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[Objective] The purpose of this study was to examine the levels of cerebrospinal fluid (CSF) apolipoprotein E (apoE) species in Alzheimer’s disease (AD) patients., [Methods] We analyzed two CSF cohorts of AD and control individuals expressing different APOE genotypes. Moreover, CSF samples from the TgF344-AD rat model were included. Samples were run in native- and SDS-PAGE under reducing or non-reducing conditions (with or without β-mercaptoethanol). Immunoprecipitation combined with mass spectrometry or western blotting analyses served to assess the identity of apoE complexes., [Results] In TgF344-AD rats expressing a unique apoE variant resembling human apoE4, a ~35-kDa apoE monomer was identified, increasing at 16.5 months compared with wild-types. In humans, apoE isoforms form disulfide-linked dimers in CSF, except apoE4, which lacks a cysteine residue. Thus, controls showed a decrease in the apoE dimer/monomer quotient in the APOE ε3/ε4 group compared with ε3/ε3 by native electrophoresis. A major contribution of dimers was found in APOE ε3/ε4 AD cases, and, unexpectedly, dimers were also found in ε4/ε4 AD cases. Under reducing conditions, two apoE monomeric glycoforms at 36 kDa and at 34 kDa were found in all human samples. In AD patients, the amount of the 34-kDa species increased, while the 36-kDa/34-kDa quotient was lower compared with controls. Interestingly, under reducing conditions, a ~100-kDa apoE complex, the identity of which was confirmed by mass spectrometry, also appeared in human AD individuals across all APOE genotypes, suggesting the occurrence of aberrantly resistant apoE aggregates. A second independent cohort of CSF samples validated these results., [Conclusion] These results indicate that despite the increase in total apoE content the apoE protein is altered in AD CSF, suggesting that function may be compromised., This work was supported by grants from the Fondo de Investigaciones Sanitarias (FIS; PI15/00665 and PI19-01359, co-funded by the Fondo Europeo de Desarrollo Regional, FEDER “Investing in your future”), from the CIBERNED (Instituto de Salud Carlos III, Spain) and from the Direcció General de Ciència I Investigació, Generalitat Valenciana (AICO/2021/308). We also acknowledge financial support from the Spanish Ministerio de Economía y Competitividad, through the “Severo Ochoa” Programme for Centres of Excellence in R&D (SEV-2017-0723). MPL is supported by a BEFPI fellowship from the Generalitat Valenciana. This study was also supported by the FIS, Contratos para la intensificación de la actividad investigadora en el SNS from Instituto de Salud Carlos III (PI14/01126, PI17/01019, PI20/01473, and INT16/00171 to JF; PI18/00435 to DA; PI14/1561, PI17/01896, and PI20/1330 to AL). This work was also supported by the CIBERNED program (Program 1, Alzheimer Disease to AL and SIGNAL study, www.signalstudy.es), the National Institutes of Health (NIA grants 1R01AG056850-01A1, R21AG056974, and R01AG061566 to JF), and Departament de Salut de la Generalitat de Catalunya, Pla Estratègic de Recerca i Innovació en Salut (SLT002/16/00408 to AL). This work was also supported by Generalitat de Catalunya (SLT006/17/00119 to JF and SLT006/17/00125 to DA). This study was also supported by the Spanish Ministry of Science, Innovation and Universities MICINN/FEDER (PID2019-107738RB-I00, to FA, a fellowship to ISD and the “Maria Maeztu Unit of Excellence Program”) and Instituto de Salud Carlos III (PI18/00893 to GS, co-funded by ERDF, “A way to make Europe”). HZ is a Wallenberg Scholar supported by grants from the Swedish Research Council (#2018-02532); the European Research Council (#681712); the Swedish State Support for Clinical Research (#ALFGBG-720931); the Alzheimer Drug Discovery Foundation (ADDF), USA (#201809-2016862); the AD Strategic Fund and the Alzheimer’s Association (#ADSF-21-831376-C, #ADSF-21-831381-C, and #ADSF-21-831377-C); the Olav Thon Foundation; the Erling-Persson Family Foundation; Stiftelsen för Gamla Tjänarinnor; Hjärnfonden, Sweden (#FO2019-0228); the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 860197 (MIRIADE); the European Union Joint Programme – Neurodegenerative Disease Research (JPND2021-00694); and the UK Dementia Research Institute at UCL (UKDRI-1003). KB is supported by the Swedish Research Council (#2017-00915); the Alzheimer Drug Discovery Foundation (ADDF), USA (#RDAPB-201809-2016615); the Swedish Alzheimer Foundation (#AF-742881); Hjärnfonden, Sweden (#FO2017-0243); the Swedish state under the agreement between the Swedish government and the County Councils, the ALF-agreement (#ALFGBG-715986); the European Union Joint Program for Neurodegenerative Disorders (JPND2019-466-236); the National Institute of Health (NIH), USA (grant #1R01AG068398-01); and the Alzheimer’s Association 2021 Zenith Award (ZEN-21-848495).

19 Pág. Centro de Investigación en Sanidad Animal (CISA), Triggering receptor expressed on myeloid cells 2 (TREM2) is an innate immune cell surface receptor that regulates microglial function and is involved in the pathophysiology of several neurodegenerative diseases. Its soluble form (sTREM2) results from shedding of the TREM2 ectodomain. The role of TREM2 in prion diseases, a group of rapidly progressive dementias remains to be elucidated. In the present study, we analysed the expression of TREM2 and its main sheddase ADAM10 in the brain of sporadic Creutzfeldt-Jakob disease (sCJD) patients and evaluated the role of CSF and plasma sTREM2 as a potential diagnostic marker of prion disease. Our data indicate that, compared to controls, TREM2 is increased in sCJD patient brains at the mRNA and protein levels in a regional and subtype dependent fashion, and expressed in a subpopulation of microglia. In contrast, ADAM10 is increased at the protein, but not the mRNA level, with a restricted neuronal expression. Elevated CSF sTREM2 is found in sCJD, genetic CJD with mutations E200K and V210I in the prion protein gene (PRNP), and iatrogenic CJD, as compared to healthy controls (HC) (AUC = 0.78-0.90) and neurological controls (AUC = 0.73-0.85), while CSF sTREM2 is unchanged in fatal familial insomnia. sTREM2 in the CSF of cases with Alzheimer's disease, and multiple sclerosis was not significantly altered in our series. CSF sTREM2 concentrations in sCJD are PRNP codon 129 and subtype-related, correlate with CSF 14-3-3 positivity, total-tau and YKL-40, and increase with disease progression. In plasma, sTREM2 is increased in sCJD compared with HC (AUC = 0.80), displaying positive correlations with plasma total-tau, neurofilament light, and YKL-40. We conclude that comparative study of TREM2 in brain and biological fluids of prion diseases reveals TREM2 to be altered in human prion diseases with a potential value in target engagement, patient stratification, and disease monitoring., We thank the HUB-ICO-IDIBELL-Biobank and the CERCA Programme of the Generalitat de Catalunya for institutional support. This study was funded by the Instituto Carlos III (Grants Number CP16/00041 and PI19/00144) to FL. This project was also funded by la Fundació La Marató de TV3 (Grants No. 201821-30, 201821-31and 201821-32 to FL, JCE and EV, respectively) and by the Fondo Europeo de Desarrollo Regional (FEDER) through the Interreg V-A España-Francia-Andorra (POCTEFA 2014–2020) programme (at 65%) to IF. AVP is supported by the Beatriu de Pinós programme (2018-BP-00129) from the Ministry of Business and nowledge of the Government of Catalonia, and cofunded by the EU Horizon 2020 programme under an MSCA grant agreement (801370). HZ is a Wallenberg Scholar supported by grants from the Swedish Research Council (#2018-02532), the European Research Council (#681712), Swedish State Support for Clinical Research (#ALFGBG-720931), the Alzheimer Drug Discovery Foundation (ADDF), USA (#201809-2016862), and the UK Dementia Research Institute at UCL. KB is supported by the Swedish Research Council (#2017-00915), the Swedish State Support for Clinical Research (ADDF), USA (#RDAPB-201809-2016615), the Swedish Alzheimer Foundation (#AF-742881), Hjärnfonden, Sweden (#FO2017-0243), the Swedish state under the agreement between the Swedish government and the County Councils, the ALF-agreement (#ALFGBG-715986), and the European Union Joint Programme for Neurodegenerative Disorders (JPND2019-466-236). AL is supported by the Ministero della Salute, Italy, for the national surveillance of Creutzfeldt-Jakob disease. This research was also supported in part by the Alberta Synergies in Alzheimer’s and Related Disorders (SynAD) programme which is funded by the Alzheimer Society of Alberta and Northwest Territories through their Hope for Tomorrow programme and the University Hospital Foundation. SynAD operates in partnership with the Neuroscience and Mental Health Institute at the University of Alberta. JADR was supported by grants from the Spanish Ministry of Science, Innovation and Universities (MICINN/FEDER) (RTI2018-099773-B-I00), the CERCA Programme, and by the Commission for Universities and Research of the Department of Innovation, Universities, and Enterprise of the Generalitat de Catalunya (SGR2017-648) and CIBERNED (CMED2018-2)

Brain changes occurring in aging can be indexed by biomarkers. We used cluster analysis to identify subgroups of cognitively unimpaired individuals (n = 99, 64–93 years) with different profiles of the cerebrospinal fluid biomarkers beta amyloid 1–42 (Aβ42), phosphorylated tau (P-tau), total tau, chitinase-3-like protein 1 (YKL-40), fatty acid binding protein 3 (FABP3), and neurofilament light (NFL). Hippocampal volume and memory were assessed across multiple follow-up examinations covering up to 6.8 years. Clustering revealed one group (39%) with more pathological concentrations of all biomarkers, which could further be divided into one group (20%) characterized by tauopathy and high FABP3 and one (19%) by brain β-amyloidosis, high NFL, and slightly higher YKL-40. The clustering approach clearly outperformed classification based on Aβ42 and P-tau alone in prediction of memory decline, with the individuals with most tauopathy and FABP3 showing more memory decline, but not more hippocampal volume change. The results demonstrate that older adults can be classified based on biomarkers beyond amyloid and tau, with improved prediction of memory decline.

Prior studies have described distinct patterns of brain gray matter and white matter alterations in Alzheimer's disease (AD) and frontotemporal lobar degeneration (FTLD), as well as differences in their cerebrospinal fluid (CSF) biomarkers profiles. We aim to investigate the relationship between early‐onset AD (EOAD) and FTLD structural alterations and CSF biomarker levels. We included 138 subjects (64 EOAD, 26 FTLD, and 48 controls), all of them with a 3T MRI brain scan and CSF biomarkers available (the 42 amino acid‐long form of the amyloid‐beta protein [Aβ42], total‐tau protein [T‐tau], neurofilament light chain [NfL], neurogranin [Ng], and 14‐3‐3 levels). We used FreeSurfer and FSL to obtain cortical thickness (CTh) and fraction anisotropy (FA) maps. We studied group differences in CTh and FA and described the “AD signature” and “FTLD signature.” We tested multiple regression models to find which CSF‐biomarkers better explained each disease neuroimaging signature. CTh and FA maps corresponding to the AD and FTLD signatures were in accordance with previous literature. Multiple regression analyses showed that the biomarkers that better explained CTh values within the AD signature were Aβ and 14‐3‐3; whereas NfL and 14‐3‐3 levels explained CTh values within the FTLD signature. Similarly, NfL levels explained FA values in the FTLD signature. Ng levels were not predictive in any of the models. Biochemical markers contribute differently to structural (CTh and FA) changes typical of AD and FTLD.

Since amyotrophic lateral sclerosis cases exhibit significant heterogeneity, we aim to investigate the association of lipid composition of plasma and CSF with amyotrophic lateral sclerosis diagnosis, its progression and clinical characteristics. Lipidome analyses would help to stratify patients on a molecular basis. For this reason, we have analysed the lipid composition of paired plasma and CSF samples from amyotrophic lateral sclerosis cases and age-matched non-amyotrophic lateral sclerosis individuals (controls) by comprehensive liquid chromatography coupled to mass spectrometry. The concentrations of neurofilament light chain—an index of neuronal damage—were also quantified in CSF samples and plasma. Amyotrophic lateral sclerosis versus control comparison, in a moderate stringency mode, showed that plasma from cases contains more differential lipids (n = 122 for raw P, Sol et al. report that in case–control approaches, plasma lipidomic profile is richer in differences than CSF. Interestingly, patients with faster progression show decreased plasmatic triacylglycerides and phospholipids, compatible with hypermetabolism. They conclude that the plasma lipidomic profile holds promise for patient stratification., Graphical Abstract Graphical Abstract

Background: Perivascular spaces (PVS) have an important role in the elimination of metabolic waste from the brain. It has been hypothesized that the enlargement of PVS (ePVS) could be affected by pathophysiological mechanisms involved in Alzheimer's disease (AD), such as abnormal levels of CSF biomarkers. However, the relationship between ePVS and these pathophysiological mechanisms remains unknown. Objective: We aimed to investigate the association between ePVS and CSF biomarkers of several pathophysiological mechanisms for AD. We hypothesized that ePVS will be associated to CSF biomarkers early in the AD continuum (i.e., amyloid positive cognitively unimpaired individuals). Besides, we explored associations between ePVS and demographic and cardiovascular risk factors. Methods: The study included 322 middle-aged cognitively unimpaired participants from the ALFA + study, many within the Alzheimer's continuum. NeuroToolKit and Elecsys® immunoassays were used to measure CSF Aβ42, Aβ40, p-tau and t-tau, NfL, neurogranin, TREM2, YKL40, GFAP, IL6, S100, and α-synuclein. PVS in the basal ganglia (BG) and centrum semiovale (CS) were assessed based on a validated 4-point visual rating scale. Odds ratios were calculated for associations of cardiovascular and AD risk factors with ePVS using logistic and multinomial models adjusted for relevant confounders. Models were stratified by Aβ status (positivity defined as Aβ42/40 < 0.071). Results: The degree of PVS significantly increased with age in both, BG and CS regions independently of cardiovascular risk factors. Higher levels of p-tau, t-tau, and neurogranin were significantly associated with ePVS in the CS of Aβ positive individuals, after accounting for relevant confounders. No associations were detected in the BG neither in Aβ negative participants. Conclusions: Our results support that ePVS in the CS are specifically associated with tau pathophysiology, neurodegeneration, and synaptic dysfunction in asymptomatic stages of the Alzheimer's continuum. The project leading to these results has received funding from “la Caixa” Foundation (ID 100010434), under agreement LCF/PR/GN17/50300004 and the Alzheimer’s Association and an international anonymous charity foundation through the TriBEKa Imaging Platform project (TriBEKa-17–519007). Additional support has been received from the Health Department of the Catalan Government (Health Research and Innovation Strategic Plan (PERIS) 2016–2020 grant# SLT002/16/00201) and the Universities and Research Secretariat, Ministry of Business and Knowledge of the Catalan Government under the grant no. 2017-SGR-892. All CRG authors acknowledge the support of the Spanish Ministry of Science, Innovation and Universities to the EMBL partnership, the Centro de Excelencia Severo Ochoa, and the CERCA Programme/Generalitat de Catalunya. NV-T is funded by a post-doctoral grant, Juan de la Cierva Programme (FJC2018-038085-I), Ministry of Science and Innovation– Spanish State Research Agency. MS-C received funding from the European Union’s Horizon 2020 Research and Innovation Program under the Marie Sklodowska-Curie action grant agreement No 752310, and currently receives funding from Instituto de Salud Carlos III (PI19/00155) and from the Spanish Ministry of Science, Innovation and Universities (Juan de la Cierva Programme grant IJC2018-037478-I). EMAU is supported by the Spanish Ministry of Science, Innovation and Universities—Spanish State Research Agency (RYC2018-026053-I). OGR is supported by the Spanish Ministry of Science, Innovation and Universities (FJCI-2017–33437). JDG is supported by the Spanish Ministry of Science and Innovation (RYC-2013–13054). KB is supported by the Swedish Research Council (#2017–00915), the Alzheimer Drug Discovery Foundation (ADDF), USA (#RDAPB-201809–2016615), the Swedish Alzheimer Foundation (#AF-742881), Hjärnfonden, Sweden (#FO2017-0243), the Swedish state under the agreement between the Swedish government and the County Councils, the ALF-agreement (#ALFGBG-715986), and European Union Joint Program for Neurodegenerative Disorders (JPND2019-466–236). HZ is a Wallenberg Scholar supported by grants from the Swedish Research Council (#2018–02532), the European Research Council (#681712), the Swedish state under the agreement between the Swedish government and the County Councils, the ALF-agreement (#ALFGBG-720931), the ADDF, USA (#201809–2016862), and the UK Dementia Research Institute at UCL.

Alpha-synucleinopathies, such as Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA), are a class of neurodegenerative diseases. A diagnosis may be challenging because clinical symptoms partially overlap, and there is currently no reliable diagnostic test available. Therefore, we aimed to identify a suitable marker protein in cerebrospinal fluid (CSF) to distinguish either between different types of alpha-synucleinopathies or between alpha-synucleinopathies and controls. In this study, the regulation of different marker protein candidates, such as alpha-synuclein (a-Syn), neurofilament light chain (NfL), glial fibrillary acidic protein (GFAP), and total tau (tau) in different types of alpha-synucleinopathies, had been analyzed by using an ultrasensitive test system called single-molecule array (SIMOA). Interestingly, we observed that CSF-NfL was significantly elevated in patients with DLB and MSA compared to patients with PD or control donors. To differentiate between groups, receiver operating characteristic (ROC) curve analysis resulted in a very good diagnostic accuracy as indicated by the area under the curve (AUC) values of 0.87-0.92 for CSF-NfL. Furthermore, we observed that GFAP and tau were slightly increased either in DLB or MSA, while a-Syn levels remained unregulated. Our study suggests NfL as a promising marker to discriminate between different types of alpha-synucleinopathies or between DLB/MSA and controls.

[cat] Els errors congènits del metabolisme són malalties d’origen genètic amb una baixa prevalença dins de la població general. Aquestes malalties solen estar causades per mutacions en gens que provoquen un mal funcionament de diversos enzims o cofactors que controlen alguna ruta metabòlica de l’organisme. Moltes d’aquestes malalties poden presentar símptomes neurològics que poden comprometre el correcte desenvolupament del sistema nerviós central (SNC), particularment durant el període neonatal i la infància. Tot i la seva baixa prevalença, cada any es descriuen nous desordres i fenotips, motiu pel qual cal seguir investigant en nous biomarcadors. Les monoamines, com la dopamina i la serotonina, són neurotransmissors que intervenen en el correcte desenvolupament de les funcions motores, perceptives, cognitives i emocionals del SNC. Diverses condicions genètiques i ambientals han demostrat que afecten de manera primària el metabolisme d’ambdós neurotransmissors o bé d’aquells cofactors essencials per la correcta biosíntesis de la dopamina i serotonina. La detecció quantitativa de metabòlits de la dopamina i serotonina (àcid homovaníl·lic (HVA), i àcid 5-hidroxiindolacètic (5-HIAA), respectivament), així com dels seus cofactors (vitamina B6 i tetrahidrobiopterina), en líquid cefaloraquidi (LCR) es considera una important prova bioquímica pel diagnòstic i seguiment de les malalties mencionades anteriorment. El LCR és un fluid biològic que s’obté mitjançant una punció lumbar, intervenció invasiva que ha de realitzar personal especialitzat i seguint un estricte protocol estandarditzat. Els pacients amb dèficit de dopamina i serotonina d’origen genètic reben tractament amb precursors d’aquests neurotransmissors. El correcte seguiment del tractament es fa per puncions lumbars successives fent que els pacients s’hagin de sotmetre varies vegades a una punció lumbar. Seria de gran utilitat poder trobar un biomarcador relacionat amb la dopamina i/o serotonina, que no estigués influenciat per factors externs, que fos un reflex del que succeeix al SNC i que pogués determinar- se en d’altres fluids biològics menys invasius, com l’orina. Per això vam avaluar la utilitat de la melatonina en orina, una hormona sintetitzada per la glàndula pineal (localitzada al cervell) a partir de la serotonina cerebral. Una vegada és sintetitzada, és alliberada a la sang perquè pugui ser metabolitzada i posteriorment eliminada per orina com a 6-sulfatoximelatonina (aMT6s). D’altra banda, tot i que l’obtenció del LCR es realitza mitjançant un procediment protocol·litzat, pot està subjecte a alteracions preanalítiques, com ara la contaminació hemàtica per punció traumàtica. Està descrit que aquesta contaminació hemàtica té efectes sobre la concentració de glucosa i de proteïnes en el LCR, però no hi ha estudis on es descrigui l’efecte en d’altres biomarcadors que s’analitzen en LCR per l’estudi de patologies neurometabòliques. Tenint en compte aquests antecedents en aquesta tesi hem desenvolupat 3 objectius: 1. Avaluar l’efecte de la contaminació hemàtica en mostres de LCR a l’hora de realitzar i interpretar les anàlisis que realitzem al laboratori: aminoàcids, metabòlits dels neurotransmissors, 5-metiltetrahidrofolat, vitamina B6, pterines (neopterina i biopterina) i tiamina. Els resultats es van publicar a la revista “Fluids and Barriers of CNS” sota el títol Effect of blood contamination of cerebrospinal fluid on amino acids, biogenic amines, pterins and vitamins (2019). 2. Establir valors de referència de l’aMT6s (metabòlit de la melatonina) en orina en població sana, per tal d’avaluar si pot ser un bon biomarcador perifèric de l’estat de la serotonina cerebral en pacients amb mutacions patològiques de gens relacionats en la biosíntesis de la serotonina. Els resultats d’aquest estudi es van publicar a la revista “Scientific Reports” sota el títol Urinary sulphatoxymelatonin as a biomarker of serotonin status in biogenic amine-deficient patients (2017). 3. Avaluar retrospectivament el fenotip clínic i els perfils bioquímics d'amines biògenes en LCR de pacients diagnosticats genèticament d’una malaltia mitocondrial. Els resultats d’aquest estudi es van publicar a la revista “Journal of Inherited Metabolism Diseases” sota el títol Cerebrospinal fluid monoamines, pterins, and folate in patients with mitochondrial diseases: systematic review and hospital experience (2018). Paral·lelament a això vam poder publicar un protocol estandarditzat de l’anàlisi de les monoamines i els seus cofactors en LCR per HPLC a la revista “Nature Protocols” (Analysis of human cerebrospinal fluid monoamines and related cofactors by HPLC. 2017).

Intraventricular hemorrhage is a common cause of morbidity and mortality in premature infants. The rupture of the germinal zone into the ventricles entails loss of neural stem cells and disturbs the normal cytoarchitecture of the region, compromising late neurogliogenesis. Here we demonstrate that neural stem cells can be easily and robustly isolated from the hemorrhagic cerebrospinal fluid obtained during therapeutic neuroendoscopic lavage in preterm infants with severe intraventricular hemorrhage. Our analyses demonstrate that these neural stem cells, although similar to human fetal cell lines, display distinctive hallmarks related to their regional and developmental origin in the germinal zone of the ventral forebrain, the ganglionic eminences that give rise to interneurons and oligodendrocytes. These cells can be expanded, cryopreserved, and differentiated in vitro and in vivo in the brain of nude mice and show no sign of tumoral transformation 6 months after transplantation. This novel class of neural stem cells poses no ethical concerns, as the fluid is usually discarded, and could be useful for the development of an autologous therapy for preterm infants, aiming to restore late neurogliogenesis and attenuate neurocognitive deficits. Furthermore, these cells represent a valuable tool for the study of the final stages of human brain development and germinal zone biology., Germinal zone neural stem cells (Gz‐NSC) are isolated from the hemorrhagic cerebrospinal fluid of preterm infants with severe intraventricular hemorrhage. These cells express ventral and posterior forebrain markers, can be differentiated and do not cause tumors. Gz‐NSC represent a valuable tool for the development of new cell therapies and the study of human Gz biology.

The elevation of neopterin in cerebrospinal fluid (CSF) has been reported in several neuroinflammatory disorders. However, it is not expected that neopterin alone can discriminate among different neuroinflammatory etiologies. We conducted an observational retrospective and case–control study to analyze the CSF biomarkers neopterin, total proteins, and leukocytes in a large cohort of pediatric patients with neuroinflammatory disorders. CSF samples from 277 subjects were included and classified into four groups: Viral meningoencephalitis, bacterial meningitis, acquired immune-mediated disorders, and patients with no-immune diseases (control group). CSF neopterin was analyzed with high-performance liquid chromatography. Microbiological diagnosis included bacterial CSF cultures and several specific real-time polymerase chain reactions. Molecular testing for multiple respiratory pathogens was also included. Antibodies against neuronal and glial proteins were tested. Canonical discriminant analysis of the three biomarkers was conducted to establish the best discriminant functions for the classification of the different clinical groups. Model validation was done by biomarker analyses in a new cohort of 95 pediatric patients. CSF neopterin displayed the highest values in the viral and bacterial infection groups. By applying canonical discriminant analysis, it was possible to classify the patients into the different groups. Validation analyses displayed good results for neuropediatric patients with no-immune diseases and for viral meningitis patients, followed by the other groups. This study provides initial evidence of a more efficient approach to promote the timely classification of patients with viral and bacterial infections and acquired autoimmune disorders. Through canonical equations, we have validated a new tool that aids in the early and differential diagnosis of these neuroinflammatory conditions.

Alzheimer&rsquo, s type dementia (AD) exhibits clinical heterogeneity, as well as differences in disease progression, as a subset of patients with a clinical diagnosis of AD progresses more rapidly (rpAD) than the typical AD of slow progression (spAD). Previous findings indicate that low cerebrospinal fluid (CSF) content of cell-free mitochondrial DNA (cf-mtDNA) precedes clinical signs of AD. We have now investigated the relationship between cf-mtDNA and other biomarkers of AD to determine whether a particular biomarker profile underlies the different rates of AD progression. We measured the content of cf-mtDNA, beta-amyloid peptide 1&ndash, 42 (A&beta, ), total tau protein (t-tau) and phosphorylated tau (p-tau) in the CSF from a cohort of 95 subjects consisting of 49 controls with a neurologic disorder without dementia, 30 patients with a clinical diagnosis of spAD and 16 patients with rpAD. We found that 37% of controls met at least one AD biomarker criteria, while 53% and 44% of subjects with spAD and rpAD, respectively, did not fulfill the two core AD biomarker criteria: high t-tau and low A&beta, in CSF. In the whole cohort, patients with spAD, but not with rpAD, showed a statistically significant 44% decrease of cf-mtDNA in CSF compared to control. When the cohort included only subjects selected by A&beta, and t-tau biomarker criteria, the spAD group showed a larger decrease of cf-mtDNA (69%), whereas in the rpAD group cf-mtDNA levels remained unaltered. In the whole cohort, the CSF levels of cf-mtDNA correlated positively with A&beta, and negatively with p-tau. Moreover, the ratio between cf-mtDNA and p-tau increased the sensitivity and specificity of spAD diagnosis up to 93% and 94%, respectively, in the biomarker-selected cohort. These results show that the content of cf-mtDNA in CSF correlates with the earliest pathological markers of the disease, A&beta, and p-tau, but not with the marker of neuronal damage t-tau. Moreover, these findings confirm that low CSF content of cf-mtDNA is a biomarker for the early detection of AD and support the hypothesis that low cf-mtDNA, together with low A&beta, and high p-tau, constitute a distinctive CSF biomarker profile that differentiates spAD from other neurological disorders.

Amyotrophic lateral sclerosis (ALS) has variable clinical course and fatal outcome. Since inflammation plays a role in the pathogenesis of ALS, chitinase-3-like protein 1 or YKL40 has been assessed as putative biomarker of disease progression. YKL40 mRNA levels are increased in anterior horn of the spinal cord (P=0.004) in sporadic ALS (sALS) cases when compared with age-matched controls. These correlate with increased mRNA expression of microglial markers AIF1 and CD68 in the spinal cord in sALS (P=0.044 and P=0.000, respectively). YKL40 mRNA and protein expression had a tendency to increase in post-mortem frontal cortex area 8 (P=0.06 and P=0.08, respectively). Yet YKL40 immunoreactivity is restricted to a subpopulation of astrocytes in these regions. YKL40 protein levels, as revealed by enzyme-linked immunosorbent assay (ELISA), are significantly increased in the CSF in sALS (n=86) compared with age-matched controls (n=21) (P=0.045). Higher levels are found in patients with fast progression when compared with patients with slow and normal progression (P=0.008 and P=0.004, respectively), and correlates with ALS-FRS-R slope (P=0.000). Additionally, increased protein levels of neurofilament light chain (NF-L) are also found in sALS (P=0.000); highest values are found in patients with fast progression when compared with cases with slow and normal progression (P=0.005 and P=0.000, respectively), and also correlate with ALS-FRS-R slope (P=0.000). Pearson's correlation test linked positively the increased levels of YKL40 with increased NF-L levels (P=0.013). These data point to YKL40 and NF-L protein levels in the CSF as a good biomarker combination of disease progression in sALS.

The exploration of accurate diagnostic markers for differential diagnosis of neurodegenerative diseases is an ongoing topic. A previous study on cerebrospinal fluid (CSF)-mitochondrial malate dehydrogenase 1 (MDH1) in sporadic Creutzfeldt&ndash, Jakob disease (sCJD) patients revealed a highly significant upregulation of MDH1. Here, we measured the CSF levels of MDH1 via enzyme-linked immunosorbent assay in a cohort of rare genetic prion disease cases, such as genetic CJD (gCJD) cases, exhibiting the E200K, V210I, P102L (Gerstmann&ndash, Str&auml, ussler&ndash, Scheinker syndrome (GSS)), or D178N (fatal familial insomnia (FFI)) mutations in the PRNP. Interestingly, we observed enhanced levels of CSF-MDH1 in all genetic prion disease patients compared to neurological controls (without neurodegeneration). While E200K and V210I carriers showed highest levels of MDH1 with diagnostic discrimination from controls of 0.87 and 0.85 area under the curve (AUC), FFI and GSS patients exhibited only moderately higher CSF-MDH1 levels than controls. An impact of the PRNP codon 129 methionine/valine (MV) genotype on the amount of MDH1 could be excluded. A correlation study of MDH1 levels with other neurodegenerative marker proteins revealed a significant positive correlation between CSF-MDH1 concentration with total tau (tau) but not with 14-3-3 in E200K, as well as in V210I patients. In conclusion, our study indicated the potential use of MDH1 as marker for gCJD patients which may complement the current panel of diagnostic biomarkers.

Identification of neurodegeneration-monitoring biomarkers would be of great clinical value for Alzheimer's disease (AD) diagnosis. Using N- or C-terminal antibodies, we studied the pro-survival synaptic effector, Kidins220, in the brain and cerebrospinal fluid (CSF) of controls and AD patients. Only the N-terminal antibody showed a positive correlation between Kidins220 and phosphorylated tau in AD brains. Using this antibody, Kidins220 was detected in CSF from AD patients where it positively correlated with CSF phosphorylated tau and tau. This study highlights the potential of Kidins220 as a CSF biomarker in AD., T.I. is funded by SAF2014-52737-P (Ministerio de Economía y Competitividad, Spain), P2010/BMD-2331-Neurodegmodels (Comunidad de Madrid, Madrid, Spain); A.L. is funded by PI11/3035 and PI14/1561 provided by FEDER (European Funds for Regional Development) and Instituto de Salud Carlos III (Ministerio de Economía y Competitividad, Spain). T.I. and A.L. are also funded by Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED) and CIBERNED cooperarive project 2013/07 (Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Spain). A.G.M is funded by a contract from CIBERNED-2013/07; O.B. is funded by the Miguel Servet Associate Investigator Project Grant (CP13/0091) and FIS (PI15/00058) provided by FEDER and Instituto de Salud Carlos III (Ministerio de Economía y Competitividad, Spain). The cost of this publication has been paid in part by FEDER funds.

Background Leucine‐rich kinase 2 (LRRK2)‐linked Parkinson's disease (PD) is clinically indistinguishable from idiopathic PD (IPD). A pleiotropic neuropathology has been recognized but the majority of studies in LRRK2 p.G2019S patients reveal Lewy‐type synucleinopathy as its principal histological substrate. To date no in vivo biomarkers of synucleinopathy have been found in LRRK2 mutation carriers. Objectives We used real‐time quaking‐induced conversion (RT‐QuIC) technique to assess the presence of alpha‐synuclein (a‐syn) aggregates in cerebrospinal fluid (CSF) of LRRK2 p.G2019S carriers. Methods CSF samples of 51 subjects were analyzed: 15 LRRK2 p.G2019S PD, 10 IPD, 16 LRRK2 p.G2019S nonmanifesting carriers (NMC) and 10 healthy controls. The presence of parkinsonism and prodromal symptoms was assessed in all study subjects. Results Forty percent (n = 6) LRRK2‐PD, and 18.8% (n = 3) LRRK2‐NMC had a positive a‐syn RT‐QuIC response. RT‐QuIC detected IPD with 90% sensitivity and 80% specificity. No clinical differences were detected between LRRK2‐PD patients with positive and negative RT‐QuIC. A positive RT‐QuIC result in LRRK2‐NMC occurred in a higher proportion of subjects meeting the Movement Disorder Society research criteria for prodromal PD. Interpretation RT‐QuIC detects a‐syn aggregation in CSF in a significant number of patients with LRRK2‐PD, but less frequently than in IPD. A small percentage of LRRK2‐NMC tested also positive. If appropriately validated in long‐term studies with large number of mutation carriers, and hopefully, postmortem or in vivo confirmation of histopathology, RT‐QuIC could contribute to the selection of candidates to receive disease modifying drugs, in particular treatments targeting a‐syn deposition.

La degeneración Lobular Frontotemporal (DLFT) es una constructo neuropatológico heterogéneo que incluye diferentes entidades neuropatológicas caracterizadas por una neurodegeneración prominente de los lóbulos frontales y temporales. A pesar de que algunos síndromes clínicos han demostrado predecir el diagnóstico neuropatológica de DLFT, la capacidad de los síndromes clínicos para predecir el diagnóstico neuropatológico es limitada. En esta tesis nos propusimos profundizar en los fundamentos fisiopatológicos y estructurales de la neurodegeneración relacionada con la DLFT mediante un estudio de biomarcadores multimodal que combinó: (i) marcadores clínicos de progresión de la enfermedad (CDR-FTLD, ALSFRS), (ii) biomarcadores en el líquido cefalorraquídeo relacionados con diferentes aspectos fisiopatológicos de la DLFT (Péptidos derivados de del procesamiento de la PPA, YKL-40 y la cadena ligera de neurofilamentos [NfL]) y (iii) el estudio mediante resonancia magnética cerebral de la macroestructura (grosor cortical) y la microestructura (difusividad media cortical) cortical en los síndromes relacionados con la DLFT (S-DLFT). Los resultados de esta tesis expanden el conocimiento previo sobre la fisiopatología de la DLFT al mostrar que (i) los niveles de sAPPβ, YKL-40 y NfL son útiles desde un punto de vista diagnóstico para la diferenciación de los S-DLFT de los pacientes con la enfermedad de Alzheimer y de los controles cognitivamente sanos; (ii) en ausencia de la fisiopatología de la enfermedad de Alzheimer, los niveles en el líquido cefalorraquídeo de los péptidos derivados del procesamiento de la PPA (Aβ1-42, Aβ1-40, Aβ1-38 y sAPPβ) se encuentran globalmente disminuidos en los S-DLFT y correlacionan con el grosor cortical a diferencia de lo observado en la enfermedad de Alzheimer donde solo se evidencia un descenso de los niveles de Aβ1-42; (iii) los niveles en el líquido cefalorraquídeo del biomarcador de actividad astroglial YKL-40 se encuentran aumentados en los S-DLFT y, en los pacientes con esclerosis lateral amiotrófica, pueden ser útiles para predecir la evolución clínica; (iv) la difusividad media cortical, un nuevo biomarcador de neuroimagen en resonancia magnética cerebral es más sensible para la detección de los primeras cambios corticales relacionados con la DLFT. En definitiva, estos hallazgos mejoran nuestro entendimiento de la neurodegeneración relacionada con la DLFT y abren nuevas puertas para el estudio personalizado de los diferentes S-DLFT. Frontotemporal lobar degeneration (FTLD), is a heterogenic pathological construct encompassing multiple neuropathological conditions primarily affecting the frontal and temporal lobes. Although multiple clinical syndromes predict the neuropathological diagnosis of FTLD, clinical-pathological correlations are far from being perfect. Cerebrospinal and imaging biomarkers represent powerful tools for the study of FTLD pathophysiology and improve the diagnostic accuracy of FTLD and its differentiation from other diseases. This thesis aims to improve our understanding of the pathophysiological and structural underpinnings of FTLD-related neurodegeneration through a multimodal biomarker approach combining: (i) clinical markers of disease progression (i.e. CDR-FTLD, ALSFRS), (ii) CSF biomarkers related to different pathophysiological aspects of FTLD (APP-derived peptides, YKL-40 and NfL) and (iii) the MRI study of cortical macrostructure (cortical thickness) and microstructure (cortical mean diffusivity) in FTLD—related syndromes (FTLD-S). We provide novel insights into the pathophysiology of FTLD by showing that: (i) CSF levels of sAPPβ, YKL-40 and NFL alone or in combination had a good diagnostic accuracy to discriminate FTLD-S from healthy controls and patients with Alzheimer’s disease (ii) In the absence of Alzheimer’s disease pathophysiology, APP-derived peptides related to the so-called amyloidogenic pathway (Aβ1-42, Aβ1-40, Aβ1-38 and sAPPβ) are globally reduced in FTLD-S and correlate with cortical macrostructure (cortical thickness); importantly this pattern of APP-derived differed from the observed in Alzheimer’s disease were a selective decrease in the CSF levels of Aβ1-42 was observed; (iii) YKL-40, a biomarker related to astroglial activity, is increased in FTLD-S and their levels may be useful for the prediction of disease progression in the ALS-FTD continuum; and (iv) cortical mean diffusivity, a novel imaging biomarker is more sensitive that cortical thickness for the study of the earliest FTLD-related neurodegeneration. These findings add to our current understanding of FTLD pathophysiology and open new doors towards precision medicine approaches in FTLD-S.

Multiple sclerosis; SERPINA3 Esclerosi múltiple; SERPINA3 Esclerosis múltiple; SERPINA3 Objective To identify biomarkers associated with progressive phases of MS and with neuroprotective potential. Methods Combined analysis of the transcriptional and proteomic profiles obtained in CNS tissue during chronic progressive phases of experimental autoimmune encephalomyelitis (EAE) with the transcriptional profile obtained during the differentiation of murine neural stem cells into neurons. Candidate biomarkers were measured by ELISA in the CSF of 65 patients with MS (29 with relapsing-remitting MS [RRMS], 20 with secondary progressive MS, and 16 with primary progressive MS [PPMS]) and 30 noninflammatory neurologic controls (NINCs). Results Integrative analysis of gene and protein expression data identified 2 biomarkers, the serine protease inhibitor Serpina3n and the calcium-binding protein S100A4, which were upregulated in chronic progressive EAE and whose expression was induced during neuronal differentiation. Immunofluorescence studies revealed a primarily neuronal expression of S100A4 and Serpina3n during EAE. CSF levels of SERPINA3, the human ortholog of murine Serpina3n, and S100A4 were increased in patients with MS compared with NINCs (SERPINA3: 1,320 vs 838.6 ng/mL, p = 0.0001; S100A4: 1.6 vs 0.8 ng/mL, p = 0.02). Within the MS group, CSF SERPINA3 levels were significantly elevated in patients with progressive forms, mainly patients with PPMS compared with patients with RRMS (1,617 vs 1,129 ng/mL, p = 0.02) and NINCs (1,617 vs 838.6 ng/mL, p = 0.0001). Of interest, CSF SERPINA3 levels significantly correlated with CSF neurofilament light chain levels only in the PPMS group (r = 0.62, p = 0.01). Conclusion These results point to a role of SERPINA3 as a biomarker associated with the progressive forms of MS, particularly PPMS. This work was supported by grants from the Fondo de Investigación Sanitaria (FIS; grant number PI17/00596), Ministry of Science and Innovation, Spain; Generalitat de Catalunya Suport Grups de Recerca (2017 SGR 0527); and the Red Española de Esclerosis Múltiple (RD16/0015/0004) funded by the FIS.

Sporadic Creutzfeldt-Jakob disease (sCJD) is the most common form of human prion disease. It is invariably fatal and displays a short clinical disease stage. The key event in sCJD is the propagation of a beta-sheet rich conformer of the physiological PrPC protein, known as PrPSc. Neuropathological disease characteristics include gliosis, neuronal loss and spongiform degeneration; disease clinical manifestations refer to mental and visual disabilities, cognitive impairment, gait or limb ataxia, myoclonus and mutism. Definite sCJD diagnosis requires post-mortem brain material histopathological examination. However, highly certain pre-mortem differential diagnosis is desired to exclude other treatable disorders and to reduce disease transmission risks. Detection and/or quantification of cerebrospinal fluid (CSF) biomarkers reflecting neuronal damage and PrPC misfolding in the diseased brain significantly enhance pre-mortem diagnosis. Previously established and newly identified biomarkers are used towards this direction. Increased CSF Neurofilament light chain (NFL) concentrations have been reported in several neurological disorders, including prion diseases. In the present study, we analyzed CSF NFL levels in two independent patient cohorts, consisting of highly suspected sCJD cases that were further classified as sCJD or non-CJD according to established diagnostic criteria. CSF NFL concentrations were increased in sCJD compared to non-CJD cases in both cohorts (area under the curve (with 95% confidence interval) equal to 0.89 (0.82 to 0.97) and 0.86 (0.77 to 0.96), respectively. CSF NFL was associated neither to age nor to sex but correlated with total-tau concentrations in both cohorts. Overall, our data provide independent validation of CSF NFL utility in sCJD differential diagnosis.

Cerebrospinal fluid (CSF) total prion protein (t-PrP) is decreased in sporadic Creutzfeldt-Jakob disease (sCJD). However, data on the comparative signatures of t-PrP across the spectrum of prion diseases, longitudinal changes during disease progression, and levels in pre-clinical cases are scarce. T-PrP was quantified in neurological diseases (ND, n = 147) and in prion diseases from different aetiologies including sporadic (sCJD, n = 193), iatrogenic (iCJD, n = 12) and genetic (n = 209) forms. T-PrP was also measured in serial lumbar punctures obtained from sCJD cases at different symptomatic disease stages, and in asymptomatic prion protein gene (PRNP) mutation carriers. Compared to ND, t-PrP concentrations were significantly decreased in sCJD, iCJD and in genetic prion diseases associated with the three most common mutations E200K, V210I (associated with genetic CJD) and D178N-129M (associated with fatal familial insomnia). In contrast, t-PrP concentrations in P102L mutants (associated with the Gerstmann-Sträussler-Scheinker syndrome) remained unaltered. In serial lumbar punctures obtained at different disease stages of sCJD patients, t-PrP concentrations inversely correlated with disease progression. Decreased mean t-PrP values were detected in asymptomatic D178-129M mutant carriers, but not in E200K and P102L carriers. The presence of low CSF t-PrP is common to all types of prion diseases regardless of their aetiology albeit with mutation-specific exceptions in a minority of genetic cases. In some genetic prion disease, decreased levels are already detected at pre-clinical stages and diminish in parallel with disease progression. Our data indicate that CSF t-PrP concentrations may have a role as a pre-clinical or early symptomatic diagnostic biomarker in prion diseases as well as in the evaluation of therapeutic interventions.

Background: Alzheimer's disease (AD) is the main medical problem in older adults with Down syndrome (DS). Studies of cerebrospinal fluid (CSF) AD biomarkers are limited and the feasibility of lumbar puncture (LP) is controversial in this population. Objective: To analyze the frequency of complications after a LP in DS. Methods: We collected data from 80 adults with DS that underwent a LP within the Down Alzheimer Barcelona Neuroimaging Initiative. Demographics, cognitive status, headache history, and presence of complications after the LP were recorded in every subject. In 53 of them (active group), this information was collected following a semi-structured and validated protocol that actively looks for complications. Other variables related to the LP procedure were also recorded. A telephone interview to the caregiver was performed 5-7 days after the procedure to ask about complications. Data from 27 subjects (clinical practice group), from whom the presence of complications was obtained in a medical follow-up visit within the three months after the LP, were also included. Results: There were no adverse events in 90% of our participants. The most frequent complication was headache (6.25%); only one subject reported a typical post-lumbar puncture headache with moderate severity that required analgesic treatment. Dizziness (3.75%) and back pain (1.25%) were also reported. All the participants that reported complications belonged to the active group. Conclusion: LP can be safely performed to study CSF biomarkers in DS. The reported complications are qualitatively similar to the general population, but are less frequently reported, even when actively searched for.

Incipient Alzheimer’s Disease (AD) is characterized by a slow onset of clinical symptoms, with pathological brain changes starting several years earlier. Consequently, it is necessary to first understand and differentiate age-related changes in brain regions in the absence of disease, and then to support early and accurate AD diagnosis. However, there is poor understanding of the initial stage of AD; seemingly healthy elderly brains lose matter in regions related to AD, but similar changes can also be found in non-demented subjects having mild cognitive impairment (MCI). By using a Linear Mixed Effects approach, we modelled the change of 166 Magnetic Resonance Imaging (MRI)-based biomarkers available at a 5-year follow up on healthy elderly control (HC, n = 46) subjects. We hypothesized that, by identifying their significant variant (vr) and quasi-variant (qvr) brain regions over time, it would be possible to obtain an age-based null model, which would characterize their normal atrophy and growth patterns as well as the correlation between these two regions. By using the null model on those subjects who had been clinically diagnosed as HC (n = 161), MCI (n = 209) and AD (n = 331), normal age-related changes were estimated and deviation scores (residuals) from the observed MRI-based biomarkers were computed. Subject classification, as well as the early prediction of conversion to MCI and AD, were addressed through residual-based Support Vector Machines (SVM) modelling. We found reductions in most cortical volumes and thicknesses (with evident gender differences) as well as in sub-cortical regions, including greater atrophy in the hippocampus. The average accuracies (ACC) recorded for men and women were: AD-HC: 94.11%, MCI-HC: 83.77% and MCI converted to AD (cAD)-MCI non-converter (sMCI): 76.72%. Likewise, as compared to standard clinical diagnosis methods, SVM classifiers predicted the conversion of cAD to be 1.9 years earlier for females (ACC:72.5%) and 1.4 years earlier for males (ACC:69.0%).

We assessed the outcome of patients with drug resistant epilepsy and neuronal antibodies who underwent epilepsy surgery. Retrospective study, information collected with a questionnaire sent to epilepsy surgery centers. Thirteen patients identified, with antibodies to GAD (8), Ma2 (2), Hu (1), LGI1 (1) or CASPR2 (1). Mean age at seizure onset: 23 years. Five patients had an encephalitic phase. Three had testicular tumors and five had autoimmune diseases. All had drug resistant temporal lobe epilepsy (median: 20 seizures/month). MRI showed unilateral temporal lobe abnormalities (mainly hippocampal sclerosis) in 9 patients, bilateral abnormalities in 3, and was normal in 1. Surgical procedures included anteromesial temporal lobectomy (10 patients), selective amygdalohippocampectomy (1), temporal pole resection (1) and radiofrequency ablation of mesial structures (1). Perivascular lymphocytic infiltrates were seen in 7/12 patients. One year outcome available in all patients, at 3 years in 9. At last visit 5/13 patients (38.5%) (with Ma2, Hu, LGI1, and 2 GAD antibodies) were in Engel's classes I or II. Epilepsy surgery may be an option for patients with drug resistant seizures associated with neuronal antibodies. Outcome seems to be worse than that expected in other etiologies, even in the presence of unilateral HS. Intracranial EEG may be required in some patients. (C) 2016 Elsevier B.V. All rights reserved.

Background: studies conducted in animal models and humans suggest the presence of a dynamic equilibrium of amyloid-β (Aβ) peptide between cerebrospinal fluid (CSF) and plasma compartments. Objective: to determine whether plasma exchange (PE) with albumin replacement was able to modify Aβ concentrations in CSF and plasma as well as to improve cognition in patients with mild-moderate Alzheimer's disease (AD). Methods: in a multicenter, randomized, patient- and rater-blind, controlled, parallel-group, phase II study, 42 AD patients were assigned (1 : 1) to PE treatment or control (sham) groups. Treated patients received a maximum of 18 PE with 5% albumin (Albutein®, Grifols) with three different schedules: two PE/weekly (three weeks), one PE/weekly (six weeks), and one PE/bi- weekly (12 weeks), plus a six-month follow-up period. Plasma and CSF Aβ1-40 and Aβ1-42 levels, as well as cognitive, functional, and behavioral measures were determined. Results: CSF Aβ1-42 levels after the last PE compared to baseline were marginally higher in PE-treated group versus controls (adjusted means of variation: 75.3 versus -45.5 pg/mL; 95% CI: -19.8, 170.5 versus 135.1, 44.2; p = 0.072). Plasma Aβ1-42 levels were lower in the PE-treated group after each treatment period (p

Aproximadamente el 85% de los pacientes con esclerosis múltiple (EM) inician la enfermedad con un síndrome clínico aislado (CIS), pero se ha descrito que entre un 30 y un 85% de los pacientes que presentan un CIS finalmente desarrollan EM clínicamente definida (EMCD). Actualmente se utilizan la presencia de alteraciones en la resonancia magnética (RM) y la detección de bandas oligoclonales en el líquido cefalorraquídeo (LCR) como marcadores pronósticos en los pacientes con CIS. Aparte de estos factores, la evidencia sobre el posible papel de otros biomarcadores moleculares ha sido controvertida por falta de validación de los resultados. Con este objetivo llevamos a cabo la presente tesis doctoral. Mediante un abordaje de proteómica en pooles de muestras de LCR de pacientes con CIS que convertían a EMCD y pacientes que permanecían como CIS se buscaron proteínas asociadas a la conversión a EM. Las proteínas que se encontraron expresadas de forma diferencial se validaron en muestras individuales mediante técnicas alternativas como el ELISA y selected reaction monitoring (SRM). De las proteínas validadas se seleccionó la chitinase 3-like 1 (CHI3L1) para su validación en una cohorte independiente y numerosa de pacientes con CIS. Además, se determinó su expresión en plasma en las diferentes formas clínicas de la enfermedad y se estudió la asociación entre un polimorfismo en el promotor del gen de la CHI3L1 y la EM. Por otro lado, se estudió la expresión de la CHI3L1 en tejido cerebral de pacientes con EM así como en las poblaciones celulares del LCR. Finalmente, se realizó el modelo animal de la enfermedad, la encefalomielitis autoinmune experimental (EAE) en ratones knockout (KO) y wild-type (WT) para la CHI3L1 y se realizaron estudios histopatológicos y ensayos de proliferación para determinar la función de la proteína en la respuesta inmune. De las proteínas encontradas en el estudio de proteómica se validaron como biomarcadores asociados a la conversión a EM la CHI3L1, la semaforina 7A y la CNDP1. La CHI3L1 además, se validó en una cohorte numerosa de pacientes con CIS, encontrándose además una correlación entre los niveles de CHI3L1 y el tiempo de conversión y el tiempo hasta alcanzar un EDSS de 3. Por otro lado, los niveles plasmáticos de CHI3L1 se encontraron incrementados en los pacientes con formas progresivas de la enfermedad respecto a los controles sanos y los pacientes con formas recurrentes. Además, se encontró una asociación entre un polimorfismo en el promotor del gen de la CHI3L1 y las formas progresivas de la enfermedad. Cuando se estudió el papel de la proteína en el modelo animal no se observaron diferencias en la gravedad del curso clínico entre los ratones WT y KO para la proteína. Tampoco se encontraron diferencias a nivel histopatológico ni en la respuesta inmune. Con los resultados obtenidos se puede concluir que la CHI3L1, semaforina 7A y CNDP1 se han validado como biomarcadores asociados con la conversión a EMCD, y que los niveles de CHI3L1 en LCR tienen además claras implicaciones pronósticas en los pacientes con CIS. A diferencia del LCR, los niveles de CHI3L1 en plasma no parecen reflejar el grado de inflamación que tiene lugar en le SNC de los pacientes con EM, y se asocian con las formas progresivas de la enfermedad. Finalmente, los estudios realizados en el modelo animal de EM no sugieren un papel inmunoregulador de la CHI3L1, en cuanto a que la eliminación del gen Brp-39 en ratones no se asoció con cambios en el curso clínico de la EAE, cambios histopatológicos, o alteraciones en las respuestas inmunitarias., In about 85% of patients with multiple sclerosis (MS) the disease starts with a clinically isolated syndrome (CIS), but only 30 to 85% of patients with CIS finally develop clinically definite MS (CDMS). At present, only magnetic resonance (MR) abnormalities and presence of oligoclonal bands (BOC) in the cerebrospinal fluid (CSF) are used as prognostic biomarkers in CIS patients. Apart from these, the role of other molecular biomarkers is still controversial due to lack of validation. This thesis was conducted with the aim to identify and validate additional molecular biomarkers associated with the conversion to MS. A proteomic study with pooled CSF samples from patients with CIS who converted to CDMS and patients who remained as CIS was performed to identify biomarkers associated with conversion to MS. Those proteins that were found differentially expressed between groups were validated in individual samples by means of alternative techniques such as ELISA and selected reaction monitoring (SRM). Among the validated proteins, chitinase 3-like 1 (CHI3L1) was selected for validation in a large cohort of CIS patients. Furthermore, CHI3L1 levels were determined in plasma from patients with different clinical forms and a polymorphism in the promoter region of the CHI3L1 gene was investigated for its potential association with the disease. Additionally, the cell source of CHI3L1 was studied in brain tissue and CSF samples from MS patients. Finally, the animal model of the disease, the experimental autoimmune encephalomyelitis (EAE) was induced in wild-type (WT) and knock-out (KO) mice for the CHI3L1 protein. Histopathological and proliferative response studies were performed to assess the role of the protein in the immune response. Amongst the proteins obtained in the proteomics study only CHI3L1, semaphorin 7A and CDNP1 were validated. Furthermore, CHI3L1 was validated in a large cohort of CIS patients. A correlation between CSF CHI3L1 levels and time to conversion and time to EDSS 3 was also found. On the other hand, plasma levels of CHI3L1 were found increased in patients with progressive forms of MS compared to healthy controls and patients with relapse-onset. Additionally there was an association between the studied polymorphism and the progressive forms of the disease. When the protein role in the disease was studied in EAE no differences were observed in the clinical course between WT and KO mice. Also, no differences were found regarding the histopathology and the immune response. In conclusion the aggregate results confirm CHI3L1, semaphorin 7A and CNDP1 as biomarkers associated with conversion to CDMS in CIS patients, and CHI3L1 CSF levels also have clear prognostic implications in CIS patients. In contrast to CSF, plasma levels of CHI3L1 are not reflecting the inflammation degree in the CNS in patients with MS, and are associated with the progressive forms of the disease. Finally, the studies performed in the animal model of MS do not suggest an immunoregulatory role of CHI3L1, since the elimination of the Brp-39 gene is not associated with changes in the clinical course, histopathology or immune response.

© 2014 Urra, Miró, Chamorro and Planas. Brain proteins are detected in the cerebrospinal fluid (CSF) and blood of stroke patients and their concentration is related to the extent of brain damage. Antibodies against brain antigens develop after stroke, suggesting a humoral immune response to the brain injury. Furthermore, induced immune tolerance is beneficial in animal models of cerebral ischemia. The presence of circulating T cells sensitized against brain antigens, and antigen presenting cells (APCs) carrying brain antigens in draining lymphoid tissue of stroke patients support the notion that stroke might induce antigen-specific immune responses. After stroke, brain proteins that are normally hidden from the periphery, inflammatory mediators, and danger signals can exit the brain through several efflux routes. They can reach the blood after leaking out of the damaged blood-brain barrier (BBB) or following the drainage of interstitial fluid to the dural venous sinus, or reach the cervical lymph nodes through the nasal lymphatics following CSF drainage along the arachnoid sheaths of nerves across the nasal submucosa. The route and mode of access of brain antigens to lymphoid tissue could influence the type of response. Central and peripheral tolerance prevents autoimmunity, but the actual mechanisms of tolerance to brain antigens released into the periphery in the presence of inflammation, danger signals, and APCs, are not fully characterized. Stroke does not systematically trigger autoimmunity, but under certain circumstances, such as pronounced systemic inflammation or infection, autoreactive T cells could escape the tolerance controls. Further investigation is needed to elucidate whether antigen-specific immune events could underlie neurological complications impairing recovery from stroke., Authors receive financial support by the Spanish Ministries of Economy MINECO (SAF2011-30492) and Health (FIS PI12/01437), and the European Community FP7 (ERA-NET program PRI-PIMNEU-2011-1342). Francesc Miró is supported by the “Red Invictus” of the Instituto de Salud Carlos III (RD12/0014/0011)

Objetivos: Determinar los motivos de consulta en el servicio de urgencias de los niños portadores de válvula de derivación de líquido cefalorraquídeo (VDL), así como sus diagnósticos al alta, y comparar el consumo de recursos que generan con el de la población general. Métodos: Se revisan los datos referentes a la sintomatología, la actuación y el diagnóstico definitivo de todas las consultas realizadas en el servicio de urgencias por parte de los niños portadores de VDL, desde el 1 de abril de 2010 al 31 de marzo de 2011 (grupo 1). Se seleccionan todas las consultas generadas en el mismo año de un grupo de pacientes no portadores de VDL inmediatamente antes de las consultas de los portadores de VDL (grupo 2). El coste económico se valora mediante el número de consultas y las exploraciones complementarias. Resultados: Se incluyen 250 consultas de portadores de VDL, que corresponden a 99 pacientes (grupo 1). Los motivos de consulta más frecuentes son los vómitos, la fiebre y la cefalea, y los diagnósticos la infección respiratoria (22,6%) y la disfunción valvular (14%). El grupo 2 queda constituido por 250 niños, que realizan 549 consultas. Los pacientes del grupo 1 reciben más exploraciones complementarias por consulta que el grupo 2 (el 48 frente al 25%; p

Cognitive reserve capacity may increase tolerance of neurodegenerative processes. However, its role regarding amyloid-B (AB 42) deposition in cognitively normal subjects is not well understood. We aimed to investigate the association between areas showing A 42-related structural changes and cognitive reserve proxies in cognitively intact subjects showing normal or abnormal AB 42 cerebrospinal fluid (CSF) concentrations. Thirty-three subjects (aged 55-85) underwent lumbar puncture and high resolution anatomical magnetic resonance imaging analyzed by voxel-based morphometry and cortical thickness procedures. Subjects with abnormal A 42 CSF levels showed significant left hippocampal atrophy and greater cortical thinning in parietal, temporal, and frontal regions (including the supramarginal and the anterior cingulate gyrus) compared to subjects with normalA 42 CSF levels. Using a multivariate general linear model, we investigated the relationship between these areas and cognitive reserve proxies. We found a significant relationship between decreased volume of the left hippocampus or decreased cortical thickness of the right supramarginal gyrus and higher cognitive reserve proxies only in the group with abnormal A 42 CSF levels. Thus, subjects with abnormal A 42 CSF levels (which may be at a higher risk of developing Alzheimer's disease) and with high scores on cognitive reserve proxies may be tolerating a more advanced neurodegenerative process in critical cortical and subcortical regions. The present results emphasize the relevance of evaluating cognitive reserve proxies, as well as the importance of using neuroimaging techniques for early diagnosis in individuals with higher reserve.

Introducción. En los neonatos con infección congénita sintomática por citomegalovirus (CMV) la neuroimagen es la mejor herramienta disponible para predecir la evolución neurológica a largo plazo. Los hallazgos en líquido cefalorraquídeo (LCR) de estos pacientes han sido escasamente estudiados. La Beta2-microglobulina (β2-m) se encuentra elevada en el LCR de los recién nacidos con infecciones del sistema nervioso central (SNC). Objetivos. (1) Determinar si la concentración de β2-m en LCR está aumentada en los neonatos con infección congénita sintomática por CMV, (2) examinar su correlación con los hallazgos de neuroimagen y (3) evaluar su valor para predecir el neurodesarrollo. Métodos. Se incluyeron todos los pacientes con infección congénita sintomática por CMV nacidos entre 1993-2009 en el Hospital Universitario La Paz, Madrid. La población control estuvo formada por un grupo de neonatos en los que se determinó la β2-m en LCR como parte de los estudios por sospecha de sepsis, meningitis o encefalitis y que finalmente presentaron cultivos estériles y estado neurológico normal al alta. La β2-m se midió mediante inmunonefelomatría. Los hallazgos de neuroimagen (ecografía, TC y/o RM) se graduaron según una escala semicuantitativa validada (grados 0-3). Todos los pacientes fueron contactados para una evaluación sistemática de su neurodesarrollo a largo plazo, que incluyó: examen neurológico y, si parálisis cerebral, graduación según el GMFCS, evaluación cognitiva mediante las escalas BSID-III, WPPSI-III o WISC-IV, según la edad, y evaluación conductual mediante el ASEBA. Se registraron la presencia de epilepsia, déficit auditivo o visual. Para los pacientes no visitados a largo plazo, se revisaron las historias de Neurología o, si no recibían seguimiento, se realizó una entrevista dirigida a los padres. Las secuelas neurológicas se clasificaron según una graduación semicuantitativa (grados 1-3). Resultados. Fueron incluidos 26 pacientes con infección congénita sintomática por CMV, 20 de ellos con determinación de β2-m en LCR y todos con neuroimagen neonatal. Tres casos fallecieron, 20 de los supervivientes fueron evaluados según el protocolo de seguimiento y en los otros tres la evaluación se hizo mediante los subrogados. La β2-m se encontró elevada con respecto a los 93 neonatos de la población control (mediana 7,2 mg/L versus 1,6 mg/L, P, SUMMARY OF DOCTORAL THESIS: “CEREBROSPINAL FLUID BETA2-MICROGLOBULIN (CSF- β2-m) IN NEWBORNS WITH SYMPTOMATIC CONGENITAL CYTOMEGALOVIRUS INFECTION (SCCI): AN ACCURATE INDICATOR OF THE SEVERITY OF CENTRAL NERVOUS SYSTEM (CNS) INVOLVEMENT” Ana Alarcón Allen Introduction. In newborns with SCCI neuroimaging is the best predictor of long-term neurodevelopment. CSF findings have seldom been described. CSF-β2-m is increased in CNS infections. Aims. (1) To determine whether CSF- β2-m is increased in newborns with SCCI, (2) to examine its correlation with neuroimaging and (3) to evaluate its ability to predict neurodevelopment. Methods. All patients with SCCI born between 1993-2009 in La Paz University Hospital (Madrid) were included. A group of newborns who underwent CSF-β2-m determination as part of a sepsis/meningitis/meningoencephalitis workup and who finally had sterile cultures and a normal neurologic status comprised the control population. β2-m was measured by immunonephelometry. Neuroimaging was graduated by a validated scoring system (grades 0-3). Follow-up assessment included: neurologic examination and, if cerebral palsy, graduation by GMFCS, cognitive evaluation by BSID-III, WPPSI-III or WISC-IV, and behavioral assessment by ASEBA. Epilepsy, hearing or visual impairment were registered. For patients not contacted, follow-up data were obtained from the Neurology clinical records or from a parental interview. Neurologic disabilities were classified (grades 1-3). Results. Twenty-six patients were included. Twenty had CSF-β2-m an all cases had neonatal neuroimaging. Three cases died, 20 surviving patients were evaluated according to our follow-up protocol and 3 had surrogate follow-up. β2-m was elevated in patients with SCCI with respect to the control group (median 7,2mg/L versus 1,6mg/L, P

El objetivo principal de esta tesis es estudiar la circulación del LCR y la patología relacionada con la misma mediante una técnica de neuroimagen no invasiva.La Resonancia Magnética inicialmente permitía realizar estudios estructurales cerebrales de alta resolución, pero no estudios funcionales o dinámicos. En la actualidad se han desarrollado nuevas secuencias de RM, por ejemplo la secuencia en contraste de fase, que anulan el tejido estático a la vez que realzan las estructuras en movimiento como son la sangre y el LCR.La secuencia de RM en contraste de fase aporta dos herramientas muy útiles para el estudio del LCR: el estudio cualitativo y el estudio cuantitativo.El estudio cualitativo permite evaluar el movimiento del LCR mediante técnica de Cine RM. La secuencia en contraste de fase adquiere varias imágenes del LCR durante el ciclo cardiaco. Si estas imágenes se visualizan secuencialmente, permitirán observar el paso del LCR a través de los distintos estrechos y agujeros de los ventrículos y en el espacio subaracnoideo.Por su parte, el estudio cuantitativo permite estudiar diferentes parámetros del movimiento del LCR en un lugar determinado. Así se puede calcular la velocidad, el flujo y el volumen de líquido que circula, por ejemplo, en el acueducto.En esta tesis se han desarrollado tres líneas de estudio. El objetivo del primer protocolo de estudio es conocer el comportamiento del LCR en personas sanas a fin de poder comprender mejor la patología que se deriva. Para ello se ha estudiado un grupo de sujetos sanos de distintas edades, lo que a su vez permite conocer si se producen cambios con la edad. El movimiento del LCR relacionado con el ciclo cardiaco vira de dirección al cambiar de sístole a diástole, siempre empezando a nivel acueductal y finalizando a nivel de las cisternas basales. Las velocidades del flujo varían intersujeto mientras que el volumen de flujo es un valor más reproducible. Estas particularidades del flujo del LCR se muestran alteradas en los pacientes con estenosis de acueducto, con malformación de Arnold Chiari tipo I y con HCA.De los resultados obtenidos del primer protocolo se deduce que el estudio cualitativo del LCR es básico para realizar el diagnóstico y establecer el tratamiento en pacientes con estenosis de acueducto y en casos de malformación de Arnold Chiari tipo I, mientras que el estudio cuantitativo es importante para el diagnóstico de los pacientes con HCA. El objetivo principal del segundo protocolo de estudio es determinar la efectividad de la cuantificación del volumen de flujo en el acueducto para el diagnóstico de HCA. Cuando el volumen de flujo diastólico es superior 91 mm3 la sensibilidad y la especificidad de este test es de 82,4% y 77,8 %. Así pues se propone realizar un estudio por Resonancia Magnética en contraste de fase en todos los pacientes con sospecha de HCA y test diagnósticos más invasivos únicamente en los casos en que exista una disociación clínica radiológica.El tercer protocolo de estudio de esta tesis tiene por objetivo evaluar la utilidad de la secuencia de Resonancia Magnética en contraste de fase en el seguimiento de pacientes intervenidos de ventriculostomía premamilar endoscópica por presentar hidrocefalia. Esta técnica permite evaluar la permeabilidad de la ventriculostomía utilizando la secuencia cualitativa, pero además, mediante la cuantificación del volumen de flujo, se puede predecir el funcionamiento de la ventriculostomía premamilar. En este estudio se demuestra que cuando el volumen de flujo absoluto (volumen de flujo sistólico y diastólico) obtenido en el primer estudio posquirúrgico es superior a 75mm3, la sensibilidad del test para determinar qué enfermos mejorarán tras el tratamiento es de 76,5 %y la especificidad es del 100%.Así pues, los resultados obtenidos en esta tesis permiten demostrar que el estudio de Resonancia Magnética en contraste de fase es una herramienta no sólo útil sino también imprescindible en el estudio de pacientes con patología de circulación del LCR., UTILITY OF MAGNETIC RESONANCE USING PHASE CONTRAST IN THE DIAGNOSIS OF CSF CIRCULATION DISORDERS.TEXT: The aim of this thesis is to know the utility of Magnetic Resonance (MR) using Phase Contrast sequences in the diagnosis of CSF circulation disorders. The CSF has a major movement from ventricular cavities and subarachnoid space to venous sinus induced by a pressure gradient between subarachnoid spaces and venous spaces. There is another type of CSF movement related to cardiac cycle. It is an in and fro type of motion. During cardiac systole the CSF have a craniocaudal movement and during cardiac diastole the direction of CSF is inverted. MR using a specific sequence denominated phase contrast allows detecting movements of fluids like CSF or blood. If the acquisition is performed triggered with cardiac movement, we can obtain a sequence of images detecting the circulation of CSF during the cardiac cycle. These images can be evaluated using cine allowing a qualitative study and also let to obtain quantifications of velocity and flow volumeTo evaluate the utility of this technique in the CSF disorders we realize tree different studies. The first study compares the flow pattern in the ventricular and subarachnoid space and the quantification of velocity and flow volume between a control group, a group with aqueductal stenosis, a group with Arnold Chiari malformation and Normal Pressure hydrocephalus (NPH) group. The results obtained demonstrated that the flow volume in the aqueduct in healthy subjects is between 20 to 80 mm3 showing an increment with age. Otherwise in patients with aqueductal stenosis and Chiari malformation, the qualitative study was very helpful in demonstrating the absence of flow in the aqueduct in the first group and the difficulty of CSF circulation in foramen magnum in the Chiari malformation. An interesting result was that in patients with NPH, the flow volume was significantly higher that the healthy subjects.The second study compare the flow volume in patients with NPH with the infusion test the Katzman used as a diagnostic tool of this syndrome. The results obtained demonstrate that the diastolic flow volume can be used as a diagnostic tool with a sensibility of 82,4% and specificity of 77,8%, using a diastolic flow volume of 91 mm3 as a threshold.The last study evaluated the MR using phase contrast in the postsurgery evaluation of patients who has undergone a premamilar endoscópica ventriculostomy (PEV). The results demonstrate that flow volume recorded in the ventriculostomy hole is an indicator of functionality of the PEV. When this value is high in the initial postsurgery exam the PEV will be open with a sensibility of 76,5 % and specificity of 100%.In conclusion: with all the results obtained in this project we conclude that MR using Phase Contrast is a essential tool to evaluate patients who suffer of CSF circulation disorders.

Axonal damage probably occurs early in the evolution of MS. Five of 38 (13%) patients had a positive assay for the neuronal 14-3-3 protein in the CSF obtained at the first clinically isolated syndrome suggestive of MS. A positive 14-3-3 assay was the only independent predictor for a shorter time to conversion to clinical definite MS (risk ratio 4.1; 95% CI 1.1 to 15) and to reach an Expanded Disability Status Scale (EDSS) > or =2 at the end of follow-up (odds ratio 14.8; 95% CI 2.86 to 76.8). The detection of the 14-3-3 protein in the CSF at the first neurologic event suggestive of MS may be a useful predictor of short-term evolution.

Objective: To analyze the MRI and CSF oligoclonal bands (OB) changes in patients with MS who underwent an autologous hematopoietic stem cell transplantation (AHSCT). Background: AHSCT is evaluated as an alternative therapy in severe MS. In previous series of AHSCT for MS, data on MRI or OB outcome were limited or not provided. Methods: Five patients with a median Kurtzke’s EDSS score of 6.5, more than two attacks, and confirmed worsening of the EDSS in the previous year received an AHSCT. Hematopoietic stem cells were mobilized with cyclophosphamide (3 g/m2) and granulocyte colony-stimulating factor (5 μg/kg/d). The graft was T cell depleted by positive CD 34+ selection. Conditioning regimen included BCNU (300 mg/m 2 ), cyclophosphamide (150 mg/kg in 3 days), and antithymocyte globulin (60 mg/kg in 4 days). MRI scans were scheduled at baseline and 1, 3, 6, and 12 months and OB analysis at baseline and 3 and 12 months post-AHSCT. Results: Four patients had a stable or improved EDSS after a median follow-up of 18 months (range, 12 to 24 months). The fifth patient’s condition deteriorated during AHSCT. She partially improved and remained stable after month 3 after AHSCT. The baseline CSF OB persisted 1 year after AHSCT. MRI studies after AHSCT showed no enhanced T1 lesions and no new or enlarging T2 lesions. The median percentage change of T2 lesion load was −11.8% (range, −26.6 to −4.0%). All patients had a decrease of corpus callosum area at 1 year (median, 12.4%; range, 7.8% to 20.5%) that did not progress in the two patients evaluated at 2 years after AHSCT. Conclusions: Although the persistence of CSF OB suggests the lymphocytes were not eliminated from the CNS, the follow-up MRI studies showed no enhanced T1 brain lesions and a reduction in the T2 lesion load that correlated with the clinical stabilization of MS after AHSCT.

Utilizando el método de tinción de la bencidina se realizó un estudio sobre las características morfológicas de la microvascularización de los tumores cerebrales; paralelamente, en la membrana corioalantoidea de embriones de pollo, se estudió la capacidad angiogénica del LCR de pacientes con tumores. La densidad vascular se observó reducida en astrocitomas benignos, oligodendrogliomas y craneo-faringiomas, ligeramente aumentada en astrocitomas de grado I I y meduloblastomas. Los astrocitomas malignos, glioblastomas y metástasis mostraron patrones vasculares muy aumentados. Los meningiomas están intensamente vascularizados y menos los neurinomas. Los LCR de pacientes con tumores primitivos del SNC o a distancia mostraron actividad angiogénica en el 93% de los casos. El 27% de los LCR controles fueron positivos observándose correlación con la edad.

Introduction Neurofilament light (NFL) levels in the cerebrospinal fluid are increased in several neurodegenerative dementias. However, their diagnostic accuracy in the differential diagnostic context is unknown. Methods Cerebrospinal fluid NFL levels were quantified in nonprimarily neurodegenerative neurological and psychiatric diseases (n = 122), mild cognitive impairment (n = 48), Alzheimer's disease (n = 108), dementia with Lewy bodies/Parkinson's disease dementia (n = 53), vascular dementia (n = 46), frontotemporal dementia (n = 41), sporadic Creutzfeldt-Jakob disease (sCJD, n = 132), and genetic prion diseases (n = 182). Results The highest NFL levels were detected in sCJD, followed by vascular dementia, frontotemporal dementia, dementia with Lewy bodies/Parkinson's disease dementia, Alzheimer's disease, and mild cognitive impairment. In sCJD, NFL levels correlated with cerebrospinal fluid tau and disease duration. NFL levels were able to differentiate sCJD from nonprimarily neurodegenerative neurological and psychiatric diseases (area under the curve = 0.99, 95% confidence interval: 0.99–1) and from the other diagnostic groups showing cognitive impairment/dementia of a non-CJD etiology (area under the curve = 0.90, 95% confidence interval: 0.87–0.92). Compared to nonprimarily neurodegenerative neurological and psychiatric diseases, NFL was also elevated in genetic prion diseases associated with the E200K, V210I, P102L, and D178N prion protein gene mutations. Discussion Increased NFL levels are a common feature in neurodegenerative dementias.

ObjectiveA high level of cerebrospinal fluid (CSF) neopterin is a marker of central nervous system inflammatory-immune mediated processes. We aimed to assess data from 606 neuropediatric patients, describing the clinical and biochemical features of those neurological disorders presenting CSF neopterin values above a new cut-off value that was defined in our laboratory.MethodsTo establish the new CSF neopterin cut-off value, we studied two groups of patients: Group 1 comprised 68 patients with meningoencephalitis, and Group 2 comprised 52 children with a confirmed peripheral infection and no central nervous system involvement. We studied 606 CSF samples from neuropediatric patients who were classified into 3 groups: genetic diagnosis (A), acquired/unknown etiologic neurologic diseases (B) and inflammatory-immune mediated processes (C).ResultsThe CSF neopterin cut-off value was 61 nmol/L. Out of 606 cases, 56 presented a CSF neopterin level above this value. Group C had significantly higher CSF neopterin, protein and leukocyte values than the other groups. Sixteen of twenty-three patients in this group had a CSF neopterin level above the cut-off, whereas three and seven patients presented increased leukocyte and protein values, respectively. A significant association was found among CSF neopterin, proteins and leukocytes in the 606 patients. White matter disturbances were associated with high CSF neopterin concentrations.ConclusionsAlthough children with inflammatory-immune mediated processes presented higher CSF neopterin values, patients with other neurological disorders also showed increased CSF neopterin concentrations. These results stress the importance of CSF neopterin analysis for the identification of inflammatory-immune mediated processes.

Melancholic depression is a biologically homogeneous clinical entity in which structural brain alterations have been described. Interestingly, reports of structural alterations in melancholia include volume increases in Cerebro-Spinal Fluid (CSF) spaces. However, there are no previous reports of CSF volume alterations using automated whole-brain voxel-wise approaches, as tissue classification algorithms have been traditionally regarded as less reliable for CSF segmentation. Here we aimed to assess CSF volumetric alterations in melancholic depression and their clinical correlates by means of a novel segmentation algorithm ('new segment', as implemented in the software Statistical Parametric Mapping-SPM8), incorporating specific features that may improve CSF segmentation. A three-dimensional Magnetic Resonance Image (MRI) was obtained from seventy patients with melancholic depression and forty healthy control subjects. Although imaging data were pre-processed with the 'new segment' algorithm, in order to obtain a comparison with previous segmentation approaches, tissue segmentation was also performed with the 'unified segmentation' approach. Melancholic patients showed a CSF volume increase in the region of the left Sylvian fissure, and a CSF volume decrease in the subarachnoid spaces surrounding medial and lateral parietal cortices. Furthermore, CSF increases in the left Sylvian fissure were negatively correlated with the reduction percentage of depressive symptoms at discharge. None of these results were replicated with the 'unified segmentation' approach. By contrast, between-group differences in the left Sylvian fissure were replicated with a non-automated quantification of the CSF content of this region. Left Sylvian fissure alterations reported here are in agreement with previous findings from non-automated CSF assessments, and also with other reports of gray and white matter insular alterations in depressive samples using automated approaches. The reliable characterization of CSF alterations may help in the comprehensive characterization of brain structural abnormalities in psychiatric samples and in the development of etiopathogenic hypotheses relating to the disorders.