Redefining migraine prevention: early treatment with anti-CGRP monoclonal antibodies enhances response in the real world.

BACKGROUND: Anti-CGRP monoclonal antibodies (anti-CGRP MAbs) are approved and available treatments for migraine prevention. Patients do not respond alike and many countries have reimbursement policies, which hinder treatments to those who might respond. This study aimed to investigate clinical factors associated with good and excellent response to anti-CGRP MAbs at 6 months. METHODS: European multicentre, prospective, real-world study, including high-frequency episodic or chronic migraine (CM) patients treated since March 2018 with anti-CGRP MAbs. We defined good and excellent responses as ≥50% and ≥75% reduction in monthly headache days (MHD) at 6 months, respectively. Generalised mixed-effect regression models (GLMMs) were used to identify variables independently associated with treatment response. RESULTS: Of the 5818 included patients, 82.3% were females and the median age was 48.0 (40.0-55.0) years. At baseline, the median of MHD was 20.0 (14.0-28.0) days/months and 72.2% had a diagnosis of CM. At 6 months (n=4963), 56.5% (2804/4963) were good responders and 26.7% (1324/4963) were excellent responders. In the GLMM model, older age (1.08 (95% CI 1.02 to 1.15), p=0.016), the presence of unilateral pain (1.39 (95% CI 1.21 to 1.60), p<0.001), the absence of depression (0.840 (95% CI 0.731 to 0.966), p=0.014), less monthly migraine days (0.923 (95% CI 0.862 to 0.989), p=0.023) and lower Migraine Disability Assessment at baseline (0.874 (95% CI 0.819 to 0.932), p<0.001) were predictors of good response (AUC of 0.648 (95% CI 0.616 to 0.680)). These variables were also significant predictors of excellent response (AUC of 0.691 (95% CI 0.651 to 0.731)). Sex was not significant in the GLMM models. CONCLUSIONS: This is the largest real-world study of migraine patients treated with anti-CGRP MAbs. It provides evidence that higher migraine frequency and greater disability at baseline reduce the likelihood of responding to anti-CGRP MAbs, informing physicians and policy-makers on the need for an earlier treatment in order to offer the best chance of treatment success.

RE-START: Exploring the effectiveness of anti-calcitonin gene-related peptide resumption after discontinuation in migraine.

BACKGROUND AND PURPOSE: According to the latest European guidelines, discontinuation of monoclonal antibodies against calcitonin gene-related peptide (anti-CGRP MAb) may be considered after 12-18 months of treatment. However, some patients may worsen after discontinuation. In this study, we assessed the response following treatment resumption. METHODS: This was a prospective study conducted in 14 Headache Units in Spain. We included patients with response to anti-CGRP MAb with clinical worsening after withdrawal and resumption of treatment. Numbers of monthly migraine days (MMD) and monthly headache days (MHD) were obtained at four time points: before starting anti-CGRP MAb (T-baseline); last month of first treatment period (T-suspension); month of restart due to worsening (T-worsening); and 3 months after resumption (T-reintroduction). The response rate to resumption was calculated. Possible differences among periods were analysed according to MMD and MHD. RESULTS: A total of 360 patients, 82% women, with a median (interquartile range [IQR]) age at migraine onset of 18 (12) years. The median (IQR) MHD at T-baseline was 20 (13) and MMD was 5 (6); at T-suspension, the median (IQR) MHD was 5 (6) and MMD was 4 (5); at T-worsening, the median (IQR) MHD was 16 (13) and MMD was 12 (6); and at T-reintroduction, the median (IQR) MHD was 8 (8) and MHD was 5 (5). In the second period of treatment, a 50% response rate was achieved by 57.4% of patients in MHD and 65.8% in MMD. Multivariate models showed significant differences in MHD between the third month after reintroduction and last month before suspension of first treatment period (p < 0.001). CONCLUSION: The results suggest that anti-CGRP MAb therapy is effective after reintroduction. However, 3 months after resumption, one third of the sample reached the same improvement as after the first treatment period.

Association of magnetic resonance imaging phenotypes and serum biomarker levels with treatment response and long-term disease outcomes in multiple sclerosis patients.

BACKGROUND AND PURPOSE: The aim was to evaluate whether magnetic resonance imaging (MRI) phenotypes defined by inflammation and neurodegeneration markers correlate with serum levels of neurofilament light chain (NfL) and glial fibrillary acidic protein (GFAP) in relapsing-remitting multiple sclerosis (RRMS) patients; and to explore the role of radiological phenotypes and biomarker levels on treatment response and long-term prognostic outcomes. METHODS: Magnetic resonance imaging scans from 80 RRMS patients were classified at baseline of interferon-beta (IFNß) treatment into radiological phenotypes defined by high and low inflammation and high and low neurodegeneration, based on the number of contrast-enhancing lesions, brain parenchymal fraction and the relative volume of non-enhancing black holes on T1-weighted images. Serum levels of NfL and GFAP were measured at baseline with single molecule array (Simoa) assays. MRI phenotypes and serum biomarker levels were investigated for their association with IFNß response, and times to second-line therapies, secondary-progressive MS (SPMS) conversion and Expanded Disability Status Scale (EDSS) 6.0. RESULTS: Mean (SD) follow-up was 17 (2.9) years. Serum NfL levels and GFAP were higher in the high inflammation (p = 0.04) and high neurodegeneration phenotypes (p = 0.03), respectively. The high inflammation phenotype was associated with poor response to IFNß treatment (p = 0.04) and with shorter time to second-line therapies (p = 0.04). In contrast, the high neurodegeneration phenotype was associated with shorter time to SPMS (p = 0.006) and a trend towards shorter time to EDSS 6.0 (p = 0.09). High serum NfL levels were associated with poor response to IFNß treatment (p = 0.004). CONCLUSIONS: Magnetic resonance imaging phenotypes defined by inflammation and neurodegeneration correlate with serum biomarker levels, and both have prognostic implications in treatment response and long-term disease outcomes.

Measuring Heat Stress for Human Health in Cities: A Low-Cost Prototype Tested in a District of Valencia, Spain.

Nowadays, the measurement of heat stress indices is of principal importance due to the escalating impact of global warming. As temperatures continue to rise, the well-being and health of individuals are increasingly at risk, which can lead to a detrimental effect on human performance and behavior. Hence, monitoring and assessing heat stress indices have become necessary for ensuring the safety and comfort of individuals. Thermal comfort indices, such as wet-bulb globe temperature (WBGT), Tropical Summer Index (TSI), and Predicted Heat Strain (PHS), as well as parameters like mean radiant temperature (MRT), are typically used for assessing and controlling heat stress conditions in working and urban environments. Therefore, measurement and monitoring of these parameters should be obtained for any environment in which people are constantly exposed. Modern cities collect and publish this relevant information following the Smart City concept. To monitor large cities, cost-effective solutions must be developed. This work presents the results of a Heat Stress Monitoring (HSM) system prototype network tested in the Benicalap-Ciutat Fallera district in Valencia, Spain. The scope of this work is to design, commission, and test a low-cost prototype that is able to measure heat stress indices. The Heat Stress Monitoring system comprises a central unit or receiver and several transmitters communicating via radiofrequency. The transmitter accurately measures wind speed, air temperature, relative humidity, atmospheric pressure, solar irradiation, and black globe temperature. The receiver has a 4G modem that sends the data to an SQL database in the cloud. The devices were tested over one year, showing that radio data transmission is reliable up to 700 m from the receiver. The system's power supply, composed of a Photovoltaic panel and Lithium-ion batteries, provided off-grid capabilities to the transmitter, with a tested backup autonomy of up to 36 days per charge. Then, indicators such as WBGT, TSI, and MRT were successfully estimated using the data collected by the devices. The material cost of a 12-point network is around EUR 2430 with a competitive price of EUR 190 per device.

(when) do counterattitudinal exemplars shift implicit racial evaluations? Replications and extensions of Dasgupta and Greenwald (2001).

Dasgupta and Greenwald (2001) demonstrated that exposure to positive Black exemplars (e.g., Colin Powell) and negative White exemplars (e.g., Jeffrey Dahmer) can reduce implicit pro-White/anti-Black evaluations, as measured by an Implicit Association Test. Here, we report seven preregistered online experiments conducted with volunteer U.S. participants (N = 6,953) that sought to replicate and probe the boundary conditions of this finding. Contrary to expectations, we found no shift in implicit racial evaluations in two close replication attempts (Experiments 1-2). Experiments 3-4 ruled out the possibility of insufficiently strong exemplar valence and subtyping as explanations for the failures to replicate. In Experiment 5, implicit racial evaluations did exhibit malleability in response to two different procedures relying on repeated evaluative pairings and evaluative statements, suggesting that they are capable of change. With insight from these studies, Experiments 6-7 were mounted with modifications to the Dasgupta and Greenwald (2001) procedure. Significant reductions in implicit pro-White/anti-Black evaluations were now observed when race, valence, and the contingency between the two were highlighted. In addition, across all experiments, the magnitude of shift in implicit racial evaluations was significantly predicted by participants' ability to recall the Black-positive and White-negative contingencies experienced during the exemplar exposure task. Together, these data suggest that exposure to counterattitudinal exemplars can shift implicit racial evaluations toward neutrality, but such malleability strongly depends on contingency awareness. We discuss implications for social cognitive theory, theoretically informed debiasing interventions, and different paths toward resolving initial replication failures. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Molecular signature associated with cladribine treatment in patients with multiple sclerosis.

Introduction: Little is known about the molecular profiling associated with the effect of cladribine in patients with multiple sclerosis (MS). Here, we aimed first to characterize the transcriptomic and proteomic profiles induced by cladribine in blood cells, and second to identify potential treatment response biomarkers to cladribine in patients with MS. Methods: Gene, protein and microRNA (miRNA) expression profiles were determined by microarrays (genes, miRNAs) and mass spectrometry (proteins) in peripheral blood mononuclear cells (PBMCs) from MS patients after in vitro treatment with cladribine in its active and inactive forms. Two bioinformatics approaches to integrate the three obtained datasets were applied: (i) a multiomics discriminant analysis (DIABLO - Data Integration Analysis for Biomarker discovery using Latent variable approaches for Omics studies); and (ii) a multi-stage integration of features selected in differential expression analysis on each dataset and then merged. Selected molecules from the in vitro study were quantified by qPCR ex vivo in PBMCs from MS patients receiving cladribine. Results: PBMCs treated in vitro with cladribine were characterized by a major downregulation of gene, protein, and miRNA expression compared with the untreated cells. An intermediate pattern between the cladribine-treated and untreated conditions was observed in PBMCs treated with cladribine in its inactive form. The differential expression analysis of each dataset led to the identification of four genes and their encoded proteins, and twenty-two miRNAs regulating their expression, that were associated with cladribine treatment. Two of these genes (PPIF and NHLRC2), and three miRNAs (miR-21-5p, miR-30b-5p, and miR-30e-5p) were validated ex vivo in MS patients treated with cladribine. Discussion: By using a combination of omics data and bioinformatics approaches we were able to identify a multiomics molecular profile induced by cladribine in vitro in PBMCs. We also identified a number of biomarkers that were validated ex vivo in PBMCs from patients with MS treated with cladribine that have the potential to become treatment response biomarkers to this drug.

Tax rules on wealth imbalance and the investigation of money laundering in Peru / Las reglas tributarias sobre desbalances patrimoniales y la investigación del lavado de activos: el caso peruano / Regras tributárias sobre desproporções patrimoniais e investigação de lavagem de dinheiro: o caso peruano

This study aims to evaluate the feasibility of using the tax methodology for determining unjustified increases in assets in money laundering cases heard in the courts. It also seeks to identify the errors made by the Tax Authority with the purpose of ascertaining whether these errors could hinder the successful application of the methodology in investigations of money laundering cases. In order to achieve these objectives, a mixed research methodology was conducted that included the analysis of rulings issued during the years 2020 and 2021 by the Peruvian Tax Court and sentences issued by the judiciary. This was complemented by semi-structured interviews with experts from the public sector, academia and accounting with relevant experience on the subject. The study found that there are some practical issues in the application of the methodology that can be overcome and that do not represent an insurmountable constraint. The study also found that the tax methodology allows for a more effective clarification of asset imbalances, and concludes that, once implementation errors have been overcome, the tax methodology can be feasibly employed to the benefit of money laundering investigations.

Esta investigación tiene como objetivos evaluar la factibilidad de utilizar la metodología tributaria de determinación de incrementos patrimoniales no justificados en la investigación de desbalances patrimoniales de casos de lavado de dinero ventilados en la vía jurisdiccional e identificar los errores que ha cometido la Administración Tributaria, a fin de establecer si estos representan problemas que pudieran dificultar la aplicación de la metodología a la investigación de casos de lavado de dinero. Con el propósito de lograr estos objetivos se condujo una investigación mixta que incluyó el análisis de las resoluciones emitidas durante los años 2020 y 2021 por el Tribunal Fiscal y sentencias emitidas por el poder judicial. Esto se complementó con entrevistas semiestructuradas a expertos del sector público, la academia y peritos contables con experiencia relevante sobre el tema. La investigación encontró que existen algunos empirismos aplicativos que pueden ser superados y que no representan ninguna restricción que no pueda ser gestionada. Se halló también que la metodología tributaria permite esclarecer de manera más efectiva los desbalances patrimoniales. La investigación concluye que, superados los errores de ejecución, es factible utilizar con ventaja la metodología tributaria en las investigaciones de lavado de dinero.

O objetivo desta pesquisa é avaliar a viabilidade do uso da metodología tributária para determinar aumentos injustificados de patrimônio na investigação de desproporções patrimoniais em casos de lavagem de dinheiro julgados nos tribunais e identificar os erros cometidos pela Administração Tributária, a fim de estabelecer se estes representam problemas que poderiam dificultar a aplicação da metodologia na investigação de casos de lavagem de dinheiro. Para atingir esses objetivos, foi realizada uma pesquisa mista que incluiu a análise de decisões emitidas durante 2020 e 2021 pelo Tribunal Tributário e sentenças emitidas pelo judiciário. Isso foi complementado por entrevistas semiestruturadas com especialistas do setor público, acadêmicos e especialistas em contabilidade com experiência relevante no assunto. A pesquisa constatou que existem alguns empirismos de aplicação que podem ser superados e que não representam nenhuma restrição que não possa ser gerenciada. Também foi constatado que a metodología tributária permite um esclarecimento mais eficaz das desproporções patrimoniais. Na pesquisa, conclui-se que, uma vez superados os erros de implementação, é viável utilizar a metodologia tributária com vantagem nas investigações de lavagem de dinheiro.

NOD mouse dorsal root ganglia display morphological and gene expression defects before and during autoimmune diabetes development.

Introduction: During the development of Autoimmune Diabetes (AD) an autoimmune attack against the Peripheral Nervous System occurs. To gain insight into this topic, analyses of Dorsal Root Ganglia (DRG) from Non-Obese Diabetic (NOD) mice were carried out. Methods: Histopathological analysis by electron and optical microscopy in DRG samples, and mRNA expression analyzes by the microarray technique in DRG and blood leukocyte samples from NOD and C57BL/6 mice were performed. Results: The results showed the formation of cytoplasmic vacuoles in DRG cells early in life that could be related to a neurodegenerative process. In view of these results, mRNA expression analyses were conducted to determine the cause and/or the molecules involved in this suspected disorder. The results showed that DRG cells from NOD mice have alterations in the transcription of a wide range of genes, which explain the previously observed alterations. In addition, differences in the transcription genes in white blood cells were also detected. Discussion: Taken together, these results indicate that functional defects are not only seen in beta cells but also in DRG in NOD mice. These results also indicate that these defects are not a consequence of the autoimmune process that takes place in NOD mice and suggest that they may be involved as triggers for its development.

msmsEDA & msmsTests: Label-Free Differential Expression by Spectral Counts.

msmsTests is an R/Bioconductor package providing functions for statistical tests in label-free LC-MS/MS data by spectral counts. These functions aim at discovering differentially expressed proteins between two biological conditions. Three tests are available: Poisson GLM regression, quasi-likelihood GLM regression, and the negative binomial of the edgeR package. The three models admit blocking factors to control for nuisance variables. To assure a good level of reproducibility a post-test filter is available, where (1) a minimum effect size considered biologically relevant, and (2) a minimum expression of the most abundant condition, may be set. A companion package, msmsEDA, proposes functions to explore datasets based on msms spectral counts. The provided graphics help in identifying outliers, the presence of eventual batch factors, and check the effects of different normalizing strategies. This protocol illustrates the use of both packages on two examples: A purely spike-in experiment of 48 human proteins in a standard yeast cell lysate; and a cancer cell-line secretome dataset requiring a biological normalization.

Serum Levels of CXCL13 Are Associated With Teriflunomide Response in Patients With Multiple Sclerosis.

BACKGROUND AND OBJECTIVES: To identify biomarkers associated with treatment response in patients with multiple sclerosis (MS) treated with the oral therapies teriflunomide, dimethyl fumarate (DMF), and fingolimod. METHODS: Serum levels of IL-6, IL-17, TNF-α, granulocyte-macrophage colony-stimulating factor, IL-10, interferon-gamma (IFN-γ) IL-1ß, and chemokine ligand 13 (CXCL13) were measured at baseline and 12 months with single molecule array (Simoa) assays in a cohort of patients with MS treated with teriflunomide (N = 19), DMF (N = 22), and fingolimod (N = 25) and classified into "no evidence of disease activity" (NEDA) and EDA patients after 1 year of treatment. RESULTS: Serum CXCL13 and TNF-α levels were significantly decreased after treatment with teriflunomide in NEDA compared with EDA patients after 1 year of treatment (p = 0.008 for both cytokines). These findings were validated in an independent cohort of patients with MS treated with teriflunomide (N = 36) and serum CXCL13, and TNF-α levels were again significantly reduced in NEDA patients (p < 0.0001 for CXCL13 and p = 0.003 for TNF-α). CXCL13, but not TNF-α, showed good performance to classify NEDA and EDA patients according to a cut-off value of 9.64 pg/mL based on the change in CXCL13 levels between baseline and 12 months, with a sensitivity of 75% and specificity of 82% in the original cohort, and sensitivity of 65.4% and specificity of 60% in the validation cohort. DISCUSSION: Altogether, these results point to CXCL13 as a treatment response biomarker to teriflunomide in relapsing-remitting patients with MS, and the change in CXCL13 levels during the first year of treatment can be used in clinical practice to identify optimal responders to teriflunomide.

Temporal and sex-dependent gene expression patterns in a renal ischemia-reperfusion injury and recovery pig model.

Men are more prone to acute kidney injury (AKI) and chronic kidney disease (CKD), progressing to end-stage renal disease (ESRD) than women. Severity and capacity to regenerate after AKI are important determinants of CKD progression, and of patient morbidity and mortality in the hospital setting. To determine sex differences during injury and recovery we have generated a female and male renal ischemia/reperfusion injury (IRI) pig model, which represents a major cause of AKI. Although no differences were found in blood urea nitrogen (BUN) and serum creatinine (SCr) levels between both sexes, females exhibited higher mononuclear infiltrates at basal and recovery, while males showed more tubular damage at injury. Global transcriptomic analyses of kidney biopsies from our IRI pig model revealed a sexual dimorphism in the temporal regulation of genes and pathways relevant for kidney injury and repair, which was also detected in human samples. Enrichment analysis of gene sets revealed five temporal and four sexual patterns governing renal IRI and recovery. Overall, this study constitutes an extensive characterization of the time and sex differences occurring during renal IRI and recovery at gene expression level and offers a template of translational value for further study of sexual dimorphism in kidney diseases.

COVID-19: epidemiología, virología y transmisibilidad / COVID-19: epidemiology, virology, and transmissibility

Hasta diciembre del 2019, seis tipos de coronavirus ya estaban identificados como generadores de enfermedad en humanos, destacándose dos brotes epidemiológicos anteriores: SARS-CoV en 2002 y MERS-CoV en 2012. El nuevo agente infeccioso que causó la pandemia de 2019 se denominó SARS-CoV-2, el que se manifiesta como un síndrome respiratorio agudo severo (CO-VID-19). Al respecto, el 30 de enero del 2020, la Organización Mundial de la Salud (OMS) decretó la emergencia sanitaria. El propósito de esta revisión fue analizar el contexto epidemio-lógico alrededor del SARS-CoV-2, mediante una búsqueda bibliográfica en las bases de datos científicas como: PubMed Central, LILACS y Google académico. Se concluyó que el SARS-CoV-2 es altamente transmisible, con una tasa de letalidad en Ecuador del 8,59%.

Six types of coronaviruses were already identified as generators of disease in humans as of 2019, with two previous epidemiological outbreaks standing out: SARS-CoV in 2002 and MERS-CoV in 2012. The new infectious agent that caused the 2019 pandemic was called SARS -CoV-2, which manifests as a severe acute respiratory syndrome (COVID-19). In this regard, on January 30, 2020, the World Health Organization decreed the health emergency. The purpose of this review was to analyze the epidemiological context around SARS-CoV-2 through a bibliographic review in scientific databases such as: PubMed Central, LILACS and Google Scholar. It was concluded that SARS-CoV-2 is highly transmissible, with a fatality rate in Ecuador of 8.59%.

Covid-19: fisiopatología, historia natural y diagnóstico / Covid-19: pathophysiology, natural history, and diagnosi

SARS CoV-2 se transmite principalmente a través de la vía aérea. La media del tiempo desde la exposición hasta la presentación de la sintomatología es de 5 días. Los pacientes infectados pueden permanecer asintomáticos o tener un amplio espectro de manifestaciones como: fiebre, tos seca y malestar general. Aproximadamente, el 20% de pacientes con la COVID-19 requiere hospitalización (10% en área general, el 5% requiere cuidados intermedios y el 5% restante ingresa a cuidados intensivos debido a su estado de gravedad). Los exámenes de laboratorio y radiológicos muestran anomalías de común presentación, pero inespecíficas. El diagnóstico es a través de la prueba de reacción en cadena de polimerasa de transcripción inversa cuando detecta SARS CoV-2. El objetivo de la revisión que se presenta es sintetizar elementos importantes relacionados con la fisiolopatología, manifestaciones clínicas y diagnóstico de la COVID-19.

SARS CoV-2 is transmitted primarily through the airway. The mean time from exposure to presentation of symptoms is 5 days. Infected patients can remain asymptomatic or have a wide spectrum of manifestations such as: fever, dry cough, and general malaise. Approximately 20% of patients with COVID-19 require hospitalization (10% in the general area, 5% require inter-mediate care, and the remaining 5% enter intensive care due to their serious condition). Labora-tory and radiological examinations show common but nonspecific abnormalities. Diagnosis is through the reverse transcription polymerase chain reaction test when it detects SARS CoV-2. This systematic review aimed to synthesize important elements related to the physiopathology, clinical manifestations, and diagnosis of COVID-19.

Identification of ZBTB18 as a novel colorectal tumor suppressor gene through genome-wide promoter hypermethylation analysis.

BACKGROUND: Cancer initiation and progression are driven by genetic and epigenetic changes. Although genome/exome sequencing has significantly contributed to the characterization of the genetic driver alterations, further investigation is required to systematically identify cancer driver genes regulated by promoter hypermethylation. RESULTS: Using genome-wide analysis of promoter methylation in 45 colorectal cancer cell lines, we found that higher overall methylation levels were associated with microsatellite instability (MSI), faster proliferation and absence of APC mutations. Because epigenetically silenced genes could represent important oncogenic drivers, we used mRNA expression profiling of colorectal cancer cell lines and primary tumors to identify a subset of 382 (3.9%) genes for which promoter methylation was negatively associated with gene expression. Remarkably, a significant enrichment in zinc finger proteins was observed, including the transcriptional repressor ZBTB18. Re-introduction of ZBTB18 in colon cancer cells significantly reduced proliferation in vitro and in a subcutaneous xenograft mouse model. Moreover, immunohistochemical analysis revealed that ZBTB18 is frequently lost or reduced in colorectal tumors, and reduced ZBTB18 expression was found to be associated with lymph node metastasis and shorter survival of patients with locally advanced colorectal cancer. CONCLUSIONS: We identified a set of 382 genes putatively silenced by promoter methylation in colorectal cancer that could significantly contribute to the oncogenic process. Moreover, as a proof of concept, we demonstrate that the epigenetically silenced gene ZBTB18 has tumor suppressor activity and is a novel prognostic marker for patients with locally advanced colorectal cancer.

CSF SERPINA3 Levels Are Elevated in Patients With Progressive MS.

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.

Putative Drivers of Aggressiveness in TCEB1-mutant Renal Cell Carcinoma: An Emerging Entity with Variable Clinical Course.

BACKGROUND: TCEB1-mutant renal cell carcinoma (RCC) is a rare variant of RCC with clear-cell features. Owing to its unique morphological and molecular features it has recently been proposed as a separate entity. Initial series suggested an indolent, early-stage phenotype. Here we expand our clinical cohort and describe a more detailed genomic analysis looking for potential drivers of aggressiveness. DESIGN, SETTING, AND PARTICIPANTS: We identified five new cases in our institutional sequencing cohort, four of whom were found to have high-stage disease (American Joint Committee on Cancer stage III/IV). Twelve previously reported cases were pooled for comparison purposes (Sato, The Cancer Genome Atlas, TRACERx Renal). OUTCOME MEASURES AND STATISTICAL ANALYSIS: We used our previously validated pipeline to analyze somatic mutations and copy number alterations (CNAs) in seven tumor samples with available data and estimated the number of cancer cells bearing each somatic mutation. The oncogenic potential of mutations was assessed using OncoKB and two other algorithms. Mann-Whitney U tests were used to evaluate differences in genomic markers between stage groups. RESULTS AND LIMITATIONS: All tumors showed biallelic inactivation of the TCEB1 gene according to a combination of somatic mutation and CNA analyses. Mutations were always found in residues involved in hydrophobic interactions with VHL. We found that high-stage tumors had additional oncogenic mutations (median 1, interquartile range [IQR] 1-1 vs 2, IQR 2-2; median difference 1, 95% confidence interval [CI] 1-1; p= 0.002) and showed whole-genome doubling events. They also seemed to have a higher burden of somatic CNAs (median fraction CNA genome 0.10, IQR 0.10-0.15 vs 0.63, IQR 0.58-0.68), however, this finding did not reach statistical significance (median difference 0.49, 95% CI 0.33-0.63; p=0.052). CONCLUSIONS: TCEB1-mutant RCC can show variable behavior ranging from very indolent to aggressive. Specific molecular events leading to high genomic instability seem to be associated with aggressiveness. This study expands the clinical spectrum of TCEB1-mutant RCC. PATIENT SUMMARY: We present four cases of aggressive TCEB1-mutant renal cell carcinoma, a rare type of kidney cancer. In-depth analysis of the genomes of these tumors revealed certain abnormalities that might explain this aggressive behavior.

A Mouse Brain-based Multi-omics Integrative Approach Reveals Potential Blood Biomarkers for Ischemic Stroke.

Stroke remains a leading cause of death and disability worldwide. Despite continuous advances, the identification of key molecular signatures in the hyper-acute phase of ischemic stroke is still a primary interest for translational research on stroke diagnosis, prognosis, and treatment. Data integration from high-throughput -omics techniques has become crucial to unraveling key interactions among different molecular elements in complex biological contexts, such as ischemic stroke. Thus, we used advanced data integration methods for a multi-level joint analysis of transcriptomics and proteomics data sets obtained from mouse brains at 2 h after cerebral ischemia. By modeling net-like correlation structures, we identified an integrated network of genes and proteins that are differentially expressed at a very early stage after stroke. We validated 10 of these deregulated elements in acute stroke, and changes in their expression pattern over time after cerebral ischemia were described. Of these, CLDN20, GADD45G, RGS2, BAG5, and CTNND2 were next evaluated as blood biomarkers of cerebral ischemia in mice and human blood samples, which were obtained from stroke patients and patients presenting stroke-mimicking conditions. Our findings indicate that CTNND2 levels in blood might potentially be useful for distinguishing ischemic strokes from stroke-mimicking conditions in the hyper-acute phase of the disease. Furthermore, circulating GADD45G content within the first 6 h after stroke could also play a key role in predicting poor outcomes in stroke patients. For the first time, we have used an integrative biostatistical approach to elucidate key molecules in the initial stages of stroke pathophysiology and highlight new notable molecules that might be further considered as blood biomarkers of ischemic stroke.

NLRP3 inflammasome as prognostic factor and therapeutic target in primary progressive multiple sclerosis patients.

Primary progressive multiple sclerosis is a poorly understood disease entity with no specific prognostic biomarkers and scarce therapeutic options. We aimed to identify disease activity biomarkers in multiple sclerosis by performing an RNA sequencing approach in peripheral blood mononuclear cells from a discovery cohort of 44 untreated patients with multiple sclerosis belonging to different clinical forms and activity phases of the disease, and 12 healthy control subjects. A validation cohort of 58 patients with multiple sclerosis and 26 healthy control subjects was included in the study to replicate the RNA sequencing findings. The RNA sequencing revealed an interleukin 1 beta (IL1B) signature in patients with primary progressive multiple sclerosis. Subsequent immunophenotyping pointed to blood monocytes as responsible for the IL1B signature observed in this group of patients. Functional experiments at baseline measuring apoptosis-associated speck-like protein containing a CARD (ASC) speck formation showed that the NOD-leucine rich repeat and pyrin containing protein 3 (NLRP3) inflammasome was overactive in monocytes from patients with primary progressive multiple sclerosis, and canonical NLRP3 inflammasome activation with a combination of ATP plus lipopolysaccharide was associated with increased IL1B production in this group of patients. Primary progressive multiple sclerosis patients with high IL1B gene expression levels in peripheral blood mononuclear cells progressed significantly faster compared to patients with low IL1B levels based on the time to reach an EDSS of 6.0 and the Multiple Sclerosis Severity Score. In agreement with peripheral blood findings, both NLRP3 and IL1B expression in brain tissue from patients with primary progressive multiple sclerosis was mainly restricted to cells of myeloid lineage. Treatment of mice with a specific NLRP3 inflammasome inhibitor attenuated established experimental autoimmune encephalomyelitis disease severity and improved CNS histopathology. NLRP3 inflammasome-specific inhibition was also effective in reducing axonal damage in a model of lipopolysaccharide-neuroinflammation using organotypic cerebellar cultures. Altogether, these results point to a role of IL1B and the NLRP3 inflammasome as prognostic biomarker and potential therapeutic target, respectively, in patients with primary progressive multiple sclerosis.

Chitinase 3-like 1 is neurotoxic in primary cultured neurons.

Chitinase 3-like 1 (CHI3L1) is known to play a role as prognostic biomarker in the early stages of multiple sclerosis (MS), and patients with high cerebrospinal fluid CHI3L1 levels have an increased risk for the development of neurological disability. Here, we investigated its potential neurotoxic effect by adding recombinant CHI3L1 in vitro to primary cultures of mouse cortical neurons and evaluating both neuronal functionality and survival by immunofluorescence. CHI3L1 induced a significant neurite length retraction after 24 and 48 hours of exposure and significantly reduced neuronal survival at 48 hours. The cytotoxic effect of CHI3L1 was neuron-specific and was not observed in mouse immune or other central nervous system cells. These results point to a selective neurotoxic effect of CHI3L1 in vitro and suggest a potential role of CHI3L1 as therapeutic target in MS patients.