Multiplexed multicolor antiviral assay amenable for high-throughput research.

To curb viral epidemics and pandemics, antiviral drugs are needed with activity against entire genera or families of viruses. Here, we develop a cell-based multiplex antiviral assay for high-throughput screening against multiple viruses at once, as demonstrated by using three distantly related orthoflaviviruses: dengue, Japanese encephalitis and yellow fever virus. Each virus is tagged with a distinct fluorescent protein, enabling individual monitoring in cell culture through high-content imaging. Specific antisera and small-molecule inhibitors are employed to validate that multiplexing approach yields comparable inhibition profiles to single-virus infection assays. To facilitate downstream analysis, a kernel is developed to deconvolute and reduce the multidimensional quantitative data to three cartesian coordinates. The methodology is applicable to viruses from different families as exemplified by co-infections with chikungunya, parainfluenza and Bunyamwera viruses. The multiplex approach is expected to facilitate the discovery of broader-spectrum antivirals, as shown in a pilot screen of approximately 1200 drug-like small-molecules.

Circuit mechanism for suppression of frontal cortical ignition during NREM sleep.

Conscious perception is greatly diminished during sleep, but the underlying circuit mechanism is poorly understood. We show that cortical ignition-a brain process shown to be associated with conscious awareness in humans and non-human primates-is strongly suppressed during non-rapid-eye-movement (NREM) sleep in mice due to reduced cholinergic modulation and rapid inhibition of cortical responses. Brain-wide functional ultrasound imaging and cell-type-specific calcium imaging combined with optogenetics showed that activity propagation from visual to frontal cortex is markedly reduced during NREM sleep due to strong inhibition of frontal pyramidal neurons. Chemogenetic activation and inactivation of basal forebrain cholinergic neurons powerfully increased and decreased visual-to-frontal activity propagation, respectively. Furthermore, although multiple subtypes of dendrite-targeting GABAergic interneurons in the frontal cortex are more active during wakefulness, soma-targeting parvalbumin-expressing interneurons are more active during sleep. Chemogenetic manipulation of parvalbumin interneurons showed that sleep/wake-dependent cortical ignition is strongly modulated by perisomatic inhibition of pyramidal neurons.

A paleo-neurologic investigation of the social brain hypothesis in frontotemporal dementia.

The social brain hypothesis posits that a disproportionate encephalization in primates enabled to adapt behavior to a social context. Also, it has been proposed that phylogenetically recent brain areas are disproportionally affected by neurodegeneration. Using structural and functional magnetic resonance imaging, the present study investigates brain-behavior associations and neural integrity of hyperspecialized and domain-general cortical social brain areas in behavioral variant frontotemporal dementia (bvFTD). The results revealed that both structure and function of hyperspecialized social areas in the middle portion of the superior temporal sulcus (STS) are compromised in bvFTD, while no deterioration was observed in domain general social areas in the posterior STS. While the structural findings adhered to an anterior-posterior gradient, the functional group differences only occurred in the hyperspecialized locations. Activity in specialized regions was associated with structural integrity of the amygdala and with social deficits in bvFTD. In conclusion, the results are in line with the paleo-neurology hypothesis positing that neurodegeneration primarily hits cortical areas showing increased specialization, but also with the compatible alternative explanation that anterior STS regions degenerate earlier, based on stronger connections to and trans-neuronal spreading from regions affected early in bvFTD.

Long term fMRI adaptation depends on adapter response in face-selective cortex.

Repetition suppression (RS) reflects a neural attenuation during repeated stimulation. We used fMRI and the subsequent memory paradigm to test the predictive coding hypothesis for RS during visual memory processing by investigating the interaction between RS and differences due to memory in category-selective cortex (FFA, pSTS, PPA, and RSC). Fifty-six participants encoded face and house stimuli twice, followed by an immediate and delayed (48 h) recognition memory assessment. Linear Mixed Model analyses with repetition, subsequent recognition performance, and their interaction as fixed effects revealed that absolute RS during encoding interacts with probability of future remembrance in face-selective cortex. This effect was not observed for relative RS, i.e. when controlled for adapter-response. The findings also reveal an association between adapter response and RS, both for short and long term (48h) intervals, after controlling for the mathematical dependence between both measures. These combined findings are challenging for predictive coding models of visual memory and are more compatible with adapter-related and familiarity accounts.

A new MRI subject position to explore simultaneous BOLD oscillations of the brain and the body.

BACKGROUND: Anatomically and physiologically, there is strong relationship between the brain and body. A new MRI platform covering both the brain and the limb would be beneficial for a more thorough understanding of the brain-body interactions. NEW METHOD: A new arm-over-head (AOH) position was developed to collect MRI of the brain and one arm simultaneously. Subject's tolerability and SNR of both the brain and limb under a serial of seven different TR (250-3000 ms) were tested. Then, blocked motor imagery tasks were performed to test the possible brain-body oscillations. RESULTS: The new MRI position provided structural images with good quality, and the AOH position had the best SNR under TR 3000 ms (p = 0.03 for the brain; p = 0.064 for the limb). Then, by using both hypothesis-free independent component analysis (ICA) and a priori seed-based functional connectivity (FC) analysis, it is demonstrated during motionless motor imagery tasks there existed possible brain-body BOLD oscillations connecting especially arm flexors to default mode, vision, and sensorimotor networks. The FC appeared at network density as low as 5%. COMPARISON WITH EXISTING METHODS: We have developed a new MRI subject position to explore the possibilities of more extensive neuronal and physiological networks. CONCLUSIONS: The results of this preliminary experiment indicate that functional brain networks might extend outside the brain. A bottom-up circulatory effect might explain this phenomenon. Nonetheless, considering the mechanism of neural top-down control and the nature of complex brain networks, the existence of a more extensive whole-body functional network is rational and possible.

Network level characteristics in the emotion recognition network after unilateral temporal lobe surgery.

The human amygdala is considered a key region for successful emotion recognition. We recently reported that temporal lobe surgery (TLS), including resection of the amygdala, does not affect emotion recognition performance (Journal of Neuroscience, 2018, 38, 9263). In the present study, we investigate the neural basis of this preserved function at the network level. We use generalized psychophysiological interaction and graph theory indices to investigate network level characteristics of the emotion recognition network in TLS patients and healthy controls. Based on conflicting emotion processing theories, we anticipated two possible outcomes: a substantial increase of the non-amygdalar connections of the emotion recognition network to compensate functionally for the loss of the amygdala, in line with basic emotion theory versus only minor changes in network level properties as predicted by psychological construction theory. We defined the emotion recognition network in the total sample and investigated group differences on five network level indices (i.e. characteristic path length, global efficiency, clustering coefficient, local efficiency and small-worldness). The results did not reveal a significant increase in the left or right temporal lobectomy group (compared to the control group) in any of the graph measures, indicating that preserved behavioural emotion recognition in TLS is not associated with a massive connectivity increase between non-amygdalar nodes at network level. We conclude that the emotion recognition network is robust and functionally able to compensate for structural damage without substantial global reorganization, in line with a psychological construction theory.

Preparation of Chitosan Molecularly Imprinted Polymers and the Recognition Mechanism for Adsorption of Alpha-Lipoic Acid.

Two effective molecularly imprinted polymers for the adsorption of alpha-lipoic acid (ALA) were synthesized by the cross-linking of chitosan with epichlorohydrin (ECH) and glutaraldehyde (GLU), respectively, in the presence of ALA as template molecules. Investigations on the molar ratios of ALA and chitosan (-NH2) in the preparation of chitosan molecularly imprinted polymers (MIPs) were carried out with a factor of ALA rebinding capabilities. The surface morphology and chemical properties of the polymers were characterized. The optimized MIPs crosslinked by ECH (MIPs-ECH) and MIPs crosslinked by GLU (MIPs-GLU) had adsorption capabilities of 12.09 mg/g and 19.72 mg/g for ALA, respectively. The adsorption behaviors of two kinds of chitosan MIPs including adsorption kinetics and isotherms were investigated in detail. Adsorption and kinetic binding experiments showed that the prepared MIPs-ECH and MIPs-GLU had selective adsorption and excellent affinity for ALA. In addition, the possible binding models between ALA and chitosan oligosaccharide were predicted by molecular dynamics simulation.

Non-overlapping and Inverse Associations Between the Sexes in Structural Brain-Trait Associations.

Personality reflects the set of psychological traits and mechanisms characteristic for an individual. The brain-trait association between personality and gray matter volume (GMv) has been well studied. However, a recent study has shown that brain structure-personality relationships are highly dependent on sex. In addition, the present study investigates the role of sex on the association between temperaments and regional GMv. Sixty-six participants (33 male) completed the Temperament and Character Inventory (TCI) and underwent structural magnetic resonance brain imaging. Mann-Whitney U tests showed a significant higher score on Novelty Seeking (NS) and Reward Dependence (RD) for females, but no significant group effects were found for Harm Avoidance (HA) and Persistence (P) score. Full factor model analyses were performed to investigate sex-temperament interaction effects on GMv. This revealed increased GMv for females in the superior temporal gyrus when linked to NS, middle temporal gyrus for HA, and the insula for RD. Males displayed increased GMv compared to females relating to P in the posterior cingulate gyrus, the medial superior frontal gyrus, and the middle cingulate gyrus, compared to females. Multiple regression analysis showed clear differences between the brain regions that correlate with female subjects and the brain correlates that correlate with male subjects. No overlap was observed between sex-specific brain-trait associations. These results increase the knowledge of the role of sex on the structural neurobiology of personality and indicate that sex differences reflect structural differences observed in the normal brain. Furthermore, sex hormones seem an important underlying factor for the found sex differences in brain-trait associations. The present study indicates an important role for sex in these brain structure-personality relationships, and implies that sex should not just be added as a covariate of no interest.

Reduced tendency to attribute mental states to abstract shapes in behavioral variant frontotemporal dementia links with cerebellar structural integrity.

Theory of mind (ToM) refers to the ability to attribute mental states to others. Behavioral variant frontotemporal dementia (bvFTD) is a neurodegenerative disorder characterized by profound deficits in social cognition, including ToM. We investigate whether bvFTD affects intention attribution tendency while viewing abstract animations and whether this might represent a primary deficit. A sample of 15 bvFTD patients and 19 matched controls were assessed on cognition and performed an implicit ToM task. They were instructed to describe what they observed in movement patterns displayed by geometrical shapes (triangles). These movement patterns either represented animacy, goal-directed actions or manipulation of mental state (ToM). The responses were scored for both accuracy and intentionality attribution. Using Voxel-Based Morphometry, we investigated the structural neuroanatomy associated with intention attribution tendency. The behavioral results revealed deficits in the bvFTD group on intentionality attribution that were specific for the ToM condition after controlling for global cognitive functioning (MMSE-score), visual attention (TMT B-score), fluid intelligence (RCPMT-score) and confrontation naming (BNT-score). In the bvFTD sample, the intention attribution tendency on the ToM-condition was associated with grey matter volume of a cluster in the cerebellum, spanning the right Crus I, Crus II, VIIIb, IX, left VIIb, IX and vermal IX and X. The results reveal a specific, primary, implicit domain-general ToM deficit in bvFTD that cannot be explained by cognitive dysfunction. Furthermore, the findings point to a contribution of the cerebellum in the social-cognitive phenotype of bvFTD.

Nanoimprinted Anisotropic Topography Preferentially Guides Axons and Enhances Nerve Regeneration.

Surface topography has a profound effect on the development of the nervous system, such as neuronal differentiation and morphogenesis. While the interaction of neurons and the surface topography of their local environment is well characterized, the neuron-topography interaction during the regeneration process remains largely unknown. To address this question, an anisotropic surface topography resembling linear grooves made from poly(ethylene-vinyl acetate) (EVA), a soft and biocompatible polymer, using nanoimprinting, is established. It is found that neurons from both the central and peripheral nervous system can survive and grow on this grooved surface. Additionally, it is observed that axons but not dendrites specifically align with these grooves. Furthermore, it is demonstrated that neurons on the grooved surface are capable of regeneration after an on-site injury. More importantly, these injured neurons have an accelerated and enhanced regeneration. Together, the data demonstrate that this anisotropic topography guides axon growth and improves axon regeneration. This opens up the possibility to study the effect of surface topography on regenerating axons and has the potential to be developed into a medical device for treating peripheral nerve injuries.

Anterior Temporal Lobectomy Impairs Neural Classification of Body Emotions in Right Superior Temporal Sulcus and Reduces Emotional Enhancement in Distributed Brain Areas without Affecting Behavioral Classification.

Humans with amygdalar lesions show proportional reductions of the emotional response to facial expressions in the fusiform face area as well as deficits in emotion recognition from facial expressions. While processing of bodily expressions shares many similarities with facial expressions, there is no substantial evidence that lesions of the amygdala result in similar behavioral and neural sequelae. We combined behavioral assessment with functional neuroimaging in a group of male and female humans with unilateral anterior temporal lobe (ATL) resections, including the amygdala (right: n = 10; left: n = 10) and 12 matched controls. The objective was to assess whether the amygdala is crucial for the recognition of body expressions and for modulatory effects on distant areas during perception of body expressions. The behavioral results revealed normal performance in both patient groups on emotion categorization of body expressions. The neuroimaging results showed that ATL patients displayed no enhanced activations in right fusiform body area and left extrastriate body area and that left ATL patients additionally displayed no enhanced activations in right posterior superior temporal sulcus and right extrastriate body area, respectively. Multivoxel pattern analysis revealed altered categorization capacity between emotional and neutral stimuli in right posterior superior temporal sulcus in right ATL patients. In addition, we also found emotional enhancement in frontal, parietal, occipital, and cingulate regions in controls. Together, our data show that the amygdala and ATLs are not necessary for recognition of dynamic body expressions, but suggest that amygdala lesions affect body emotion processing in distant brain areas.SIGNIFICANCE STATEMENT For humans, information from emotional expressions of others is crucial to support social interactions. The majority of emotion studies has focused on facial expressions; however, in daily life, we also use information from body postures and body movement. Visual processing of body expressions relies on a brain network, including body-specific visual areas and visuomotor areas. Even though the importance of the amygdala and its modulatory effects on distant brain regions have been documented, it remains unclear whether the amygdala plays a crucial role in emotional body processing. By combining behavioral and neuroimaging data in patients with amygdalar lesions, we provide further evidence for its modulatory effect on distant areas during the perception of body expressions.

Gray Matter Volume of a Region in the Thalamic Pulvinar Is Specifically Associated with Novelty Seeking.

Personality reflects the set of psychological traits and mechanisms characteristic for an individual. Geno-neuro-biologically inspired personality accounts have proposed a set of temperaments and characters that jointly compose personality profiles. The present study addresses the link between neurobiology and personality and investigates the association between temperament traits and regional gray matter volume. Furthermore, the specificity of these associations as well as the underlying components that drive the association are addressed. One hundred and four participants completed the Temperament and Character Inventory (TCI) and underwent structural magnetic resonance brain imaging. The participants included premanifest carriers of Huntington's disease, as this population is associated with temperament-related neuropsychiatric symptoms. Whole brain voxel-based multiple regression analyses on gray matter volume revealed a significant specific positive correlation between a region in the left thalamic pulvinar and novelty seeking score, controlled for the other traits (Pheight < 0.05, FWE-corrected). No significant associations were observed for the other temperament traits. Region of interest analyses showed that this association is driven by the subscale NS2: impulsiveness. The results increase the knowledge of the structural neurobiology of personality and indicate that individual differences in novelty seeking reflect the structural differences observed in the brain in an area that is widely and densely connected, which is in line with the typically domain-general behavioral influence of personality traits on a wide range of affective, perceptual, mnemotic, executive, and other cognitive functions.

Studying emotion theories through connectivity analysis: Evidence from generalized psychophysiological interactions and graph theory.

Psychological construction models of emotion state that emotions are variable concepts constructed by fundamental psychological processes, whereas according to basic emotion theory, emotions cannot be divided into more fundamental units and each basic emotion is represented by a unique and innate neural circuitry. In a previous study, we found evidence for the psychological construction account by showing that several brain regions were commonly activated when perceiving different emotions (i.e. a general emotion network). Moreover, this set of brain regions included areas associated with core affect, conceptualization and executive control, as predicted by psychological construction models. Here we investigate directed functional brain connectivity in the same dataset to address two questions: 1) is there a common pathway within the general emotion network for the perception of different emotions and 2) if so, does this common pathway contain information to distinguish between different emotions? We used generalized psychophysiological interactions and information flow indices to examine the connectivity within the general emotion network. The results revealed a general emotion pathway that connects neural nodes involved in core affect, conceptualization, language and executive control. Perception of different emotions could not be accurately classified based on the connectivity patterns from the nodes of the general emotion pathway. Successful classification was achieved when connections outside the general emotion pathway were included. We propose that the general emotion pathway functions as a common pathway within the general emotion network and is involved in shared basic psychological processes across emotions. However, additional connections within the general emotion network are required to classify different emotions, consistent with a constructionist account.

Investigating hyperoxic effects in the rat brain using quantitative susceptibility mapping based on MRI phase.

PURPOSE: To test whether susceptibility imaging can detect microvenous oxygen saturation changes, induced by hyperoxia, in the rat brain. METHODS: A three-dimensional gradient-echo with a flow compensation sequence was used to acquire T2*-weighted images of rat brains during hyperoxia and normoxia. Quantitative susceptibility mapping (QSM) and QSM-based microvenous oxygenation venography were computed from gradient-echo (GRE) phase images and compared between the two conditions. Pulse oxygen saturation (SpO2 ) in the cortex was examined and compared with venous oxygen saturation (SvO2 ) estimated by QSM. Oxygen saturation change calculated by a conventional Δ R2* map was also compared with the ΔSvO2 estimated by QSM. RESULTS: Susceptibilities of five venous and tissue regions were quantified separately by QSM. Venous susceptibility was reduced by nearly 10%, with an SvO2 shift of 10% during hyperoxia. A hyperoxic effect, confirmed by SpO2 measurement, resulted in an SvO2 increase in the cortex. The ΔSvO2 between hyperoxia and normoxia was consistent with what was estimated by the Δ R2* map in five regions. CONCLUSION: These findings suggest that a quantitative susceptibility map is a promising technique for SvO2 measurement. This method may be useful for quantitatively investigating oxygenation-dependent functional MRI studies. Magn Reson Med 77:592-602, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

Chitosan in Molecularly-Imprinted Polymers: Current and Future Prospects.

Chitosan is widely used in molecular imprinting technology (MIT) as a functional monomer or supporting matrix because of its low cost and high contents of amino and hydroxyl functional groups. The various excellent properties of chitosan, which include nontoxicity, biodegradability, biocompatibility, and attractive physical and mechanical performances, make chitosan a promising alternative to conventional functional monomers. Recently, chitosan molecularly-imprinted polymers have gained considerable attention and showed significant potential in many fields, such as curbing environmental pollution, medicine, protein separation and identification, and chiral-compound separation. These extensive applications are due to the polymers' desired selectivity, physical robustness, and thermal stability, as well as their low cost and easy preparation. Cross-linkers, which fix the functional groups of chitosan around imprinted molecules, play an important role in chitosan molecularly-imprinted polymers. This review summarizes the important cross-linkers of chitosan molecularly-imprinted polymers and illustrates the cross-linking mechanism of chitosan and cross-linkers based on the two glucosamine units. Finally, some significant attempts to further develop the application of chitosan in MIT are proposed.

Single-frequency excitation wideband MRI (SE-WMRI).

PURPOSE: In this study, single-frequency excitation wideband magnetic resonance imaging (MRI) (SE-WMRI) was proposed to obtain high-quality accelerated images by reducing phase-encoding steps while applying separation gradients. METHODS: A zig-zag k-space trajectory with reduced phase-encoding steps and an increased readout sampling rate was proposed. A unique gradient design with buffer intervals near the trajectory turns was employed to avoid undersampling and image artifacts. A gridding method and Fourier transform were used for image reconstruction. Quantitative analysis was performed on phantom images to investigate the characteristics of the acceleration method. RESULTS: The proposed method showed evident improvements in the accelerated phantom images, substantially reducing the ringing and blurring artifacts found using previous methods. Furthermore, the accelerated images exhibited the same signal-to-noise ratio as standard imaging. The accelerated in vivo experiment also produced the same quality as standard imaging. CONCLUSIONS: The proposed SE-WMRI method can effectively remove image artifacts and acquire images of higher temporal or spatial resolutions with less compromise.

Common neural correlates of emotion perception in humans.

Whether neuroimaging findings support discriminable neural correlates of emotion categories is a longstanding controversy. Two recent meta-analyses arrived at opposite conclusions, with one supporting (Vytal and Hamann []: J Cogn Neurosci 22:2864-2885) and the other opposing this proposition (Lindquist et al. []: Behav Brain Sci 35:121-143). To obtain direct evidence regarding this issue, we compared activations for four emotions within a single fMRI design. Angry, happy, fearful, sad and neutral stimuli were presented as dynamic body expressions. In addition, observers categorized motion morphs between neutral and emotional stimuli in a behavioral experiment to determine their relative sensitivities. Brain-behavior correlations revealed a large brain network that was identical for all four tested emotions. This network consisted predominantly of regions located within the default mode network and the salience network. Despite showing brain-behavior correlations for all emotions, muli-voxel pattern analyses indicated that several nodes of this emotion general network contained information capable of discriminating between individual emotions. However, significant discrimination was not limited to the emotional network, but was also observed in several regions within the action observation network. Taken together, our results favor the position that one common emotional brain network supports the visual processing and discrimination of emotional stimuli.

Fractal analysis of spontaneous fluctuations of the BOLD signal in the human brain networks.

PURPOSE: To investigate what extent brain regions are continuously interacting during resting-state, independent component analyses (ICA) was applied to analyze resting-state functional MRI (RS-fMRI) data. According to the analyzed results, it was surprisingly found that low frequency fluctuations (LFFs), which belong to the 1/f signal (a signal with power spectrum whose power spectral density is inversely proportional to the frequency), have been classified into groups using ICA; furthermore, the spatial distributions of these groups within the brain were found to resemble the spatial distributions of different networks, which manifests that the signal characteristics of RS LFFs are distinct across networks. In our work, we applied the 1/f model in the fractal analyses to further investigate this distinction. MATERIALS AND METHODS: Twenty healthy participants got involved in this study. They were scanned to acquire the RS-fMRI data. The acquired data were first processed with ICA to obtain the networks of the resting brain. Afterward, the blood-oxygenation level dependent (BOLD) signals of these networks were processed with the fractal analyses for obtaining the fractal parameter α. RESULTS: α was found to significantly vary across networks, which reveals that the fractal characteristic of LFFs differs across networks. According to prior literatures, this difference could be brought by the discrepancy of hemodynamic response amplitude (HRA) between networks. Hence, in our work, we also performed the computational simulation to discover the relationship between α and HRA. Based on the simulation results, HRA is highly linear-correlated with the fractal characteristics of LFFs which is revealed by α. CONCLUSION: Our results support that the origin of RS-fMRI signals contains arterial fluctuations. Hence, in addition to the commonly used method such as synchrony analysis and power spectral analysis, another approach, the fractal analysis, is suggested for acquiring the information of hemodynamic responses by means of RS-fMRI data.

Brain deactivation in the outperformance in bimodal tasks: an FMRI study.

While it is known that some individuals can effectively perform two tasks simultaneously, other individuals cannot. How the brain deals with performing simultaneous tasks remains unclear. In the present study, we aimed to assess which brain areas corresponded to various phenomena in task performance. Nineteen subjects were requested to sequentially perform three blocks of tasks, including two unimodal tasks and one bimodal task. The unimodal tasks measured either visual feature binding or auditory pitch comparison, while the bimodal task required performance of the two tasks simultaneously. The functional magnetic resonance imaging (fMRI) results are compatible with previous studies showing that distinct brain areas, such as the visual cortices, frontal eye field (FEF), lateral parietal lobe (BA7), and medial and inferior frontal lobe, are involved in processing of visual unimodal tasks. In addition, the temporal lobes and Brodmann area 43 (BA43) were involved in processing of auditory unimodal tasks. These results lend support to concepts of modality-specific attention. Compared to the unimodal tasks, bimodal tasks required activation of additional brain areas. Furthermore, while deactivated brain areas were related to good performance in the bimodal task, these areas were not deactivated where the subject performed well in only one of the two simultaneous tasks. These results indicate that efficient information processing does not require some brain areas to be overly active; rather, the specific brain areas need to be relatively deactivated to remain alert and perform well on two tasks simultaneously. Meanwhile, it can also offer a neural basis for biofeedback in training courses, such as courses in how to perform multiple tasks simultaneously.