RESUMEN
Persistence is an attribute of long-term memories (LTM) that has recently caught researcher's attention in search for mechanisms triggered by experience that assure memory perdurability. Up-to-date, scarce evidence of relationship between reconsolidation and persistence has been described. Here, we characterized hippocampal ERK participation in LTM reconsolidation and persistence using an inhibitory avoidance task (IA) at different time points. Intra-dorsal-hippocampal (dHIP) administration of an ERK inhibitor (PD098059, PD, 1.0µg/hippocampus) 3h after retrieval did not affect reconsolidation of a strong IA, when tested 24h apart. However, the same manipulation impaired performance when animals were tested at 7d, regardless of the training's strength; and being specific to memory reactivation. To the best of our knowledge, this is the first report showing that persistence might be triggered after memory reactivation involving an ERK/MAPK-dependent process.
Asunto(s)
Reacción de Prevención/fisiología , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Hipocampo/metabolismo , Consolidación de la Memoria/fisiología , Memoria a Largo Plazo/fisiología , Recuerdo Mental/fisiología , Inhibidores de Proteínas Quinasas/farmacología , Animales , Reacción de Prevención/efectos de los fármacos , Quinasas MAP Reguladas por Señal Extracelular/antagonistas & inhibidores , Flavonoides/administración & dosificación , Flavonoides/farmacología , Hipocampo/efectos de los fármacos , Consolidación de la Memoria/efectos de los fármacos , Memoria a Largo Plazo/efectos de los fármacos , Recuerdo Mental/efectos de los fármacos , Ratones , Inhibidores de Proteínas Quinasas/administración & dosificación , Factores de TiempoRESUMEN
Reconsolidation has been defined as the process of memory stabilization after retrieval involving, among others, gene expression regulation and post-translational modifications. Many of these mechanisms are shared with memory consolidation. Here, we studied hippocampal ERK participation on memory reconsolidation of an inhibitory avoidance task in CF-1 mice. We found a retrieval-induced cytosolic ERK2 activation in the hippocampus (HIP) 15 min after memory reactivation, and an inhibition at 45 min. PD098059, a MEK1/2 (MAPK/ERK kinase) inhibitor, administered in the HIP immediately after retrieval impaired memory in a dose-dependent fashion. However, infusions of the highest dose of PD098059 performed 40 min after retrieval enhanced memory in mice trained with a weaker footshock. These results suggest for the first time that ERK2 is involved in memory reconsolidation in a biphasic fashion. Furthermore, the inhibition of ERK could either impair or enhance mice performance depending on ERK state of activation.
Asunto(s)
Reacción de Prevención/fisiología , Sistema de Señalización de MAP Quinasas/fisiología , Memoria/fisiología , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Procesamiento Proteico-Postraduccional/fisiología , Animales , Hipocampo/metabolismo , Masculino , Ratones , FosforilaciónRESUMEN
An adult male farmer with chronic active hepatitis and cirrhosis despite previous circulating anti-HBs antibodies was studied. No markers of other hepatotropic viral infection were observed. HBV DNA was detected in serum by PCR and was characterized further by restriction fragment length polymorphism (RFLP) and sequencing of cloned PCR products derived from the S gene. The HBV DNA was ascribed to genotype F, and single-strand conformational polymorphism (SSCP) demonstrated the co-circulation of multiple quasispecies. Some of the variants exhibited changes located within the neutralizing "a" determinant, located between amino acids 124-147 of the S protein. Within this region, two clones showed either C124R or C124Y mutations. Other mutations were Q129R, C138R, C139R, and S140T (one clone each). Outside the "a" determinant several substitutions were documented. The high degree of the quasispecies variability was probably linked to the severity of the infection. Most members of the patient's family were infected with HBV, all with genotype F.
Asunto(s)
Anticuerpos contra la Hepatitis B/análisis , Antígenos de Superficie de la Hepatitis B/análisis , Virus de la Hepatitis B/genética , Hepatitis B Crónica/virología , Sustitución de Aminoácidos , Anticuerpos contra la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/inmunología , Hepatitis B Crónica/inmunología , Hepatitis B Crónica/patología , Humanos , Neoplasias Hepáticas/virología , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Mutación , Linaje , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo Conformacional Retorcido-Simple , Análisis de Secuencia de ADNRESUMEN
Typing of hepatitis C virus (HCV) isolates from Argentine patients was performed by using different methodologies in a population of 243 patients. HCV subtype was assigned based upon restriction fragment length polymorphism (RFLP). HCV RNA genomes obtained from serum samples were classified as belonging to clade 1 (53.5%), 2 (23. 0%), or 3 (8.6%); 14.8% of samples showed HCV mixed infections, more frequently implying different subtypes within the same clade. In addition to RFLP typing, phylogenetic relatedness among sequences from both 5' untranslated region (n = 50) and nonstructural 5B coding region (n = 15) was established.
Asunto(s)
Hepacivirus/genética , Regiones no Traducidas 5'/química , Adolescente , Adulto , Anciano , Niño , Preescolar , Femenino , Genotipo , Hepacivirus/clasificación , Humanos , Hígado/fisiopatología , Masculino , Persona de Mediana Edad , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Estudios Retrospectivos , Abuso de Sustancias por Vía Intravenosa/complicacionesRESUMEN
GBV-C/HGV RNA was investigated in serum samples from 70 HIV(+) intravenous drug users (IVDU), as well as from 200 blood donors from Buenos Aires, Argentina. Viral RNA was demonstrated in 21 IVDU by reverse transcription-nested PCR of the 5' UTR. c-DNA amplified products were analyzed and their sequences compared with those downloaded from GenBank. A phylogenetic tree based on 171 sequences demonstrated the presence of three major genogroups, including two subgroups, within local samples, i.e. group 1 (n=1), 2a (n=11), 2b (n=4) and 3 (n=5). These results agreed entirely with those obtained by a novel RFLP (J. Clin. Microbiol. 37, 1340-1347, 1999) of the same 5' UTR amplicons. As expected, GBV-C/HGV RNA prevalence was significantly higher among IVDU than among blood donors (P<0.0001), although within the latter group an unexpectedly high rate was also detected, since 11 of 200 sera (5.5%) proved positive. These viral isolates were ascribed either to subgroup 2a (n=5), subgroup 2b (n=5) or genogroup 3 (n=1). Briefly, this partial view of GBV-C/HGV molecular epidemiology in Argentina shows: (i) different rates of GBV-C/HGV infection within both IVDU and blood donors; (ii) a high prevalence of viral RNA among blood donors; and (iii) a predominant circulation of genogroup 2, with minor contribution of groups 3 and 1.
Asunto(s)
Donantes de Sangre , Flaviviridae/genética , Infecciones por VIH/complicaciones , Abuso de Sustancias por Vía Intravenosa/complicaciones , Adulto , Antígenos Virales/genética , Argentina , Femenino , Flaviviridae/aislamiento & purificación , Pruebas Genéticas , Variación Genética , Infecciones por VIH/virología , Humanos , Masculino , Glicoproteínas de Membrana/genética , Proteínas del Envoltorio Viral/genéticaRESUMEN
A phylogenetic tree based on 150 5' untranslated region sequences deposited in GenBank database allowed segregation of the sequences into three major groups, including two subgroups, i.e., 1, 2a, 2b, and 3, supported by bootstrap analysis. Restriction site analysis of these sequences predicted that HinfI and either AatII or AciI could be used for genomic typing with 99.4% accuracy. cDNA sequencing and subsequent alignment of 21 Argentine GB virus C/hepatitis G virus strains confirmed restriction fragment length polymorphism patterns theoretically predicted. This method may be useful for a rapid screening of samples when either epidemiological or transmission studies of this agent are carried out.
Asunto(s)
Regiones no Traducidas 5'/química , Flaviviridae/clasificación , Polimorfismo de Longitud del Fragmento de Restricción , Secuencia de Bases , ADN Complementario/química , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Neste trabalho é apresentado o projeto e desenvolvimento de um espectrofluorímetro para a obtenção de espectros de fluorescência e refletância difusa de tecidos biológicos em um tempo inferior a 1s. Para acessar a região da diagnose, o sistema utiliza um catéter a fibra óptica para excitação do tecido e captação da fluorescência emitida. O sistema é desenvolvido para aplicação em procedimentos clínicos in vivo, onde o tempo de realização do experimento é objetivo de extrema importância
Abstract- This work presents the project and development of a spectrofluorimeter in order to obtain the fluorescence spectra and diffuse reflectance from biological tissues with period of time below 1 s. The diagnose region is reached with an optical fiber catheter for tissue excitation and collection of the emitted fluorescence. This system is developed for in vivo clinicai applications, where duration of the experimental procedure is a very important parameter