Congress
Copyright © 2017 John Libbey Eurotext. Downloaded by an anonymous user on 21/04/2017.
8th Latin American Congress on Epilepsy Abstracts
Platform session 1 Thursday, 18 September, 2014 001 Identificac¸ão de variac¸ões no número de cópias em pacientes com malformac¸ões do córtex cerebral -------------------------------------------------------------------------Torres FR1 , Mazutti M1 , Guerreiro M1 , Montenegro MA1 , dos Santos AC2 , Terra VC2 , Sakamoto AS2 , Cendes F1 , Lopes-Cendes I1 UNICAMP/Faculty of Medical Sciences, Campinas, Brazil, 2 USP/Faculty of Medical Sciences, Ribeirão Preto, Brazil
doi:10.1684/epd.2014.0694
1
Objectivo: Pacientes com malformac¸ões do córtex cerebral (MCC) frequentemente apresentam crises epiléticas refratárias ao tratamento clínico. Avanc¸os em genética molecular têm levado a um melhor entendimento dos mecanismos envolvidos na etiologia das MCC. No entanto, mutac¸ões por alterac¸ões de sequência nucleotídica não são identificadas em um número significativo de pacientes. Estudos recentes mostram que variac¸ões genômicas estruturais, também conhecidas por variac¸ões no número de cópias genômicas (CNVs) estão envolvidas em um grande número de doenc¸as neurológicas. O objetivo desse estudo foi investigar se CNVs estão envolvidas em diferentes tipos de MCC. Métodos: Foram utilizados SNP-microarrays de alta resoluc¸ão CytoScan® HD para investigar the presenc¸a of CNVs em um grupo de 52 pacientes com MCC, incluído heterotopia nodular periventricular, espectro lisencefalia-heterotopia subcortical em banda, esquizencefalia e polimicrogiria. Para avaliar o possível impacto patogênico das CNVs identificadas foram consultados o Database of Genomic Variants (DGV) e o The International Standards for Cytogenomic Arrays Con-
sortium (ISCA). Os genes contidos nas CNVs foram pesquisados no Gene Ontology Terms (Go). Resultados: Nós identificamos um total de 31 novas CNVs, das quais 19 tem potencial patogênico. As CNVs identificadas tem tamanho médio de 245 kb, variando de 113 a 600 kb. Cada CNV contem aproximadamente dois genes. As CNVs com potencial deletério afetam genes envolvidos no transporte vesicular (TSNARE1), organizac¸ão do citoesqueleto de actina (DAAM1, ACTR6), direcionamento dos axônios e migrac¸ão neuronal (PLXNA1, KIRREL3, ARX, DCX). Vários genes afetados pelas CNVs tem papéis em vias moleculares relacionadas as MCC. Conclusões: Mutac¸ões deletérias em genes que controlam mitose, transporte mediado por vesículas, organizac¸ão do citoesqueleto e migrac¸ão neuronal como WDR62, AFGRF2, FLN1 e LIS1 foram descritos em pacientes com MCC. Portanto, os SNP-arrays se mostram como uma importante ferramenta para identificar alterac¸ões genéticas envolvidas com MCC e genes candidatos envolvidos com a patogênese destas doenc¸as.
Abnormal copy number variations identified in patients with malformations of cerebral cortex -------------------------------------------------------------------------Torres FR1 , Mazutti M1 , Guerreiro M1 , Montenegro MA1 , dos Santos AC2 , Terra VC2 , Sakamoto AS2 , Cendes F1 , Lopes-Cendes I1 1
UNICAMP/Faculty of Medical Sciences, Campinas, Brazil, 2 USP/Faculty of Medical Sciences, Ribeirão Preto, Brazil
Objective: Patients with malformations of cerebral cortex (MCC) often suffer from seizures refractory to antiepileptic drugs. Advances in molecular biology have led to a better understanding of MCC etiology. However, the genetic cause still remains unidentified
Epileptic Disord, Vol. 16, Abstracts 8th LACE, Buenos Aires (Argentina), 17th-20th September 2014
1
Copyright © 2017 John Libbey Eurotext. Downloaded by an anonymous user on 21/04/2017.
Congress
in the majority of patients. Recent studies have implicated large, rare copy number variations (CNVs) in a range of neurodevelopmental disorders. The aim of this study was search for deleterious CNVs in patients with MCC. Methods: We used a high resolution SNP array platform (CytoScan® HD) to investigate CNVs in a cohort of 52 patients with MCC, including lissencephaly spectrum, nodular periventricular heterotopia, polymicrogyria and schizencephaly. To assess the clinical significance of the insertions/deletions found, we checked CNVs in the Database of Genomic Variants (DGV) and The International Standards For Cytogenomic Arrays Consortium (ISCA). Genes located within the rare CNVs were subjected for specific Gene Ontology Terms (Go). Results: We detected 31 rare CNVs, including 19 potentially pathogenic variants. Average size of CNVs was 245 kb, ranging from 113 kb to 600 kb. Each CNVcontains approximately two genes. Potentially pathogenic CNVs, according to DGV and ISCA databases, contain genes involved in cell division (NCAPG2, HAUS7), vesicle mediated transport (TSNARE1), actin cytoskeleton organization (DAAM1, ACTR6) and axon guidance or neuronal migration (PLXNA1, KIRREL3, ARX, DCX). Several genes that reside in regions spanned by rare and potentially pathogenic CNVs described here are related to molecular pathways involved with MCC. Conclusions: Deleterious mutations in genes controlling mitosis, vesicle mediated transport, cytoskeleton organization and neuronal migration, such as WDR62, AFGRF2, FLN1 and LIS1 have been reported in patients with MCC. Therefore, SNP arrays have shown to be a powerful tool for identifying the genetic abnormalities causing MCC. In addition, this approach can identify candidate genes involved in MCC pathogenesis.
002 Ensamble de modelos computacionales para ˜ de fármacos para el tratamiento asistir el diseno de la epilepsia refractaria -------------------------------------------------------------------------Gantner ME1 , Bruno-Blanch LE1 , Talevi A1 1
National University of La Plata, Biological Sciences, La Plata, Argentina
de la epilepsia refractaria y otros desórdenes del sistema nervioso central. Presentamos el desarrollo de un ensamble de modelos computacionales no lineales capaz de discriminar entre sustratos y no sustratos de BCRP. Métodos: Se generó una base de datos de 262 sustratos y no sustratos de BCRP. Los clasificadores no lineales se obtuvieron utilizando el algoritmo de árboles de decisión J48. Los 12 mejores clasificadores se validaron y se ensamblaron calculando el ranking promedio. Resultados: Experimentos computacionales realizados demostraron que el ensamble presenta la mejor capacidad para discriminar entre sustratos y no sustratos de BCRP. Conclusiones: El ensamble desarrollado es una her˜ de ramienta de gran utilidad para asistir el diseno fármacos destinados al tratamiento de la epilepsia refractaria. Referencias: 1. Nakanishi H, Yonezawa A, Matsubara K, Yano I. Impact of P-glycoprotein and breast cancer resistance protein on the brain distribution of antiepileptic drugs in knockout mouse models. Eur J Pharmacol 2013 ;710: 20-8. 2. Tucker TG1 , Milne AM, Fournel-Gigleux S, Fenner KS, Coughtrie MW. Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistanceassociated protein 2 in human liver and duodenum. Biochem Pharmacol 2012; 83: 279-85. 3. Dauchy S1 , Dutheil F, Weaver RJ, Chassoux F, Daumas-Duport C, Couraud PO, et al. ABC transporters, cytochromes P450 and their main transcription factors: expression at the human blood-brain barrier. J Neurochem 2008; 107: 1518-28. 4. Uchida Y1 , Ohtsuki S, Katsukura Y, Ikeda C, Suzuki T, Kamiie J, et al. Quantitative targeted absolute proteomics of human blood-brain barrier transporters and receptors. J Neurochem 2011; 117: 333-45.
Computational model ensemble to assist drug design for the treatment of refractory epilepsy -------------------------------------------------------------------------Gantner ME1 , Bruno-Blanch LE1 , Talevi A1
Objetivo: La epilepsia refractaria se encuentra asociada a la sobreexpresión de transportadores de la familia ABC tales como la glicoproteína P (Pgp) y la proteína de resistencia del cáncer de mama (BCRP) [1]. Reportes indican que BCRP es el transportador más expresado en el intestino [2] y la barrera hematoencefálica [3, 4] de tejidos sanos, limitando la absorción oral y la biodisponibilidad de sus sustratos en el cerebro. El reconocimiento temprano de éstos es por tanto ˜ esencial para disenar fármacos para el tratamiento
2
1
National University of La Plata, Biological Sciences, La Plata, Argentina
Objective: Refractory epilepsy has been associated with an overexpression of ABC (ATP-binding cassette) transporters such as P-glycoprotein (Pgp) and Breast Cancer Resistance Protein (BCRP) [1]. A number of reports indicate that BCRP is the most abundantly expressed ABC efflux transporter throughout the intestine [2] and the blood-brain barrier [3, 4] of healthy tissues, limiting oral absorption and
Epileptic Disord, Vol. 16, Abstracts 8th LACE, Buenos Aires (Argentina), 17th-20th September 2014
Copyright © 2017 John Libbey Eurotext. Downloaded by an anonymous user on 21/04/2017.
brain bioavailability of its substrates. Early recognition of BCRP substrates is thus essential to design novel therapeutics for the treatment of refractory epilepsy and other central nervous system conditions linked to BCRP-mediated multidrug resistance issues. Therefore, we present the development of an ensemble of nonlinear computational models capable of discriminating between BCRP substrates and nonsubstrates. Methods: A dataset of 262 substrates and nonsubstrates of BCRP was compiled from literature. J48 decision tree-inducing algorithm was used to obtain independent nonlinear classifiers and the 12 decision trees with the best classification performance were validated and combined using rank average classifier ensemble. Results: According to the computational experiments conducted, the resulting ensemble has the best capacity to discriminate between BCRP substrates and non-substrates. Conclusions: The ensemble developed is a potentially valuable tool to assist the discovery of new drugs for the treatment of refractory epilepsy. References: 1. Nakanishi H, Yonezawa A, Matsubara K, Yano I. Impact of P-glycoprotein and breast cancer resistance protein on the brain distribution of antiepileptic drugs in knockout mouse models. Eur J Pharmacol 2013; 710: 20-8. 2. Tucker TG1 , Milne AM, Fournel-Gigleux S, Fenner KS, Coughtrie MW. Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistanceassociated protein 2 in human liver and duodenum. Biochem Pharmacol 2012; 83: 279-85. 3. Dauchy S1 , Dutheil F, Weaver RJ, Chassoux F, Daumas-Duport C, Couraud PO, et al. ABC transporters, cytochromes P450 and their main transcription factors: expression at the human blood-brain barrier. J Neurochem 2008; 107: 1518-28. 4. Uchida Y1 , Ohtsuki S, Katsukura Y, Ikeda C, Suzuki T, Kamiie J, et al. Quantitative targeted absolute proteomics of human blood-brain barrier transporters and receptors. J Neurochem 2011; 117: 333-45.
003 Monitorización electroencefalográfica continua en unidades de pacientes críticos -------------------------------------------------------------------------Uribe R1 , Acevedo K1 , Moya P1 , Krakowiak MJ1 , Mesa T1 , Santin J1 , Godoy J1 1
Pontificia Universidad Católica de Chile, Santiago, Chile
Introducción: El monitoreo electroencefalográfico continuo (mEEGc) corresponde a la principal herramienta diagnóstica de crisis epilépticas subclínicas
y del estatus epiléptico no convulsivo (EENC) en las unidades de paciente crítico (UPC). Métodos: Nosotros evaluamos de forma prospectiva los monitoreos realizados durante los últimos 9 meses en las UPC pediátricas y de adulto. Resultados: Se analizaron 38 mEEGc, correspondientes a 36 pacientes (19 hombres). La edad promedio ˜ ˜ fue 50 anos (rango: entre 5 meses y 88 anos). La indicación de monitorización más frecuente fue compromiso de conciencia (53%) y luego la presencia de crisis epilépticas objetivadas por personal de la UPC (42%). En promedio las monitorizaciones duraron 85 horas (4-479). Se encontró SENC en 13 pacientes (34%) y crisis subclínicas en 10 (26%). En promedio la duración de los SENC fue de 79 horas (11-400). Ocho pacientes cumplen criterios de SENC súperrefractario (correspondientes al 62% de los SENC). Como tratamiento, se utilizó anestésicos en 13 casos y en promedio se usaron 3 antiepilépticos (0-10). En el 79% de los casos el monitoreo ejerció un cambio en la conducta en el tratante. La mortalidad general en el seguimiento fue de 18% y el 40% de los pacientes finaliza con secuelas neurológicas graves. Conclusiones: El mEEGc es una herramienta fundamental en el manejo de las UPC y permite el diagnóstico certero de SENC y crisis subclínicas, en especial en pacientes comprometidos de conciencia y sin claros signos de crisis. Estos hallazgos se asocian a una importante mortalidad o la presencia de secuelas neurológicas graves futura.
Continuous EEG monitoring in critical care -------------------------------------------------------------------------Uribe R1 , Acevedo K1 , Moya P1 , Krakowiak MJ1 , Mesa T1 , Santin J1 , Godoy J1 1
Pontificia Universidad Católica de Chile, Santiago, Chile
Introduction: Continuous electroencephalographic monitoring (cEEGm) is the main tool for diagnosis in subclinical seizures and non-convulsive status epilepticus (NCSE), in the intensive care units (ICU). Methods: We prospectively evaluated the cEEGm conducted between April 2012 and January 2013 in the adult and pediatric ICU. Results: We analyzed 38 cEEGm, corresponding to 36 patients (19 men). Mean age was 50 years (range: 5 months to 88 years). The most common indication for monitoring was impaired consciousness (53%), seconded by the presence of seizures observed by an ICU staff (42%). On average, monitoring lasted 85 hours (range: 4 to 479 hours). We found NCSE in 13 patients (34%) and subclinical seizures in 10 (26%). The mean duration of NCSE was 79 hours (range: 11 to 400 hours). Eight patients met criteria for superrefractory NCSE (62% of all NCES). The treatment of
Epileptic Disord, Vol. 16, Abstracts 8th LACE, Buenos Aires (Argentina), 17th-20th September 2014
3
Congress
Copyright © 2017 John Libbey Eurotext. Downloaded by an anonymous user on 21/04/2017.
choice was anesthetics in 13 patients and the association of 3 antiepileptic drugs was commonly used (range: 0 to 10 drugs). In 79% of the patients, cEEGm motivated a change in the treatment plan. Overall mortality was 18%, and severe neurological sequelae were observed in 40% of patients. Conclusions: The cEEGm is a fundamental tool in the management of ICU patients; it allows accurate diagnosis of NCSE and subclinical epileptic activity, especially in patients with impaired consciousness and no clear evidence of seizures. These findings are associated with significant mortality and future neurologic sequelae.
004 Inmunorreactividad para nestina en el giro dentado de pacientes con esclerosis hipocampal -------------------------------------------------------------------------D’Alessio L1,2,3 , Konopka H4 , Escobar E4 , Gori MB4 , Solis P5 , Seoane E3 , Kochen S4 1 Cell Biology and Neuroscience E de Robertis Institute, CONICET, Buenos Aires University, Buenos Aires, Argentina, 2 Epilepsy Center, Ramos Mejía Hospital, Buenos Aires, Argentina, 3 Neurosurgery Center, Ramos Mejía Hospital, Buenos Aires, Argentina, 4 Cell Biology and Neuroscience E de Robertis Institute, Buenos Aires, Argentina, 5 Hospital Ramos Mejia, Buenos Aires, Argentina
Objetivo: La nestina es una proteína que se expresa en las células progenitoras neurales (NPCs) y en el giro dentado (DG) identifica neuronas proliferativas y células de nueva generación. El objetivo de este estudio fue el de determinar la inmunoreactividad para nestina en el hipocampo de pacientes con epilepsia del lóbulo temporal (TLE) y esclerosis del hipocampo (HE) que recibieron tratamiento quirúrgico. Métodos: Se estudiaron secciones de hipocampos de 16 pacientes con HE y TLE resistente que fueron procesadas mediante inmunoperoxidasa para nestina. Material de archivo de 8 hipocampos post mortem normales fueron procesados simultáneamente. Se determinó mediante análisis de imagen computarizado (imagen J) el área total reactiva para nestina, el número total de células positivas (NC) por campo y el valor medio de grises (MGV). Se utilizó el test de Student como prueba estadística. Resultados: Se encontraron NC en giro dentado (DG), capas piramidales (CA1- CA4) y en las áreas subpiales (SZ). En el DG del tejido epiléptico, las NC positivas presentaron dispersión celular, localización ectópica, aumento del soma y del área reactiva (p
Copyright © 2017 John Libbey Eurotext. Downloaded by an anonymous user on 21/04/2017.
8th Latin American Congress on Epilepsy Abstracts
Platform session 1 Thursday, 18 September, 2014 001 Identificac¸ão de variac¸ões no número de cópias em pacientes com malformac¸ões do córtex cerebral -------------------------------------------------------------------------Torres FR1 , Mazutti M1 , Guerreiro M1 , Montenegro MA1 , dos Santos AC2 , Terra VC2 , Sakamoto AS2 , Cendes F1 , Lopes-Cendes I1 UNICAMP/Faculty of Medical Sciences, Campinas, Brazil, 2 USP/Faculty of Medical Sciences, Ribeirão Preto, Brazil
doi:10.1684/epd.2014.0694
1
Objectivo: Pacientes com malformac¸ões do córtex cerebral (MCC) frequentemente apresentam crises epiléticas refratárias ao tratamento clínico. Avanc¸os em genética molecular têm levado a um melhor entendimento dos mecanismos envolvidos na etiologia das MCC. No entanto, mutac¸ões por alterac¸ões de sequência nucleotídica não são identificadas em um número significativo de pacientes. Estudos recentes mostram que variac¸ões genômicas estruturais, também conhecidas por variac¸ões no número de cópias genômicas (CNVs) estão envolvidas em um grande número de doenc¸as neurológicas. O objetivo desse estudo foi investigar se CNVs estão envolvidas em diferentes tipos de MCC. Métodos: Foram utilizados SNP-microarrays de alta resoluc¸ão CytoScan® HD para investigar the presenc¸a of CNVs em um grupo de 52 pacientes com MCC, incluído heterotopia nodular periventricular, espectro lisencefalia-heterotopia subcortical em banda, esquizencefalia e polimicrogiria. Para avaliar o possível impacto patogênico das CNVs identificadas foram consultados o Database of Genomic Variants (DGV) e o The International Standards for Cytogenomic Arrays Con-
sortium (ISCA). Os genes contidos nas CNVs foram pesquisados no Gene Ontology Terms (Go). Resultados: Nós identificamos um total de 31 novas CNVs, das quais 19 tem potencial patogênico. As CNVs identificadas tem tamanho médio de 245 kb, variando de 113 a 600 kb. Cada CNV contem aproximadamente dois genes. As CNVs com potencial deletério afetam genes envolvidos no transporte vesicular (TSNARE1), organizac¸ão do citoesqueleto de actina (DAAM1, ACTR6), direcionamento dos axônios e migrac¸ão neuronal (PLXNA1, KIRREL3, ARX, DCX). Vários genes afetados pelas CNVs tem papéis em vias moleculares relacionadas as MCC. Conclusões: Mutac¸ões deletérias em genes que controlam mitose, transporte mediado por vesículas, organizac¸ão do citoesqueleto e migrac¸ão neuronal como WDR62, AFGRF2, FLN1 e LIS1 foram descritos em pacientes com MCC. Portanto, os SNP-arrays se mostram como uma importante ferramenta para identificar alterac¸ões genéticas envolvidas com MCC e genes candidatos envolvidos com a patogênese destas doenc¸as.
Abnormal copy number variations identified in patients with malformations of cerebral cortex -------------------------------------------------------------------------Torres FR1 , Mazutti M1 , Guerreiro M1 , Montenegro MA1 , dos Santos AC2 , Terra VC2 , Sakamoto AS2 , Cendes F1 , Lopes-Cendes I1 1
UNICAMP/Faculty of Medical Sciences, Campinas, Brazil, 2 USP/Faculty of Medical Sciences, Ribeirão Preto, Brazil
Objective: Patients with malformations of cerebral cortex (MCC) often suffer from seizures refractory to antiepileptic drugs. Advances in molecular biology have led to a better understanding of MCC etiology. However, the genetic cause still remains unidentified
Epileptic Disord, Vol. 16, Abstracts 8th LACE, Buenos Aires (Argentina), 17th-20th September 2014
1
Copyright © 2017 John Libbey Eurotext. Downloaded by an anonymous user on 21/04/2017.
Congress
in the majority of patients. Recent studies have implicated large, rare copy number variations (CNVs) in a range of neurodevelopmental disorders. The aim of this study was search for deleterious CNVs in patients with MCC. Methods: We used a high resolution SNP array platform (CytoScan® HD) to investigate CNVs in a cohort of 52 patients with MCC, including lissencephaly spectrum, nodular periventricular heterotopia, polymicrogyria and schizencephaly. To assess the clinical significance of the insertions/deletions found, we checked CNVs in the Database of Genomic Variants (DGV) and The International Standards For Cytogenomic Arrays Consortium (ISCA). Genes located within the rare CNVs were subjected for specific Gene Ontology Terms (Go). Results: We detected 31 rare CNVs, including 19 potentially pathogenic variants. Average size of CNVs was 245 kb, ranging from 113 kb to 600 kb. Each CNVcontains approximately two genes. Potentially pathogenic CNVs, according to DGV and ISCA databases, contain genes involved in cell division (NCAPG2, HAUS7), vesicle mediated transport (TSNARE1), actin cytoskeleton organization (DAAM1, ACTR6) and axon guidance or neuronal migration (PLXNA1, KIRREL3, ARX, DCX). Several genes that reside in regions spanned by rare and potentially pathogenic CNVs described here are related to molecular pathways involved with MCC. Conclusions: Deleterious mutations in genes controlling mitosis, vesicle mediated transport, cytoskeleton organization and neuronal migration, such as WDR62, AFGRF2, FLN1 and LIS1 have been reported in patients with MCC. Therefore, SNP arrays have shown to be a powerful tool for identifying the genetic abnormalities causing MCC. In addition, this approach can identify candidate genes involved in MCC pathogenesis.
002 Ensamble de modelos computacionales para ˜ de fármacos para el tratamiento asistir el diseno de la epilepsia refractaria -------------------------------------------------------------------------Gantner ME1 , Bruno-Blanch LE1 , Talevi A1 1
National University of La Plata, Biological Sciences, La Plata, Argentina
de la epilepsia refractaria y otros desórdenes del sistema nervioso central. Presentamos el desarrollo de un ensamble de modelos computacionales no lineales capaz de discriminar entre sustratos y no sustratos de BCRP. Métodos: Se generó una base de datos de 262 sustratos y no sustratos de BCRP. Los clasificadores no lineales se obtuvieron utilizando el algoritmo de árboles de decisión J48. Los 12 mejores clasificadores se validaron y se ensamblaron calculando el ranking promedio. Resultados: Experimentos computacionales realizados demostraron que el ensamble presenta la mejor capacidad para discriminar entre sustratos y no sustratos de BCRP. Conclusiones: El ensamble desarrollado es una her˜ de ramienta de gran utilidad para asistir el diseno fármacos destinados al tratamiento de la epilepsia refractaria. Referencias: 1. Nakanishi H, Yonezawa A, Matsubara K, Yano I. Impact of P-glycoprotein and breast cancer resistance protein on the brain distribution of antiepileptic drugs in knockout mouse models. Eur J Pharmacol 2013 ;710: 20-8. 2. Tucker TG1 , Milne AM, Fournel-Gigleux S, Fenner KS, Coughtrie MW. Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistanceassociated protein 2 in human liver and duodenum. Biochem Pharmacol 2012; 83: 279-85. 3. Dauchy S1 , Dutheil F, Weaver RJ, Chassoux F, Daumas-Duport C, Couraud PO, et al. ABC transporters, cytochromes P450 and their main transcription factors: expression at the human blood-brain barrier. J Neurochem 2008; 107: 1518-28. 4. Uchida Y1 , Ohtsuki S, Katsukura Y, Ikeda C, Suzuki T, Kamiie J, et al. Quantitative targeted absolute proteomics of human blood-brain barrier transporters and receptors. J Neurochem 2011; 117: 333-45.
Computational model ensemble to assist drug design for the treatment of refractory epilepsy -------------------------------------------------------------------------Gantner ME1 , Bruno-Blanch LE1 , Talevi A1
Objetivo: La epilepsia refractaria se encuentra asociada a la sobreexpresión de transportadores de la familia ABC tales como la glicoproteína P (Pgp) y la proteína de resistencia del cáncer de mama (BCRP) [1]. Reportes indican que BCRP es el transportador más expresado en el intestino [2] y la barrera hematoencefálica [3, 4] de tejidos sanos, limitando la absorción oral y la biodisponibilidad de sus sustratos en el cerebro. El reconocimiento temprano de éstos es por tanto ˜ esencial para disenar fármacos para el tratamiento
2
1
National University of La Plata, Biological Sciences, La Plata, Argentina
Objective: Refractory epilepsy has been associated with an overexpression of ABC (ATP-binding cassette) transporters such as P-glycoprotein (Pgp) and Breast Cancer Resistance Protein (BCRP) [1]. A number of reports indicate that BCRP is the most abundantly expressed ABC efflux transporter throughout the intestine [2] and the blood-brain barrier [3, 4] of healthy tissues, limiting oral absorption and
Epileptic Disord, Vol. 16, Abstracts 8th LACE, Buenos Aires (Argentina), 17th-20th September 2014
Copyright © 2017 John Libbey Eurotext. Downloaded by an anonymous user on 21/04/2017.
brain bioavailability of its substrates. Early recognition of BCRP substrates is thus essential to design novel therapeutics for the treatment of refractory epilepsy and other central nervous system conditions linked to BCRP-mediated multidrug resistance issues. Therefore, we present the development of an ensemble of nonlinear computational models capable of discriminating between BCRP substrates and nonsubstrates. Methods: A dataset of 262 substrates and nonsubstrates of BCRP was compiled from literature. J48 decision tree-inducing algorithm was used to obtain independent nonlinear classifiers and the 12 decision trees with the best classification performance were validated and combined using rank average classifier ensemble. Results: According to the computational experiments conducted, the resulting ensemble has the best capacity to discriminate between BCRP substrates and non-substrates. Conclusions: The ensemble developed is a potentially valuable tool to assist the discovery of new drugs for the treatment of refractory epilepsy. References: 1. Nakanishi H, Yonezawa A, Matsubara K, Yano I. Impact of P-glycoprotein and breast cancer resistance protein on the brain distribution of antiepileptic drugs in knockout mouse models. Eur J Pharmacol 2013; 710: 20-8. 2. Tucker TG1 , Milne AM, Fournel-Gigleux S, Fenner KS, Coughtrie MW. Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistanceassociated protein 2 in human liver and duodenum. Biochem Pharmacol 2012; 83: 279-85. 3. Dauchy S1 , Dutheil F, Weaver RJ, Chassoux F, Daumas-Duport C, Couraud PO, et al. ABC transporters, cytochromes P450 and their main transcription factors: expression at the human blood-brain barrier. J Neurochem 2008; 107: 1518-28. 4. Uchida Y1 , Ohtsuki S, Katsukura Y, Ikeda C, Suzuki T, Kamiie J, et al. Quantitative targeted absolute proteomics of human blood-brain barrier transporters and receptors. J Neurochem 2011; 117: 333-45.
003 Monitorización electroencefalográfica continua en unidades de pacientes críticos -------------------------------------------------------------------------Uribe R1 , Acevedo K1 , Moya P1 , Krakowiak MJ1 , Mesa T1 , Santin J1 , Godoy J1 1
Pontificia Universidad Católica de Chile, Santiago, Chile
Introducción: El monitoreo electroencefalográfico continuo (mEEGc) corresponde a la principal herramienta diagnóstica de crisis epilépticas subclínicas
y del estatus epiléptico no convulsivo (EENC) en las unidades de paciente crítico (UPC). Métodos: Nosotros evaluamos de forma prospectiva los monitoreos realizados durante los últimos 9 meses en las UPC pediátricas y de adulto. Resultados: Se analizaron 38 mEEGc, correspondientes a 36 pacientes (19 hombres). La edad promedio ˜ ˜ fue 50 anos (rango: entre 5 meses y 88 anos). La indicación de monitorización más frecuente fue compromiso de conciencia (53%) y luego la presencia de crisis epilépticas objetivadas por personal de la UPC (42%). En promedio las monitorizaciones duraron 85 horas (4-479). Se encontró SENC en 13 pacientes (34%) y crisis subclínicas en 10 (26%). En promedio la duración de los SENC fue de 79 horas (11-400). Ocho pacientes cumplen criterios de SENC súperrefractario (correspondientes al 62% de los SENC). Como tratamiento, se utilizó anestésicos en 13 casos y en promedio se usaron 3 antiepilépticos (0-10). En el 79% de los casos el monitoreo ejerció un cambio en la conducta en el tratante. La mortalidad general en el seguimiento fue de 18% y el 40% de los pacientes finaliza con secuelas neurológicas graves. Conclusiones: El mEEGc es una herramienta fundamental en el manejo de las UPC y permite el diagnóstico certero de SENC y crisis subclínicas, en especial en pacientes comprometidos de conciencia y sin claros signos de crisis. Estos hallazgos se asocian a una importante mortalidad o la presencia de secuelas neurológicas graves futura.
Continuous EEG monitoring in critical care -------------------------------------------------------------------------Uribe R1 , Acevedo K1 , Moya P1 , Krakowiak MJ1 , Mesa T1 , Santin J1 , Godoy J1 1
Pontificia Universidad Católica de Chile, Santiago, Chile
Introduction: Continuous electroencephalographic monitoring (cEEGm) is the main tool for diagnosis in subclinical seizures and non-convulsive status epilepticus (NCSE), in the intensive care units (ICU). Methods: We prospectively evaluated the cEEGm conducted between April 2012 and January 2013 in the adult and pediatric ICU. Results: We analyzed 38 cEEGm, corresponding to 36 patients (19 men). Mean age was 50 years (range: 5 months to 88 years). The most common indication for monitoring was impaired consciousness (53%), seconded by the presence of seizures observed by an ICU staff (42%). On average, monitoring lasted 85 hours (range: 4 to 479 hours). We found NCSE in 13 patients (34%) and subclinical seizures in 10 (26%). The mean duration of NCSE was 79 hours (range: 11 to 400 hours). Eight patients met criteria for superrefractory NCSE (62% of all NCES). The treatment of
Epileptic Disord, Vol. 16, Abstracts 8th LACE, Buenos Aires (Argentina), 17th-20th September 2014
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choice was anesthetics in 13 patients and the association of 3 antiepileptic drugs was commonly used (range: 0 to 10 drugs). In 79% of the patients, cEEGm motivated a change in the treatment plan. Overall mortality was 18%, and severe neurological sequelae were observed in 40% of patients. Conclusions: The cEEGm is a fundamental tool in the management of ICU patients; it allows accurate diagnosis of NCSE and subclinical epileptic activity, especially in patients with impaired consciousness and no clear evidence of seizures. These findings are associated with significant mortality and future neurologic sequelae.
004 Inmunorreactividad para nestina en el giro dentado de pacientes con esclerosis hipocampal -------------------------------------------------------------------------D’Alessio L1,2,3 , Konopka H4 , Escobar E4 , Gori MB4 , Solis P5 , Seoane E3 , Kochen S4 1 Cell Biology and Neuroscience E de Robertis Institute, CONICET, Buenos Aires University, Buenos Aires, Argentina, 2 Epilepsy Center, Ramos Mejía Hospital, Buenos Aires, Argentina, 3 Neurosurgery Center, Ramos Mejía Hospital, Buenos Aires, Argentina, 4 Cell Biology and Neuroscience E de Robertis Institute, Buenos Aires, Argentina, 5 Hospital Ramos Mejia, Buenos Aires, Argentina
Objetivo: La nestina es una proteína que se expresa en las células progenitoras neurales (NPCs) y en el giro dentado (DG) identifica neuronas proliferativas y células de nueva generación. El objetivo de este estudio fue el de determinar la inmunoreactividad para nestina en el hipocampo de pacientes con epilepsia del lóbulo temporal (TLE) y esclerosis del hipocampo (HE) que recibieron tratamiento quirúrgico. Métodos: Se estudiaron secciones de hipocampos de 16 pacientes con HE y TLE resistente que fueron procesadas mediante inmunoperoxidasa para nestina. Material de archivo de 8 hipocampos post mortem normales fueron procesados simultáneamente. Se determinó mediante análisis de imagen computarizado (imagen J) el área total reactiva para nestina, el número total de células positivas (NC) por campo y el valor medio de grises (MGV). Se utilizó el test de Student como prueba estadística. Resultados: Se encontraron NC en giro dentado (DG), capas piramidales (CA1- CA4) y en las áreas subpiales (SZ). En el DG del tejido epiléptico, las NC positivas presentaron dispersión celular, localización ectópica, aumento del soma y del área reactiva (p
