The activation mechanism of Irga6, an interferon-inducible GTPase contributing to mouse resistance against Toxoplasma gondii

Background: The interferon-inducible immunity-related GTPases (IRG proteins/p47 GTPases) are a distinctive family of GTPases that function as powerful cell-autonomous resistance factors. The IRG protein, Irga6 (IIGP1), participates in the disruption of the vacuolar membrane surrounding the intracellular parasite, Toxoplasma gondii, through which it communicates with its cellular hosts. Some aspects of the protein's behaviour have suggested a dynamin-like molecular mode of action, in that the energy released by GTP hydrolysis is transduced into mechanical work that results in deformation and ultimately rupture of the vacuolar membrane. Results: Irga6 forms GTP-dependent oligomers in vitro and thereby activates hydrolysis of the GTP substrate. In this study we define the catalytic G-domain interface by mutagenesis and present a structural model, of how GTP hydrolysis is activated in Irga6 complexes, based on the substrate-twinning reaction mechanism of the signal recognition particle (SRP) and its receptor (SRalpha). In conformity with this model, we show that the bound nucleotide is part of the catalytic interface and that the 3'hydroxyl of the GTP ribose bound to each subunit is essential for trans-activation of hydrolysis of the GTP bound to the other subunit. We show that both positive and negative regulatory interactions between IRG proteins occur via the catalytic interface. Furthermore, mutations that disrupt the catalytic interface also prevent Irga6 from accumulating on the parasitophorous vacuole membrane of T. gondii, showing that GTP-dependent Irga6 activation is an essential component of the resistance mechanism. Conclusions: The catalytic interface of Irga6 defined in the present experiments can probably be used as a paradigm for the nucleotide-dependent interactions of all members of the large family of IRG GTPases, both activating and regulatory. Understanding the activation mechanism of Irga6 will help to explain the mechanism by which IRG proteins exercise their resistance function. We find no support from sequence or G-domain structure for the idea that IRG proteins and the SRP GTPases have a common phylogenetic origin. It therefore seems probable, if surprising, that the substrate-assisted catalytic mechanism has been independently evolved in the two protein families

Brain Vital Signs in Elite Ice Hockey: Towards Characterizing Objective and Specific Neurophysiological Reference Values for Concussion Management.

Background: Prior concussion studies have shown that objective neurophysiological measures are sensitive to detecting concussive and subconcussive impairments in youth ice-hockey. These studies monitored brain vital signs at rink-side using a within-subjects design to demonstrate significant changes from pre-season baseline scans. However, practical clinical implementation must overcome inherent challenges related to any dependence on a baseline. This requires establishing the start of normative reference data sets. Methods: The current study collected specific reference data for N = 58 elite, youth, male ice-hockey players and compared these with a general reference dataset from N = 135 of males and females across the lifespan. The elite hockey players were recruited to a select training camp through CAA Hockey, a management agency for players drafted to leagues such as the National Hockey League (NHL). The statistical analysis included a test-retest comparison to establish reliability, and a multivariate analysis of covariance to evaluate differences in brain vital signs between groups with age as a covariate. Findings: Test-retest assessments for brain vital signs evoked potentials showed moderate-to-good reliability (Cronbach's Alpha > 0.7, Intraclass correlation coefficient > 0.5) in five out of six measures. The multivariate analysis of covariance showed no overall effect for group (p = 0.105), and a significant effect of age as a covariate was observed (p < 0.001). Adjusting for the effect of age, a significant difference was observed in the measure of N100 latency (p = 0.022) between elite hockey players and the heterogeneous control group. Interpretation: The findings support the concept that normative physiological data can be used in brain vital signs evaluation in athletes, and should additionally be stratified for age, skill level, and experience. These can be combined with general norms and/or individual baseline assessments where appropriate and/or possible. The current results allow for brain vital sign evaluation independent of baseline assessment, therefore enabling objective neurophysiological evaluation of concussion management and cognitive performance optimization in ice-hockey

Developing Brain Vital Signs: Initial Framework for Monitoring Brain Function Changes over Time

Clinical assessment of brain function relies heavily on indirect behavior-based tests. Unfortunately, behavior-based assessments are subjective and therefore susceptible to several confounding factors. Event-related brain potentials (ERPs), derived from electroencephalography (EEG), are often used to provide objective, physiological measures of brain function. Historically, ERPs have been characterized extensively within research settings, with limited but growing clinical applications. Over the past 20 years, we have developed clinical ERP applications for the evaluation of functional status following serious injury and/or disease. This work has identified an important gap: the need for a clinically accessible framework to evaluate ERP measures. Crucially, this enables baseline measures before brain dysfunction occurs, and might enable the routine collection of brain function metrics in the future much like blood pressure measures today. Here, we propose such a framework for extracting specific ERPs as potential “brain vital signs.” This framework enabled the translation/transformation of complex ERP data into accessible metrics of brain function for wider clinical utilization. To formalize the framework, three essential ERPs were selected as initial indicators: (1) the auditory N100 (Auditory sensation); (2) the auditory oddball P300 (Basic attention); and (3) the auditory speech processing N400 (Cognitive processing). First step validation was conducted on healthy younger and older adults (age range: 22–82 years). Results confirmed specific ERPs at the individual level (86.81–98.96%), verified predictable age-related differences (P300 latency delays in older adults, p &lt; 0.05), and demonstrated successful linear transformation into the proposed brain vital sign (BVS) framework (basic attention latency sub-component of BVS framework reflects delays in older adults, p &lt; 0.05). The findings represent an initial critical step in developing, extracting, and characterizing ERPs as vital signs, critical for subsequent evaluation of dysfunction in conditions like concussion and/or dementia

Brain Vital Signs: Expanding From the Auditory to Visual Modality

The critical need for rapid objective, physiological evaluation of brain function at point-of-care has led to the emergence of brain vital signs—a framework encompassing a portable electroencephalography (EEG) and an automated, quick test protocol. This framework enables access to well-established event-related potential (ERP) markers, which are specific to sensory, attention, and cognitive functions in both healthy and patient populations. However, all our applications to-date have used auditory stimulation, which have highlighted application challenges in persons with hearing impairments (e.g., aging, seniors, dementia). Consequently, it has become important to translate brain vital signs into a visual sensory modality. Therefore, the objectives of this study were to: 1) demonstrate the feasibility of visual brain vital signs; and 2) compare and normalize results from visual and auditory brain vital signs. Data were collected from 34 healthy adults (33 ± 13 years) using a 64-channel EEG system. Visual and auditory sequences were kept as comparable as possible to elicit the N100, P300, and N400 responses. Visual brain vital signs were elicited successfully for all three responses across the group (N100: F = 29.8380, p &lt; 0.001; P300: F = 138.8442, p &lt; 0.0001; N400: F = 6.8476, p = 0.01). Initial auditory-visual comparisons across the three components showed attention processing (P300) was found to be the most transferrable across modalities, with no group-level differences and correlated peak amplitudes (rho = 0.7, p = 0.0001) across individuals. Auditory P300 latencies were shorter than visual (p &lt; 0.0001) but normalization and correlation (r = 0.5, p = 0.0033) implied a potential systematic difference across modalities. Reduced auditory N400 amplitudes compared to visual (p = 0.0061) paired with normalization and correlation across individuals (r = 0.6, p = 0.0012), also revealed potential systematic modality differences between reading and listening language comprehension. This study provides an initial understanding of the relationship between the visual and auditory sequences, while importantly establishing a visual sequence within the brain vital signs framework. With both auditory and visual stimulation capabilities available, it is possible to broaden applications across the lifespan

Brain Vital Signs: Expanding From the Auditory to Visual Modality

The critical need for rapid objective, physiological evaluation of brain function at point-of-care has led to the emergence of brain vital signs—a framework encompassing a portable electroencephalography (EEG) and an automated, quick test protocol. This framework enables access to well-established event-related potential (ERP) markers, which are specific to sensory, attention, and cognitive functions in both healthy and patient populations. However, all our applications to-date have used auditory stimulation, which have highlighted application challenges in persons with hearing impairments (e.g., aging, seniors, dementia). Consequently, it has become important to translate brain vital signs into a visual sensory modality. Therefore, the objectives of this study were to: 1) demonstrate the feasibility of visual brain vital signs; and 2) compare and normalize results from visual and auditory brain vital signs. Data were collected from 34 healthy adults (33 ± 13 years) using a 64-channel EEG system. Visual and auditory sequences were kept as comparable as possible to elicit the N100, P300, and N400 responses. Visual brain vital signs were elicited successfully for all three responses across the group (N100:&nbsp;F&nbsp;= 29.8380,&nbsp;p&nbsp;&lt; 0.001; P300:&nbsp;F&nbsp;= 138.8442,&nbsp;p&nbsp;&lt; 0.0001; N400:&nbsp;F&nbsp;= 6.8476,&nbsp;p&nbsp;= 0.01). Initial auditory-visual comparisons across the three components showed attention processing (P300) was found to be the most transferrable across modalities, with no group-level differences and correlated peak amplitudes (rho = 0.7,&nbsp;p&nbsp;= 0.0001) across individuals. Auditory P300 latencies were shorter than visual (p&nbsp;&lt; 0.0001) but normalization and correlation (r&nbsp;= 0.5,&nbsp;p&nbsp;= 0.0033) implied a potential systematic difference across modalities. Reduced auditory N400 amplitudes compared to visual (p&nbsp;= 0.0061) paired with normalization and correlation across individuals (r&nbsp;= 0.6,&nbsp;p&nbsp;= 0.0012), also revealed potential systematic modality differences between reading and listening language comprehension. This study provides an initial understanding of the relationship between the visual and auditory sequences, while importantly establishing a visual sequence within the brain vital signs framework. With both auditory and visual stimulation capabilities available, it is possible to broaden applications across the lifespan. The critical need for rapid objective, physiological evaluation of brain function at point-of-care has led to the emergence of brain vital signs—a framework encompassing a portable electroencephalography (EEG) and an automated, quick test protocol. This framework enables access to well-established event-related potential (ERP) markers, which are specific to sensory, attention, and cognitive functions in both healthy and patient populations. However, all our applications to-date have used auditory stimulation, which have highlighted application challenges in persons with hearing impairments (e.g., aging, seniors, dementia). Consequently, it has become important to translate brain vital signs into a visual sensory modality. Therefore, the objectives of this study were to: 1) demonstrate the feasibility of visual brain vital signs; and 2) compare and normalize results from visual and auditory brain vital signs. Data were collected from 34 healthy adults (33 ± 13 years) using a 64-channel EEG system. Visual and auditory sequences were kept as comparable as possible to elicit the N100, P300, and N400 responses. Visual brain vital signs were elicited successfully for all three responses across the group (N100:&nbsp;F&nbsp;= 29.8380,&nbsp;p&nbsp;&lt; 0.001; P300:&nbsp;F&nbsp;= 138.8442,&nbsp;p&nbsp;&lt; 0.0001; N400:&nbsp;F&nbsp;= 6.8476,&nbsp;p&nbsp;= 0.01). Initial auditory-visual comparisons across the three components showed attention processing (P300) was found to be the most transferrable across modalities, with no group-level differences and correlated peak amplitudes (rho = 0.7,&nbsp;p&nbsp;= 0.0001) across individuals. Auditory P300 latencies were shorter than visual (p&nbsp;&lt; 0.0001) but normalization and correlation (r&nbsp;= 0.5,&nbsp;p&nbsp;= 0.0033) implied a potential systematic difference across modalities. Reduced auditory N400 amplitudes compared to visual (p&nbsp;= 0.0061) paired with normalization and correlation across individuals (r&nbsp;= 0.6,&nbsp;p&nbsp;= 0.0012), also revealed potential systematic modality differences between reading and listening language comprehension. This study provides an initial understanding of the relationship between the visual and auditory sequences, while importantly establishing a visual sequence within the brain vital signs framework. With both auditory and visual stimulation capabilities available, it is possible to broaden applications across the lifespan

Age, education and intellectual quotient influences : structural equation modeling on the study of Benton Visual Retention Test (BVRT)

This study searched for sociodemographic influences on visual memory and visuoconstructive ability in healthy and clinical samples evaluated with Benton Visual Retention Test (BVRT) in two studies. In Study 1, we searched for changes related to age in children, adolescents, adults and elderly on the performance of the BVRT. In Study 2, we investigated the relations among age, years of education and intellectual quotient (IQ) on the performance of the BVRT using Structural Equation Modeling (SEM). Participants were 624 individuals aged between six and 89 years old (M = 25.40; SD = 22.34) from the normatization and evidence validity studies at Brazil. We used a sociodemographic questionnaire, BVRT and IQ measure was estimated. Study 1 has shown a performance similar to the developmental graphics with a U-inverted pattern in relation to age: An increase of the visual memory ability in the children and adolescent groups as age increases, a tendency of a decrease in the performance in the adult group that intensifies in the elderly group. Study 2 found that the model for the BVRT performance tested by SEM denoted satisfactory goodness-of-fit indexes, χ2/gl = 2.67, p < .001; CFI = .92; TLI = .93; RMSEA = .004, 90% CI = [.03, .05];WLSMV = 1.79, and corroborated the theoretical assumption. The SEM model confirmed in this study highlight the strong role of years of education in the prediction of BVRT scores

Brain vital signs: Auditory to visual translation

An objective measure is greatly needed to monitor the impacts of injury or disease on our brain health. In order to provide such a measure, the brain vital sign framework utilizes an established, non-invasive, and physiology-based technology, Electroencephalogram (EEG), along with a rapid auditory sequence used to elicit and assess specific markers of cognitive function (marked by event-related potentials (ERPs)). To date, applications for brain vital signs have included evaluation of cognitive function in healthy and patient populations. To expand the applications, this study aims to translate the established rapid auditory sequence to a visual based assessment. The objectives are to: 1) demonstrate the viability of visual brain vital signs assessment and 2) examine the differences between the two modalities. EEG data was collected in 30 healthy adults (33±14yrs) and analyzed at central electrodes. Similar to the interlaced auditory sequence, the visual sequence utilized an oddball paradigm (standard vs. deviant stimuli) to evoke a sensory (N100) and attention (P300) response, and a word pair paradigm (congruent vs. incongruent stimuli) to evoke a semantic language response (N400). Comparison of mean amplitudes between stimuli revealed the targeted ERPs were successfully evoked in the visual modality at a group-level as expected (N100: p &lt; 0.001; P300: p &lt; 0.0001; N400: p = 0.0105). Attention processing (P300) was found to be the most transferrable across modalities, with no group-level differences and correlated peak amplitudes (rho = 0.7, p =0.0001) across individuals. Auditory P300 latencies were shorter than visual (p &lt; 0.0001) but normalization and correlation (r =0.5, p = 0.0033) implied a potential systematic difference across modalities. Reduced auditory N400 amplitudes compared to visual (p = 0.0061) paired with normalization and correlation across individuals (r = 0.6, p = 0.0012), also revealed potential systematic modality differences between reading and listening language comprehension. This study provides initial understanding of the relationship between the visual and auditory sequences, while importantly establishing a visual sequence within the brain vital signs framework as a potential translational tool to monitor brain health over the human lifespan in broader populations, such as those with hearing impairments, congenital or due to injury or aging

The activation mechanism of Irga6, an interferon-inducible GTPase contributing to mouse resistance against Toxoplasma gondii

Background: The interferon-inducible immunity-related GTPases (IRG proteins/p47 GTPases) are a distinctive family of GTPases that function as powerful cell-autonomous resistance factors. The IRG protein, Irga6 (IIGP1), participates in the disruption of the vacuolar membrane surrounding the intracellular parasite, Toxoplasma gondii, through which it communicates with its cellular hosts. Some aspects of the protein%27s behaviour have suggested a dynamin-like molecular mode of action, in that the energy released by GTP hydrolysis is transduced into mechanical work that results in deformation and ultimately rupture of the vacuolar membrane. Results: Irga6 forms GTP-dependent oligomers in vitro and thereby activates hydrolysis of the GTP substrate. In this study we define the catalytic G-domain interface by mutagenesis and present a structural model, of how GTP hydrolysis is activated in Irga6 complexes, based on the substrate-twinning reaction mechanism of the signal recognition particle (SRP) and its receptor (SR alpha). In conformity with this model, we show that the bound nucleotide is part of the catalytic interface and that the 3%27hydroxyl of the GTP ribose bound to each subunit is essential for trans-activation of hydrolysis of the GTP bound to the other subunit. We show that both positive and negative regulatory interactions between IRG proteins occur via the catalytic interface. Furthermore, mutations that disrupt the catalytic interface also prevent Irga6 from accumulating on the parasitophorous vacuole membrane of T. gondii, showing that GTP-dependent Irga6 activation is an essential component of the resistance mechanism. Conclusions: The catalytic interface of Irga6 defined in the present experiments can probably be used as a paradigm for the nucleotide-dependent interactions of all members of the large family of IRG GTPases, both activating and regulatory. Understanding the activation mechanism of Irga6 will help to explain the mechanism by which IRG proteins exercise their resistance function. We find no support from sequence or G-domain structure for the idea that IRG proteins and the SRP GTPases have a common phylogenetic origin. It therefore seems probable, if surprising, that the substrate-assisted catalytic mechanism has been independently evolved in the two protein families