Cortico-striatal-thalamic-cortical Research Articles

Abnormal structure and function of white matter in obsessive-compulsive disorder

BackgroundAbnormal structure and function of gray matter (GM) have been discovered in the cortico-striatal-thalamic-cortical (CSTC) circuit in obsessive-compulsive disorder (OCD). The GM structure and function may be influenced by the structure and function of the white matter (WM). Therefore, it is crucial to explore the characteristics of WM in OCD. MethodsDiffusion tensor imaging and resting-state functional magnetic resonance imaging data of 52 patients with OCD and 39 healthy controls (HCs) were collected. The tract-based spatial statistics, amplitude of low-frequency fluctuations (ALFF), and structural-functional coupling approaches were utilized to explore the WM structure and function. Furthermore, the relationship between the abnormal WM structure and function and clinical symptoms of OCD was investigated using Pearson's correlation. Support vector machine was performed to evaluate whether patients with OCD could be identified with the changed WM structure and function. ResultsCompared to HCs, the lower fractional anisotropy (FA) values of four clusters including the superior corona radiata, anterior corona radiata, right superior longitudinal fasciculus, corpus callosum, left posterior corona radiata, fornix, and the right anterior limb of internal capsule, reduced ALFF/FA ratio in the left anterior thalamic radiation (ATR), and the decreased functional connectivity between the left ATR and the left dorsal lateral prefrontal cortex within CSTC circuit at rest were observed in OCD. The decreased ALFF/FA ratio in the left ATR negatively correlated with Yale-Brown Obsessive-Compulsive Scale obsessive thinking scores and Hamilton Anxiety Rating Scale scores in OCD. Furthermore, the features that combined the abnormal WM structure and function performed best in distinguishing OCD from HCs with the appropriate accuracy (0.80), sensitivity (0.82), as well as specificity (0.80). ConclusionCurrent research discovered changed WM structure and function in OCD. Furthermore, abnormal WM structural-functional coupling may lead to aberrant GM connectivity within the CSTC circuit at rest in OCD.Trial Registration: Study on the mechanism of brain network in obsessive-compulsive disorder with multi-model magnetic resonance imaging (ChiCTR-COC-17013301).

Thalamo-cortical circuits associated with trait- and state-repetitive negative thinking in major depressive disorder

BackgroundRepetitive negative thinking (RNT), often referred to as rumination in the mood disorders literature, is a symptom dimension associated with poor prognosis and suicide in major depressive disorder (MDD). Given the transdiagnostic nature of RNT, this study aimed to evaluate the hypothesis that neurobiological substrates of RNT in MDD may share the brain mechanisms underlying obsessions, particularly those involving cortico-striatal-thalamic-cortical (CSTC) circuits. MethodsThirty-nine individuals with MDD underwent RNT induction during fMRI. Trait-RNT was measured by the Ruminative Response Scale (RRS) and state-RNT was measured by a visual analogue scale. We employed a connectome-wide association analysis examining the association between RNT intensity with striatal and thalamic connectivity. ResultsA greater RRS score was associated with hyperconnectivity of the right mediodorsal thalamus with prefrontal cortex, including lateral orbitofrontal cortex, along with Wernicke's area and posterior default mode network nodes (t = 4.66–6.70). A greater state-RNT score was associated with hyperconnectivity of the right laterodorsal thalamus with bilateral primary sensory and motor cortices, supplementary motor area, and Broca's area (t = 4.51–6.57). Unexpectedly, there were no significant findings related to the striatum. ConclusionsThe present results suggest RNT in MDD is subserved by abnormal connectivity between right thalamic nuclei and cortical regions involved in both visceral and higher order cognitive processing. Emerging deep-brain neuromodulation methods may be useful to establish causal relationships between dysfunction of right thalamic-cortical circuits and RNT in MDD.

1H-MRS of the anterior cingulate cortex in obsessive-compulsive disorder: metabolic abnormalities in pgACC - controlled study

IntroductionObsessive-compulsive disorder (OCD) is connected with increased activity in cortico-striatal-thalamic-cortical (CSTC) loop. The anterior cingulate cortex (ACC) is a part of this loop and plays a role in error detection and monitoring and processing of conflicting information, core OCD clinical signs. This area also contains a high density of Von Economo neurons. Biochemistry in this area is closely connected with the pathophysiology of OCD.ObjectivesDecreased concentrations of total N-acetylaspartate (tNAA) have been reported in ACC in patients with OCD compared to healthy controls (HC), with increase after successful treatment. Findings by other metabolites: choline-containing compounds (tCho), total creatine (tCr) and myo-inositol are not consistent. Differences in levels of tNAA, tCho, tCr would correlate with the severity of symptoms measured by Y-BOCS. In the comparison in the subgroup of patients with/ without medication, there will be differences in levels of metabolites.Methods54 patients diagnosed with OCD according to ICD-10 and DSM-IV criteria, and 54 HC matched for age and sex were included in the study. They underwent MRI and MR Spectroscopy on a 3T Magnetom Prisma scanner (Siemens, Germany) equipped with a 64-channel volume head coil (Fig. 2). After spectral quality control, 28 OCD and 28 HC subjects were included in the statistical analysis. OCD subjects were interviewed using the Y-BOCS to evaluate the severity of the symptoms. Patients enrolled in the study were without medication at least 5 days before MRI or on a stable dosage of SSRI antidepressants. To assess the intergroup differences Wilcoxon Rank Sum test or Kruskal-Wallis test was used as appropriate. The correlation between metabolite levels and clinical characteristics was assessed by Spearman’s rank correlation coefficient. The statistics were calculated using R, version 4.1.1.ResultsWe found no differences in levels of tNAA in ACC in OCD vs. HC (p=0,21; see Tab.1, Fig.1). We found significantly increased level of tCho, tCr and Ins in OCD vs. HC (p=0,03; p=0,004 resp.; p=0,017 resp.). tCr levels corelated negatively with YBOCS compulsions subscale (p=0,046; cor=-0,38). tCho levels showed a trend to negative correlation with Y-BOCS compulsions subscale (p=0,067; cor=-0,35). Analysis on the subgroup with (13 subjects, 46,43 %) and without (13 subjects, 46,43 %) stable SSRI medication did not reveal significant differences.Image:Image 2:ConclusionsOur study found difference in ACC by OCD patients compared to HC, mainly increased tCho, tCr and Ins. Also, the study shows a significant correlation between the severity of compulsions and tCr levels. We can see this trend also in the correlation of the tCho and Y-BOCS compulsions subscale. Similar tNAA level by OCD and HC groups could indicates correctly adjusted medication or stable state by enrolled patients.Disclosure of InterestNone Declared

Invasive and Non-invasive Neurostimulation for OCD.

It becomes increasingly clear that (non-)invasive neurostimulation is an effective treatment for obsessive-compulsive disorder (OCD). In this chapter we review the available evidence on techniques and targets, clinical results including a meta-analysis, mechanisms of action, and animal research. We focus on deep brain stimulation (DBS), but also cover non-invasive neurostimulation including transcranial magnetic stimulation (TMS). Data shows that most DBS studies target the ventral capsule/ventral striatum (VC/VS), with an overall 76% response rate in treatment-refractory OCD. Also TMS holds clinical promise. Increased insight in the normalizing effects of neurostimulation on cortico-striatal-thalamic-cortical (CSTC) loops - through neuroimaging and animal research - provides novel opportunities to further optimize treatment strategies. Advancing clinical implementation of neurostimulation techniques is essential to ameliorate the lives of the many treatment-refractory OCD patients.

Modeling essential connections in obsessive–compulsive disorder patients using functional MRI

ObjectObsessive–compulsive disorder (OCD) is a mental disease in which people experience uncontrollable and repetitive thoughts or behaviors. Clinical diagnosis of OCD is achieved by using neuropsychological assessment metrics, which are often subjectively affected by psychologists and patients. In this study, we propose a classification model for OCD diagnosis using functional MR images.MethodsUsing functional connectivity (FC) matrices calculated from brain region of interest (ROI) pairs, a novel Riemann Kernel principal component analysis (PCA) model is employed for feature extraction, which preserves the topological information in the FC matrices. Hierarchical features are then fed into an ensemble classifier based on the XGBoost algorithm. Finally, decisive features extracted during classification are used to investigate the brain FC variations between patients with OCD and healthy controls.ResultsThe proposed algorithm yielded a classification accuracy of 91.8%. Additionally, the well‐known cortico–striatal–thalamic–cortical (CSTC) circuit and cerebellum were found as highly related regions with OCD. To further analyze the cerebellar‐related function in OCD, we demarcated cerebellum into three subregions according to their anatomical and functional property. Using these three functional cerebellum regions as seeds for brain connectivity computation, statistical results showed that patients with OCD have decreased posterior cerebellar connections.ConclusionsThis study provides a new and efficient method to characterize patients with OCD using resting‐state functional MRI. We also provide a new perspective to analyze disease‐related features. Despite of CSTC circuit, our model‐driven feature analysis reported cerebellum as an OCD‐related region. This paper may provide novel insight to the understanding of genetic etiology of OCD.

An MRI Study of the Metabolic and Structural Abnormalities in Obsessive-Compulsive Disorder.

Obsessive-compulsive disorder (OCD) is a neuropsychiatric illness characterized by obsessions and/or compulsions. Its pathophysiology is still not well understood but it is known that the cortico-striatal-thalamic-cortical (CSTC) circuitry plays an important role. Here, we used a multi-method MRI approach combining proton magnetic resonance spectroscopy (H1-MRS) and diffusion tensor imaging (DTI) techniques to investigate both the metabolic and the microstructural white matter (WM) changes of the anterior cingulate cortex (ACC) in OCD patients as compared to healthy controls. Twenty-three OCD patients and 21 age-, sex-, and education-matched healthy volunteers participated in the study. Our 1H-MRS findings show increased levels of Glx in ACC in OCD. Further, significantly lower fractional anisotropy (FA) values were observed in OCD patients’ left cingulate bundle (CB) as compared to healthy controls. Finally, there was a negative correlation between FA in the left CB and level of obsessions, as well as the duration of the illness. Our findings reinforce the involvement of CSTC bundles in pathophysiology of OCD, pointing to a specific role of glutamate (glutamine) and WM integrity.

Developmentally divergent sexual dimorphism in the cortico-striatal-thalamic-cortical psychosis risk pathway.

Structural and functional cortico-striatal-thalamic-cortical (CSTC) circuit abnormalities have been observed in schizophrenia and the clinical high-risk state. However, this circuit is sexually dimorphic and changes across neurodevelopment. We examined effects of sex and age on structural and functional properties of the CSTC circuit in a large sample of youth with and without psychosis spectrum symptoms (PSS) from the Philadelphia Neurodevelopmental Cohort. T1-weighted and resting-state functional MRI scans were collected on a 3T Siemens scanner, in addition to participants' cognitive and psychopathology data. After quality control, the total sample (aged 11-21) was n = 1095 (males = 485, females = 610). Structural subdivisions of the striatum and thalamus were identified using the MAGeT Brain segmentation tool. Functional seeds were segmented based on brain network connectivity. Interaction effects among PSS group, sex, and age on striatum, thalamus, and subdivision volumes were examined. A similar model was used to test effects on functional connectivity of the CSTCcircuit. A sex by PSS group interaction was identified, whereby PSS males had higher volumes and PSS females had lower volumes in striatal and thalamic subdivisions. Reduced functional striato-cortical connectivity was found in PSS youth, primarily driven by males, whereby younger male PSS youth also exhibited thalamo-cortical hypo-connectivity (compared to non-PSS youth), vs. striato-cortical hyper-connectivity in older male PSS youth (compared to non-PSS youth). Youth with PSS demonstrate sex and age-dependent differences in striatal and thalamic subdivision structure and functional connectivity. Further efforts at biomarker discovery and early therapeutic intervention targeting the CSTCcircuit in psychosis should consider effects of sex and age.

Aberrant resting-state functional connectivity of salience network in first-episode schizophrenia.

The disruption of salience network (SN) has been consistently found in patients with schizophrenia and thought to give rise to specific symptoms. However, the functional dysconnectivity pattern of SN remains unclear in first-episode schizophrenia (FES). Sixty-five patients with FES and sixty-six health controls (HC) were enrolled in this study and underwent resting-state functional magnetic resonance imaging (rs-fMRI). The eleven regions of interest (ROIs) within SN were derived from the peaks of the group independent component analysis (gICA). Seed-based whole-brain functional connectivity (FC) analyses were performed with all SN ROIs as the seeds. Both hyper- and hypo-connectivity of SN were found in the FES. Specifically, the increased FC mainly existed between the SN and cortico-cerebellar sub-circuit and prefrontal cortex, while the reduced FC mainly existed within cortico-striatal-thalamic-cortical (CSTC) sub-circuit. Our findings suggest that FES is associated with pronounced dysregulation of SN, characterized prominently by hyperconnectivity of SN-prefrontal cortex and cerebellum, as well as hypoconnectivity of CSTC sub-circuit of the SN.

Combined resting state functional magnetic resonance imaging and diffusion tensor imaging study in patients with idiopathic restless legs syndrome

ObjectiveRestless legs syndrome (RLS) is a common neurological disorder characterized by an urge to move the legs along with paraesthesia deep within them. In this study, we aimed to use diffusion tensor imaging (DTI) and regional homogeneity (ReHo) to investigate the changes in regional spontaneous brain activity change for RLS patients against age- and gender-matched normal control (NC) subjects. MethodsA total of 35 RLS patients and 27 age- and gender-matched NC subjects were recruited for group comparison research that used DTI and ReHo techniques. DTI was analysed by FSL and tract-based spatial statistics (TBSS) software to measure the values of fractional anisotropy (FA) or mean diffusivity (MD) in brain regions. Statistical Parametric Mapping 8 (SPM8) was used for data preprocessing and Data Processing Assistant for Resting-State fMRI (DPARSF) toolbox was used for ReHo calculation. For multiple comparison correction, the AlphaSim program implemented in AFNI was used to control the false-positive rate (corrected p < 0.05). ResultsThere was no significant difference between the iRLS and NC groups in age or gender. In the one-sample t-test, both the NC and RSL groups showed increased ReHo in the bilateral posterior cingulate/precuneus cortex compared to the groups' global means, indicating that the default mode network was at rest. The RLS group showed a smaller cluster size than the NC group. In the two-sample t-test, the RLS group showed increased ReHo in the bilateral middle frontal gyrus, anterior cingulate cortex, caudate nucleus, insula, thalamus, putamen and left posterior cingulate cortex compared to the NC group. The statistical analysis of DTI images did not show any difference between the two groups. TBSS group comparison did not reveal any difference in FA or mean diffusivity (MD) of any brain region. ConclusionRLS patients showed that greater ReHo within the striatum, thalamus and the limbic system, which implies that the emotional processing, motion control and cognition in the cortico-striatal-thalamic-cortical (CSTC) loop may be the site of dysfunction in RLS patients. This finding may provide imaging evidence to explore the pathophysiology of RLS. On the other hand, we did not see any change in the microstructure in the DTI analysis for RLS patients when compared to the NC group, which suggests a metabolic impairment.

A Framework for Understanding the Emerging Role of Corticolimbic-Ventral Striatal Networks in OCD-Associated Repetitive Behaviors.

Significant interest in the mechanistic underpinnings of obsessive-compulsive disorder (OCD) has fueled research on the neural origins of compulsive behaviors. Converging clinical and preclinical evidence suggests that abnormal repetitive behaviors are driven by dysfunction in cortico-striatal-thalamic-cortical (CSTC) circuits. These findings suggest that compulsive behaviors arise, in part, from aberrant communication between lateral orbitofrontal cortex (OFC) and dorsal striatum. An important body of work focused on the role of this network in OCD has been instrumental to progress in the field. Disease models focused primarily on these regions, however, fail to capture an important aspect of the disorder: affective dysregulation. High levels of anxiety are extremely prevalent in OCD, as is comorbidity with major depressive disorder. Furthermore, deficits in processing rewards and abnormalities in processing emotional stimuli are suggestive of aberrant encoding of affective information. Accordingly, OCD can be partially characterized as a disease in which behavioral selection is corrupted by exaggerated or dysregulated emotional states. This suggests that the networks producing OCD symptoms likely expand beyond traditional lateral OFC and dorsal striatum circuit models, and highlights the need to cast a wider net in our investigation of the circuits involved in generating and sustaining OCD symptoms. Here, we address the emerging role of medial OFC, amygdala, and ventral tegmental area projections to the ventral striatum (VS) in OCD pathophysiology. The VS receives strong innervation from these affect and reward processing regions, and is therefore poised to integrate information crucial to the generation of compulsive behaviors. Though it complements functions of dorsal striatum and lateral OFC, this corticolimbic-VS network is less commonly explored as a potential source of the pathology underlying OCD. In this review, we discuss this network’s potential role as a locus of OCD pathology and effective treatment.

Abnormal striatal resting-state functional connectivity in adolescents with obsessive–compulsive disorder

Neuroimaging research has implicated abnormalities in cortico-striatal-thalamic-cortical (CSTC) circuitry in pediatric obsessive–compulsive disorder (OCD). In this study, resting-state functional magnetic resonance imaging (R-fMRI) was used to investigate functional connectivity in the CSTC circuitry in adolescents with OCD. Imaging was obtained with the Human Connectome Project (HCP) scanner using newly developed pulse sequences which allow for higher spatial and temporal resolution. Fifteen adolescents with OCD and 13 age- and gender-matched healthy controls (ages 12–19) underwent R-fMRI on the 3T HCP scanner. Twenty-four minutes of resting-state scans (two consecutive 12-min scans) were acquired. We investigated functional connectivity of the striatum using a seed-based, whole brain approach with anatomically-defined seeds placed in the bilateral caudate, putamen, and nucleus accumbens. Adolescents with OCD compared with controls exhibited significantly lower functional connectivity between the left putamen and a single cluster of right-sided cortical areas including parts of the orbitofrontal cortex, inferior frontal gyrus, insula, and operculum. Preliminary findings suggest that impaired striatal connectivity in adolescents with OCD in part falls within the predicted CSTC network, and also involves impaired connections between a key CSTC network region (i.e., putamen) and key regions in the salience network (i.e., insula/operculum). The relevance of impaired putamen-insula/operculum connectivity in OCD is discussed.

A preliminary study of functional connectivity of medication naïve children with obsessive–compulsive disorder

Evidence suggests that obsessive-compulsive disorder (OCD) is associated with a dysfunction in the cortico-striatal-thalamic-cortical (CSTC) circuitry. Resting state functional connectivity magnetic resonance imaging (rs-fcMRI) allows measurements of resting state networks (RSNs), brain networks that are present at 'rest'. However, although OCD has a typical onset during childhood or adolescence, only two other studies have performed rs-fcMRI comparisons of RSNs in children and adolescents with OCD against healthy controls. In the present study, we performed resting state functional magnetic resonance imaging using a 3 Tesla MRI, in 11 medication-naïve children and adolescents with OCD and 9 healthy controls. In contrast to previous studies that relied on a priori determination of RSNs, we determined resting state functional connectivity with a data-driven independent component analysis (ICA). Consistent with previous reports in healthy adults, we identified 13 RSNs. Case-control un-adjusted statistical significance (p<0.05) was found for two networks. Firstly, increased connectivity (OCD>control) in the right section of Brodmann area 43 of the auditory network; Secondly, decreased connectivity in the right section of Brodmann area 8 and Brodmann area 40 in the cingulate network. Our preliminary findings of case-control differences in RSNs lend further support to the CSTC hypothesis of OCD, as well as implicating other regions of the brain outside of the CSTC.

Cortico-Striatal-Thalamic-Cortical (CSTC) Circuits, Tourette's Disorder, and OCD

Cortico-Striatal-Thalamic-Cortical (CSTC) Circuits, Tourette's Disorder, and OCD

Structural equation modeling and principal component analysis of gray matter volumes in major depressive and bipolar disorders: Differences in latent volumetric structure

Abnormalities of the cortico-striatal-thalamic-cortical (CSTC) and the limbic-cortico-striatal-thalamic-cortical (LCSTC) circuits have been hypothesized in mood disorders. We performed principal component analysis (PCA) to identify latent volumetric systems on regional brain volumes and correlated these patterns with clinical characteristics; further, we performed exploratory structural equation modeling (SEM) to test a priori hypotheses about the organization among the structures comprising the CSTC and LCSTC circuits, and to investigate differences among subjects with bipolar disorder (BD), major depressive disorder (MDD), and healthy controls (HC). Participants included 45 BD and 31 MDD patients, and 72 HC. Regional MR brain volumes were used to calculate patterns of volumetric covariance. The identified latent volumetric systems were related to the depression severity and the duration of illness. BD differed from HC on the estimated parameters describing the paths of cortico-striatal, thalamo-striatal and intrastriatal loops of the CSTC circuit, and the paths between anterior and posterior cingulate cortex (PCC), and hippocampus to amygdala of the LCSTC circuit. MDD differed from HC on the paths between putamen and thalamus, and PCC to hippocampus. This study provides evidence to suggest different organizational patterns among structures within the CSTC and LCSTC circuits for BD, MDD, and HC, which may point to structural abnormalities underlying mood disorders.

Internalizing Versus Externalizing Comorbidity: Neural Circuit Hypothesis

The aim of the present study was to review and update historical controversies in relation to the limbic system in light of advances in the neuroanatmy and neurobiology of cortico-striatal-thalamic-cortical (CSTC) neural circuits. It is hypothesized that observed comorbidities between internalizing and externalizing childhood syndromes may be better understood in terms of excess communication between subcortical brain circuits. A neuroanatomical understanding of reciprocal and non-reciprocal connections between modular circuits provides a basis for understanding previously puzzling aspects of internalizing, externalizing comorbidity. Function depends on the degree of cooperative integration of goal orientation, Pavlovian stimulus response, and operant processing in cortical-subcortical circuits, and interventions may be targeted at any of these functions.

Latent volumetric structure of the human brain: Exploratory factor analysis and structural equation modeling of gray matter volumes in healthy children and adults

Previous studies have investigated patterns of volumetric covariance (i.e. intercorrelation) among brain regions. Methodological issues, however, have limited the validity and generalizability of findings from these prior studies. Additionally, patterns of volumetric covariance have often been assumed to reflect the presence of structural networks, but this assumption has never been tested formally. We identified patterns of volumetric covariance, correlated these patterns with behavioral measures, and tested the hypothesis that the observed patterns of covariance reflect the presence of underlying networks. Specifically, we performed factor analysis on regional brain volumes of 99 healthy children and adults, and we correlated factor scores with scores on the Stroop Word-Color Interference Test. We identified four latent volumetric systems in each hemisphere: dorsal cortical, limbic, posterior, and basal ganglia. The positive correlation of the right posterior system with Stroop scores suggested that larger latent volumes are detrimental to inhibitory control. We also applied Structural Equation Modeling (SEM) to our dataset (n = 107) to test whether a model based on the anatomical pathways within cortico-striatal-thalamic-cortical (CSTC) circuits accounts for the covariances observed in our sample. The degree to which SEM predicted volumetric covariance in the CSTC circuit depended on whether we controlled for age and whole brain volume in the analyses. Removing the effects of age worsened the fit of the model, pointing to a possible developmental component in establishing connections within CSTC circuits. These modeling techniques may prove useful in the future for the study of structural networks in disease populations.

The psychobiology of obsessive-compulsive disorder: how important is the role of disgust?

Psychobiologic models of obsessive-compulsive disorder (OCD) have focused on cortico-striatal-thalamic-cortical (CTSC) circuits, noting normal function in cognitive and motoric procedural strategies. Such models have relied on the classification of OCD as an anxiety disorder, seldom exploring other relevant emotions. Based on the hypothesis that a central emotion in OCD is disgust, the authors review the literature on its psychobiology and its relevance to current models of OCD. There are important parallels between the psychobiology of OCD and that of disgust. Obsessive- compulsive disorder may be conceptualized in terms of a false contamination alarm in which disgust plays a crucial organizing or embodying role, not only at a basic brain level, but also in terms of the psychosocial aspects of the disorder. Just as psychobiologic models of panic disorder and post- traumatic stress disorder have been strengthened by the inclusion of preclinical work on amygdala-mediated fear conditioning, so findings on disgust and its mediating CSTC circuits may generate useful hypotheses for OCD research.

The cognitive-affective neuroscience of obsessive-compulsive disorder.

There is substantial evidence that obsessive-compulsive disorder (OCD) is mediated by specific cortico-striatal- thalamic-cortical (CTSC) circuits. Here we discuss very recent publications that address the following questions: How does damage to CSTC circuitry come about?; What are the neurochemical systems involved in mediating this circuitry?; and What are the implications of such damage for understanding the pathogenesis and management of OCD? A cognitive-affective neuroscience perspective is helpful in advancing our understanding of the role of these circuits in OCD and the dysfunctional procedural strategies that appear to characterize this disorder. Furthermore, this model is becoming integrated with a range of data including brain imaging, genetic, immunologic, and neurochemical findings.