Level Of Central Nervous System Research Articles

Identification of CNS-specific biomarkers for monitoring the progression of multiple sclerosis

Abstract There are currently no reliable methods for assessing the progression of multiple sclerosis (MS) from the relapsing-remitting to the secondary progressive form. This gap in knowledge hinders the ability for therapeutic intervention and results in continued relapses and physiological deterioration. To begin to address the urgent need for biomarkers of progressive MS we investigated proteome changes over the disease course of progressive experimental autoimmune encephalomyelitis (EAE) in NOD mice as a preclinical model of the disease. Our lab has pioneered a high-throughput quantitative proteomic technique, which we have previously used to quantify expression levels of central nervous system (CNS) proteins over the course of monophasic EAE in C57BL/6 mice, producing a predictive protein biomarker fingerprint for clinical relapses. In this study, we utilized this established proteomic technique as well as bioinformatics tools to prioritize key proteins whose expression level in the CNS correlated specifically with the progressive phase of disease in the NOD EAE model. Bioinformatics analysis identified 11 proteins with differential expression in the CNS of mice across the time course of NOD EAE, three of which are CNS specific. We are seeking to detect corollary changes in these three CNS-specific proteins in the serum, pointing to a minimally invasive means of monitoring disease progress and measuring drug efficacy. Our studies will provide a proof-of-concept for identifying homologous human biomarkers to guide treatment in individual patients. Furthermore, our results may provide insights into mechanisms that contribute to disease pathology and offer additional therapeutic targets for slowing the progression of MS.

Characterization of the Effects of L-4-Chlorokynurenine on Nociception in Rodents.

These studies show that systemic administration of the prodrug 4-Cl-KYN produces high central nervous system levels of 7-Cl-KYNA, a potent and highly selective antagonist of the NMDA receptor. Compared with other drugs tested, 4-Cl-KYN has robust antinociceptive effects with a better side effect profile, highlighting its potential for treating hyperpathic pain states.

Drug prescription appropriateness in the elderly: an Italian study.

PurposeCorrect drug prescription in the elderly is a difficult task that requires careful survey of the current pharmacological therapies. In this article, we reviewed the drug prescriptions provided to 860 persons aged 65 years or over, residing in a small city of Lombardy, Italy.MethodsSubjects were recruited from a local nursing home, the Pavia and Vigevano Neuropsychological Center for Alzheimer’s Disease, general practitioners’ offices, and the local University of the Third Age. For each patient, the amount of potentially inappropriate prescriptions (PIPs), sedative and anticholinergic load (SL and AL, respectively), and drug–drug interactions were evaluated.ResultsWidespread polypharmacy, giving rise to 10.06% of PIPs in the whole collection of prescriptions, was observed. In particular, PIPs mainly concern drugs acting at the central nervous system level, mostly benzodiazepines and antipsychotics. Moreover, approximately one-fourth of the subjects had an elevated SL and approximately one-tenth a high AL. Drug–drug interactions were frequent (266 requiring medical attention), up to five for each single patient. Of concern was the underuse of antidementia drugs: only 20 patients received a cholinesterase inhibitor or memantine, although 183 patients were potentially suitable for this treatment.ConclusionThese results demonstrate the need to develop novel strategies aimed at improving the quality of drug prescription.

Adaptive and Behavioral Changes in Kynurenine 3-Monooxygenase Knockout Mice: Relevance to Psychotic Disorders

BackgroundKynurenine 3-monooxygenase converts kynurenine to 3-hydroxykynurenine, and its inhibition shunts the kynurenine pathway—which is implicated as dysfunctional in various psychiatric disorders—toward enhanced synthesis of kynurenic acid, an antagonist of both α7 nicotinic acetylcholine and N-methyl-D-aspartate receptors. Possibly as a result of reduced kynurenine 3-monooxygenase activity, elevated central nervous system levels of kynurenic acid have been found in patients with psychotic disorders, including schizophrenia. MethodsIn the present study, we investigated adaptive—and possibly regulatory—changes in mice with a targeted deletion of Kmo (Kmo–/–) and characterized the kynurenine 3-monooxygenase–deficient mice using six behavioral assays relevant for the study of schizophrenia. ResultsGenome-wide differential gene expression analyses in the cerebral cortex and cerebellum of these mice identified a network of schizophrenia- and psychosis-related genes, with more pronounced alterations in cerebellar tissue. Kynurenic acid levels were also increased in these brain regions in Kmo–/– mice, with significantly higher levels in the cerebellum than in the cerebrum. Kmo–/– mice exhibited impairments in contextual memory and spent less time than did controls interacting with an unfamiliar mouse in a social interaction paradigm. The mutant animals displayed increased anxiety-like behavior in the elevated plus maze and in a light/dark box. After a D-amphetamine challenge (5 mg/kg, intraperitoneal), Kmo–/– mice showed potentiated horizontal activity in the open field paradigm. ConclusionsTaken together, these results demonstrate that the elimination of Kmo in mice is associated with multiple gene and functional alterations that appear to duplicate aspects of the psychopathology of several neuropsychiatric disorders.

Cranial dural permeability of inflammatory nociceptive mediators: Potential implications for animal models of migraine.

Background Application of inflammatory mediators to the cranial dura has been used as a method to activate and sensitize neurons in the meningeal sensory pathway in preclinical behavioral studies of headache mechanisms. However, the relatively high concentrations and volumes used in these studies raise the question of whether the applied agents might pass through the dura to act directly on central neurons, thus bypassing the dural afferent pathway. Methods We used a radiolabeling approach to quantify the meningeal permeability of two of the inflammatory mediators, 5-HT and PGE2, when applied to the cranial dura as part of an inflammatory mixture used in preclinical headache models. Results Both agents could be detected in samples taken four hours after dural application in the cerebrospinal fluid (CSF) and, in measurements made only for PGE2, in the central nervous system (CNS) as well. Based on our measurements, we made estimates of the CSF and CNS levels that would be attained with the higher concentrations and volumes of 5HT and PGE2 that were exogenously applied in previous pre-clinical headache studies. These estimated levels were comparable to or larger than normal endogenous levels, potentially large enough to have physiological effects. Conclusions The finding that the cranial meninges are permeable to the two tested inflammatory mediators PGE2 and 5-HT raises some uncertainty about whether the behavioral changes observed in prior pre-clinical headache studies with these as well as other agents can be attributed entirely to the activation of dural nociceptors, particularly when the agents are applied at concentrations several orders of magnitude above physiological levels.

Insomnia in Chinese Medicine: The Heart of the Matter.

Chronic insomnia affects a significant proportion of the general population worldwide, and is associated with several serious medical conditions. From the Western scientific literature, hyper-arousal (on the cognitive-emotional, behavioral, autonomic, or central nervous system level) is a final common pathway involved in its pathogenesis. However, from a Chinese medicine (CM) perspective, it is the Heart, capitalized to denote the functional system as described in CM theory, that is the key organ involved in insomnia due to its role as the "seat of consciousness." This article explores how insomnia is understood from the CM perspective, in particular the role of the Heart, and some of the neurophysiological evidence that supports these ancient theoretical understandings. The potential role of the vagus nerve and its relationship with the (biomedical) heart and CM Heart is also examined. Finally, some of the evidence in association with mechanisms of action of acupuncture in insomnia, in particular its impact on cardiovascular variables associated with insomnia, is presented, along with findings of systematic reviews.

EP 108. EEG-based arousal markers for individualized treatment in obsessive compulsive disorder

Objectives Cognitive behavioral therapy (CBT), selective serotonin reuptake inhibitors (SSRIs) and their combination are effective in the treatment of obsessive–compulsive disorder (OCD). Based on findings of increased levels of central nervous system (CNS)-arousal in OCD, aim of this study was to test the hypothesis that electroencephalogram (EEG)-based CNS- arousal markers differ for responders compared to non-responders to a therapy. Further goal was to identify specific response- predictors for either CBT, SSRI treatment or a combined therapy. Methods CNS-arousal during a fifteen-minute resting state condition was assessed by means of electroencephalogram (EEG)-vigilance stages using VIGALL (Vigilance Algorithm Leipzig) from 51 unmedicated patients suffering from OCD. Clinical Global Impression (CGI) and Yale-Brown Obsessive Compulsive Scale (YBOCS) were used to assess response or non-response after three to six months following therapy with either CBT ( n = 22), SSRI ( n = 12) or combination of both ( n = 17). Results Responders showed significant lower amounts of high CNS-arousal stage W (ANOVA F = 4.8; p 0.02 ). The percentage of W-stages also negatively correlated with YBOCS changes after treatment ( r = −0.32, p 0.03 ). Further, CNS-arousal as assessed by amounts of vigilance stages also allowed discrimination between response to one of the three different treatment arms CBT, SSRIs or the combination of both. Discussion The results of lower CNS-arousal in responders in comparison to non-responders in treatment of OCD suggest, that EEG-based markers might be useful for individualizing treatment in OCD, e.g. for decisions of early augmentation. Especially the fact that CNS-arousal markers were different for responders to either CBT, SSRI or combined therapy underlines the possible predictive value of EEG-based markers. Prospective studies in this field are strongly warranted.

Predictors of central vestibular disorders from videonystagmography tests

Abstract The diagnosis of central vestibular lesion is challenging and sometimes there is an overlap in symptoms and signs with a peripheral vestibular lesion. In some selected cases, dizziness is the only presenting symptom and in other patients, mild neurological symptoms as numbness are ignored. Videonystagmography (VNG) is considered a useful method for diagnosing vertigo of peripheral origin; however, not all the patients with central vertigo can be diagnosed easily. Benign paroxysmal positional vertigo and central positional vertigo share common criteria. The aim of the present study is to assess the usefulness of different VNG tests as predictors of central vestibular disorders, to determine the criteria that differentiate central positional nystagmus from the peripheral type, and to attempt to relate the abnormality in different VNG tests to certain central nervous system (CNS) levels. A retrospective study was carried out on 51 patients with possible central vestibular disorders from VNG tests battery and were referred for an MRI for further assessment. According to MRI results, the patients were divided into group A (31 patients), the group with manifest MRI findings, and group B, the group with free MRI (20 patients). Different VNG tests were compared between both groups. Three predictors of CNS lesion by VNG were determined: fixation index (FI), oculomotor tests, and central positional nystagmus; there was a statistically significant difference between both groups in FI, oculomotor tests, and central positional nystagmus, and apogeotropic criteria or nystagmus in multiple plains. There was no relation between any of those predictors and specific levels in the CNS. VNG tests are a good diagnostic tool to differentiate between peripheral and central vestibular lesions. The inclusion of FI and positional tests to the oculomotor tests increases the sensitivity of the VNG. In some cases, it is difficult to distinguish between benign paroxysmal positional vertigo and central positional vertigo; apogeotropic nystagmus and nystagmus in multiple plains should raise the suspicion of CNS lesion. Anterior circulation ischemia may lead to chronic vertigo symptoms. Central vestibular vertigo could be caused by dysfunction or excitation of various structures in the CNS including the vestibular cortex.

PTU-132 Autonomic Function Scores and Sleep Disturbance Scores in FGID Patients with and Without Upper Limb Temperature Dissociation

Introduction Background Central autonomic dysfunction may result in a unilateral dissociation of temperature sensation, and is increasingly being investigated in FGIDs. The autonomic function score (AFS) and sleep disturbance score (SDS) are accepted measures in aiding the diagnosis of autonomic dysfunction. Aims To assess and compare the AFS and SDS of consecutive FGID patients attending a specialist clinic, with and without upper limb temperature dissociation. Methods Clinic attenders with a FGID diagnosis were sequentially invited to undertake the temperature sensation test. This comprised holding a bag of ice in both hands for 30 seconds and stating if they perceived the temperature to be either unilaterally colder or bilaterally equally cold. Unilaterally colder was considered to be evidence of upper limb temperature sensory dissociation. Patients had their total AFS calculated as follows: headaches, dizziness, sweating and sleep disturbance were severity scored using a Likert scale of 0–3, the sum of which is the AFS. The mean total AFS and mean sleep disturbance score were compared between patients with and without temperature dissociation. Results 173 patients completed the AFS and 7 were excluded from the analysis for incomplete data leaving 166. 115 patients (69%) had upper limb temperature dissociation (group 1) compared to 51 (31%) with normal temperature sensation bilaterally (group 2). The mean total AFS in group 1 was 6.35 [SD 3.17] compared with 5.01 [SD 2.87] in group 2 [P = 0.009]. Mean sleep disturbance scores were not significantly different between the groups. Conclusion FGID patients with upper limb temperature dissociation appear to have a significantly higher autonomic function score than those without temperature dissociation. This seems to suggest that the cause of the temperature dissociation may well be due to autonomic dysfunction acting at a central nervous system level. These results are an interesting observation from our clinic but are likely to be limited by inherent methodological biases in the data collection. Further investigation of autonomic function scores and temperature dissociation are required in formal and appropriately powered clinical research studies to validate these findings. Disclosure of Interest None Declared

A randomized controlled study to evaluate the effect of bexarotene on amyloid-β and apolipoprotein E metabolism in healthy subjects

IntroductionWe conducted a phase Ib proof of mechanism trial to determine whether bexarotene (Targretin) increases central nervous system (CNS) apolipoprotein E (apoE) levels and alters Aβ metabolism in normal healthy individuals with the APOE ε3/ε3 genotype. MethodsWe used stable isotope labeling kinetics (SILK-ApoE and SILK-Aβ) to measure the effect of bexarotene on the turnover rate of apoE and Aβ peptides and stable isotope spike absolute quantitation (SISAQ) to quantitate their concentrations in the cerebrospinal fluid (CSF). Normal subjects were treated for 3 days with bexarotene (n = 3 women, 3 men, average 32 years old) or placebo (n = 6 women, average 30.2 years old) before administration of C13-leucine and collection of plasma and CSF over the next 48 hours. Bexarotene concentrations in plasma and CSF were also measured. ResultsOral administration of bexarotene resulted in plasma levels of 1 to 2 μM; however, only low nM levels were found in CSF. Bexarotene increased CSF apoE by 25% but had no effect on metabolism of Aβ peptides. DiscussionBexarotene has poor CNS penetration in normal human subjects. Drug treatment resulted in a modest increase in CSF apoE levels. The absence of an effect on Aβ metabolism is likely reflective of the low CNS levels of bexarotene achieved. This study documents the utility of SILK-ApoE technology in measuring apoE kinetics in humans. Trial RegistrationThis trial is registered at clinicaltrials.gov (NCT02061878).

Several synthetic progestins disrupt the glial cell specific-brain aromatase expression in developing zebra fish

The effects of some progestins on fish reproduction have been recently reported revealing the hazard of this class of steroidal pharmaceuticals. However, their effects at the central nervous system level have been poorly studied until now. Notwithstanding, progesterone, although still widely considered primarily a sex hormone, is an important agent affecting many central nervous system functions. Herein, we investigated the effects of a large set of synthetic ligands of the nuclear progesterone receptor on the glial-specific expression of the zebrafish brain aromatase (cyp19a1b) using zebrafish mechanism-based assays. Progesterone and 24 progestins were first screened on transgenic cyp19a1b-GFP zebrafish embryos. We showed that progesterone, dydrogesterone, drospirenone and all the progesterone-derived progestins had no effect on GFP expression. Conversely, all progestins derived from 19-nortesterone induced GFP in a concentration-dependent manner with EC50 ranging from the low nM range to hundreds nM. The 19-nortestosterone derived progestins levonorgestrel (LNG) and norethindrone (NET) were further tested in a radial glial cell context using U251-MG cells co-transfected with zebrafish ER subtypes (zfERα, zfERβ1 or zfERβ2) and cyp19a1b promoter linked to luciferase. Progesterone had no effect on luciferase activity while NET and LNG induced luciferase activity that was blocked by ICI 182,780. Zebrafish-ERs competition assays showed that NET and LNG were unable to bind to ERs, suggesting that the effects of these compounds on cyp19a1b require metabolic activation prior to elicit estrogenic activity. Overall, we demonstrate that 19-nortestosterone derived progestins elicit estrogenic activity by inducing cyp19a1b expression in radial glial cells. Given the crucial role of radial glial cells and neuro-estrogens in early development of brain, the consequences of exposure of fish to these compounds require further investigation.

Identifying biomarkers for monitoring progression of multiple sclerosis

Abstract There are currently no reliable methods for assessing the progression of multiple sclerosis (MS) from the relapsing-remitting to the secondary progressive form. This gap in knowledge hinders the ability for therapeutic intervention and ultimately results in continued relapses and physiological deterioration. To begin to address the urgent need for biomarkers of progressive MS we investigated proteome changes over the disease course of progressive experimental autoimmune encephalomyelitis (EAE) in NOD mice as a preclinical model of the disease. Our lab has pioneered a novel high-throughput quantitative proteomic technique which we used to quantify expression levels of central nervous system (CNS) proteins over the course of monophasic EAE in C57.BL/6 mice. We utilized bioinformatics tools to prioritize key proteins whose expression level correlated specifically with the progressive phase of disease in the NOD EAE model. Importantly, we were able to detect corollary changes in these CNS-specific proteins in the serum, pointing to a minimally invasive means of monitoring disease progress and measuring drug efficacy. Our studies will provide a proof-of-concept for identifying homologous human biomarkers to guide treatment in individual patients. Furthermore, our results may provide insights into mechanisms that contribute to disease pathology and offer additional therapeutic targets for slowing the progression of MS.

Music as a driver for brain plasticity

Sensory-motor skills of musicians have some specific qualities: learning begins at an early age in a playful atmosphere. Routines for stereotyped movements are rehearsed for extended periods of time with gradually increasing degrees of complexity. Via auditory feedback, the motor performance is extremely controllable by both, performer and audience. All movements are strongly linked to emotions, – pleasure or anxiety –, processed by the limbic system. These specific circumstances seem to play an important role for plastic adaptation at several levels of the central nervous system. There is a dark side to the increasing specialization and prolonged training of modern musicians, namely loss of control and degradation of skilled hand movements, a disorder referred to as musicians’ cramp or focal dystonia. Neuroimaging studies point to dysfunctional (or maladaptive) neuroplasticity as its cause. I will present new data on functional plasticity in musicians. It seems that early training leads to clearly more efficient networks as compared to late start of training. With respect to Musician’s dystonia, there are different categories of motor disturbances either linked to psychological traits, or to basal ganglia dysfunction. I will present a model accounting for our findings concerning the triggering factors of musician’s dystonia.

Neuroimmune Crosstalk in CNS Disorders: The Histamine Connection.

The neuroimmune system represents a dense network of biochemical signals associated with neurotransmitters, neuropeptides, neurohormones, cytokines, chemokines, and growth factors synthesized in neurons, glial cells and immune cells, to maintain systemic homeostasis. Endogenous and/or exogenous, noxious stimuli in any tissue are captured by sensor cells to inform the brain; likewise, signals originating at the central nervous system (CNS) level are transmitted to peripheral immune effectors which react to central stimuli. This multidirectional information system makes it possible for the CNS to respond to peripheral damage and for alterations in brain function to be reflected in peripheral immune changes. Different CNS disorders, such as anxiety, depression, psychosis, stroke, Alzheimer's disease, Parkinson's disease, attention-deficit hyperactivity disorder, migraine, epilepsy, vascular dementia, mental retardation, cerebrovascular encephalopathy, multiple sclerosis, brain tumors, cranial nerve neuropathies, mental retardation and post-traumatic brain injury exhibit changes in CD3, CD4, CD7, HLA-DR, CD25, CD28, and CD56 immune markers. Histamine is an important pleiotropic factor in neuroimmune regulation. This biogenic amine shows age-and sex-dependent changes in the CNS, and is significantly altered, together with interleukin- 1β and TNF-α, in Alzheimer's disease and other neurodegenerative disorders in which neuroinflammation appears to be an aggravating phenotype. Therapeutic intervention to halt progression of deleterious neuroinflammatory reactions in CNS disorders is a major challenge for molecular pharmacology in the future.

N-acetyl proline-glycine-proline: implications for neurological disorders.

N-acetyl proline-glycine-proline: implications for neurological disorders.

Therapeutical Neurotargeting via Magnetic Nanocarrier: Implications to Opiate-Induced Neuropathogenesis and NeuroAIDS.

Magnetite (Fe3O4) is the most commonly and extensively explored magnetic nanoparticles (MNPs) for drug-targeting and imaging in the field of biomedicine. Nevertheless, its potential application as safe and effective drug-carrier for CNS (Central Nervous System) anomalies is very limited. Previous studies have shown an entangled epidemic of opioid use and HIV infection and increased neuropathogenesis. Opiate such as morphine, heroine, etc. are used frequently as recreational drugs. Existing treatments to alleviate the action of opioid are less effective at CNS level due to impermeability of therapeutic molecules across brain barriers. Thus, development of an advanced nanomedicine based approach may pave the way for better treatment strategies. We herein report magnetic nanoformulation of a highly selective and potent morphine antagonist, CTOP (D-Pen-Cys-Tyr-DTrp-Orn-Thr-Pen-Thr-NH2), which is impenetrable to the brain. MNPs, synthesized in size range from 25 to 40 nm, were characterized by Transmission electron microscopy and assembly of MNPs-CTOP nanoformulations were confirmed by FTIR spectroscopy and fluorescent detection. Flow-cytometry analysis showed that biological efficacy of this nanoformulation in prevention of morphine induced apoptosis in peripheral blood mononuclear cells remains equivalent to that of free CTOP. Similarly, confocal microscopy reveals comparable efficacy of free and MNPs bound CTOP in protecting modulation of neuronal dendrite and spine morphology during morphine exposure and morphine-treated HIV infection. Further, typical transmigration assay showed increased translocation of MNPs across in vitro blood-brain barrier upon exposure of external magnetic force where barrier integrity remains unaltered. Thus, the developed nanoformulation could be effective in targeting brain by application of external magnetic force to treat morphine addiction in HIV patients.

Affective Modulation of Brain and Autonomic Responses in Patients With Fibromyalgia.

Emotional dysregulation and abnormal processing of affective information are thought to play a significant role for the maintenance of pain in fibromyalgia. The motivational priming hypothesis states that negative emotions could increase pain via activation of the aversive system, thus leading to an affective modulation of defensive reflexes. Nevertheless, little is known about peripheral and central correlates of affective reflex modulation in fibromyalgia. Thirty patients with fibromyalgia and 30 healthy individuals were asked to view three video clips from a self-perspective to induce specific mood states. Video clips consisted of the same virtual walk through different locations of a park under three affective environments (unpleasant, pleasant, and neutral). Startle eyeblink reflex and heart rate response elicited by abrupt startle noises, as well as heart rate variability and electroencephalography (EEG) oscillations were recorded when participants were passively viewing the virtual environments. Patients with fibromyalgia rated all environments as more negative and arousing than did healthy controls (p values < .05). Nevertheless, startle eyeblink reflex and heart rate response were lower in patients with fibromyalgia than in healthy controls when viewing all three environments (p values < .05). Patients with fibromyalgia also displayed lower heart rate variability, as well as higher EEG power (2-22 Hz) during all environments than did healthy controls (p values < .05). Patients with fibromyalgia were characterized by relevant deficits in affective modulation of startle and cardiac responses, heart rate variability, and EEG power spectra in response to sustained induction of affective states. These findings suggest an alteration of emotional and attentional aspects of information processing at subjective, autonomic, and central nervous system levels.

Homocysteine and Neuropsychiatric Disease

Elevated homocysteine (HCY) is a risk factor for a variety of vascular and neuropsychiatric pathologies. Although long recognized as a contributor to disease, and as an amino acid that is destructive to cell integrity and DNA, agents that were developed specifically to lower homocysteine were not available until very recently. Those agents are mainly composed of B vitamins in their metabolized forms, as they function as coenzymes in the pathway of homocysteine degradation. It is no surprise that hyperhomocysteinemia is most often the result of low B6, B9, or B12 levels; however, blood levels may not always reflect levels in the central nervous system (CNS), where HCY metabolism is most critical. HCY levels may also be normal in the periphery but not reflect CNS levels, where vascular, neuronal, and DNA damage is occurring. This article discusses the basics of HCY, its proposed mechanisms of toxicity, and the clinical consequences of hyperhomocysteinemia. [ Psychiatr Ann . 2015;45(9):463–468.]

Post-transcriptional modifications caused by TDP-43 mutations in mouse and man.

TDP43 is a ubiquitously expressed prevalently nuclear protein involved in RNA splicing, RNA stability and miRNA processing. Amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD) and inclusion body myopathy (IBM) are characterized by TDP43 being depleted from the nucleus and accumulating in cytoplasmic inclusions, which are the pathological hallmark of the disease – these diseases are also defined as “TDP43 proteinopathies”. Mutations in TDP43 have been found to be causative of a proportion of ALS cases reinforcing the primary importance of this molecule in disease pathogenesis. The pathogenic mechanism by which TDP43 acts is unclear, and both loss of nuclear function (LOF) and gain of function (GOF) mechanisms have been proposed. We here discuss how we used muscle tissue, which provides high quality RNA of patient disease tissue, to investigate the consequences of TDP43 mislocalization. Further, we characterize two novel mouse TDP43 mutant lines, carrying ENU-induced mutations in order to study the effects of TDP43 mutations expressed at physiological levels in the mammalian central nervous system. One mutation (deltaRNA) strongly reduces the RNA-binding capacity of the protein; the second mutation (C-TERM) is in the glycine-rich C-terminal domain where the majority of human pathogenic mutations are found. Our results underline the importance of studying models with physiological expression levels of TDP43 mutations and shed light on the different effects on RNA metabolism caused by the TDP43 loss and gain of function.

Cerebrospinal fluid markers reveal intrathecal inflammation in progressive multiple sclerosis

The management of complex patients with neuroimmunological diseases is hindered by an inability to reliably measure intrathecal inflammation. Currently implemented laboratory tests developed >40 years ago either are not dynamic or fail to capture low levels of central nervous system (CNS) inflammation. Therefore, we aimed to identify and validate biomarkers of CNS inflammation in 2 blinded, prospectively acquired cohorts of untreated patients with neuroimmunological diseases and embedded controls, with the ultimate goal of developing clinically useful tools. Because biomarkers with maximum utility reflect immune phenotypes, we included an assessment of cell specificity in purified primary immune cells. Biomarkers were quantified by optimized electrochemiluminescent immunoassays. Among markers with cell-specific secretion, soluble CD27 is a validated biomarker of intrathecal T-cell activation, with an area under the receiver operating characteristic curve of 0.97. Comparing the quantities of cerebrospinal fluid (CSF) immune cells and their respective cell-specific soluble biomarkers (released by CSF cells as well as their counterparts in CNS tissue) provided invaluable information about stationary CNS immune responses, previously attainable via brain biopsy only. Unexpectedly, progressive and relapsing-remitting multiple sclerosis (MS) patients have comparable numbers of activated intrathecal T and B cells, which are preferentially embedded in CNS tissue in the former group. The cell-specific biomarkers of intrathecal inflammation may improve diagnosis and management of neuroimmunological diseases and provide pharmacodynamic markers for future therapeutic developments in patients with intrathecal inflammation that is not captured by imaging, such as in progressive MS.