Effects of Experiencing CS-US Pairings on Instructed Fear Reversal.

Fear learning allows us to identify and anticipate aversive events and adapt our behavior accordingly. This is often thought to rely on associative learning mechanisms where an initially neutral conditioned stimulus (CS) is repeatedly paired with an aversive unconditioned stimulus (US), eventually leading to the CS also being perceived as aversive and threatening. Importantly, however, humans also show verbal fear learning. Namely, they have the ability to change their responses to stimuli rapidly through verbal instructions about CS-US pairings. Past research on the link between experience-based and verbal fear learning indicated that verbal instructions about a reversal of CS-US pairings can fully override the effects of previously experienced CS-US pairings, as measured through fear ratings, skin conductance, and fear-potentiated startle. However, it remains an open question whether such instructions can also annul learned CS representations in the brain. Here, we used a fear reversal paradigm (female and male participants) in conjunction with representational similarity analysis of fMRI data to test whether verbal instructions fully override the effects of experienced CS-US pairings in fear-related brain regions or not. Previous research suggests that only the right amygdala should show lingering representations of previously experienced threat ("pavlovian trace"). Unexpectedly, we found evidence for the residual effect of prior CS-US experience to be much more widespread than anticipated, in the amygdala but also cortical regions like the dorsal anterior cingulate or dorsolateral prefrontal cortex. This finding shines a new light on the interaction of different fear learning mechanisms, at times with unexpected consequences.SIGNIFICANCE STATEMENT Humans are able to learn about aversive stimuli both from experience (i.e., repeated pairings of conditioned stimulus (CS) and unconditioned stimulus (US; pavlovian conditioning), and from verbal instructions about stimulus pairings. Understanding how experience-based and verbal learning processes interact is key for understanding the cognitive and neural underpinnings of fear learning. We tested whether prior aversive experiences (CS-US pairings) affected subsequent verbal learning, searching for lingering threat signals after verbal instructions reversed a CS from being threatening to being safe. While past research suggested such threat signals can only be found in the amygdala, we found evidence to be much more widespread, including the medial and lateral PFC. This highlights how experience-based and verbal learning processes interact to support adaptive behavior.

Adaptive coding of stimulus information in human frontoparietal cortex during visual classification.

The neural mechanisms of how frontal and parietal brain regions support flexible adaptation of behavior remain poorly understood. Here, we used functional magnetic resonance imaging (fMRI) and representational similarity analysis (RSA) to investigate frontoparietal representations of stimulus information during visual classification under varying task demands. Based on prior research, we predicted that increasing perceptual task difficulty should lead to adaptive changes in stimulus coding: task-relevant category information should be stronger, while task-irrelevant exemplar-level stimulus information should become weaker, reflecting a focus on the behaviorally relevant category information. Counter to our expectations, however, we found no evidence for adaptive changes in category coding. We did find weakened coding at the exemplar-level within categories however, demonstrating that task-irrelevant information is de-emphasized in frontoparietal cortex. These findings reveal adaptive coding of stimulus information at the exemplar-level, highlighting how frontoparietal regions might support behavior even under challenging conditions.

Targeting Mitochondria with ClpP Agonists as a Novel Therapeutic Opportunity in Breast Cancer.

Breast cancer is the most frequently diagnosed malignancy worldwide and the leading cause of cancer mortality in women. Despite the recent development of new therapeutics including targeted therapies and immunotherapy, triple-negative breast cancer remains an aggressive form of breast cancer, and thus improved treatments are needed. In recent decades, it has become increasingly clear that breast cancers harbor metabolic plasticity that is controlled by mitochondria. A myriad of studies provide evidence that mitochondria are essential to breast cancer progression. Mitochondria in breast cancers are widely reprogrammed to enhance energy production and biosynthesis of macromolecules required for tumor growth. In this review, we will discuss the current understanding of mitochondrial roles in breast cancers and elucidate why mitochondria are a rational therapeutic target. We will then outline the status of the use of mitochondria-targeting drugs in breast cancers, and highlight ClpP agonists as emerging mitochondria-targeting drugs with a unique mechanism of action. We also illustrate possible drug combination strategies and challenges in the future breast cancer clinic.

Manipulating free will beliefs using online video games.

Research in social psychology and experimental philosophy has investigated lay people's free will beliefs (FWB). Using different approaches (i.e. experimental manipulations and vignette studies), they investigated how FWB relate to other concepts, and whether changing FWB has an impact on downstream processes such as social behavior. However, both approaches have shortcomings. While experimental manipulations used in social psychology suffer from demand effects, vignettes used in experimental philosophy are often highly abstract. Across two pre-registered studies, we developed a new approach by merging them in an online video game setting. Using this novel, experience-based FWB manipulation, we found that decreasing FWB impacted variables such as perceived control and responsibility in both studies. While the experience-based manipulation influenced participants' beliefs in free will within the context of the experience ("Within the context of the scenario, would the agent believe in free will?") in the first study, this manipulation effect did not transfer to participants' general FWB ("Do you believe in free will?") in the second study. Overall, our findings suggest a way forward in studying laypeople's beliefs in free will.

Manipulating Belief in Free Will and Its Downstream Consequences: A Meta-Analysis.

Ever since some scientists and popular media put forward the idea that free will is an illusion, the question has risen what would happen if people stopped believing in free will. Psychological research has investigated this question by testing the consequences of experimentally weakening people's free will beliefs. The results of these investigations have been mixed, with successful experiments and unsuccessful replications. This raises two fundamental questions: Can free will beliefs be manipulated, and do such manipulations have downstream consequences? In a meta-analysis including 145 experiments (95 unpublished), we show that exposing individuals to anti-free will manipulations decreases belief in free will and increases belief in determinism. However, we could not find evidence for downstream consequences. Our findings have important theoretical implications for research on free will beliefs and contribute to the discussion of whether reducing people's belief in free will has societal consequences.

Flotillin-2 regulates epidermal growth factor receptor activation, degradation by Cbl-mediated ubiquitination, and cancer growth.

Epidermal growth factor receptor (EGFR) signaling is frequently dysregulated in various cancers. The ubiquitin ligase Casitas B-lineage lymphoma proto-oncogene (Cbl) regulates degradation of activated EGFR through ubiquitination and acts as an adaptor to recruit proteins required for trafficking. Here, we used stable isotope labeling with amino acids in cell culture mass spectrometry to compare Cbl complexes with or without epidermal growth factor (EGF) stimulation. We identified over a hundred novel Cbl interactors, and a secondary siRNA screen found that knockdown of Flotillin-2 (FLOT2) led to increased phosphorylation and degradation of EGFR upon EGF stimulation in HeLa cells. In PC9 and H441 cells, FLOT2 knockdown increased EGF-stimulated EGFR phosphorylation, ubiquitination, and downstream signaling, reversible by EGFR inhibitor erlotinib. CRISPR knockout (KO) of FLOT2 in HeLa cells confirmed EGFR downregulation, increased signaling, and increased dimerization and endosomal trafficking. Furthermore, we determined that FLOT2 interacted with both Cbl and EGFR. EGFR downregulation upon FLOT2 loss was Cbl dependent, as coknockdown of Cbl and Cbl-b restored EGFR levels. In addition, FLOT2 overexpression decreased EGFR signaling and growth. Overexpression of wildtype (WT) FLOT2, but not the soluble G2A FLOT2 mutant, inhibited EGFR phosphorylation upon EGF stimulation in HEK293T cells. FLOT2 loss induced EGFR-dependent proliferation and anchorage-independent growth. Lastly, FLOT2 KO increased tumor formation and tumor volume in nude mice and NSG mice, respectively. Together, these data demonstrated that FLOT2 negatively regulated EGFR activation and dimerization, as well as its subsequent ubiquitination, endosomal trafficking, and degradation, leading to reduced proliferation in vitro and in vivo.

The impact of free will beliefs on implicit learning.

A growing number of studies demonstrate that belief in free will (FWB) is dynamic, and can be reduced experimentally. Most of these studies assume that doing so has beneficial effects on behavior, as FWBs are thought to subdue unwanted automatic processes (e.g. racial stereotypes). However, relying on automatic processes can sometimes be advantageous, for instance during implicit learning (e.g. detecting and exploiting statistical regularities in the environment). In this registered report, we tested whether experimentally reducing FWBs positively affected implicit motor learning. We hypothesized that reducing FWBs would lead to both faster and stronger implicit learning, as measured using the alternating serial reaction time (ASRT) task. While we did show a manipulation effect on free will beliefs, there was no detectable effect on implicit learning processes. This finding adds to the growing body of evidence that free will belief manipulations do not meaningfully affect downstream behavior.

Relating free will beliefs and attitudes.

Most people believe in free will, which is foundational for our sense of agency and responsibility. Past research demonstrated that such beliefs are dynamic, and can be manipulated experimentally. Much less is known about free will attitudes (FWAs; do you value free will?), whether they are equally dynamic, and about their relation to free will beliefs (FWBs). If FWAs were strongly positive, people might be reluctant to revise their beliefs even in the face of strong evidence to do so. In this registered report, we developed a novel measure of FWAs and directly related FWBs and attitudes for the first time. We found FWBs and attitudes to be positively related, although to a lesser degree than determinism or dualism beliefs/attitudes. Nevertheless, an experimental manipulation technique aimed at reducing FWBs (Crick text) showed remarkably specific effects on FWBs only, and no effects on FWAs. Overall, these results provide valuable new insights into laypeople's views on free will by including a novel measure of FWAs. They also provide evidence for the validity of a common experimental technique that has been rightfully criticized in the literature lately.

The Representations of Chinese Characters: Evidence from Sublexical Components.

Little research has been done about the neural substrate of the sublexical level of Chinese word recognition. In particular, it is unclear how radicals participate in Chinese word processing. We compared two measures of radical combinability, position-general radical combinability (GRC) and position-specific radical combinability (SRC) depending on whether the position of the radical is taken into account. We selected characters with embedded target radicals that had different GRC and SRC measures. These measures were used as predictors in a parametric modulation analysis and a multivariate representational similarity analysis. Human participants with native Mandarin speakers (17 males and 24 females) were asked to read words in search of animal words. Results showed that SRC is a better predictor than GRC in decoding the neural patterns. Whole-brain analysis indicated that SRC is encoded bilaterally in the inferior frontal gyrus (IFG, pars opercularis, and pars triangularis), the middle frontal gyrus (MFG), and a region on the border of the superior parietal lobule and the inferior parietal lobule (SPL/IPL). Region-of-interest-based RSA confirmed the results of the whole-brain analysis. Furthermore, we observed a correlation of another sublexical variable, logographeme composition, with bilateral activity in SPL. Logographemes refer to the basic stroke combinations that form radicals and characters. Finally, we observed involvement of bilateral cerebellum activity in Chinese word recognition. Our findings confirm the importance of sublexical components (SRC and logographeme composition) in Chinese word recognition and also confirm that Chinese word recognition involves more bilateral processing than word recognition in alphabetical languages.SIGNIFICANCE STATEMENT Chinese is a logographic language. However, characters contain informative subword components (radicals, logographemes, and strokes). We investigated whether the position of the radical is important. We presented carefully selected words and looked where brain activity correlated with subword information. Results indicate that position-dependent radicals predict brain encoding in a network of regions associated with Chinese word recognition, including higher order regions such as bilateral IFG, MFG, and SPL/IPL. Logographeme composition had an effect as well. Our findings provide strong evidence (1) for the importance of position-specific radical information and logographemes in Chinese word recognition, (2) that current brain imaging techniques are best suited to study these, and (3) that confirms the interactive nature of Chinese character recognition.

Dynamic computation of value signals via a common neural network in multi-attribute decision-making.

Studies in decision neuroscience have identified robust neural representations for the value of choice options. However, overall values often depend on multiple attributes, and it is not well understood how the brain evaluates different attributes and integrates them to combined values. In particular, it is not clear whether attribute values are computed in distinct attribute-specific regions or within the general valuation network known to process overall values. Here, we used a functional magnetic resonance imaging choice task in which abstract stimuli had to be evaluated based on variations of the attributes color and motion. The behavioral data showed that participants responded faster when overall values were high and attribute value differences were low. On the neural level, we did not find that attribute values were systematically represented in areas V4 and V5, even though these regions are associated with attribute-specific processing of color and motion, respectively. Instead, attribute values were associated with activity in the posterior cingulate cortex, ventral striatum and posterior inferior temporal gyrus. Furthermore, overall values were represented in dorsolateral and ventromedial prefrontal cortex, and attribute value differences in dorsomedial prefrontal cortex, which suggests that these regions play a key role for the neural integration of attribute values.

Neural response to aggressive and positive interactions in violent offenders and nonviolent individuals.

BACKGROUND: Due to its severe negative consequences, human violence has been targeted by a vast number of studies. Yet, neurobiological mechanisms underlying violence are still widely unclear and it seems necessary to aim for high ecological validity to learn about mechanisms contributing to violence in real life. METHODS: The present functional magnetic resonance imaging (fMRI) study investigated the neurofunction of individuals with a history of violent offenses compared with that of controls using a laboratory paradigm requesting individuals to empathically engage in videos depicting provocative aggressive and positive social interactions from a first-person perspective. RESULTS: The contrast of aggressive vs. positive scenarios revealed midbrain activation patterns associated with caudal periaqueductal gray (PAG) in violent offenders; In controls, the rostral PAG was involved. Additionally, only in controls, this contrast revealed an involvement of the amygdaloidal complex. Moreover, in violent offenders the contrast of positive vs. aggressive situations revealed an involvement of areas in the insula, post-central gyrus and anterior cingulate cortex. CONCLUSIONS: Our results support findings on the differential role of PAG subdivisions in response to threat and point to altered processing of positive social interactions in violent offenders. They further support the notion that changes in PAG recruitment might contribute to violent individuals "taking action" instead of freezing in case of threatening situations.

Fatty acid synthase mediates high glucose-induced EGFR activation in oral dysplastic keratinocytes.

BACKGROUND: Recent studies point to the epidermal growth factor receptor (EGFR) as a critical mediator of type 2 diabetes mellitus (T2DM)-induced renal, cardiac, and ocular complications. T2DM is considered a systemic contributing factor in oral carcinogenesis. Similarly, increased EGFR gene copy number and protein expression strongly predict tumor progression. Yet, the impact of hyperglycemia on EGFR activity in oral potentially malignant disorders remains unclear. We recently reported that fatty acid synthase (FASN), a key de novo lipogenic enzyme, mediates EGFR activation in nicotine-treated oral dysplastic keratinocytes. While in non-malignant tissues FASN expression is extremely low, it is frequently upregulated in several cancers, including oral squamous cell carcinoma. The present study was carried out to investigate whether high glucose conditions trigger pro-oncogenic responses in oral dysplastic keratinocytes via FASN-mediated EGFR activation. METHODS: Cell viability and migration of oral dysplastic keratinocytes were evaluated when exposed to normal (5 mM) or high (20 mM) glucose conditions in the presence of FASN and EGFR inhibitors. Western blotting was also performed to assess changes in FASN protein expression and EGFR activation. RESULTS: Oral dysplastic keratinocytes exposed to high glucose led to EGFR activation in a FASN-dependent manner. Likewise, high glucose significantly enhanced cell viability and migration in a FASN/EGFR-mediated fashion. Notably, EGFR inhibition by the anti-EGFR monoclonal antibody cetuximab significantly reduced the proliferation of FASN-overexpressing oral dysplastic keratinocytes. CONCLUSION: These novel findings suggest that FASN may act as a key targetable metabolic regulator of glucose-induced EGFR oncogenic signaling in oral potentially malignant disorders.

Frontoparietal action-oriented codes support novel instruction implementation.

A key aspect of human cognitive flexibility concerns the ability to convert complex symbolic instructions into novel behaviors. Previous research proposes that this transformation is supported by two neurocognitive states: an initial declarative maintenance of task knowledge, and an implementation state necessary for optimal task execution. Furthermore, current models predict a crucial role of frontal and parietal brain regions in this process. However, whether declarative and procedural signals independently contribute to implementation remains unknown. We report the results of an fMRI experiment in which participants executed novel instructed stimulus-response associations. We then used a pattern-tracking procedure to quantify the contribution of format-unique signals during instruction implementation. This revealed independent procedural and declarative representations of novel S-Rs in frontoparietal areas, prior to execution. Critically, the degree of procedural activation predicted subsequent behavioral performance. Altogether, our results suggest an important contribution of frontoparietal regions to the neural architecture that regulates cognitive flexibility.

Shared Neural Representations of Cognitive Conflict and Negative Affect in the Medial Frontal Cortex.

Influential theories of Medial Frontal Cortex (MFC) function suggest that the MFC registers cognitive conflict as an aversive signal, but no study directly tested this idea. Instead, recent studies suggested that nonoverlapping regions in the MFC process conflict and affect. In this preregistered human fMRI study (male and female), we used MVPAs to identify which regions respond similarly to conflict and aversive signals. The results reveal that, of all conflict- and value-related regions, only the ventral pre-supplementary motor area (or dorsal anterior cingulate cortex) showed a shared neural pattern response to different conflict and affect tasks. These findings challenge recent conclusions that conflict and affect are processed independently, and provide support for integrative views of MFC function.SIGNIFICANCE STATEMENT Multiple theories propose that the MFC, and the dorsal ACC in particular, integrates information related to suboptimal outcomes from different psychological domains (e.g., cognitive control and negative affect) with the aim of adaptively steering behavior. In contrast to recent studies in the field, we provide evidence for the idea that cognitive control and negative affect are integrated in the MFC by showing that a classification algorithm trained on discerning cognitive control (conflict vs no conflict) can predict affect (negative vs positive) in the voxel pattern response of the dorsal ACC/pre-SMA.

The contribution of phonological information to visual word recognition: Evidence from Chinese phonetic radicals.

Lateralization is a critical characteristic of language production and also plays a role in visual word recognition. However, the neural mechanisms underlying the interactions between visual input and spoken word representations are still unclear. We investigated the contribution of sub-lexical phonological information in visual word processing by exploiting the fact that Chinese characters can contain phonetic radicals in either the left or right half of the character. FMRI data were collected while 39 Chinese participants read words in search of target color words. On the basis of whole-brain analysis and three laterality analyses of regions of interest, we argue that visual information from centrally presented Chinese characters is split in the fovea and projected to the contralateral visual cortex, from which phonological information can be extracted rapidly if the character contains a phonetic radical. Extra activation, suggestive of more effortful processing, is observed when the phonetic radical is situated in the left half of the character and therefore initially sent to the visual cortex in the right hemisphere that is less specialized for language processing. Our results are in line with the proposal that phonological information helps written word processing by means of top-down feedback.

Cultural pressure and biased responding in free will attitudes.

Whether you believe free will exists has profound effects on your behaviour, across different levels of processing, from simple motor action to social cognition. It is therefore important to understand which specific lay theories are held in the general public and why. Past research largely focused on investigating free will beliefs (FWB, 'Do you think free will exists?'), but largely ignored a second key aspect: free will attitudes (FWA, 'Do you like/value will?'). Attitudes are often independently predictive of behaviour, relative to beliefs, yet we currently know very little about FWAs in the general public. One key issue is whether such attitudes are subject to biased, socially desirable responding. The vast majority of the general public strongly believes in the existence of free will, which might create cultural pressure to value free will positively as well. In this registered report, we used a very large (N = 1100), open available dataset measuring implicit and explicit attitudes towards free will and determinism to address this issue. Our results indicate that both explicit and implicit attitudes towards free will are more positive than attitudes towards determinism. We also show that people experience cultural pressure to value free will, and to devalue determinism. Yet, we found no strong evidence that this cultural pressure affected either implicit or explicit attitudes in this dataset.

Investigating the effect of trustworthiness on instruction-based reflexivity.

Unlike other species, humans are capable of rapidly learning new behavior from a single instruction. While previous research focused on the cognitive processes underlying the rapid, automatic implementation of instructions, the fundamentally social nature of instruction following has remained largely unexplored. Here, we investigated whether instructor trustworthiness modulates instruction implementation using both explicit and reflexive measures. In a first preregistered study, we validated a new paradigm to manipulate the perceived trustworthiness of two different virtual characters and showed that such a manipulation reliably induced implicit associations between the virtual characters and trustworthiness attributes. Moreover, we show that trustworthy instructors are followed more frequently and faster. In two additional preregistered experiments, we tested if trustworthiness towards the instructor influenced the cognitive processes underlying instruction implementation. While we show that verbally conveyed instructions led to automatic instruction implementation, this effect was not modulated by the trustworthiness of the instructor. Thus, we succeeded to design and validate a novel trustworthiness manipulation (Experiment 1) and to create a social variant of the instruction-based reflexivity paradigm (Experiments 2 and 3). However, this instruction-based reflexivity effect was not modulated by the instructors' trustworthiness.

Rethinking the Characterization of Nanoscale Field-Effect Transistors: A Universal Approach.

Standard methods for calculating transport parameters in nanoscale field-effect transistors (FETs), namely carrier concentration and mobility, require a linear connection between the gate voltage and channel conductance; however, this is often not the case. One reason often overlooked is that shifts in chemical and electric potential can partially compensate each other, commonly referred to as quantum capacitance. In nanoscale FETs, capacitance is often unmeasurable and an analytical formula is required, which assumes the conducting channel as metallic and common methods of determining threshold voltage no longer couple properly into transport equations. As present and future FET structures become smaller and have increased channel-gate coupling, this issue will render standard methods impossible to use. This work discusses the validity of common methods of characterization for nanoscale FETs, develops a universal model to determine transport properties by only measuring the threshold voltage of an FET and presents a new parameter to easily classify FETs as either quantum capacitance-limited or metallic approximated charge transport. Also considered in this work is electrical hysteresis from trap states and, in combination with the proposed universal model, novel techniques are introduced to measure and remove the errors associated with these effects often ignored in literature.

Are all behavioral reward benefits created equally? An EEG-fMRI study.

Reward consistently boosts performance in cognitive tasks. Although many different reward manipulations exist, systematic comparisons are lacking. Reward effects on cognitive control are usually studied using monetary incentive delay (MID; cue-related reward information) or stimulus-reward association (SRA; target-related reward information) tasks. While for MID tasks, evidence clearly implicates reward-triggered global increases in proactive control, it is unclear how reward effects arise in SRA tasks, and in how far such mechanisms overlap during task preparation and target processing. Here, we address these questions with simultaneous EEG-fMRI using a Stroop task with four different block types. In addition to MID and SRA blocks, we used an SRA-task modification with reward-irrelevant cues (C-SRA) and regular reward-neutral Stroop-task blocks. Behaviorally, we observed superior performance for all reward conditions compared to Neutral, and more pronounced reward effects in the SRA and C-SRA blocks, compared to MID blocks. The fMRI data showed similar reward effects in value-related areas for events that signaled reward availability (MID cues and (C-)SRA targets), and comparable reward modulations in cognitive-control regions for all targets regardless of block type. This result pattern was echoed by the EEG data, showing clear markers of valuation and cognitive control, which only differed during task preparation, whereas reward-related modulations during target processing were again comparable across block types. Yet, considering only cue-related fMRI data, C-SRA cues triggered preparatory control processes beyond reward-unrelated MID cues, without simultaneous modulations in typical reward areas, implicating enhanced task preparation that is not directly driven by a concurrent neural reward-anticipation response.

Outcome contingency selectively affects the neural coding of outcomes but not of tasks.

Value-based decision-making is ubiquitous in every-day life, and critically depends on the contingency between choices and their outcomes. Only if outcomes are contingent on our choices can we make meaningful value-based decisions. Here, we investigate the effect of outcome contingency on the neural coding of rewards and tasks. Participants performed a reversal-learning paradigm in which reward outcomes were contingent on trial-by-trial choices, and performed a 'free choice' paradigm in which rewards were random and not contingent on choices. We hypothesized that contingent outcomes enhance the neural coding of rewards and tasks, which was tested using multivariate pattern analysis of fMRI data. Reward outcomes were encoded in a large network including the striatum, dmPFC and parietal cortex, and these representations were indeed amplified for contingent rewards. Tasks were encoded in the dmPFC at the time of decision-making, and in parietal cortex in a subsequent maintenance phase. We found no evidence for contingency-dependent modulations of task signals, demonstrating highly similar coding across contingency conditions. Our findings suggest selective effects of contingency on reward coding only, and further highlight the role of dmPFC and parietal cortex in value-based decision-making, as these were the only regions strongly involved in both reward and task coding.