ABSTRACT
La ingestión de alcohol se ha vinculado con cambios característicos en la actividad EEG, y estos cambios dependen de diversos factores; si bien se reconoce en la literatura una amplia variabilidad de diseños experimentales, la gran mayoría de estos se han centrado en reportar el efecto del alcohol en sujetos alcohólicos con antecedentes de consumo de dosis casi siempre altas y frecuentes, y en un menor porcentaje, el efecto del alcohol cuando hay un consumo agudo de dosis bajas de alcohol. El presente proyecto registró la actividad eléctrica cerebral de la atención implicada en la conducción con el equipo BCI (brain control interface) EPOC, bajo el efecto de 0,300 g de alcohol, correspondiente a un porcentaje de 0,02 % BAC (blood alcohol content) en prueba de alcoholímetro, y a su vez en ausencia de alcohol mediante un diseño pre-experimental con preprueba-postprueba, con un solo grupo de 30 estudiantes universitarios entre 18 y 45 años de edad. Los resultados mostraron que el alcohol en dosis bajas logra generar cambios en la dinámica de las ondas, disminuyendo la amplitud de ondas rápidas como alfa (9-13Hz) y beta (14-30 Hz), específicamente en zonas asociadas a los lóbulos frontales implicadas en tareas de atención sostenida en conducción.
The ingestion of alcohol has been linked to characteristic changes in EEG activity, and these changes depend on several factors. Previous research has been conducted with a variety of experimental designed, but most have focused on reporting the effect of alcohol consumption in subjects with a history of alcohol abuse, and a few have reported the effects of lower doses of alcohol. This project recorded brain activity related to attention in a driving situation with an emotiv EPOP brain control interface (BCI) device after ingestion of 0,300 g of alcohol (0,02 % BAC) or none in a pre-experiemental pre-test and post-test design with 30 college students aged 18-45. Results suggest that lower doses of alcohol change wave dynamics, reducing the amplitude of fast alpha (9-13Hz) and beta (14-30Hz) waves in frontal lobe zones involved in sustained attention in driving.
ABSTRACT
We examined the epigenetic mechanisms involved in human T-cell lymphotropic virus type 1 (HTLV-1) Tax expression. Blockade of histone deacetylation with trichostatin A (TSA) resulted in Tax upregulation. Using a chromatin immunoprecipitation (ChIP) assay, we verified local histone hyperacetylation at the HTLV-1 LTR in response to TSA. In agreement, HDAC3 transfection led to reductions in both Tax expression and histone acetylation. HDAC3 mutations and deletions spanning the catalytic site had variable ability to repress Tax, but HDAC activity was not essential for repression. Immunoprecipitation studies revealed that Tax co-exists in a complex containing both histone deacetylase 1 (HDAC1) and 3 (HDAC3). Our results suggest that HDACs may actively participate in the repression of HTLV-1 Tax transcription.
Subject(s)
Gene Expression Regulation , Genes, pX/genetics , Histone Deacetylases/physiology , Repressor Proteins , Transcription, Genetic , Acetylation , Cells, Cultured , Chromatin Immunoprecipitation , Enzyme Inhibitors/pharmacology , Histone Deacetylase Inhibitors , Histone Deacetylases/pharmacology , Histones , Human T-lymphotropic virus 1/metabolism , Humans , Hydroxamic Acids/pharmacology , Mutation , Promoter Regions, Genetic , Sequence Deletion , Transcriptional ActivationABSTRACT
To effectively direct targeted repression, the class I histone deacetylases (HDACs) associate with many important regulatory proteins. In this paper we describe the molecular characterization of a member of the Jumonji domain 2 (JMJD2) family of proteins, and demonstrate its binding to both class I HDACs and the retinoblastoma protein (pRb). JMJD2 proteins are characterized by the presence of two leukemia-associated protein/plant homeodomain (LAP/PHD) zinc fingers, one JmjN, one JmjC (containing an internal retinoblastoma-binding protein 2 (RBBP2)-like sequence), and two Tudor domains. The first member of this group, JMJD2A, is widely expressed in human tissues and cell lines, and high endogenous expression of JMJD2A mRNA was found in several cell types, including human T-cell lymphotropic virus 1 (HTLV-1)-infected cell lines. JMJD2A and JMJD2B exhibit cell type-specific responses to the HDAC inhibitor trichostatin A. We show that the JMJD2A protein associates in vivo with pRb and class I HDACs, and mediates repression of E2F-regulated promoters. In HTLV-1 virus-infected cells, we find that JMJD2A binds to the viral Tax protein. Antibodies to JMJD2A recognize the native protein but also a half-sized protein fragment, the latter up-regulated in THP-1 cells during the G(2)/M phase of the cell cycle. The ability of JMJD2A to associate with pRb and HDACs and potentiate pRb-mediated repression of E2F-regulated promoters implies an important role for this protein in cell proliferation and oncogenesis.
Subject(s)
DNA-Binding Proteins/genetics , Transcription Factors/genetics , Amino Acid Sequence , Conserved Sequence , DNA Primers , DNA, Complementary/genetics , Electroporation , Gene Library , Histone Deacetylases/genetics , Humans , Jumonji Domain-Containing Histone Demethylases , Liver/embryology , Molecular Sequence Data , Neoplasm Proteins/genetics , Oxidoreductases, N-Demethylating , Phylogeny , Polymerase Chain Reaction , Recombinant Fusion Proteins/metabolism , TransfectionABSTRACT
Multiple molecular defects trigger cell death in amyotrophic lateral sclerosis (ALS). Among these, altered transcriptional activity may perturb many cellular functions, leading to a cascade of secondary pathological effects. We showed that pharmacological treatment, using the histone deacetylase inhibitor sodium phenylbutyrate, significantly extended survival and improved both the clinical and neuropathological phenotypes in G93A transgenic ALS mice. Phenylbutyrate administration ameliorated histone hypoacetylation observed in G93A mice and induced expression of nuclear factor-kappaB (NF-kappaB) p50, the phosphorylated inhibitory subunit of NF-kappaB (pIkappaB) and beta cell lymphoma 2 (bcl-2), but reduced cytochrome c and caspase expression. Curcumin, an NF-kappaB inhibitor, and mutation of the NF-kappaB responsive element in the bcl-2 promoter, blocked butyrate-induced bcl-2 promoter activity. We provide evidence that the pharmacological induction of NF-kappaB-dependent transcription and bcl-2 gene expression is neuroprotective in ALS mice by inhibiting programmed cell death. Phenylbutyrate acts to phosphorylate IkappaB, translocating NF-kappaB p50 to the nucleus, or to directly acetylate NF-kappaB p50. NF-kappaB p50 transactivates bcl-2 gene expression. Up-regulated bcl-2 blocks cytochrome c release and subsequent caspase activation, slowing motor neuron death. These transcriptional and post-translational pathways ultimately promote motor neuron survival and ameliorate disease progression in ALS mice. Phenylbutyrate may therefore provide a novel therapeutic approach for the treatment of patients with ALS.
Subject(s)
Amyotrophic Lateral Sclerosis/physiopathology , Apoptosis/genetics , Gene Expression/drug effects , Phenylbutyrates/pharmacology , Acetylation/drug effects , Amyotrophic Lateral Sclerosis/genetics , Amyotrophic Lateral Sclerosis/mortality , Amyotrophic Lateral Sclerosis/pathology , Animals , Caspase Inhibitors , Cell Survival/drug effects , Curcumin/pharmacology , Cytochromes c/antagonists & inhibitors , Disease Progression , Histones/metabolism , Male , Mice , Mice, Transgenic , Motor Neurons/drug effects , NF-kappa B/metabolism , NF-kappa B p50 Subunit , Promoter Regions, Genetic/drug effects , Protein Precursors/metabolism , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , Spinal Cord/drug effects , Spinal Cord/metabolism , Spinal Cord/pathologyABSTRACT
We demonstrate that the histone deacetylase (HDAC) inhibitor drug trichostatin A (TSA) reduces spinal cord inflammation, demyelination, neuronal and axonal loss and ameliorates disability in the relapsing phase of experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis (MS). TSA up-regulates antioxidant, anti-excitotoxicity and pro-neuronal growth and differentiation mRNAs. TSA also inhibits caspase activation and down-regulates gene targets of the pro-apoptotic E2F transcription factor pathway. In splenocytes, TSA reduces chemotactic, pro-Th1 and pro-proliferative mRNAs. A transcriptional imbalance in MS may contribute to immune dysregulation and neurodegeneration, and we identify HDAC inhibition as a transcriptional intervention to ameliorate this imbalance.
Subject(s)
Encephalomyelitis, Autoimmune, Experimental/drug therapy , Gene Expression Regulation/drug effects , Hydroxamic Acids/therapeutic use , Protein Synthesis Inhibitors/therapeutic use , Animals , Cell Death/drug effects , Cells, Cultured , Cerebral Cortex/cytology , Cytokines/genetics , Cytokines/metabolism , Disease Models, Animal , Drug Administration Schedule , Drug Interactions , Embryo, Mammalian , Encephalomyelitis, Autoimmune, Experimental/chemically induced , Encephalomyelitis, Autoimmune, Experimental/genetics , Encephalomyelitis, Autoimmune, Experimental/pathology , Female , Gene Expression Profiling/methods , Glycoproteins , Immunohistochemistry/methods , Mice , Mice, Inbred C57BL , Myelin-Oligodendrocyte Glycoprotein , Neurons/drug effects , Oligonucleotide Array Sequence Analysis/methods , Peptide Fragments , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction/methods , Severity of Illness Index , Spleen/drug effects , Spleen/metabolism , Tetrazolium Salts , Thiazoles , Time FactorsABSTRACT
We performed microarray analysis of peripheral blood mononuclear cells (PBMCs) from multiple sclerosis (MS) patients and detected a profile of immune cell activation, autoantigen upregulation, and enhanced E2F pathway transcription. Accordingly, E2f1-deficient mice manifested only mild disability upon induction of experimental autoimmune encephalomyelitis (EAE). Furthermore, PBMCs from Avonex-treated patients had lower expression of E2F targets. The profile was enriched in genes known to harbor MS-associated polymorphisms, or localized to MS susceptibility chromosomal regions. Our study shows that PBMC microarrays reflect MS pathobiology that can be validated in the EAE model.
Subject(s)
Cell Cycle Proteins , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Gene Expression Profiling , Leukocytes, Mononuclear/metabolism , Multiple Sclerosis, Relapsing-Remitting/genetics , Multiple Sclerosis, Relapsing-Remitting/metabolism , Oligonucleotide Array Sequence Analysis , Signal Transduction/genetics , Transcription Factors/genetics , Transcription Factors/metabolism , Adolescent , Adult , Animals , Chromosome Mapping , DNA-Binding Proteins/deficiency , DNA-Binding Proteins/physiology , E2F Transcription Factors , E2F1 Transcription Factor , Encephalomyelitis, Autoimmune, Experimental/genetics , Encephalomyelitis, Autoimmune, Experimental/immunology , Encephalomyelitis, Autoimmune, Experimental/metabolism , Female , Gene Expression Profiling/methods , Gene Expression Profiling/statistics & numerical data , Humans , Interferon beta-1a , Interferon-beta/therapeutic use , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/immunology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Middle Aged , Multigene Family/drug effects , Multiple Sclerosis, Relapsing-Remitting/drug therapy , Multiple Sclerosis, Relapsing-Remitting/immunology , Oligonucleotide Array Sequence Analysis/methods , Oligonucleotide Array Sequence Analysis/statistics & numerical data , Signal Transduction/drug effects , Signal Transduction/immunology , Transcription Factors/deficiency , Transcription Factors/physiologyABSTRACT
Little is known about global gene expression patterns in the human neurodegenerative disease amyotrophic lateral sclerosis (ALS). To address this, we used high-density oligonucleotide microarray technology to compare expression levels of approximately 6,800 genes in postmortem spinal cord gray matter obtained from individuals with ALS as well as normal individuals. Using Fisher discriminant analysis (FDA) and leave-one-out cross-validation (LOOCV), we discerned an ALS-specific signature. Moreover, it was possible to distinguish familial ALS (FALS) from sporadic ALS (SALS) gene expression profiles. Characterization of the specific genes significantly altered in ALS uncovered a pro-inflammatory terminal state. Moreover, we found alterations in genes involved in mitochondrial function, oxidative stress, excitotoxicity, apoptosis, cytoskeletal architecture, RNA transcription and translation, proteasomal function, and growth and signaling. It is apparent from this study that DNA microarray analysis and appropriate bioinformatics can reveal distinct phenotypic changes that underlie the terminal stages of neurodegeneration in ALS.