host-directed cellular functions of an orphan effector molecule that promotes survival in macrophages, providing both an upstream cause and detailed picture of lysosome failure in foamy macrophages.

Returning a smile: Initiating a social interaction with a facial emotional expression influences the evaluation of the expression received in return.

Face-to-face social interactions are characterized by the reciprocal exchange of facial emotions between interaction partners. Typically, facial emotional expressions have been studied in passive observation paradigms, while interactive mechanisms remain unknown. In the current study we investigate how sending a facial emotional expression influenced the evaluation of an emotional expression received in return. Sixty-eight participants were cued to direct a facial emotional expression (happy, angry, neutral) towards a virtual agent in front of them. The virtual agent then responded with either the same or another emotional expression (happy, angry). Evaluation of the response expressions was measured via ratings of valence and arousal as well as EMG recordings of the M. corrugator supercilii and the M. zygomaticus major. Results revealed a significant interaction between the emotion of the initial facial expression and the response expression. Valence of happy response expressions were increased when participants had initially displayed a smile compared to a neutral expression or a frown. This was also reflected in the EMG responses. Initiating an interaction with a smile increased Zygomaticus activation for happy relative to angry response expressions compared to when the interaction was initiated with a frown. In contrast, no interplay of the initial and the response expression was observed in the Corrugator. These findings demonstrate that smiling or frowning at another person can modulate socio-emotional processing of subsequent social cues. Therefore, the present study highlights the interactive nature of facial emotional expressions.

The ABCD stop signal data: Response to Bissett et al.

This paper responds to a recent critique by Bissett et al. of the fMRI Stop task used in the Adolescent Brain Cognitive Development℠ Study (ABCD Study®). The critique focuses primarily on a task design feature related to race model assumptions (i.e., that the Go and Stop processes are fully independent). In response, we note that the race model is quite robust against violations of its assumptions. Most importantly, while Bissett raises conceptual concerns with the task we focus here on analyzes of the task data and conclude that the concerns appear to have minimal impact on the neuroimaging data (the validity of which do not rely on race model assumptions) and have far less of an impact on the performance data than the critique suggests. We note that Bissett did not apply any performance-based exclusions to the data they analyzed, a number of the trial coding errors they flagged were already identified and corrected in ABCD annual data releases, a number of their secondary concerns reflect sensible design decisions and, indeed, their own computational modeling of the ABCD Stop task suggests the problems they identify have just a modest impact on the rank ordering of individual differences in subject performance.

Experimental validation of state equations and dynamic route maps for phase change memristive devices.

Phase Change Memory (PCM) is an emerging technology exploiting the rapid and reversible phase transition of certain chalcogenides to realize nanoscale memory elements. PCM devices are being explored as non-volatile storage-class memory and as computing elements for in-memory and neuromorphic computing. It is well-known that PCM exhibits several characteristics of a memristive device. In this work, based on the essential physical attributes of PCM devices, we exploit the concept of Dynamic Route Map (DRM) to capture the complex physics underlying these devices to describe them as memristive devices defined by a state-dependent Ohm's law. The efficacy of the DRM has been proven by comparing numerical results with experimental data obtained on PCM devices.

Face masks reduce interpersonal distance in virtual reality.

During the COVID-19 pandemic several behavioral measures have been implemented to reduce viral transmission. While these measures reduce the risk of infections, they may also increase risk behavior. Here, we experimentally investigate the influence of face masks on physical distancing. Eighty-four participants with or without face masks passed virtual agents in a supermarket environment to reach a target while interpersonal distance was recorded. Agents differed in wearing face masks and age (young, elderly). In addition, situational constraints varied in whether keeping a distance of 1.5 m required an effortful detour or not. Wearing face masks (both self and other) reduced physical distancing. This reduction was most prominent when keeping the recommended distance was effortful, suggesting an influence of situational constraints. Similarly, increased distances to elderly were only observed when keeping a recommended distance was effortless. These findings highlight contextual constraints in compensation behavior and have important implications for safety policies.

GABARAP sequesters the FLCN-FNIP tumor suppressor complex to couple autophagy with lysosomal biogenesis.

Adaptive changes in lysosomal capacity are driven by the transcription factors TFEB and TFE3 in response to increased autophagic flux and endolysosomal stress, yet the molecular details of their activation are unclear. LC3 and GABARAP members of the ATG8 protein family are required for selective autophagy and sensing perturbation within the endolysosomal system. Here, we show that during the conjugation of ATG8 to single membranes (CASM), Parkin-dependent mitophagy, and Salmonella-induced xenophagy, the membrane conjugation of GABARAP, but not LC3, is required for activation of TFEB/TFE3 to control lysosomal capacity. GABARAP directly binds to a previously unidentified LC3-interacting motif (LIR) in the FLCN/FNIP tumor suppressor complex and mediates sequestration to GABARAP-conjugated membrane compartments. This disrupts FLCN/FNIP GAP function toward RagC/D, resulting in impaired substrate-specific mTOR-dependent phosphorylation of TFEB. Thus, the GABARAP-FLCN/FNIP-TFEB axis serves as a molecular sensor that coordinates lysosomal homeostasis with perturbations and cargo flux within the autophagy-lysosomal network.

Extrahepatic Surgery in Cirrhosis Significantly Increases Portal Pressure in Preclinical Animal Models.

Background: Liver cirrhosis is a relevant comorbidity with increasing prevalence. Postoperative decompensation and development of complications in patients with cirrhosis remains a frequent clinical problem. Surgery has been discussed as a precipitating event for decompensation and complications of cirrhosis, but the underlying pathomechanisms are still obscure. The aim of this study was to analyze the role of abdominal extrahepatic surgery in cirrhosis on portal pressure and fibrosis in a preclinical model. Methods: Compensated liver cirrhosis was induced using tetrachlormethane (CCL4) inhalation and bile duct ligation (BDL) models in rats, non-cirrhotic portal hypertension by partial portal vein ligation (PPVL). Intestinal manipulation (IM) as a model of extrahepatic abdominal surgery was performed. 2 and 7 days after IM, portal pressure was measured in-vivo. Hydroxyproline measurements, Sirius Red staining and qPCR measurements of the liver were performed for evaluation of fibrosis development and hepatic inflammation. Laboratory parameters of liver function in serum were analyzed. Results: Portal pressure was significantly elevated 2 and 7 days after IM in both models of cirrhosis. In the non-cirrhotic model the trend was the same, while not statistically significant. In both cirrhotic models, IM shows strong effects of decompensation, with significant weight loss, elevation of liver enzymes and hypoalbuminemia. 7 days after IM in the BDL group, Sirius red staining and hydroxyproline levels showed significant progression of fibrosis and significantly elevated mRNA levels of hepatic inflammation compared to the respective control group. A progression of fibrosis was not observed in the CCL4 model. Conclusion: In animal models of cirrhosis with continuous liver injury (BDL), IM increases portal pressure, and development of fibrosis. Perioperative portal pressure and hence inflammation processes may be therapeutic targets to prevent post-operative decompensation in cirrhosis.

Angry facial expressions bias towards aversive actions.

Social interaction requires fast and efficient processing of another person's intentions. In face-to-face interactions, aversive or appetitive actions typically co-occur with emotional expressions, allowing an observer to anticipate action intentions. In the present study, we investigated the influence of facial emotions on the processing of action intentions. Thirty-two participants were presented with video clips showing virtual agents displaying a facial emotion (angry vs. happy) while performing an action (punch vs. fist-bump) directed towards the observer. During each trial, video clips stopped at varying durations of the unfolding action, and participants had to recognize the presented action. Naturally, participants' recognition accuracy improved with increasing duration of the unfolding actions. Interestingly, while facial emotions did not influence accuracy, there was a significant influence on participants' action judgements. Participants were more likely to judge a presented action as a punch when agents showed an angry compared to a happy facial emotion. This effect was more pronounced in short video clips, showing only the beginning of an unfolding action, than in long video clips, showing near-complete actions. These results suggest that facial emotions influence anticipatory processing of action intentions allowing for fast and adaptive responses in social interactions.

On the Thermal Models for Resistive Random Access Memory Circuit Simulation.

Resistive Random Access Memories (RRAMs) are based on resistive switching (RS) operation and exhibit a set of technological features that make them ideal candidates for applications related to non-volatile memories, neuromorphic computing and hardware cryptography. For the full industrial development of these devices different simulation tools and compact models are needed in order to allow computer-aided design, both at the device and circuit levels. Most of the different RRAM models presented so far in the literature deal with temperature effects since the physical mechanisms behind RS are thermally activated; therefore, an exhaustive description of these effects is essential. As far as we know, no revision papers on thermal models have been published yet; and that is why we deal with this issue here. Using the heat equation as the starting point, we describe the details of its numerical solution for a conventional RRAM structure and, later on, present models of different complexity to integrate thermal effects in complete compact models that account for the kinetics of the chemical reactions behind resistive switching and the current calculation. In particular, we have accounted for different conductive filament geometries, operation regimes, filament lateral heat losses, the use of several temperatures to characterize each conductive filament, among other issues. A 3D numerical solution of the heat equation within a complete RRAM simulator was also taken into account. A general memristor model is also formulated accounting for temperature as one of the state variables to describe electron device operation. In addition, to widen the view from different perspectives, we deal with a thermal model contextualized within the quantum point contact formalism. In this manner, the temperature can be accounted for the description of quantum effects in the RRAM charge transport mechanisms. Finally, the thermometry of conducting filaments and the corresponding models considering different dielectric materials are tackled in depth.

The time course of speaker-specific language processing.

Listeners are sensitive to a speaker's individual language use and generate expectations for particular speakers. It is unclear, however, how such expectations affect online language processing. In the present EEG study, we presented thirty-two participants with auditory sentence stimuli of two speakers. Speakers differed in their use of two particular syntactic structures, easy subject-initial SOV structures and more difficult object-initial OSV structures. One speaker, the SOV-Speaker, had a high proportion of SOV sentences (75%) and a low proportion of OSV sentences (25%), and vice-versa for the OSV-Speaker. Participants were exposed to the speakers' individual language use in a training session followed by a test session on the consecutive day. ERP-results show that early stages of sentence processing are driven by syntactic processing only and are unaffected by speaker-specific expectations. In a late stage, however, an interaction between speaker and syntax information was observed. For the SOV-Speaker condition, the classical P600-effect reflected the effort of processing difficult and unexpected sentence structures. For the OSV-Speaker condition, both structures elicited different responses on frontal electrodes, possibly indexing effort to switch from a local speaker model to a global model of language use. Overall, the study identifies distinct neural mechanisms related to speaker-specific expectations.

On Local Activity and Edge of Chaos in a NaMLab Memristor.

Local activity is the capability of a system to amplify infinitesimal fluctuations in energy. Complex phenomena, including the generation of action potentials in neuronal axon membranes, may never emerge in an open system unless some of its constitutive elements operate in a locally active regime. As a result, the recent discovery of solid-state volatile memory devices, which, biased through appropriate DC sources, may enter a local activity domain, and, most importantly, the associated stable yet excitable sub-domain, referred to as edge of chaos, which is where the seed of complexity is actually planted, is of great appeal to the neuromorphic engineering community. This paper applies fundamentals from the theory of local activity to an accurate model of a niobium oxide volatile resistance switching memory to derive the conditions necessary to bias the device in the local activity regime. This allows to partition the entire design parameter space into three domains, where the threshold switch is locally passive (LP), locally active but unstable, and both locally active and stable, respectively. The final part of the article is devoted to point out the extent by which the response of the volatile memristor to quasi-static excitations may differ from its dynamics under DC stress. Reporting experimental measurements, which validate the theoretical predictions, this work clearly demonstrates how invaluable is non-linear system theory for the acquirement of a comprehensive picture of the dynamics of highly non-linear devices, which is an essential prerequisite for a conscious and systematic approach to the design of robust neuromorphic electronics. Given that, as recently proved, the potassium and sodium ion channels in biological axon membranes are locally active memristors, the physical realization of novel artificial neural networks, capable to reproduce the functionalities of the human brain more closely than state-of-the-art purely CMOS hardware architectures, should not leave aside the adoption of resistance switching memories, which, under the appropriate provision of energy, are capable to amplify the small signal, such as the niobium dioxide micro-scale device from NaMLab, chosen as object of theoretical and experimental study in this work.

Widespread Recommendations Can Change Our Habits of Hand-Washing and Physical Distance During the COVID-19 Pandemic.

Background: Habits and behaviors in everyday life currently need to be modified as quickly as possible due to the COVID-19 pandemic. Two of the most effective tools to prevent infection seem to be regular and thorough hand-washing and physical distancing during interpersonal interactions. Method: Two hundred and eighty-four participants completed a short survey to investigate how previous habits regarding hand-washing and physical distancing have changed in the general population as a function of the current pandemic and the thereby increased information and constant recommendations regarding these behaviors. Results: Participants aged 51 and older reported a greater change in everyday hand-washing behavior than younger participants. In addition, participants aged 31 and older selected significantly greater distances to have a conversation than younger participants. However, that was not the case if participants had to actively stop their conversational partner from approaching. Conclusion: Participants aged 51 years and older seem to be well aware of their at-risk status during the current pandemic and might therefore be willing to change their behavior more strongly than younger survey participants. Nevertheless, they seem to struggle with enforcing the current rules towards others. The group aged between 31 and 50 years, however, reports a comparable level of fear, but no corresponding change in hand-washing behavior. Future surveys should try to provide more insight into why this might be the case.

Autophagy Assays for Biological Discovery and Therapeutic Development.

Autophagy is a lysosome-dependent intracellular degradation system required for various physiological processes and can be dysregulated in human disease. To understand its biological significance and underlying mechanisms, measuring autophagic activity (i.e., autophagic flux) is critical. However, navigating which assays to use, and when, is complicated and at times the results are often interpreted inappropriately. This review will summarize both advantages and disadvantages of currently available methods to monitor autophagy. In addition, we discuss how these assays should be used in high-throughput screens to identify autophagy-modulating drugs and genes and the general features needed for biomarkers to assess autophagy in humans.