RESUMO
Adapting to particular features of a haptic shape, for example, the slant of a surface, affects how a subsequently touched shape is perceived (aftereffect). Previous studies showed that this adaptation is largely based on our proprioceptive sense of hand posture, yet the influence of vision on haptic shape adaptation has been relatively unexplored. Here, using a slant-adaptation paradigm, we investigated whether visual information affects haptic adaptation and, if so, how. To this end, we varied the available visual cues during the adaptation period. This process ranged from providing visual information only about the slant of the surface, or the reference frame in which it is presented, to only providing visual information about the location of the fingertips. Additionally, we tested several combinations of these visual cues. We show that, as soon as the visual information can be used as a spatial reference to link the own fingertip position to the surface slant, haptic adaptation is very much reduced. This result means that, under these viewing conditions, vision dominates touch and is one reason why we do not easily adapt to haptic shape in our daily life, because we usually have visual information about both hand and object available simultaneously.
Assuntos
Tecnologia Háptica , Percepção do Tato , Humanos , Mãos , Sinais (Psicologia) , PosturaRESUMO
Inclusive design does not stop at removing physical obstacles such as staircases. It also involves identifying architectural features that impose sensory burdens, such as repetitive visual patterns that are known to potentially cause dizziness or visual discomfort. In order to assess their influence on human gait and its stability, three repetitive patterns-random dots, repetitive stripes, and repetitive waves (Lisbon pattern)-were displayed in a coloured and greyscale variant in a virtual reality (VR) environment. The movements of eight participants were recorded using a motion capture system and electromyography (EMG). During all test conditions, a significant increase in the muscular activity of leg flexor muscles was identified just before touchdown. Further, an increase in the activity of laterally stabilising muscles during the swing phase was observed for all of the test conditions. The lateral and vertical centre of mass (CoM) deviation was statistically evaluated using a linear mixed model (LMM). The patterns did cause a significant increase in the CoM excursion in the vertical direction but not in the lateral direction. These findings are indicative of an inhibited and more cautious gait style and a change in control strategy. Furthermore, we quantified the induced discomfort by using both algorithmic estimates and self-reports. The Fourier-based methods favoured the greyscaled random dots over repetitive stripes. The colour metric favoured the striped pattern over the random dots. The participants reported that the wavey Lisbon pattern was the most disruptive. For architectural and structural design, this study indicates (1) that highly repetitive patterns should be used with care in consideration of their impact on the human visuomotor system and its behavioural effects and (2) that coloured patterns should be used with greater caution than greyscale patterns.
Assuntos
Eletromiografia , Marcha , Realidade Virtual , Caminhada , Humanos , Caminhada/fisiologia , Masculino , Marcha/fisiologia , Feminino , Adulto , Músculo Esquelético/fisiologia , Adulto Jovem , Fenômenos Biomecânicos/fisiologia , AlgoritmosRESUMO
The coherent experience of the self and the world depends on the ability to integrate vs. segregate sensory information. Optimal temporal integration between the senses is mediated by oscillatory properties of neural activity. Previous research showed reduced temporal sensitivity to multisensory events in schizotypy, a personality trait linked to schizophrenia. Here we used the tactile-induced Double-Flash-Illusion (tDFI) to investigate the tactile-to-visual temporal sensitivity in schizotypy, as indexed by the temporal window of illusion (TWI) and its neural underpinnings. We measured EEG oscillations within the beta band, recently shown to correlate with the tDFI. We found individuals with higher schizotypal traits to have wider TWI and slower beta waves accounting for the temporal window within which they perceive the illusion. Our results indicate reduced tactile-to-visual temporal sensitivity to mediate the effect of slowed oscillatory beta activity on schizotypal personality traits. We conclude that slowed oscillatory patterns might constitute an early marker for psychosis proneness.
Assuntos
Ilusões , Esquizofrenia , Transtorno da Personalidade Esquizotípica , Percepção Auditiva , Humanos , Personalidade , TatoRESUMO
It is well established that both an increase in reactive oxygen species (ROS: i.e. O2â¢-, H2O2 and OHâ¢), as well as protein aggregation, accompany ageing and proteinopathies such as Parkinson's and Alzheimer's disease. However, it is far from clear whether there is a causal relation between the two. This review describes how protein aggregation can be affected both by redox signalling (downstream of H2O2), as well as by ROS-induced damage, and aims to give an overview of the current knowledge of how redox signalling affects protein aggregation and vice versa. Redox signalling has been shown to play roles in almost every step of protein aggregation and amyloid formation, from aggregation initiation to the rapid oligomerization of large amyloids, which tend to be less toxic than oligomeric prefibrillar aggregates. We explore the hypothesis that age-associated elevated ROS production could be part of a redox signalling-dependent-stress response in an attempt to curb protein aggregation and minimize toxicity.
Assuntos
Oxirredução , Agregados Proteicos , Transdução de Sinais , Envelhecimento , Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/química , Esclerose Lateral Amiotrófica/metabolismo , Animais , Humanos , Peróxido de Hidrogênio/química , Camundongos , Oxigênio/química , Doença de Parkinson/metabolismo , Ligação Proteica , Dobramento de Proteína , Proteostase , Espécies Reativas de OxigênioRESUMO
Humans can learn and store multiple visuomotor mappings (dual-adaptation) when feedback for each is provided alternately. Moreover, learned context cues associated with each mapping can be used to switch between the stored mappings. However, little is known about the associative learning between cue and required visuomotor mapping, and how learning generalises to novel but similar conditions. To investigate these questions, participants performed a rapid target-pointing task while we manipulated the offset between visual feedback and movement end-points. The visual feedback was presented with horizontal offsets of different amounts, dependent on the targets shape. Participants thus needed to use different visuomotor mappings between target location and required motor response depending on the target shape in order to "hit" it. The target shapes were taken from a continuous set of shapes, morphed between spiky and circular shapes. After training we tested participants performance, without feedback, on different target shapes that had not been learned previously. We compared two hypotheses. First, we hypothesised that participants could (explicitly) extract the linear relationship between target shape and visuomotor mapping and generalise accordingly. Second, using previous findings of visuomotor learning, we developed a (implicit) Bayesian learning model that predicts generalisation that is more consistent with categorisation (i.e. use one mapping or the other). The experimental results show that, although learning the associations requires explicit awareness of the cues' role, participants apply the mapping corresponding to the trained shape that is most similar to the current one, consistent with the Bayesian learning model. Furthermore, the Bayesian learning model predicts that learning should slow down with increased numbers of training pairs, which was confirmed by the present results. In short, we found a good correspondence between the Bayesian learning model and the empirical results indicating that this model poses a possible mechanism for simultaneously learning multiple visuomotor mappings.
Assuntos
Retroalimentação Sensorial/fisiologia , Modelos Neurológicos , Desempenho Psicomotor/fisiologia , Algoritmos , Teorema de Bayes , Biologia Computacional , HumanosRESUMO
When repeatedly switching between two visuomotor mappings, e.g. in a reaching or pointing task, adaptation tends to speed up over time. That is, when the error in the feedback corresponds to a mapping switch, fast adaptation occurs. Yet, what is learned, the relative error or the absolute mappings? When switching between mappings, errors with a size corresponding to the relative difference between the mappings will occur more often than other large errors. Thus, we could learn to correct more for errors with this familiar size (Error Learning). On the other hand, it has been shown that the human visuomotor system can store several absolute visuomotor mappings (Mapping Learning) and can use associated contextual cues to retrieve them. Thus, when contextual information is present, no error feedback is needed to switch between mappings. Using a rapid pointing task, we investigated how these two types of learning may each contribute when repeatedly switching between mappings in the absence of task-irrelevant contextual cues. After training, we examined how participants changed their behaviour when a single error probe indicated either the often-experienced error (Error Learning) or one of the previously experienced absolute mappings (Mapping Learning). Results were consistent with Mapping Learning despite the relative nature of the error information in the feedback. This shows that errors in the feedback can have a double role in visuomotor behaviour: they drive the general adaptation process by making corrections possible on subsequent movements, as well as serve as contextual cues that can signal a learned absolute mapping.
Assuntos
Adaptação Fisiológica/fisiologia , Função Executiva/fisiologia , Retroalimentação Sensorial/fisiologia , Aprendizagem/fisiologia , Desempenho Psicomotor/fisiologia , Adulto , Sinais (Psicologia) , Humanos , Adulto JovemRESUMO
In case of delayed visual feedback during visuomotor tasks, like in some sluggish computer games, humans can modulate their behavior to compensate for the delay. However, opinions on the nature of this compensation diverge. Some studies suggest that humans adapt to feedback delays with lasting changes in motor behavior (aftereffects) and a recalibration of time perception. Other studies have shown little or no evidence for such semipermanent recalibration in the temporal domain. We hypothesize that predictability of the reference signal (target to be tracked) is necessary for semipermanent delay adaptation. To test this hypothesis, we trained participants with a 200 ms visual feedback delay in a visually guided manual tracking task, varying the predictability of the reference signal between conditions, but keeping reference motion and feedback delay constant. In Experiment 1, we focused on motor behavior. Only training in the predictable condition brings about all of the adaptive changes and aftereffects expected from delay adaptation. In Experiment 2, we used a synchronization task to investigate perceived simultaneity (perceptuomotor learning). Supporting the hypothesis, participants recalibrated subjective visuomotor simultaneity only when trained in the predictable condition. Such a shift in perceived simultaneity was also observed in Experiment 3, using an interval estimation task. These results show that delay adaptation in motor control can modulate the perceived temporal alignment of vision and kinesthetically sensed movement. The coadaptation of motor prediction and target prediction (reference extrapolation) seems necessary for such genuine delay adaptation. This offers an explanation for divergent results in the literature.
Assuntos
Adaptação Fisiológica/fisiologia , Retroalimentação Sensorial/fisiologia , Desempenho Psicomotor/fisiologia , Adolescente , Adulto , Feminino , Humanos , Masculino , Percepção do Tempo , Visão Ocular/fisiologia , Adulto JovemRESUMO
We experience the world mostly in a multisensory fashion using a combination of all of our senses. Depending on the modality we can select different exploration strategies for extracting perceptual information. For instance, using touch we can enclose an object in our hand to explore parts of the object in parallel. Alternatively, we can trace the object with a single finger to explore its parts in a serial fashion. In this study we investigated whether the exploration mode (parallel vs. serial) affects the way sensory signals are combined. To this end, participants visually and haptically explored surfaces that varied in roll angle and indicated which side of the surface was perceived as higher. In Experiment 1, the exploration mode was the same for both modalities (i.e., both parallel or both serial). In Experiment 2, we introduced a difference in exploration mode between the two modalities (visual exploration was parallel while haptic exploration was serial or vice versa). The results showed that visual and haptic signals were combined in a statistically optimal fashion only when the exploration modes were the same. In case of an asymmetry in the exploration modes across modalities, integration was suboptimal. This indicates that spatial-temporal discrepancies in the acquisition of information in the two senses (i.e., haptic and visual) can lead to the breakdown of sensory integration.
Assuntos
Orientação , Percepção do Tato/fisiologia , Percepção Visual/fisiologia , Retroalimentação , Feminino , Humanos , Funções Verossimilhança , Masculino , Imagens de Fantasmas , Adulto JovemRESUMO
Adaptation to specific visuomotor mappings becomes faster when switching back and forth between them. What is learned when repeatedly switching between the visuomotor mappings: the absolute mappings or the relative shift between the mappings? To test this, we trained participants in a rapid pointing task using a unique color cue as context for each mapping between pointing location and visual feedback. After extensive training, participants adapted to a new mapping using a neutral contextual cue. For catch trials (a change in cue and no visual feedback) different adaptation performances are predicted depending on how the mappings are encoded. When encoding an absolute mapping for each cue, participants would fall back to the mapping associated with the cue irrespective of the state they are currently in. In contrast, when a shift in mapping is encoded for the cue, pointing performance will shift relative to the current mapping by an amount equal to the difference between the previously learned mappings. Results indicate that the contextual cues signal absolute visuomotor mappings rather than relative shifts between mappings.
Assuntos
Adaptação Fisiológica , Sinais (Psicologia) , Retroalimentação Sensorial/fisiologia , Aprendizagem/fisiologia , Memória/fisiologia , Desempenho Psicomotor/fisiologia , Humanos , Estimulação Luminosa/métodos , Tempo de Reação , Adulto JovemRESUMO
A deficient sense of self, typically observed in schizophrenia spectrum disorders, is often accompanied by abnormalities in bodily perception and awareness. These abnormalities are seemingly among the most powerful predictive factors for the onset of schizophrenic illnesses. According to the hypothesis of the psychosis continuum, high schizotypal traits in the general population may be characterized by a progressive sense of detachment from one's lived body. Building upon previous research that found an abnormal Body Structural Representation (BSR) in individuals with schizophrenia, this study aims to extend these findings to schizotypy. To investigate this, we utilized the Finger Localization Task (FLT), in which participants must identify the finger touched by the experimenter, and the In Between Task (IBT), in which two fingers are touched and participants must specify the number of fingers in between the two stimulated fingers. We found that individuals with high schizotypy were significantly less accurate than individuals with low schizotypy in determining the spatial configuration of their own fingers relative to each other. Most significantly, performances on both tasks were negatively correlated with the score on the Dissociative Experiences Scale (DES). These findings support the hypothesis that the progressive loss of one's sense of self is associated with abnormal bodily experiences and dissociative symptomatology which may represent a potential marker for schizophrenia proneness.
Assuntos
Transtornos Psicóticos , Esquizofrenia , Transtorno da Personalidade Esquizotípica , Imagem Corporal , Transtornos Dissociativos , HumanosRESUMO
Redox signaling is controlled by the reversible oxidation of cysteine thiols, a post-translational modification triggered by H2O2 acting as a second messenger. However, H2O2 actually reacts poorly with most cysteine thiols and it is not clear how H2O2 discriminates between cysteines to trigger appropriate signaling cascades in the presence of dedicated H2O2 scavengers like peroxiredoxins (PRDXs). It was recently suggested that peroxiredoxins act as peroxidases and facilitate H2O2-dependent oxidation of redox-regulated proteins via disulfide exchange reactions. It is unknown how the peroxiredoxin-based relay model achieves the selective substrate targeting required for adequate cellular signaling. Using a systematic mass-spectrometry-based approach to identify cysteine-dependent interactors of the five human 2-Cys peroxiredoxins, we show that all five human 2-Cys peroxiredoxins can form disulfide-dependent heterodimers with a large set of proteins. Each isoform displays a preference for a subset of disulfide-dependent binding partners, and we explore isoform-specific properties that might underlie this preference. We provide evidence that peroxiredoxin-based redox relays can proceed via two distinct molecular mechanisms. Altogether, our results support the theory that peroxiredoxins could play a role in providing not only reactivity but also selectivity in the transduction of peroxide signals to generate complex cellular signaling responses.
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The perception of a bistable stimulus as one or the other interpretation can be biased by prior presentations of that stimulus. Such learning effects have been found to be long lasting even after small amounts of training. The effectiveness of training may be influenced by preexposure to the ambiguous stimulus. Here we investigate the role of preexposure for learning a position-dependent perceptual bias. We used rotating Necker Cubes as the bistable stimuli, which were presented at two locations: above or below fixation. On training trials, additional depth cues disambiguated the rotation direction contingent on the location. On test trials, the rotating cube was presented without disambiguation cues. Without preexposure to the ambiguous stimulus, subjects learned to perceive the cube to be rotating in the trained direction for both locations. However, subjects that were preexposed to the ambiguous stimulus did not learn the trained percept-location contingency, even though the preexposure was very short compared to the subsequent training. Preexposure to the disambiguated stimulus did not interfere with learning. This indicates a fundamental difference between ambiguous test and disambiguated training trials for learning a perceptual bias. In short, small variations in paradigm can have huge effects for the learning of perceptual biases for ambiguous stimuli.
Assuntos
Sinais (Psicologia) , Aprendizagem/fisiologia , Percepção Visual/fisiologia , Humanos , Memória/fisiologia , Estimulação Luminosa/métodosRESUMO
In our daily life, we often interact with objects using both hands raising the question the question to what extent information between the hands is shared. It has, for instance, been shown that curvature adaptation aftereffects can transfer from the adapted hand to the non-adapted hand. However, this transfer only occurred for dynamic exploration, e.g. by moving a single finger over a surface, but not for static exploration when keeping static contact with the surface and combining the information from different parts of the hand. This raises the question to what extent adaptation to object shape is shared between the hands when both hands are used in static fashion simultaneously and the object shape estimates require information from both hands. Here we addressed this question in three experiments using a slant adaptation paradigm. In Experiment 1 we investigated whether an aftereffect of static bimanual adaptation occurs at all and whether it transfers to conditions in which one hand was moving. In Experiment 2 participants adapted either to a felt slanted surface or simply be holding their hands in mid-air at similar positions, to investigate to what extent the effects of static bimanual adaptation are posture-based rather than object based. Experiment 3 further explored the idea that bimanual adaptation is largely posture based. We found that bimanual adaptation using static touch did lead to aftereffects when using the same static exploration mode for testing. However, the aftereffect did not transfer to any exploration mode that included a dynamic component. Moreover, we found similar aftereffects both with and without a haptic surface. Thus, we conclude that static bimanual adaptation is of proprioceptive nature and does not occur at the level at which the object is represented.
Assuntos
Mãos/fisiologia , Desempenho Psicomotor , Percepção do Tato/fisiologia , Adaptação Fisiológica , Adulto , Feminino , Humanos , Masculino , Equilíbrio Postural/fisiologia , Tato , Adulto JovemRESUMO
The tumor suppressor p16INK4A induces cell cycle arrest and senescence in response to oncogenic transformation and is therefore frequently lost in cancer. p16INK4A is also known to accumulate under conditions of oxidative stress. Thus, we hypothesized it could potentially be regulated by reversible oxidation of cysteines (redox signaling). Here we report that oxidation of the single cysteine in p16INK4A in human cells occurs under relatively mild oxidizing conditions and leads to disulfide-dependent dimerization. p16INK4A is an all α-helical protein, but we find that upon cysteine-dependent dimerization, p16INK4A undergoes a dramatic structural rearrangement and forms aggregates that have the typical features of amyloid fibrils, including binding of diagnostic dyes, presence of cross-ß sheet structure, and typical dimensions found in electron microscopy. p16INK4A amyloid formation abolishes its function as a Cyclin Dependent Kinase 4/6 inhibitor. Collectively, these observations mechanistically link the cellular redox state to the inactivation of p16INK4A through the formation of amyloid fibrils.
Assuntos
Inibidor p16 de Quinase Dependente de Ciclina/química , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Cisteína/química , Amiloide/química , Ciclo Celular , Senescência Celular , Inibidor p16 de Quinase Dependente de Ciclina/genética , Células HEK293 , Humanos , Modelos Moleculares , Oxirredução , Multimerização Proteica , Estrutura Secundária de ProteínaRESUMO
When interacting with virtual environments, feedback delays between making a movement and seeing the visual consequences of that movement are detrimental for the subjective quality of the VR experience. Here we used standard measures of subjective experiences such as ownership, agency and presence to investigate whether prolonged exposure to the delay, and thus the possibility to adapt to it, leads to the recovery of the qualitative experience of VR. Participants performed a target-tracking task in a Virtual Reality environment. We measured the participants' tracking performance in terms of spatial and temporal errors with respect to the target in both No-Delay and Delay conditions. Additionally, participants rated their sense of "ownership" of holding a virtual tool, agency and presence on each trial using sliding scales. These single trial ratings were compared to the results of the more traditional questionnaires for ownership and agency and presence for both No-Delay and Delay conditions. We found that the participants' sliding scales ratings corresponded very well to the scores obtained from the traditional questionnaires. Moreover, not only did participants behaviourally adapt to the delay, their ratings of ownership and agency significantly improved with prolonged exposure to the delay. Together the results suggest a tight link between the ability to perform a behavioural task and the subjective ratings of ownership and agency in virtual reality.
Assuntos
Retroalimentação , Percepção , Realidade Virtual , Adaptação Psicológica , Adulto , Feminino , Humanos , Cinésica , Masculino , Pesquisa Qualitativa , Autoeficácia , Fatores de Tempo , Adulto JovemRESUMO
We have investigated the role of saccades and fixation positions in two perceptual rivalry paradigms (slant rivalry and Necker cube) and in two binocular rivalry paradigms (grating and house-face rivalry), and we compared results obtained from two different voluntary control conditions (natural viewing and hold percept). We found that for binocular rivalry, rather than for perceptual rivalry, there is a marked positive temporal correlation between saccades and perceptual flips at about the moment of the flip. Across different voluntary control conditions the pattern of temporal correlation did not change (although the amount of correlation did frequently, but not always, change), indicating that subjects do not use different temporal eye movement schemes to exert voluntary control. Analysis of the fixation positions at about the moment of the flips indicates that the fixation position by itself does not determine the percept but that subjects prefer to fixate at different positions when asked to hold either of the different percepts.
Assuntos
Movimentos Sacádicos/fisiologia , Disparidade Visual/fisiologia , Visão Binocular/fisiologia , Percepção de Profundidade/fisiologia , Fixação Ocular/fisiologia , Humanos , Reconhecimento Visual de Modelos/fisiologia , Estimulação Luminosa/métodos , PsicofísicaRESUMO
Particularly promising studies on visual awareness exploit a generally used perceptual bistability phenomenon, "binocular rivalry"--in which the two eyes' images alternately dominate--because it can dissociate the visual input from the perceptual output. To successfully study awareness, it is crucial to know the extent to which eye movements alter the input. Although there is convincing evidence that perceptual alternations can occur without eye movements, the literature on their exact role is mixed. Moreover, recent work has demonstrated that eye movements, first, correlate positively with perceptual alternations in binocular rivalry, and second, often accompany covert attention shifts (that were previously thought to be purely mental). Here, we asked whether eye movements cause perceptual alternations, and if so, whether it is either the execution of the eye movement or the resulting retinal image change that causes the alternation. Subjects viewed repetitive line patterns, enabling a distinction of saccades that did produce foveal image changes from those that did not. Subjects reported binocular rivalry alternations. We found that, although a saccade is not essential to initiate percept changes, the foveal image change resulting from a (micro)saccade is a deciding factor for percept dominance. We conclude that the foveal image must change to have a saccade cause a change in awareness. This sheds new light on the interaction between spatial attention shifts and perceptual alternations.
Assuntos
Conscientização , Retina/fisiologia , Movimentos Sacádicos/fisiologia , Disparidade Visual/fisiologia , Visão Binocular/fisiologia , Percepção Visual/fisiologia , Fixação Ocular/fisiologia , Fóvea Central/fisiologia , HumanosRESUMO
Humans combine redundant multisensory estimates into a coherent multimodal percept. Experiments in cue integration have shown for many modality pairs and perceptual tasks that multisensory information is fused in a statistically optimal manner: observers take the unimodal sensory reliability into consideration when performing perceptual judgments. They combine the senses according to the rules of Maximum Likelihood Estimation to maximize overall perceptual precision. This tutorial explains in an accessible manner how to design optimal cue integration experiments and how to analyse the results from these experiments to test whether humans follow the predictions of the optimal cue integration model. The tutorial is meant for novices in multisensory integration and requires very little training in formal models and psychophysical methods. For each step in the experimental design and analysis, rules of thumb and practical examples are provided. We also publish Matlab code for an example experiment on cue integration and a Matlab toolbox for data analysis that accompanies the tutorial online. This way, readers can learn about the techniques by trying them out themselves. We hope to provide readers with the tools necessary to design their own experiments on optimal cue integration and enable them to take part in explaining when, why and how humans combine multisensory information optimally.
Assuntos
Teorema de Bayes , Encéfalo/fisiologia , Sinais (Psicologia) , Modelos Neurológicos , Percepção/fisiologia , Células Receptoras Sensoriais/fisiologia , Humanos , Funções Verossimilhança , Teoria Psicológica , Projetos de PesquisaRESUMO
Human touch is an inherently active sense: to estimate an object's shape humans often move their hand across its surface. This way the object is sampled both in a serial (sampling different parts of the object across time) and parallel fashion (sampling using different parts of the hand simultaneously). Both the serial (moving a single finger) and parallel (static contact with the entire hand) exploration modes provide reliable and similar global shape information, suggesting the possibility that this information is shared early in the sensory cortex. In contrast, we here show the opposite. Using an adaptation-and-transfer paradigm, a change in haptic perception was induced by slant-adaptation using either the serial or parallel exploration mode. A unified shape-based coding would predict that this would equally affect perception using other exploration modes. However, we found that adaptation-induced perceptual changes did not transfer between exploration modes. Instead, serial and parallel exploration components adapted simultaneously, but to different kinaesthetic aspects of exploration behaviour rather than object-shape per se. These results indicate that a potential combination of information from different exploration modes can only occur at down-stream cortical processing stages, at which adaptation is no longer effective.
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To investigate the underlying causes of chemoresistance in malignant pleural mesothelioma, we have studied mesothelioma cell lines as 3D spheroids, which acquire increased chemoresistance compared to 2D monolayers. We asked whether the gene expression of 3D spheroids would reveal mechanisms of resistance. To address this, we measured gene expression of three mesothelioma cell lines, M28, REN and VAMT, grown as 2D monolayers and 3D spheroids. A total of 209 genes were differentially expressed in common by the three cell lines in 3D (138 upregulated and 71 downregulated), although a clear resistance pathway was not apparent. We then compared the list of 3D genes with two publicly available datasets of gene expression of 56 pleural mesotheliomas compared to normal tissues. Interestingly, only three genes were increased in both 3D spheroids and human tumors: argininosuccinate synthase 1 (ASS1), annexin A4 (ANXA4) and major vault protein (MVP); of these, ASS1 was the only consistently upregulated of the three genes by qRT-PCR. To measure ASS1 protein expression, we stained 2 sets of tissue microarrays (TMA): one with 88 pleural mesothelioma samples and the other with additional 88 pleural mesotheliomas paired with matched normal tissues. Of the 176 tumors represented on the two TMAs, ASS1 was expressed in 87 (50%; staining greater than 1 up to 3+). For the paired samples, ASS1 expression in mesothelioma was significantly greater than in the normal tissues. Reduction of ASS1 expression by siRNA significantly sensitized mesothelioma spheroids to the pro-apoptotic effects of bortezomib and of cisplatin plus pemetrexed. Although mesothelioma is considered by many to be an ASS1-deficient tumor, our results show that ASS1 is elevated at the mRNA and protein levels in mesothelioma 3D spheroids and in human pleural mesotheliomas. We also have uncovered a survival role for ASS1, which may be amenable to targeting to undermine mesothelioma multicellular resistance.