Organoid microphysiological system preserves pancreatic islet function within 3D matrix.

Three-dimensional (3D) multicellular organoids recapitulate the native complexities of human tissue better than traditional cellular monolayers. As organoids are insufficiently supported using standard static culture, microphysiological systems (MPSs) provide a key enabling technology to maintain organoid physiology in vitro. Here, a polydimethylsiloxane-free MPS that enables continuous dynamic culture and serial in situ multiparametric assessments was leveraged to culture organoids, specifically human and rodent pancreatic islets, within a 3D alginate hydrogel. Computational modeling predicted reduced hypoxic stress and improved insulin secretion compared to static culture. Experimental validation via serial, high-content, and noninvasive assessments quantitatively confirmed that the MPS platform retained organoid viability and functionality for at least 10 days, in stark contrast to the acute decline observed overnight under static conditions. Our findings demonstrate the importance of a dynamic in vitro microenvironment for the preservation of primary organoid function and the utility of this MPS for in situ multiparametric assessment.

Platelet adhesion and human umbilical vein endothelial cell cytocompatibility of biodegradable segmented polyurethanes prepared with 4,4'-methylene bis(cyclohexyl isocyanate), poly(caprolactone) diol and butanediol or dithioerythritol as chain extenders.

Biodegradable segmented polyurethanes were prepared with poly(caprolactone) diol as a soft segment, 4,4'-methylene bis(cyclohexyl isocyanate) (HMDI) and either butanediol or dithioerythritol as chain extenders. Platelet adhesion was similar in all segmented polyurethanes studied and not different from Tecoflex® although an early stage of activation was observed on biodegradable segmented polyurethane prepared with dithioerythritol. Relative viability was higher than 80% on human umbilical vein endothelial cells in contact with biodegradable segmented polyurethane extracts after 1, 2 and 7 days. Furthermore, both biodegradable segmented polyurethane materials supported human umbilical vein endothelial cell adhesion, spreading, and viability similar to Tecoflex® medical-grade polyurethane. These biodegradable segmented polyurethanes represent promising materials for cardiovascular applications.

Human fear-related motor neurocircuitry.

Using functional magnetic resonance imaging and an experimental paradigm of instructed fear, we observed a striking pattern of decreased activity in primary motor cortex with increased activity in dorsal basal ganglia during anticipation of aversive electrodermal stimulation in 42 healthy participants. We interpret this pattern of activity in motor neurocircuitry in response to cognitively-induced fear in relation to evolutionarily-conserved responses to threat that may be relevant to understanding normal and pathological fear in humans.

Extending animal models of fear conditioning to humans.

A goal of fear and anxiety research is to understand how to treat the potentially devastating effects of anxiety disorders in humans. Much of this research utilizes classical fear conditioning, a simple paradigm that has been extensively investigated in animals, helping outline a brain circuitry thought to be responsible for the acquisition, expression and extinction of fear. The findings from non-human animal research have more recently been substantiated and extended in humans, using neuropsychological and neuroimaging methodologies. Research across species concur that the neural correlates of fear conditioning include involvement of the amygdala during all stages of fear learning, and prefrontal areas during the extinction phase. This manuscript reviews how animal models of fear are translated to human behavior, and how some fears are more easily acquired in humans (i.e., social-cultural). Finally, using the knowledge provided by a rich animal literature, we attempt to extend these findings to human models targeted to helping facilitate extinction or abolishment of fears, a trademark of anxiety disorders, by discussing efficacy in modulating the brain circuitry involved in fear conditioning via pharmacological treatments or emotion regulation cognitive strategies.

Fear of losing money? Aversive conditioning with secondary reinforcers.

Money is a secondary reinforcer that acquires its value through social communication and interaction. In everyday human behavior and laboratory studies, money has been shown to influence appetitive or reward learning. It is unclear, however, if money has a similar impact on aversive learning. The goal of this study was to investigate the efficacy of money in aversive learning, comparing it with primary reinforcers that are traditionally used in fear conditioning paradigms. A series of experiments were conducted in which participants initially played a gambling game that led to a monetary gain. They were then presented with an aversive conditioning paradigm, with either shock (primary reinforcer) or loss of money (secondary reinforcer) as the unconditioned stimulus. Skin conductance responses and subjective ratings indicated that potential monetary loss modulated the conditioned response. Depending on the presentation context, the secondary reinforcer was as effective as the primary reinforcer during aversive conditioning. These results suggest that stimuli that acquire reinforcing properties through social communication and interaction, such as money, can effectively influence aversive learning.

Perceptions of moral character modulate the neural systems of reward during the trust game.

Studies of reward learning have implicated the striatum as part of a neural circuit that guides and adjusts future behavior on the basis of reward feedback. Here we investigate whether prior social and moral information about potential trading partners affects this neural circuitry. Participants made risky choices about whether to trust hypothetical trading partners after having read vivid descriptions of life events indicating praiseworthy, neutral or suspect moral character. Despite equivalent reinforcement rates for all partners, participants were persistently more likely to make risky choices with the 'good' partner. As expected from previous studies, activation of the caudate nucleus differentiated between positive and negative feedback, but only for the 'neutral' partner. Notably, it did not do so for the 'good' partner and did so only weakly for the 'bad' partner, suggesting that prior social and moral perceptions can diminish reliance on feedback mechanisms in the neural circuitry of trial-and-error reward learning.

An fMRI study of reward-related probability learning.

The human striatum has been implicated in processing reward-related information. More recently, activity in the striatum, particularly the caudate nucleus, has been observed when a contingency between behavior and reward exists, suggesting a role for the caudate in reinforcement-based learning. Using a gambling paradigm, in which affective feedback (reward and punishment) followed simple, random guesses on a trial by trial basis, we sought to investigate the role of the caudate nucleus as reward-related learning progressed. Participants were instructed to make a guess regarding the value of a presented card (if the value of the card was higher or lower than 5). They were told that five different cues would be presented prior to making a guess, and that each cue indicated the probability that the card would be high or low. The goal was to learn the contingencies and maximize the reward attained. Accuracy, as measured by participant's choices, improved throughout the experiment for cues that strongly predicted reward, while no change was observed for unpredictable cues. Event-related fMRI revealed that activity in the caudate nucleus was more robust during the early phases of learning, irrespective of contingencies, suggesting involvement of this region during the initial stages of trial and error learning. Further, the reward feedback signal in the caudate nucleus for well-learned cues decreased as learning progressed, suggesting an evolving adaptation of reward feedback expectancy as a behavior-outcome contingency becomes more predictable.

A double dissociation in the affective modulation of startle in humans: effects of unilateral temporal lobectomy.

In the present study we report a double dissociation between right and left medial temporal lobe damage in the modulation of fear responses to different types of stimuli. We found that right unilateral temporal lobectomy (RTL) patients, in contrast to control subjects and left temporal lobectomy (LTL) patients, failed to show potentiated startle while viewing negative pictures. However, the opposite pattern of impairment was observed during a stimulus that patients had been told signaled the possibility of shock. Control subjects and RTL patients showed potentiated startle while LTL patients failed to show potentiated startle. We hypothesize that the right medial temporal lobe modulates fear responses while viewing emotional pictures, which involves exposure to (emotional) visual information and is consistent with the emotional processing traditionally ascribed to the right hemisphere. In contrast, the left medial temporal lobe modulates fear responses when those responses are the result of a linguistic/cognitive representation acquired through language, which, like other verbally mediated material, generally involves the left hemisphere. Additional evidence from case studies suggests that, within the medial temporal lobe, the amygdala is responsible for this modulation.

Methods for developmental studies of fear conditioning circuitry.

Psychophysiologic studies use air puff as an aversive stimulus to document abnormal fear conditioning in children of parents with anxiety disorders. This study used functional magnetic resonance imaging (fMRI) to examine changes in amygdala activity during air-puff conditioning among adults. Blood oxygen level-dependent (BOLD) signal was monitored in seven adults during 16 alternating presentations of two different colored lights (CS+ vs. CS-), one of which was consistently paired with an aversive air puff. A region-of-interest analysis demonstrated differential change in BOLD signal in the right but not left amygdala across CS+ versus CS- viewing. The amygdala is engaged by pairing of a light with an air puff. Given that prior studies relate air-puff conditioning to risk for anxiety in children, these methods may provide an avenue for directly studying the developmental neurobiology of fear conditioning.

Lesions of the human amygdala impair enhanced perception of emotionally salient events.

Commensurate with the importance of rapidly and efficiently evaluating motivationally significant stimuli, humans are probably endowed with distinct faculties and maintain specialized neural structures to enhance their detection. Here we consider that a critical function of the human amygdala is to enhance the perception of stimuli that have emotional significance. Under conditions of limited attention for normal perceptual awareness-that is, the attentional blink-we show that healthy observers demonstrate robust benefits for the perception of verbal stimuli of aversive content compared with stimuli of neutral content. In contrast, a patient with bilateral amygdala damage has no enhanced perception for such aversive stimulus events. Examination of patients with either left or right amygdala resections shows that the enhanced perception of aversive words depends specifically on the left amygdala. All patients comprehend normally the affective meaning of the stimulus events, despite the lack of evidence for enhanced perceptual encoding of these events in patients with left amygdala lesions. Our results reveal a neural substrate for affective influences on perception, indicating that similar neural mechanisms may underlie the affective modulation of both recollective and perceptual experience.

Functional neuroimaging and episodic memory: a perspective.

One area of research that has significantly benefited from the recent development of functional neuroimaging techniques is the study of memory. In this review we explore what has been learned about the neural basis of normal memory function using these techniques. We focus on episodic memory, which is characterized by the ability to consciously recollect memories for facts or events. We highlight three neuroanatomical regions that have been linked to episodic memory: The hippocampal complex, the prefrontal cortex and the amygdala. For each of these regions, we discuss the behavioral methods of assessment and specific episodic memory processes, particularly encoding and retrieval. Finally, we briefly comment on the potential clinical applications for this research and other memory systems.

Activation of the left amygdala to a cognitive representation of fear.

We examined the neural substrates involved when subjects encountered an event linked verbally, but not experientially, to an aversive outcome. This instructed fear task models a primary way humans learn about the emotional nature of events. Subjects were told that one stimulus (threat) represents an aversive event (a shock may be given), whereas another (safe) represents safety (no shock will be given). Using functional magnetic resonance imaging (fMRI), activation of the left amygdala was observed in response to threat versus safe conditions, which correlated with the expression of the fear response as measured by skin conductance. Additional activation observed in the insular cortex is proposed to be involved in conveying a cortical representation of fear to the amygdala. These results suggest that the neural substrates that support conditioned fear across species have a similar but somewhat different role in more abstract representations of fear in humans.

Performance on indirect measures of race evaluation predicts amygdala activation.

We used fMRI to explore the neural substrates involved in the unconscious evaluation of Black and White social groups. Specifically, we focused on the amygdala, a subcortical structure known to play a role in emotional learning and evaluation. In Experiment 1, White American subjects observed faces of unfamiliar Black and White males. The strength of amygdala activation to Black-versus-White faces was correlated with two indirect (unconscious) measures of race evaluation (Implicit Association Test [IAT] and potentiated startle), but not with the direct (conscious) expression of race attitudes. In Experiment 2, these patterns were not obtained when the stimulus faces belonged to familiar and positively regarded Black and White individuals. Together, these results suggest that amygdala and behavioral responses to Black-versus-White faces in White subjects reflect cultural evaluations of social groups modified by individual experience.

Contribution of the anteromedial temporal lobes to the evaluation of facial emotion.

Amygdala damage can result in impairments in evaluating facial expressions largely specific to fear. In contrast, right-hemisphere cortical lesions result in a more global deficit in facial emotion evaluation. This study addressed these 2 contrasting findings by investigating amygdala and adjacent cortical contributions to the evaluation of facial emotion in 12 patients with right and 11 patients with left unilateral anteromedial temporal lobectomy (RTL and LTL, respectively) and 23 normal controls. RTL but not LTL patients revealed impaired intensity ratings that included but were not exclusive to fear, with the most severe deficits confined to expressions related to affective states of withdrawal-avoidance. This suggests that affective hemispheric specializations in cortical function may extend to subcortical limbic regions. In addition, the right amygdala and adjacent cortex may be part of a neural circuit representing facial expressions of withdrawal.

Expression without recognition: contributions of the human amygdala to emotional communication.

A growing body of evidence from humans and other animals suggests the amygdala may be a critical neural substrate for emotional processing. In particular, recent studies have shown that damage to the human amygdala impairs the normal appraisal of social signals of emotion, primarily those of fear. However, effective social communication depends on both the ability to receive (emotional appraisal) and the ability to send (emotional expression) signals of emotional state. Although the role of the amygdala in the appraisal of emotion is well established, its importance for the production of emotional expressions is unknown. We report a case study of a patient with bilateral amygdaloid damage who, despite a severe deficit in interpreting facial expressions of emotion including fear, exhibits an intact ability to express this and other basic emotions. This dissociation suggests that a single neural module does not support all aspects of the social communication of emotional state.

Recognition of facial emotion in nine individuals with bilateral amygdala damage.

Findings from several case studies have shown that bilateral amygdala damage impairs recognition of emotions in facial expressions, especially fear. However, one study did not find such an impairment, and, in general, comparison across studies has been made difficult because of the different stimuli and tasks employed. In a collaborative study to facilitate such comparisons, we report here the recognition of emotional facial expressions in nine subjects with bilateral amygdala damage, using a sensitive and quantitative assessment. Compared to controls, the subjects as a group were significantly impaired in recognizing fear, although individual performances ranged from severely impaired to essentially normal. Most subjects were impaired on several negative emotions in addition to fear, but no subject was impaired in recognizing happy expressions. An analysis of response consistency showed that impaired recognition of fear could not be attributed simply to mistaking fear for another emotion. While it remains unclear why some subjects with amygdala damage included here are not impaired on our task, the results overall are consistent with the idea that the amygdala plays an important role in triggering knowledge related to threat and danger signaled by facial expressions.

Memory deficits for implicit contextual information in amnesic subjects with hippocampal damage.

The role of the hippocampus and adjacent medial temporal lobe structures in memory systems has long been debated. Here we show in humans that these neural structures are important for encoding implicit contextual information from the environment. We used a contextual cuing task in which repeated visual context facilitates visual search for embedded target objects. An important feature of our task is that memory traces for contextual information were not accessible to conscious awareness, and hence could be classified as implicit. Amnesic patients with medial temporal system damage showed normal implicit perceptual/skill learning but were impaired on implicit contextual learning. Our results demonstrate that the human medial temporal memory system is important for learning contextual information, which requires the binding of multiple cues.

Intact recognition of vocal expressions of fear following bilateral lesions of the human amygdala.

A recent case study found that bilateral damage to the amygdala impairs the normal appraisal of vocal expressions of fear. However, the single source of evidence for this auditory emotion-processing impairment is from a patient with extra-amygdaloid damage that may include the basal ganglia, which have been shown to be important for prosody evaluation. In this study we provide evidence of preserved evaluation of vocal expressions of fear in a female patient (S.P.) with bilateral damage to the amygdala but with intact basal ganglia. This same patient has previously been shown to be impaired in the evaluation of facial expressions, including fear. These results indicate that the analysis of nonverbal signals of fear from different input channels are dissociable, being at least partially dependent on different brain structures. We suggest that the amygdala, in conjunction with the basal ganglia, may support the normal appraisal of auditory signals of danger.