MK2 Phosphorylates RIPK1 to Prevent TNF-Induced Cell Death.

TNF is an inflammatory cytokine that upon binding to its receptor, TNFR1, can drive cytokine production, cell survival, or cell death. TNFR1 stimulation causes activation of NF-κB, p38α, and its downstream effector kinase MK2, thereby promoting transcription, mRNA stabilization, and translation of target genes. Here we show that TNF-induced activation of MK2 results in global RIPK1 phosphorylation. MK2 directly phosphorylates RIPK1 at residue S321, which inhibits its ability to bind FADD/caspase-8 and induce RIPK1-kinase-dependent apoptosis and necroptosis. Consistently, a phospho-mimetic S321D RIPK1 mutation limits TNF-induced death. Mechanistically, we find that phosphorylation of S321 inhibits RIPK1 kinase activation. We further show that cytosolic RIPK1 contributes to complex-II-mediated cell death, independent of its recruitment to complex-I, suggesting that complex-II originates from both RIPK1 in complex-I and cytosolic RIPK1. Thus, MK2-mediated phosphorylation of RIPK1 serves as a checkpoint within the TNF signaling pathway that integrates cell survival and cytokine production.

Chimeric antigen receptor dendritic cells targeted delivery of a single tumoricidal factor for cancer immunotherapy.

BACKGROUND: Chimeric antigen receptor (CAR)-T cells have been used to treat blood cancers by producing a wide variety of cytokines. However, they are not effective in treating solid cancers and can cause severe side-effects, including cytokine release syndrome. TNFα is a tumoricidal cytokine, but it markedly increases the protein levels of cIAP1 and cIAP2, the members of inhibitor of apoptosis protein (IAP) family of E3 ubiquitin ligase that limits caspase-induced apoptosis. Degradation of IAP proteins by an IAP antagonist does not effectively kill cancer cells but enables TNFα to strongly induce cancer cell apoptosis. It would be a promising approach to treat cancers by targeted delivery of TNFα through an inactive adoptive cell in combination with an IAP antagonist. METHODS: Human dendritic cells (DCs) were engineered to express a single tumoricidal factor, TNFα, and a membrane-anchored Mucin1 antibody scFv, named Mucin 1 directed DCs expressing TNFα (M-DCsTNF). The efficacy of M-DCsTNF in recognizing and treating breast cancer was tested in vitro and in vivo. RESULTS: Mucin1 was highly expressed on the surface of a wide range of human breast cancer cell lines. M-DCsTNF directly associated with MDA-MB-231 cells in the bone of NSG mice. M-DCsTNF plus an IAP antagonist, SM-164, but neither alone, markedly induce MDA-MB-231 breast cancer cell apoptosis, which was blocked by TNF antibody. Importantly, M-DCsTNF combined with SM-164, but not SM-164 alone, inhibited the growth of patient-derived breast cancer in NSG mice. CONCLUSION: An adoptive cell targeting delivery of TNFα combined with an IAP antagonist is a novel effective approach to treat breast cancer and could be expanded to treat other solid cancers. Unlike CAR-T cell, this novel adoptive cell is not activated to produce a wide variety of cytokines, except for additional overexpressed TNF, and thus could avoid the severe side effects such as cytokine release syndrome.

Effect of experimental modulation of mood on exertional dyspnoea in chronic obstructive pulmonary disease.

BACKGROUND AND OBJECTIVE: Dyspnoea is a debilitating symptom in individuals with chronic obstructive pulmonary disease (COPD) and a range of other chronic cardiopulmonary diseases and is often associated with anxiety and depression. The present study examined the effect of visually-induced mood shifts on exertional dyspnoea in individuals with COPD. METHODS: Following familiarization, 20 participants with mild to severe COPD (age 57-79 years) attended three experimental sessions on separate days, performing two 5-min treadmill exercise tests separated by a 30-min interval on each day. During each exercise test, participants viewed either a positive, negative or neutral set of images sourced from the International Affective Picture System (IAPS) and rated dyspnoea or leg fatigue (0-10). Heart rate (HR) and peripheral oxygen saturation (SpO2 ) were measured at 1-min intervals during each test. Mood valence ratings were obtained using Self-Assessment Manikin (SAM) scale (1-9). RESULTS: Mood valence ratings were significantly higher when viewing positive (end-exercise mean ± SEM = 7.6 ± 0.3) compared to negative IAPS images (2.4 ± 0.3, p < 0.001). Dyspnoea intensity (mean ± SEM = 5.8 ± 0.4) and dyspnoea unpleasantness (5.6 ± 0.3) when viewing negative images were significantly higher compared to positive images (4.2 ± 0.4, p = 0.004 and 3.4 ± 0.5, p = 0.003). Eighty-five percent of participants (n = 17) met the minimal clinically important difference (MCID) criteria for both dyspnoea intensity and unpleasantness. HR, SpO2 and leg fatigue did not differ significantly between conditions. CONCLUSION: These findings indicate that the negative affective state worsens dyspnoea in COPD, thereby suggesting strategies aimed at reducing the likelihood of negative mood or improving the mood may be effective in managing morbidity associated with dyspnoea in COPD.

A look through dark-colored glasses: The Dark Tetrad and affective processing.

The Dark Tetrad personality traits (i.e., narcissism, Machiavellianism, psychopathy, and sadism) have been continuously linked to various deficits in affective reactivity. The aim of this study was to examine the relationship of the Dark Tetrad and processing of emotional pictures. A total of 144 participants (56.9% female, Mage = 22.18, SDage = 2.26) completed measures of the Dark Tetrad, and rated pictures selected form the International Affective Picture System (IAPS), classified in accordance with their norms into highly arousing positive and negative. Affective processing measures included participants' valence and arousal ratings, while cognitive processing was measured my means of the response latency for each response. The results showed that the dark traits were related only to valence, but not arousal ratings. Higher narcissism and lower sadism were associated with more positive valence ratings of positive pictures, and higher sadism was associated with more positive ratings of negative pictures. Moreover, higher Machiavellianism predicted faster assessment of valence and arousal of both positive and negative pictures, and higher sadism predicted slower assessment of negative pictures' valence. Obtained results indicate that deficiencies in affective processing are more pronounced in sadism compared to other dark traits, while Machiavellianism is associated with advantages in cognitive processing, highlighting their significance and uniqueness in the Dark Tetrad constellation.

Dissociating representations of affect and motion in visual cortices.

While a delicious dessert being presented to us may elicit strong feelings of happiness and excitement, the same treat falling slowly away can lead to sadness and disappointment. Our emotional response to the item depends on its visual motion direction. Despite this importance, it remains unclear whether (and how) cortical areas devoted to decoding motion direction represents or integrates emotion with perceived motion direction. Motion-selective visual area V5/MT+ sits, both functionally and anatomically, at the nexus of dorsal and ventral visual streams. These pathways, however, differ in how they are modulated by emotional cues. The current study was designed to disentangle how emotion and motion perception interact, as well as use emotion-dependent modulation of visual cortices to understand the relation of V5/MT+ to canonical processing streams. During functional magnetic resonance imaging (fMRI), approaching, receding, or static motion after-effects (MAEs) were induced on stationary positive, negative, and neutral stimuli. An independent localizer scan was conducted to identify the visual-motion area V5/MT+. Through univariate and multivariate analyses, we demonstrated that emotion representations in V5/MT+ share a more similar response profile to that observed in ventral visual than dorsal, visual structures. Specifically, V5/MT+ and ventral structures were sensitive to the emotional content of visual stimuli, whereas dorsal visual structures were not. Overall, this work highlights the critical role of V5/MT+ in the representation and processing of visually acquired emotional content. It further suggests a role for this region in utilizing affectively salient visual information to augment motion perception of biologically relevant stimuli.

Inhibitor of apoptosis proteins antagonist SM164 ameliorates experimental MPO-ANCA-associated vasculitis via enhancing fatty acid oxidation in neutrophils.

OBJECTIVES: Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a group of life-threatening autoimmune diseases. Inhibitors of apoptosis proteins (IAPs) are a class of molecules engaged in cell death and inflammation, interventions of which are proven effective in a number of inflammatory diseases. Here we tested whether targeting IAPs could ameliorate AAV and explored the potential mechanism. METHODS: We collected 19 kidney specimens from patients with myeloperoxidase (MPO)-AAV to investigate the expression of IAPs. The IAP pan-inhibitor SM164 was used to treat the experimental autoimmune vasculitis (EAV) rat model of AAV. RNA sequencing of renal cortex and enrichment analysis were developed to interpret gene expression. Functional experiments were performed to investigate the role of SM164 on neutrophils and endothelial cells. RESULTS: The expression of three IAPs (cIAP1, cIAP2 and XIAP) was upregulated in kidneys of AAV patients compared with normal controls. SM164 dramatically reduced renal injury in EAV rats. Transcriptomic analysis revealed prominent alterations in fatty acid oxidation and respiratory burst following SM164 treatment. Functional studies demonstrated that SM164 inhibited neutrophil activation induced by MPO-ANCA positive IgG or serum from MPO-AAV patients, and such inhibitory effect was abolished by gene silencing or pharmacological inhibition of fatty acid oxidation. SM164 also inhibited the adhesion of neutrophils to endothelial cells with little effect on the endothelial injury induced by serum from MPO-AAV patients. CONCLUSION: Inhibition of IAPs with SM164 played a protective role in AAV through enhancing intracellular fatty acid oxidation in neutrophils.

Electrophysiological normative responses to emotional, neutral, and cigarette-related images.

To create reproducible emotional probes, affective scientists rely on sets of standardized pictures that are normed using subjective ratings of valence and emotional arousal. However, when emotional responses are investigated using neurophysiological measures, it might be more appropriate to select pictures integrating information from normative subjective reports and normative neurophysiological responses. Here, we provide electrophysiological normative responses for 323 emotional pictures (215 from the IAPS) covering a wide range of categories (erotica, romantic, appetizing foods, landscapes, people engaged in mundane activities, household objects, disgusting objects, accidents, sad people, violence, mutilations, and cigarette-related contents). Event-related potentials (ERPs) and subjective ratings of pleasure and emotional arousal were collected from 763 individuals (52% females, 41% white) aged between 18 and 65 (mean = 43). For each image, the mean amplitude of the late positive potential (LPP, an electrophysiological index of motivational relevance) and the mean subjective ratings of valence and arousal were calculated. We validated our procedure by showing that the subjective ratings of valence and arousal from this sample were highly correlated to the IAPS' published norms (Pearson r = .97 for pleasure and r = .82 for emotional arousal). LPP responses and subjective ratings of emotional arousal also were correlated (Pearson r = .61), but some categories reported being significantly more arousing than neutral (i.e., food, landscapes, and unpleasant objects) did not evoke LPPs significantly different from those evoked by neutral pictures. Researchers interested in probing the brain's affective systems can use these electrophysiological normative responses to create emotional probes that evoke reliable neuroaffective responses.

Evolutionary novelty in the apoptotic pathway of aphids.

Apoptosis, a conserved form of programmed cell death, shows interspecies differences that may reflect evolutionary diversification and adaptation, a notion that remains largely untested. Among insects, the most speciose animal group, the apoptotic pathway has only been fully characterized in Drosophila melanogaster, and apoptosis-related proteins have been studied in a few other dipteran and lepidopteran species. Here, we studied the apoptotic pathway in the aphid Acyrthosiphon pisum, an insect pest belonging to the Hemiptera, an earlier-diverging and distantly related order. We combined phylogenetic analyses and conserved domain identification to annotate the apoptotic pathway in A. pisum and found low caspase diversity and a large expansion of its inhibitory part, with 28 inhibitors of apoptosis (IAPs). We analyzed the spatiotemporal expression of a selected set of pea aphid IAPs and showed that they are differentially expressed in different life stages and tissues, suggesting functional diversification. Five IAPs are specifically induced in bacteriocytes, the specialized cells housing symbiotic bacteria, during their cell death. We demonstrated the antiapoptotic role of these five IAPs using heterologous expression in a tractable in vivo model, the Drosophila melanogaster developing eye. Interestingly, IAPs with the strongest antiapoptotic potential contain two BIR and two RING domains, a domain association that has not been observed in any other species. We finally analyzed all available aphid genomes and found that they all show large IAP expansion, with new combinations of protein domains, suggestive of evolutionarily novel aphid-specific functions.

A Systematic Review of International Affective Picture System (IAPS) around the World.

Standardized Emotion Elicitation Databases (SEEDs) allow studying emotions in laboratory settings by replicating real-life emotions in a controlled environment. The International Affective Pictures System (IAPS), containing 1182 coloured images as stimuli, is arguably the most popular SEED. Since its introduction, multiple countries and cultures have validated this SEED, making its adoption on the study of emotion a worldwide success. For this review, 69 studies were included. Results focus on the discussion of validation processes by combining self-report and physiological data (Skin Conductance Level, Heart Rate Variability and Electroencephalography) and self-report only. Cross-age, cross-cultural and sex differences are discussed. Overall, IAPS is a robust instrument for emotion elicitation around the world.

Influence of Perinatal Factors on Gene Expression of IAPs Family and Main Factors of Pluripotency: OCT4 and SOX2 in Human Breast Milk Stem Cells-A Preliminary Report.

Due to their therapeutic potential, mesenchymal stem cells are the subject of intensive research on the use of their potential in the treatment of, among others, neurodegenerative diseases or immunological diseases. They are among the newest in the field of medicine. The presented study aimed to evaluate the expression of eight genes from the IAP family and the gene regulating IAP-XAF1-in stem cells derived from human milk, using the qPCR method. The relationships between the expression of genes under study and clinical data, such as maternal age, maternal BMI, week of pregnancy in which the delivery took place, bodyweight of the newborn, the number of pregnancies and deliveries, and the time elapsed since delivery, were also analyzed. The research was carried out on samples of human milk collected from 42 patients hospitalized in The Clinic of Obstetrics and Perinatology of the Independent Public Clinical Hospital No. 4, in Lublin. The conducted research confirmed the expression of the following genes in the tested material: NAIP, BIRC2, BIRC3, BIRC5, BIRC6, BIRC8, XIAP, XAF1, OCT4 and SOX2. Moreover, several dependencies of the expression of individual genes on the maternal BMI (BIRC5, XAF1 and NAIP), the time since childbirth (BIRC5, BIRC6, XAF1 and NAIP), the number of pregnancies and deliveries (BIRC2, BIRC5, BIRC6 and XAF1), the manner of delivery (XAF1 and OCT4), preterm labor (BIRC6 and NAIP) were demonstrated. Additionally, we found positive relationships between gene expression of BIRC7, BIRC8 and XAF1 and the main factors of pluripotency: SOX2 and OCT4. This work is the first to investigate the expression of genes from the IAPs family in mother's milk stem cells.

Decoding the temporal dynamics of affective scene processing.

Natural images containing affective scenes are used extensively to investigate the neural mechanisms of visual emotion processing. Functional fMRI studies have shown that these images activate a large-scale distributed brain network that encompasses areas in visual, temporal, and frontal cortices. The underlying spatial and temporal dynamics, however, remain to be better characterized. We recorded simultaneous EEG-fMRI data while participants passively viewed affective images from the International Affective Picture System (IAPS). Applying multivariate pattern analysis to decode EEG data, and representational similarity analysis to fuse EEG data with simultaneously recorded fMRI data, we found that: (1) ∼80 ms after picture onset, perceptual processing of complex visual scenes began in early visual cortex, proceeding to ventral visual cortex at ∼100 ms, (2) between ∼200 and ∼300 ms (pleasant pictures: ∼200 ms; unpleasant pictures: ∼260 ms), affect-specific neural representations began to form, supported mainly by areas in occipital and temporal cortices, and (3) affect-specific neural representations were stable, lasting up to ∼2 s, and exhibited temporally generalizable activity patterns. These results suggest that affective scene representations in the brain are formed temporally in a valence-dependent manner and may be sustained by recurrent neural interactions among distributed brain areas.

Inhibiting the inhibitors: Targeting anti-apoptotic proteins in cancer and therapy resistance.

The cytotoxic effect of anti-cancer drugs relies on their ability to induce programmed cell death known as apoptosis. Evading apoptosis is a common characteristic of cancer cells and it is linked to both carcinogenesis and anticancer drug resistance. To escape apoptosis, cancer cells often express high levels of anti-apoptotic proteins and become "addicted "to them for their survival. Consequently, anti-apoptotic proteins have emerged as attractive druggable targets for the development of cancer therapeutics. In this review we focus on two major anti-apoptotic protein families: IAPs (Inhibitor of Apoptosis) proteins and Bcl-2 (B-cell lymphoma-2) family members. We also discuss insights into the regulation of these proteins by natural antagonists, which has provided the conceptual basis for developing novel anti-cancer drugs. Significantly, the pro-apoptotic protein ARTS (apoptosis-related protein in the TGF-ß signaling pathway; Sept4_i2) acts as a dual antagonist of both X-linked inhibitor of apoptosis protein (XIAP) and Bcl-2. Because upregulation of anti-apoptotic proteins in response to cancer therapy contributes to drug resistance, targeted inhibition of these proteins is expected to enhance the efficacy of chemotherapy. Finally, we discuss the role of proteasome-mediated degradation in the regulation of apoptosis, and how this mechanism can be harnessed to develop small molecules that stimulate degradation of anti-apoptotic proteins for cancer therapy. This strategy has the potential to overcome drug resistance more effectively than mere inhibition. Therefore, this approach may allow use of lower drug concentrations and thereby reduce cytotoxicity and untoward side effects.

Chemosensitivity to HM90822, a novel synthetic IAP antagonist, is determined by p-AKT-inducible XIAP phosphorylation in human pancreatic cancer cells.

Inhibitor of apoptosis proteins (IAPs) are overexpressed in the majority of cancers and prevent apoptosis by inhibiting caspases. IAPs have therefore attracted considerable attention as potential targets for anticancer therapy. Here, we demonstrated that HM90822 (abbreviated HM822; a new synthetic IAP antagonist) induced apoptotic cell death via proteasome-dependent degradation of BIR2/3 domain-containing IAPs in human pancreatic cancer cells. HM822 inhibited the expression of XIAP and cIAP1/2 proteins in Panc-1 and BxPC-3 cells, which are sensitive to HM822. HM822 also induced IAP ubiquitination and promoted proteasome-dependent IAP degradation. However, cells expressing phospho-XIAP (Ser87) and AKT exhibited resistance to HM822. In other words, the overexpression of AKT-CA (constitutive active form for AKT) or AKT-WT induced resistance to HM822. In addition, in Panc-1 xenograft and orthotopic mouse models, we revealed that tumor growth was suppressed by the administration of HM822. Taken together, these results suggest that HM822 induces apoptosis through ubiquitin/proteasome-dependent degradation of BIR3 domain-containing IAPs. These findings suggest that phospho-XIAP and phospho-AKT may be used as biomarkers for predicting the efficacy of HM822 in pancreatic cancer patients.

Deactivation of default-mode network and early suppression of decision-making areas during retrieval period by high-arousing emotions improves performance in verbal working memory task.

Emotions affect many aspects of cognition (attention, decision-making, problem solving, conflict resolution, task switching, social cognition, etc.), but the cortical areas or networks through which these effects are achieved are still debatable. In the present study, the effect of emotion on cognition was studied in healthy young individuals (n = 56). Emotions were induced using high-arousing negative, positive, and low-arousing neutral pictures from the International Affective Picture System (IAPS). Sternberg's verbal working memory task was administered at baseline and after each emotion exposure, while high-density EEG was recorded. Cortical sources were calculated using sLORETA in the 500-ms window (for every 100 ms bin) before the response and were compared with baseline. Though the number of correct responses were comparable, reaction times after emotion exposure reduced significantly. Source analysis revealed significant deactivation of default mode network (DMN) areas as well as early deactivation of decision-making areas during Sternberg's task performed after both the negative and positive emotions. This early deactivation, much before the response was made, when compared with baseline suggests that tasks performed under high-arousing emotional states may help in making decisions earlier or faster. We conclude that the exposure to high-arousing emotional stimuli improves verbal working memory by helping in directing the attentional resources toward the task, thus decreasing the decision-making time and further suppressing the DMN areas.

Emotional visual stimuli affect the evaluation of tactile stimuli presented on the arms but not the related electrodermal responses.

Considering the wealth of recent studies on affective touch, to date, little research addressed the role of the other sensory modalities in the modulation of hedonic tactile perception. Here, we investigated the behavioral and electrodermal signature of the interaction between simultaneously presented visual and tactile stimuli. In three experiments, participants were presented with emotional pictures (international affective picture system; IAPS), while their forearm was gently stroked by means of different tactile textures (i.e., sandpaper, satin, tinfoil, abrasive sponge, and skin-to-skin contact). In Experiment 1, the participants evaluated the pleasantness of the tactile stimulation received, while in Experiment 2 they evaluated the pictures emotional valence. In Experiment 3 the participants rated the pleasantness, the smoothness and the softness of the textures; skin conductance responses (SCRs) were also measured. In sum, the results revealed that while the visual valence ratings were not modulated by the tactile stimulation, the hedonic and sensory tactile ratings were modulated by the visual presentation of both positively and negatively valenced pictures, as well as by neutral pictures. The modulatory effects occurring during visuo-tactile interactions might thus be not necessarily reciprocal. Moreover, the SCRs were not differently affected by the visuo-tactile or tactile conditions of stimulus presentation, suggesting a dissociation between behavioral and electrodermal effects in multisensory interactions.

Reprogramming Tumor Associated Macrophage Phenotype by a Polysaccharide from Ilex asprella for Sarcoma Immunotherapy.

We report here the discovery of an acidic polysaccharide, namely IAPS-2, from the root of Ilex asprella, with anti-tumor activity via a repolarizing tumor associated macrophages (TAMs) phenotype. We obtained IAPS-2 polysaccharide from this herb based on acidity and found that IAPS-2 expressed the activity of promoting the secretion of anti-tumor cytokines in macrophages. Furthermore, we evaluated its anti-tumor effect on TAM cells, through the activation of nuclear factor-κB (NF-κB) and signal transducer and activator of transcription (STAT) signaling. In particular, in the tumor murine model, IAPS-2 demonstrated that it could significantly inhibit the growth of tumors via modulating the function of TAMs and increase the animal survival rate. In summary, IAPS-2, with a clearly illustrated chemical composition, potent anti-tumor activity, and a solid mechanism of action, may be developed into a valuable therapeutic tool for cancer immunotherapy.

IAPs and cell migration.

Inhibitors of apoptosis (IAPs) constitute a family of cell signaling regulators controlling several fundamental biological processes such as innate immunity, inflammation, cell death, cell proliferation, and cell differentiation. Increasing evidence from in vivo and in vitro studies indicate a function for IAPs in the modulation of invasive and migratory properties of cells. Here, we present and discuss the mechanisms whereby IAPs can control cell migration.

Divergent influences of anterior cingulate cortex GABA concentrations on the emotion circuitry.

Neuroimaging research has revealed that emotion processing recruits a widespread neural network including the dorsal anterior cingulate cortex (dACC), hippocampus, and amygdala. Recent studies have started to investigate the role of the primary inhibitory neurotransmitter γ-aminobutyric acid (GABA) on brain function, but little is known about the influences of GABA on this emotion circuitry. Using magnetic resonance spectroscopy, we investigated the role of GABA levels in the dACC on emotion processing by presenting emotional and neutral pictures to 68 healthy male participants during functional magnetic resonance imaging. Results revealed opposing associations of dACC GABA levels and neural activity. GABA levels were positively correlated with blood oxygen level dependent (BOLD) responses to emotional stimuli in the amygdala and to emotional and neutral stimuli in the hippocampus. In contrast, GABA levels were negatively correlated with BOLD responses for the comparison between positive and negative stimuli in the dACC. Our results suggest positive influences of dACC GABA on BOLD responses in the hippocampus and amygdala, and negative influences on BOLD responses in the dACC that are dependent on emotional valence.

Cell death: From initial concepts to pathways to clinical applications - Personal reflections of a clinical researcher.

Cell death research has come a long way from the description of cellular phenomena to identification of pathways to the role of apoptosis in tissue homeostasis in the pathophysiology of diseases and the delineation of targets for therapeutic intervention. In this very personal view of a clinical researcher in the field from the very beginning to recent clinical application, essential elements of cell death research over the past decades are seen from a perspective of development of science and medicine.

Machiavellian emotion regulation in a cognitive reappraisal task: An fMRI study.

Affective coldness is one of the main features of Machiavellianism. Recent studies have revealed that Machiavellians are emotionally detached and that this "affective blunting" is associated with intense feelings, emotional instability, negative emotions, and difficulty in enduring distress. We used brain-imaging techniques to investigate emotion regulation in Machiavellianism at a neuropsychological level. We used situations in which participants were required to demonstrate emotional flexibility to explore the controversy surrounding the fact that Machiavellianism is associated with both cold-mindedness and emotional instability. Participants performed a reappraisal task in which emotionally evocative pictures (from the International Affective Picture System) were presented in different contexts (negative, positive, and neutral). They were asked to interpret a scenario according to its title and to reinterpret it according to another context created by a new title (e.g., negatively labeled pictures shifted to positively labeled ones). During task performance, Machiavellians showed increased activation of brain regions associated with emotion generation-for example, the amygdala and insula. This indicates that Machiavellian individuals are able to be involved emotionally in social situations. Increased activation in the temporal and parahippocampal regions during reappraisal suggests that Machiavellians use semantic-perceptual processes to construct alternative interpretations of the same situation and have enhanced memory for emotional stimuli. Furthermore, they seem to possess an intense awareness that leads them to shift attention from external to internal information to detect environmental changes. These cognitive processes may enable them to adjust their behavior quickly. This study supports the flexibility hypothesis of Machiavellianism and suggests that Machiavellians' approach to emotion regulation is linked to their rational mode of thinking.