RESUMO
BACKGROUND: The smallpox vaccine is associated with more serious adverse events than any other live attenuated vaccine in use today. Although studies have examined serum cytokine levels in primary vaccine recipients at 1 and 3-5 weeks after vaccination with the smallpox vaccine, serial measurements have not been performed, and studies in revaccinated subjects have not been conducted. METHODS: We analyzed cytokine responses in both primary vaccine recipients and revaccinated subjects every other day for 2 weeks after vaccination. RESULTS: Primary vaccine recipients had maximal levels of granulocyte-colony-stimulating factor on days 6-7 after vaccination; peak levels of tumor necrosis factor (TNF)-alpha, soluble TNF receptor 1, interferon (IFN)-gamma, IFN-inducible protein-10 (IP-10), interleukin (IL)-6, and tissue inhibitor of metalloproteinases-1 on days 8-9 after vaccination; peak levels of soluble TNF receptor 2 and monokine induced by IFN-gamma (MIG) on days 10-11 after vaccination; and peak levels of granulocyte-macrophage-colony-stimulating factor on days 12-13 after vaccination. Primary vaccine recipients were significantly more likely to have higher peak levels of IFN-gamma, IP-10, and MIG after vaccination than were revaccinated subjects. Primary vaccine recipients were significantly more likely to have fatigue, lymphadenopathy, and headache, as well as a longer duration of these symptoms and more hours missed from work, compared with revaccinated subjects. CONCLUSIONS: The increased frequency and duration of symptoms observed in primary vaccine recipients, compared with revaccinated subjects, paralleled the increases in serum cytokine levels in these individuals. TRIAL REGISTRATION: Clinicaltrials.gov identifier NCT00325975.
Assuntos
Citocinas/sangue , Vacina Antivariólica/farmacologia , Adulto , Quimiocina CXCL10/sangue , Feminino , Fator Estimulador de Colônias de Granulócitos/sangue , Humanos , Interferon gama/sangue , Interferons/sangue , Interleucinas/sangue , Cinética , Masculino , Pessoa de Meia-Idade , Monocinas/sangue , Receptores do Fator de Necrose Tumoral/sangue , Vacina Antivariólica/administração & dosagem , Vacina Antivariólica/efeitos adversos , Vacina Antivariólica/imunologia , Fator de Necrose Tumoral alfa/sangue , Adulto JovemRESUMO
Mutations in MECP2 cause the autism-spectrum disorder Rett syndrome. MeCP2 is predicted to bind to methylated promoters and silence transcription. However, the first large-scale mapping of neuronal MeCP2-binding sites on 26.3 Mb of imprinted and nonimprinted loci revealed that 59% of MeCP2-binding sites are outside of genes and that only 6% are in CpG islands. Integrated genome-wide promoter analysis of MeCP2 binding, CpG methylation, and gene expression revealed that 63% of MeCP2-bound promoters are actively expressed and that only 6% are highly methylated. These results indicate that the primary function of MeCP2 is not the silencing of methylated promoters.
Assuntos
Regulação da Expressão Gênica , Proteína 2 de Ligação a Metil-CpG/metabolismo , Síndrome de Rett/genética , Sítios de Ligação , Imunoprecipitação da Cromatina , Ilhas de CpG , Metilação de DNA , Inativação Gênica , Humanos , Proteína 2 de Ligação a Metil-CpG/genética , Neurônios/metabolismo , Regiões Promotoras GenéticasRESUMO
BACKGROUND: Maternally-derived duplications that include the imprinted region on the proximal long arm of chromosome 15 underlie a complex neurobehavioral disorder characterized by cognitive impairment, seizures and a substantial risk for autism spectrum disorders1. The duplications most often take the form of a supernumerary pseudodicentric derivative chromosome 15 [der(15)] that has been called inverted duplication 15 or isodicentric 15 [idic(15)], although interstitial rearrangements also occur. Similar to the deletions found in most cases of Angelman and Prader Willi syndrome, the duplications appear to be mediated by unequal homologous recombination involving low copy repeats (LCR) that are found clustered in the region. Five recurrent breakpoints have been described in most cases of segmental aneuploidy of chromosome 15q11-q13 and previous studies have shown that most idic(15) chromosomes arise through BP3:BP3 or BP4:BP5 recombination events. RESULTS: Here we describe four duplication chromosomes that show evidence of atypical recombination events that involve regions outside the common breakpoints. Additionally, in one patient with a mosaic complex der(15), we examined homologous pairing of chromosome 15q11-q13 alleles by FISH in a region of frontal cortex, which identified mosaicism in this tissue and also demonstrated pairing of the signals from the der(15) and the normal homologues. CONCLUSION: Involvement of atypical BP in the generation of idic(15) chromosomes can lead to considerable structural heterogeneity.
Assuntos
Aberrações Cromossômicas , Cromossomos Humanos Par 15/genética , Duplicação Gênica , Isocromossomos/genética , Síndrome de Angelman/genética , Southern Blotting , Encéfalo/ultraestrutura , Linhagem Celular , Cromossomos Artificiais Bacterianos , Metilação de DNA , Feminino , Genótipo , Humanos , Hibridização in Situ Fluorescente , Cariotipagem , Masculino , Síndrome de Prader-Willi/genéticaRESUMO
Synopsis The literature pertaining to the rehabilitation of ice hockey players seeking to return to sport after anterior cruciate ligament reconstruction (ACLR) is currently limited. The purpose of this clinical commentary was to present a criterion-based progression for return to ice hockey for athletes after ACLR. First, we review pertinent literature and provide previously published guidelines on general rehabilitation after ACLR. Then, we present a 4-phase, on-ice skating progression with objective criteria to initiate each phase. During the early on-ice phase, the athlete is reintroduced to specific demands, including graded exposure to forward, backward, and crossover skating. In the intermediate on-ice phase, the emphasis shifts to developing power and introducing anticipated changes of direction within a controlled environment. During the late on-ice phase, the focus progresses to developing anaerobic endurance and introducing unanticipated changes of direction, but still without other players or contact. Finally, once objective return-to-sport criteria are met, noncontact team drills, outnumbered and even-numbered drills, practices, scrimmages, and games are progressively reintroduced during the return-to-sport phase. Recommendations for off-ice strength and conditioning exercises complement the on-ice progression. Additionally, we apply the return-to-hockey progression framework to a case report of a female collegiate defensive ice hockey player who returned to sport successfully after ACLR. This criterion-based return-to-hockey progression may guide rehabilitation specialists managing athletes returning to ice hockey after ACLR. J Orthop Sports Phys Ther 2017;47(5):324-333. Epub 29 Mar 2017. doi:10.2519/jospt.2017.7245.
Assuntos
Lesões do Ligamento Cruzado Anterior/reabilitação , Reconstrução do Ligamento Cruzado Anterior/reabilitação , Terapia por Exercício/métodos , Hóquei/lesões , Volta ao Esporte , Lesões do Ligamento Cruzado Anterior/cirurgia , Humanos , Guias de Prática Clínica como Assunto , Recuperação de Função FisiológicaRESUMO
BACKGROUND: More than 85% of Rett syndrome (RTT) patients have heterozygous mutations in the X-linked MECP2 gene which encodes methyl-CpG-binding protein 2, a transcriptional repressor that binds methylated CpG sites. Because MECP2 is subject to X chromosome inactivation (XCI), girls with RTT express either the wild type or mutant MECP2 in each of their cells. To test the hypothesis that MECP2 mutations result in genome-wide transcriptional deregulation and identify its target genes in a system that circumvents the functional mosaicism resulting from XCI, we performed gene expression profiling of pure populations of untransformed T-lymphocytes that express either a mutant or a wild-type allele. METHODS: Single T lymphocytes from a patient with a c.473C>T (p.T158M) mutation and one with a c.1308-1309delTC mutation were subcloned and subjected to short term culture. Gene expression profiles of wild-type and mutant clones were compared by oligonucleotide expression microarray analysis. RESULTS: Expression profiling yielded 44 upregulated genes and 77 downregulated genes. We compared this gene list with expression profiles of independent microarray experiments in cells and tissues of RTT patients and mouse models with Mecp2 mutations. These comparisons identified a candidate MeCP2 target gene, SPOCK1, downregulated in two independent microarray experiments, but its expression was not altered by quantitative RT-PCR analysis on brain tissues from a RTT mouse model. CONCLUSION: Initial expression profiling from T-cell clones of RTT patients identified a list of potential MeCP2 target genes. Further detailed analysis and comparison to independent microarray experiments did not confirm significantly altered expression of most candidate genes. These results are consistent with other reported data.
Assuntos
Perfilação da Expressão Gênica , Linfócitos/metabolismo , Síndrome de Rett/genética , Animais , Células Cultivadas , Células Clonais/metabolismo , Modelos Animais de Doenças , Regulação da Expressão Gênica , Humanos , Proteína 2 de Ligação a Metil-CpG/genética , Proteína 2 de Ligação a Metil-CpG/isolamento & purificação , Camundongos , Análise em Microsséries , Proteínas Mutantes/genética , Proteínas Mutantes/isolamento & purificação , Proteoglicanas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Síndrome de Rett/imunologiaRESUMO
MDMA (3,4-methylenedioxymethamphetamine) induces thermogenesis in a mitochondrial uncoupling protein 3-dependent manner. There is evidence that this hyperthermia is mediated in part by the lipolytic release of free fatty acids, that subsequently activate uncoupling protein 3 in skeletal muscle mitochondria. We hypothesize that atrial natriuretic peptide (ANP), a strong lipolytic mediator, may contribute to the induction and maintenance of MDMA-induced thermogenesis. The specific aims of this study were to (1) determine if ANP is released following MDMA administration, and (2) use the ANP receptor antagonist, Anantin, to ascertain the role of ANP in MDMA-induced hyperthermia. ANP levels were measured in plasma at baseline, 10, 20, 30 and 60 min following MDMA (40 mg/kg, sc) administration in 16 male Sprague-Dawley rats. A robust increase in ANP was seen within 10 min of MDMA administration. ANP levels returned to baseline at 20 min and then gradually rose over the 60 min monitoring period. The administration of Anantin (40 mg, ip), 15 min before and after MDMA, significantly attenuated the MDMA-induced hyperthermia. We conclude that ANP signaling contributes to the hyperthermia induced by MDMA.
Assuntos
Fator Natriurético Atrial/sangue , Febre/induzido quimicamente , Alucinógenos/toxicidade , N-Metil-3,4-Metilenodioxianfetamina/toxicidade , Pirogênios/toxicidade , Animais , Antipiréticos/uso terapêutico , Fator Natriurético Atrial/antagonistas & inibidores , Regulação da Temperatura Corporal/efeitos dos fármacos , Febre/sangue , Febre/prevenção & controle , Alucinógenos/antagonistas & inibidores , Masculino , N-Metil-3,4-Metilenodioxianfetamina/antagonistas & inibidores , Peptídeos Cíclicos/uso terapêutico , Pirogênios/antagonistas & inibidores , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Receptores do Fator Natriurético Atrial/antagonistas & inibidores , Transdução de Sinais/efeitos dos fármacos , Simpatomiméticos/antagonistas & inibidores , Simpatomiméticos/toxicidade , Fatores de TempoRESUMO
Mutations within the gene encoding methyl CpG binding protein 2 (MECP2) cause the autism-spectrum neurodevelopmental disorder Rett Syndrome (RTT). MECP2 recruits histone deacetylase to methylated DNA and acts as a long-range regulator of methylated genes. Despite ubiquitous MECP2 expression, the phenotype of RTT and the Mecp2-deficient mouse is largely restricted to the postnatal brain. Since Mecp2-deficient mice have a defect in neuronal maturation, we sought to understand how MECP2/Mecp2 mutations globally affect histone modifications during postnatal brain development by an immunofluorescence approach. Using an antibody specific to acetylated histone H3 lysine 9 (H3K9ac), a bright punctate nuclear staining pattern was observed as MECP2 expression increased in early postnatal neuronal nuclei. As neurons matured in juvenile and adult brain samples, the intensity of H3K9ac staining was reduced. Mecp2-deficient mouse and RTT cerebral neurons lacked this developmental reduction in H3K9ac staining compared to age-matched controls, resulting in a significant increase in neuronal nuclei with bright H3K9ac punctate staining. In contrast, trimethylated histone H3 lysine 9 (H3K9me3) localized to heterochromatin independent of MeCP2, but showed significantly reduced levels in Mecp2 deficient mouse and RTT brain. Autism brain with reduced MECP2 expression displayed similar histone H3 alterations as RTT brain. These observations suggest that MeCP2 regulates global histone modifications during a critical postnatal stage of neuronal maturation. These results have implications for understanding the molecular pathogenesis of RTT and autism in which MECP2 mutation or deficiency corresponds with arrested neurodevelopment.
Assuntos
Histonas/metabolismo , Lisina/metabolismo , Proteína 2 de Ligação a Metil-CpG/fisiologia , Neurônios/química , Acetilação , Animais , Imunofluorescência , Histonas/química , Proteína 2 de Ligação a Metil-CpG/genética , Camundongos , Camundongos Endogâmicos C57BL , Fenótipo , Síndrome de Rett/genéticaRESUMO
Rett syndrome (RTT), caused by mutations in MECP2 (encoding methyl CpG binding protein 2), and Angelman syndrome (AS), caused by maternal deficiency of chromosome 15q11-13, are autism-spectrum neurodevelopmental disorders. MeCP2 is a transcriptional repressor of methylated genes, but MECP2 mutation does not directly affect the imprinted expression of genes within 15q11-13. We tested a potential role for MeCP2 in the homologous pairing of imprinted 15q11-13 alleles in human brain tissue and differentiated neurons by fluorescence in situ hybridization (FISH). FISH analysis of control cerebral samples demonstrated a significant increase in homologous pairing specific to chromosome 15 from infant to juvenile brain samples. Significant and specific deficiencies in the percentage of paired chromosome 15 alleles were observed in RTT, AS and autism brain samples when compared with normal controls. SH-SY5Y neuroblastoma cells also showed a significant and specific increase in the percentage of chromosome 15q11-13 paired alleles following induced differentiation in vitro. Transfection with a methylated oligonucleotide decoy specifically blocked binding of MeCP2 to the SNURF/SNRPN promoter within 15q11-13 and significantly lowered the percentage of paired 15q11-13 alleles in SH-SY5Y cells. These combined results suggest a role for MeCP2 in chromosome organization in the developing brain and provide a potential mechanistic association between several related neurodevelopmental disorders.