Comparing RNA extraction methods to face the variations in RNA quality using two human biological matrices.

BACKGROUND: Nucleic acids, RNA among them, are widely used in biomedicine and Biotechnology. Because of their susceptibility to degradation by RNases, the handling and extraction process of RNA from cells and tissues require specialized personnel and standardized methods to guarantee high purity and integrity. Due to the diversity of techniques found in the market, a comparative study between different RNA extraction methods is useful to facilitate the best choice for the researcher or in research service platforms such as biobanks to see the traceability of the samples. METHODS AND RESULTS: In this study, we have compared seven different RNA extraction methods: manual (TRIzol™), semiautomated (QIAGEN™, Bio-Rad, Monarch®, and Canvax™), and fully automated (QIAcube™ and Maxwell®) processes, from two biological matrices: human Jurkat T cells and peripheral blood mononuclear cells (PBMC). Results showed marked differences in the RNA quality and functionality according to the method employed for RNA extraction and the matrix used. DISCUSSION: QIAcube™ and semi-automated extraction methods were perceived as the best options because of their lower variability, good functionality, and lower cost (P < 0.001). These data contribute to facilitating researchers or research service platforms (Biobanks) in decision-making practices and emphasize the relevance of the selection of the RNA extraction method in each experimental procedure or traceability study to guarantee both quality standards and its reproducibility.

Environmental exposure to pesticides and Amyotrophic Lateral Sclerosis in the South of Spain.

OBJECTIVE: To determine if there is a relationship between environmental exposure to pesticides and the prevalence of Amyotrophic Lateral Sclerosis (ALS) in Andalusia. METHOD: We carried out a case-control study using the logistic regression method to verify the relationship between the prevalence of ALS in the area exposed to pesticides versus the unexposed area, through the Odds Ratio statistical test. RESULTS: The study population consisted of 519 individuals diagnosed with ALS between January 2016 and December 2018 according to the CMBD (Minimum Basic Data Set) as cases. In the control group, we have 8,384,083 individuals obtained from data from the National Institute of Statistics (INE). The Odds Ratio (OR) was used as a measure of association between cases and controls, obtaining an OR between 0.76 and 1.08 for the confidence interval of the CI (95%). CONCLUSIONS: Despite the existence of various studies that suggest a possible association between environmental exposure to pesticides and the risk of Amyotrophic Lateral Sclerosis, our analysis of the Andalusian population did not find significant evidence of this association.

Functional outcomes of patients with NORSE and FIRES treated with immunotherapy: A systematic review.

OBJECTIVE: To determine the frequency of good functional outcomes in patients with NORSE and FIRES treated with immunotherapy. METHODS: We performed a systematic search of the MedLine and EMBASE databases to gather studies including at least 5 patients with NORSE or FIRES and at least one patient treated with immunotherapy, and reporting functional outcomes. Good functional outcome was defined as a modified Rankin Scale (mRS) score ≤ 2 (or an equivalent measure) at the last available follow-up assessment. Only patients with known functional outcomes were included in the analysis. RESULTS: We analyzed 16 studies including a total of 161 patients with NORSE. Six studies were carried out only with FIRES patients (n = 64). Of the 161 patients with NORSE, 141 (87.5%) received immunotherapy. Outcome data were available for 135, 56 of whom (41.4%) achieved good functional outcomes. Twenty-four of the 58 patients with FIRES treated with immunotherapy and for whom outcome data were available achieved good functional outcomes (41.3%). Mortality rates in patients with NORSE and FIRES treated with immunotherapy were 20/121 (16.5%) and 6/58 (10.3%), respectively. By type of immunotherapy, good functional outcomes were achieved in 36/89 patients receiving glucocorticoids (40.4%), 27/71 patients receiving IV immunoglobulins (38%), 11/37 patients treated with plasma exchange (29.7%), 5/17 patients receiving rituximab (29.4%), and 2/13 patients receiving cyclophosphamide (15.3%). CONCLUSION: Despite the lack of randomised clinical trials, immunotherapy is frequently prescribed to patients with NORSE and FIRES. However, rates of functional dependence and mortality remain high in these patients. Second-line therapies achieved lower rates of good outcomes, probably because they were administered to patients with more severe, refractory disease.

Environmental exposure to pesticides and Amyotrophic Lateral Sclerosis in the South of Spain.

OBJECTIVE: To determine if there is a relationship between environmental exposure to pesticides and the prevalence of Amyotrophic Lateral Sclerosis (ALS) in Andalusia. METHOD: We carried out a case-control study using the logistic regression method to verify the relationship between the prevalence of ALS in the area exposed to pesticides versus the unexposed area, through the Odds Ratio statistical test. RESULTS: The study population consisted of 519 individuals diagnosed with ALS between January 2016 and December 2018 according to the CMBD (Minimum Basic Data Set) as cases. In the control group, we have 8,384,083 individuals obtained from data from the National Institute of Statistics (INE). The Odds Ratio (OR) was used as a measure of association between cases and controls, obtaining an OR between 0.76 and 1.08 for the confidence interval of the CI (95%). CONCLUSIONS: Despite the existence of various studies that suggest a possible association between environmental exposure to pesticides and the risk of Amyotrophic Lateral Sclerosis, our analysis of the Andalusian population did not find significant evidence of this association.

Case fatality of COVID-19 in patients with neurodegenerative dementia. / Análisis de letalidad por COVID-19 en pacientes con demencia neurodegenerativa.

INTRODUCTION: The elderly population is the group most threatened by COVID-19, with the highest mortality rates. This study aims to analyse the case fatality of COVID-19 in a cohort of patients with degenerative dementia. METHODS: We conducted a descriptive case-control study of a sample of patients diagnosed with primary neurodegenerative dementia. RESULTS: Twenty-four of the 88 patients with COVID-19 included in the study died: 10/23 (43.4%) patients diagnosed with dementia and 14/65 (21.5%) controls; this difference was statistically significant. DISCUSSION: Our results suggest that case fatality of COVID-19 is significantly higher among patients with primary degenerative dementia than in other patients with similar mean ages and comorbidities.

Miller-Fisher syndrome after SARS-CoV-2 infection.

INTRODUCTION: On March 11th, 2020, the WHO declared the SARS-Cov-2 pandemic. Syndromes have been detected in relation to COVID-19 such as encephalitis, acute necrotizing hemorrhagic encephalopathy and cerebrovascular complications. There are also cases of peripheral nervous system involvement. METHODS: Our case would be the 3rd patient with MFS associated with COVID-19 as far as we know. RESULTS: We present a 51 years old female diagnosed with MFS two weeks after COVID-19. RTPCR to SARS-CoV-2 was negative but IgG was positive. CONCLUSION: Most of the cases were mild or moderate with typical signs and symptoms. All were treated with IV immunoglobulin with good response in most cases. Despite the short evolution time of the cases surviving the current pandemic, the description of cases of post-infectious neurological syndromes suggests that this is probably not an infrequent complication in the subacute stage of Covid-19 disease.

Impact of SARS-CoV-2 infection on neurodegenerative and neuropsychiatric diseases: a delayed pandemic? / Influencia de la infección SARS-CoV-2 sobre enfermedades neurodegenerativas y neuropsiquiátricas: ¿una pandemia demorada?

INTRODUCTION: SARS-CoV-2 was first detected in December 2019 in the Chinese city of Wuhan and has since spread across the world. At present, the virus has infected over 1.7 million people and caused over 100 000 deaths worldwide. Research is currently focused on understanding the acute infection and developing effective treatment strategies. In view of the magnitude of the epidemic, we conducted a speculative review of possible medium- and long-term neurological consequences of SARS-CoV-2 infection, with particular emphasis on neurodegenerative and neuropsychiatric diseases of neuroinflammatory origin, based on the available evidence on neurological symptoms of acute SARS-CoV-2 infection. DEVELOPMENT: We systematically reviewed the available evidence about the pathogenic mechanisms of SARS-CoV-2 infection, the immediate and lasting effects of the cytokine storm on the central nervous system, and the consequences of neuroinflammation for the central nervous system. CONCLUSIONS: SARS-CoV-2 is a neuroinvasive virus capable of triggering a cytokine storm, with persistent effects in specific populations. Although our hypothesis is highly speculative, the impact of SARS-CoV-2 infection on the onset and progression of neurodegenerative and neuropsychiatric diseases of neuroinflammatory origin should be regarded as the potential cause of a delayed pandemic that may have a major public health impact in the medium to long term. Cognitive and neuropsychological function should be closely monitored in COVID-19 survivors.

[Case fatality of COVID-19 in patients with neurodegenerative dementia]. / Análisis de letalidad por COVID-19 en pacientes con demencia neurodegenerativa.

Introduction: The elderly population is the group most threatened by COVID-19, with the highest mortality rates. This study aims to analyse the case fatality of COVID-19 in a cohort of patients with degenerative dementia. Methods: We conducted a descriptive case-control study of a sample of patients diagnosed with primary neurodegenerative dementia. Results: Twenty-four of the 88 patients with COVID-19 included in the study died: 10/23 (43.4%) patients diagnosed with dementia and 14/65 (21.5%) controls; this difference was statistically significant. Discussion: Our results suggest that case fatality of COVID-19 is significantly higher among patients with primary degenerative dementia than in other patients with similar mean ages and comorbidities.

[Myasthenia gravis induced by inmuno checkpoints inhibitors: first case report secondary to avelumab therapy and review of published cases]. / Miastenia grave inducida por tratamiento con inhibidores del punto de control inmunologico: primer caso secundario a avelumab y revision de casos previamente publicados.

INTRODUCTION: The inmuno checkpoints inhibitors are new revolutionary treatment for many neoplastic diseases in advanced stadium. There are described several types of neurological complications induced by nivolumab: polyneuropathy, seizures, radiculitis and myasthenia gravis disease. CASE REPORT: A 65 years old man with metastatic lung adenocarcinoma who presented myasthenia gravis disease induced by avelumab therapy with good response to treatment with pyridostigmine and withdrawal of avelumab. CONCLUSIONS: The exact mechanism by which this drug induces myasthenia gravis is still unknown and there is probably a different pathophysiological process to idiopathic myasthenia gravis. An important fact is the variability in the time of onset of myasthenia gravis after initiating treatment with inmuno checkpoints inhibitors. From the clinical point of view, most of the reported cases appeared with a generalized form of myasthenia gravis with bulbar involvement and later developed ophthalmoparesis and fluctuating palpebral ptosis. Our case as well as the review of the previous literature can be useful to alert the clinical neurologist about the possibility of the development of immune-mediated cases of this nature induced by the treatment with avelumab in clinical practice as well as to guide its clinical, prognostic and clinical characteristics.

TITLE: Miastenia grave inducida por tratamiento con inhibidores del punto de control inmunologico: primer caso secundario a avelumab y revision de casos previamente publicados.Introduccion. Los farmacos inhibidores del punto de control inmunologico han supuesto una revolucion en el tratamiento de varios procesos neoplasicos en estadio avanzado. Sin embargo, se han descrito numerosas complicaciones neurologicas, entre las que se encuentran polineuropatias, crisis epilepticas, radiculitis y miastenia grave. Caso clinico. Varon de 65 años con adenocarcinoma de pulmon en estadio IV en tratamiento con avelumab que desarrolla una miastenia grave ocular seropositiva, con buena respuesta a la piridostigmina y retirada de la medicacion. Conclusiones. El mecanismo exacto por el cual el avelumab induce una miastenia grave aun se desconoce, y probablemente existe un proceso fisiopatologico diferente al de la miastenia grave idiopatica. Un dato importante es la variabilidad en cuanto al tiempo de aparicion de la miastenia grave despues de iniciar el tratamiento con inhibidores del punto de control inmunologico. Desde el punto de vista clinico, la mayoria de los casos descritos comenzo con una forma de miastenia grave generalizada con afectacion bulbar que posteriormente desarrollo oftalmoparesia y ptosis palpebral fluctuante. Este caso, asi como la revision de la bibliografia, puede ser util para alertar al neurologo clinico sobre la posibilidad del desarrollo de cuadros inmunomediados de esta naturaleza inducidos por el tratamiento con avelumab en la practica clinica y para orientar sus caracteristicas clinicas, pronosticas y de tratamiento.

[Epidemiology of epilepsy in Spain and Latin America]. / Epidemiologia de la epilepsia en España y Latinoamerica.

INTRODUCTION: The connection between Spain and Latin America on the cultural, ethnic and commercial levels has been very important over the last five centuries, and this accounts for the existence of a common identity that can condition the epidemiology of chronic diseases with genetic and environmental determinants, such as epilepsy. In the last 15 years significant changes have come about in the economic development and the healthcare conditions in these countries as well as the migratory flows among them that may have brought about changes in the previous epidemiological situation. We present an exhaustive review of the epidemiological studies describing the status of epilepsy in Spain and Latin America. DEVELOPMENT: A bibliographic search was conducted of descriptive epidemiology studies about epilepsy in Spain and in each of the countries of Latin America. The methodology and quality of each study are reviewed and data on prevalence and incidence are extracted for each country. A total of 796 studies are evaluated, of which 55 (48 on prevalence and seven on incidence) meet eligibility criteria. CONCLUSIONS: There is no evidence of a variation in the epidemiological situation of epilepsy in Latin America. Some prevalence and incidence rates are still higher than in western countries. This difference is especially apparent in countries where cysticercosis is endemic and is inversely proportional to the wealth of the country, measured by the per capita gross domestic product. There is no evidence of any change in the epidemiology of epilepsy in Spain despite the migratory flows of countries with a high prevalence of epilepsy in recent years.

TITLE: Epidemiologia de la epilepsia en España y Latinoamerica.Introduccion. La conexion entre España y Latinoamerica en los ultimos cinco siglos ha sido muy importante desde el punto de vista cultural, etnico y comercial, lo que justifica la existencia de una identidad comun que puede condicionar la epidemiologia de enfermedades cronicas con determinantes geneticos y medioambientales, como la epilepsia. En los ultimos quince años se han producido cambios significativos en el desarrollo economico y de condiciones sanitarias en estos paises, asi como flujos migratorios entre ellos que pueden haber variado la situacion epidemiologica previa. Planteamos una revision exhaustiva de los estudios de epidemiologia descriptiva de la epilepsia en España y Latinoamerica. Desarrollo. Busqueda bibliografica de los estudios de epidemiologia descriptiva sobre epilepsia en España y cada uno de los paises de Latinoamerica. Se revisan su metodologia y su calidad, y se extraen los datos de prevalencia e incidencia por pais. Se evaluan 796 estudios, de los cuales 55 (48 de prevalencia y siete de incidencia) cumplen los requisitos de inclusion. Conclusiones. No existe evidencia de una variacion de la situacion epidemiologica de la epilepsia en Latinoamerica. Siguen existiendo tasas de prevalencia e incidencia mas altas que en los paises occidentales. Esta diferencia es especialmente evidente en paises endemicos para cisticercosis y esta inversamente relacionada con la riqueza del pais medida por el producto interior bruto per capita. No existe evidencia de cambio en la epidemiologia de la epilepsia en España a pesar de los flujos migratorios de paises con alta prevalencia de epilepsia en los ultimos anos.

EPICON consensus: recommendations for proper management of switching to eslicarbazepine acetate in epilepsy. / Consenso Delphi EPICON: recomendaciones sobre el manejo adecuado del cambio a acetato de eslicarbazepina en epilepsia.

INTRODUCTION: The objective of the EPICON Project is to develop a set of recommendations on how to adequately switch from carbamazepine (CBZ) and oxcarbazepine (OXC) to eslicarbazepine acetate (ESL) in some patients with epilepsy. METHODS: A steering committee drafted a questionnaire of 56 questions regarding the transition from CBZ or OXC to ESL in clinical practice (methodology and change situation). The questionnaire was then distributed to 54 epilepsy experts in 2 rounds using the Delphi method. An agreement/disagreement consensus was defined when a median ≥ 7 points or ≤ 3 was achieved, respectively, and a relative interquartile range ≤ 0.40. We analysed the results obtained to reach our conclusions. RESULTS: Our main recommendations were the following: switching from CBZ to ESL must be carried out over a period of 1 to 3 weeks with a CBZ:ESL dose ratio of 1:1.3 and is recommended for patients who frequently forget to take their medication, those who work rotating shifts, polymedicated patients, subjects with cognitive problems, severe osteoporosis-osteopaenia, dyslipidaemia, or liver disease other than acute liver failure, as well as for men with erectile dysfunction caused by CBZ. The transition from OXC to ESL can take place overnight with an OXC:ESL dose ratio of 1:1 and it is recommended for patients who frequently forget to take their medication, those who work rotating shifts, polymedicated patients, or those with cognitive problems. The transition was not recommended for patients with prior rash due to CBZ or OXC use. CONCLUSION: The EPICON Project offers a set of recommendations about the clinical management of switching from CBZ or OXC to ESL, using the Delphi method.

[Treatment adherence with levetiracetam: a non-interventionist retrospective observation-based study]. / Adhesion al tratamiento con levetiracetam: estudio observacional retrospectivo no intervencionista.

INTRODUCTION: Treatment adherence is a factor that is affecting the effectiveness of antiepileptic drugs. Levetiracetam is a drug whose effectiveness and safety is well established and is available in different oral formulations (granulates in sachets, tablets, oral solution), but information on treatment adherence/compliance with these oral formulations is limited. AIM: To determine treatment adherence with levetiracetam formulations (granulates in sachets, tablets) in adult and elderly people. PATIENTS AND METHODS: Retrospective observational non-interventionist study. During the three months before the study patients should be treated with levetiracetam (granulates in sachets or tablets), either alone or in combination. Compliance tests (Green-Morisky modified test) as well as a satisfaction questionnaire investigator and patient/caregiver is assessed. Data were analyzed using SPSS v. 21.0 program. RESULTS: A total of 466 patients completed the study. The average age was 52.75 ± 19.17 years old and the average over 65 years of 72.79 ± 6.15. 55.4% were men. Compliance is related to the variable 'pharmaceutical formulation' and with age (p = 0.031). The risk of non-compliance was 86.4% higher among those taking tablets versus granulates in sachets of levetiracetam (odds ratio: 1.864). Likewise, the non-compliance was greater among patients over 65 years. The non-compliance was related to attributing of memory failures. CONCLUSIONS: Patients older are more difficult to compliance. The granulates levetiracetam in sachets improves compliance.

TITLE: Adhesion al tratamiento con levetiracetam: estudio observacional retrospectivo no intervencionista.Introduccion. La adhesion al tratamiento es un factor que esta repercutiendo en la eficacia de los farmacos antiepilepticos. El levetiracetam es un principio activo cuya eficacia y seguridad estan ampliamente demostradas y que esta disponible en diferentes formulaciones orales (sobres, comprimidos y solucion oral), pero la informacion sobre la adhesion/cumplimiento del tratamiento con estas formulaciones orales es escasa. Objetivo. Conocer la adhesion al tratamiento con formulaciones de levetiracetam (granulado en sobre y comprimidos) en adultos y ancianos. Pacientes y metodos. Estudio observacional retrospectivo no intervencionista. Durante los tres meses previos al estudio debia estar en tratamiento con levetiracetam (granulado en sobre o comprimidos), bien en monoterapia o en combinacion. Se valoraron los tests de cumplimiento (cuestionario de Morisky-Green modificado), asi como un cuestionario de satisfaccion del investigador y del paciente/cuidador. Los datos se analizaron con el programa SPSS v. 21.0. Resultados. Un total de 466 pacientes completaron el estudio. La edad media fue de 52,75 ± 19,17 años, y la media en los mayores de 65 años fue de 72,79 ± 6,15 años. Un 55,4% eran hombres. El cumplimiento se relaciona con la variable 'formulacion galenica' y con la edad (p = 0,031). El riesgo de incumplimiento es un 86,4% mayor entre los que toman comprimidos frente a granulado de levetiracetam (odds ratio: 1,864). Asimismo, el incumplimiento es mayor entre los pacientes mayores de 65 años. El incumplimiento se relaciona con la atribucion de fallos de memoria. Conclusiones. Los pacientes mas mayores presentan mas dificultades en el cumplimiento. El granulado de levetiracetam en sobres favorece el cumplimiento.

Validation of a short useful questionnaire in Spanish for the epidemiological screening of epilepsy in Spain. EPIBERIA Questionnaire.

INTRODUCTION: There is a major gap in knowledge about the epidemiology of epilepsy in Mediterranean countries. The EPIBERIA group was formed with the aim of promoting the conducting of epidemiological studies in this region in order to improve this situation. This paper deals with the validation of a brief questionnaire for screening patients with epilepsy in the general population. METHODS: We selected an English-language questionnaire previously validated by the Ottman group. It was translated, modified to suit the characteristics of the Spanish population, and administered to a sample of 200 patients (93 epileptics and 107 non-epileptic patient controls) sampled consecutively from 5 epilepsy units in different cities in Spain. Both groups were homogeneous in demographic variables and the control group was representative of the general population. RESULTS: We obtained a sensitivity of 100% and a specificity of 74.77% for the least rigorous correction model for the questionnaire, with a sensitivity of 94.62% and a specificity of 99.07% for the most stringent correction model. The PPV ranged from 7.48% for the first case to 69.49% in the second, assuming an epilepsy prevalence of 2%. CONCLUSIONS: The questionnaire EPIBERIA is a valid Spanish tool for epilepsy screening in the general population in Spain.

Cajal's first steps in scientific research.

More than 125 years ago, Santiago Ramón y Cajal was able to draft and prove the neuron doctrine, and later, to develop prophetic theories about neural function and plasticity, many of which have been proven by current neuroscience. It was chance that made Cajal, during his doctorate studies, have his first contact with histology and force him to study the then current theories about pathogenesis of inflammation. Thus, he gained knowledge of the vascular hypothesis, by Julius Cohnheim, a German pathologist who, opposing the opinion of his teacher and father of cellular pathology, Rudolf Virchow, made leukocytes the protagonists of inflammation, given their ability to develop ameboid movements directed by chemical signals. Cohnheim's chemotactic theory deeply influenced Cajal's conception of biology. So, the basic postulates of chemotaxis can be identified at different moments in Cajal's research, from the description of the "growth cone" in embryonic neuroblasts, the origin of the neurotrophic theory, to the proposal of the pathophysiological mechanisms of neuronal plasticity. From Cajal's point of view, the neurons move during their development and also adapt to different external circumstances. Chemical endogenous substances can stimulate this movement in a similar way to leukocytes during the process of inflammation.

Antiepileptic treatment in patients with epilepsy and other comorbidities.

BACKGROUND: A high number of patients with epilepsy have comorbidities. The type of comorbidity is an important factor in deciding on the most suitable treatment, including that for acute epileptic seizures and chronic antiepileptic treatment. Evidence-based criteria should guide the selection of the appropriate antiepileptic drugs given specific comorbidities. METHODS: We performed a comprehensive search of the scientific literature on epilepsy treatment in patients with the following comorbidities: heart disease, lung disease, liver disease, kidney disease, porphyria, organ transplantation, thyroid disease, metabolic disorder, infection, mental disability, psychiatric disorder, cognitive impairment, stroke, and brain tumour. RESULTS: Most of the studies were case series and retrospective analyses. No randomised controlled trials specifically designed for this type of clinical situation were identified. The level of scientific evidence to guide clinical decisions is therefore low. CONCLUSIONS: In this review we make recommendations based on the best scientific evidence available for treating epilepsy in patients with other comorbidities, including the treatment of epileptic seizures in acute situations as well as chronic antiepileptic treatment. When no scientific evidence is available, our recommendations are based on pharmacokinetic criteria and tolerability of antiepileptic drugs, using accumulated experience and the consensus of the members of the Andalusian Epilepsy Society.