Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 20
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
J Cereb Blood Flow Metab ; 40(2): 298-313, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30398083

RESUMO

Neurovascular coupling (through which local cerebral blood flow changes in response to neural activation are mediated) is impaired in many diseases including diabetes. Current preclinical rodent models of neurovascular coupling rely on invasive surgery and instrumentation, but transgenic zebrafish coupled with advances in imaging techniques allow non-invasive quantification of cerebrovascular anatomy, neural activation, and cerebral vessel haemodynamics. We therefore established a novel non-invasive, non-anaesthetised zebrafish larval model of neurovascular coupling, in which visual stimulus evokes neuronal activation in the optic tectum that is associated with a specific increase in red blood cell speed in tectal blood vessels. We applied this model to the examination of the effect of glucose exposure on cerebrovascular patterning and neurovascular coupling. We found that chronic exposure of zebrafish to glucose impaired tectal blood vessel patterning and neurovascular coupling. The nitric oxide donor sodium nitroprusside rescued all these adverse effects of glucose exposure on cerebrovascular patterning and function. Our results establish the first non-mammalian model of neurovascular coupling, offering the potential to perform more rapid genetic modifications and high-throughput screening than is currently possible using rodents. Furthermore, using this zebrafish model, we reveal a potential strategy to ameliorate the effects of hyperglycemia on cerebrovascular function.

2.
Dis Model Mech ; 12(9)2019 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-31481433

RESUMO

Diabetes is associated with dysfunction of the neurovascular unit, although the mechanisms of this are incompletely understood and currently no treatment exists to prevent these negative effects. We previously found that the nitric oxide (NO) donor sodium nitroprusside (SNP) prevents the detrimental effect of glucose on neurovascular coupling in zebrafish. We therefore sought to establish the wider effects of glucose exposure on both the neurovascular unit and on behaviour in zebrafish, and the ability of SNP to prevent these. We incubated 4-days post-fertilisation (dpf) zebrafish embryos in 20 mM glucose or mannitol for 5 days until 9 dpf, with or without 0.1 mM SNP co-treatment for 24 h (8-9 dpf), and quantified vascular NO reactivity, vascular mural cell number, expression of a klf2a reporter, glial fibrillary acidic protein (GFAP) and transient receptor potential cation channel subfamily V member 4 (TRPV4), as well as spontaneous neuronal activation at 9 dpf, all in the optic tectum. We also assessed the effect on light/dark preference and locomotory characteristics during free-swimming studies. We find that glucose exposure significantly reduced NO reactivity, klf2a reporter expression, vascular mural cell number and TRPV4 expression, while significantly increasing spontaneous neuronal activation and GFAP expression (all in the optic tectum). Furthermore, when we examined larval behaviour, we found that glucose exposure significantly altered light/dark preference and high and low speed locomotion while in light. Co-treatment with SNP reversed all these molecular and behavioural effects of glucose exposure. Our findings comprehensively describe the negative effects of glucose exposure on the vascular anatomy, molecular phenotype and function of the optic tectum, and on whole-organism behaviour. We also show that SNP or other NO donors may represent a therapeutic strategy to ameliorate the complications of diabetes on the neurovascular unit.This article has an associated First Person interview with the first author of the paper.

3.
EMBO Rep ; 20(8): e47047, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31379129

RESUMO

We identify a novel endothelial membrane behaviour in transgenic zebrafish. Cerebral blood vessels extrude large transient spherical structures that persist for an average of 23 min before regressing into the parent vessel. We term these structures "kugeln", after the German for sphere. Kugeln are only observed arising from the cerebral vessels and are present as late as 28 days post fertilization. Kugeln do not communicate with the vessel lumen and can form in the absence of blood flow. They contain little or no cytoplasm, but the majority are highly positive for nitric oxide reactivity. Kugeln do not interact with brain lymphatic endothelial cells (BLECs) and can form in their absence, nor do they perform a scavenging role or interact with macrophages. Inhibition of actin polymerization, Myosin II, or Notch signalling reduces kugel formation, while inhibition of VEGF or Wnt dysregulation (either inhibition or activation) increases kugel formation. Kugeln represent a novel Notch-dependent NO-containing endothelial organelle restricted to the cerebral vessels, of currently unknown function.

4.
Nat Commun ; 10(1): 453, 2019 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-30692543

RESUMO

Venous endothelial cells are molecularly and functionally distinct from their arterial counterparts. Although veins are often considered the default endothelial state, genetic manipulations can modulate both acquisition and loss of venous fate, suggesting that venous identity is the result of active transcriptional regulation. However, little is known about this process. Here we show that BMP signalling controls venous identity via the ALK3/BMPR1A receptor and SMAD1/SMAD5. Perturbations to TGF-ß and BMP signalling in mice and zebrafish result in aberrant vein formation and loss of expression of the venous-specific gene Ephb4, with no effect on arterial identity. Analysis of a venous endothelium-specific enhancer for Ephb4 shows enriched binding of SMAD1/5 and a requirement for SMAD binding motifs. Further, our results demonstrate that BMP/SMAD-mediated Ephb4 expression requires the venous-enriched BMP type I receptor ALK3/BMPR1A. Together, our analysis demonstrates a requirement for BMP signalling in the establishment of Ephb4 expression and the venous vasculature.


Assuntos
Receptores de Proteínas Morfogenéticas Ósseas Tipo I/genética , Proteínas Morfogenéticas Ósseas/genética , Regulação da Expressão Gênica no Desenvolvimento , Transdução de Sinais/genética , Veias/metabolismo , Animais , Animais Geneticamente Modificados , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/metabolismo , Proteínas Morfogenéticas Ósseas/metabolismo , Células Endoteliais/metabolismo , Camundongos Knockout , Camundongos Transgênicos , Receptor EphB4/genética , Receptor EphB4/metabolismo , Proteína Smad1/genética , Proteína Smad1/metabolismo , Proteína Smad5/genética , Proteína Smad5/metabolismo , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo , Veias/embriologia , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
5.
Appl Plant Sci ; 5(1)2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28090406

RESUMO

PREMISE OF THE STUDY: The search for resistance/tolerance to the devastating citrus huanglongbing disease (syn. HLB or citrus greening) is generating an increasing number of new plants of diverse genetic makeup. As the increasing number of new plants require more space, resources, and time, the need for faster and more efficient HLB screening tests becomes crucial. METHODS AND RESULTS: The leaf-disc grafting system described here consists in replacing a disc of leaf tissue with a similar disc from an infected plant. This can be performed in young seedlings not yet big enough to endure other types of grafting. Graft success and infection rates average approximately 80%. CONCLUSIONS: We describe the successful adaptation of leaf-disc grafting as a powerful screening tool for HLB. The system requires minimal plant material and can be performed in seedlings at a very young age with increased efficiency in terms of time, space, and resources.

6.
PLoS One ; 10(10): e0141611, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26506092

RESUMO

INTRODUCTION AND OBJECTIVES: The zinc-finger transcription factor Krϋppel-like factor 2 (KLF2) transduces blood flow into molecular signals responsible for a wide range of responses within the vasculature. KLF2 maintains a healthy, quiescent endothelial phenotype. Previous studies report a range of phenotypes following morpholino antisense oligonucleotide-induced klf2a knockdown in zebrafish. Targeted genome editing is an increasingly applied method for functional assessment of candidate genes. We therefore generated a stable klf2a mutant zebrafish and characterised its cardiovascular and haematopoietic development. METHODS AND RESULTS: Using Transcription Activator-Like Effector Nucleases (TALEN) we generated a klf2a mutant (klf2ash317) with a 14bp deletion leading to a premature stop codon in exon 2. Western blotting confirmed loss of wild type Klf2a protein and the presence of a truncated protein in klf2ash317 mutants. Homozygous klf2ash317 mutants exhibit no defects in vascular patterning, survive to adulthood and are fertile, without displaying previously described morphant phenotypes such as high-output cardiac failure, reduced haematopoetic stem cell (HSC) development or impaired formation of the 5th accessory aortic arch. Homozygous klf2ash317 mutation did not reduce angiogenesis in zebrafish with homozygous mutations in von Hippel Lindau (vhl), a form of angiogenesis that is dependent on blood flow. We examined expression of three klf family members in wildtype and klf2ash317 zebrafish. We detected vascular expression of klf2b (but not klf4a or biklf/klf4b/klf17) in wildtypes but found no differences in expression that might account for the lack of phenotype in klf2ash317 mutants. klf2b morpholino knockdown did not affect heart rate or impair formation of the 5th accessory aortic arch in either wildtypes or klf2ash317 mutants. CONCLUSIONS: The klf2ash317 mutation produces a truncated Klf2a protein but, unlike morpholino induced klf2a knockdown, does not affect cardiovascular development.


Assuntos
Sistema Cardiovascular/crescimento & desenvolvimento , Sistema Hematopoético/crescimento & desenvolvimento , Fatores de Transcrição Kruppel-Like/genética , Morfogênese/genética , Proteínas de Peixe-Zebra/genética , Animais , Regulação da Expressão Gênica no Desenvolvimento , Genótipo , Humanos , Fatores de Transcrição Kruppel-Like/antagonistas & inibidores , Fatores de Transcrição Kruppel-Like/biossíntese , Morfolinos/genética , Mutação , Transdução de Sinais , Peixe-Zebra/genética , Peixe-Zebra/crescimento & desenvolvimento , Proteínas de Peixe-Zebra/antagonistas & inibidores , Proteínas de Peixe-Zebra/biossíntese
7.
FEMS Microbiol Lett ; 358(1): 14-20, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25053267

RESUMO

The Flavobacterium psychrophilum gliding motility N (GldN) protein was investigated to determine its ability to elicit antibody responses and provide protective immunity in rainbow trout Oncorhynchus mykiss (Walbaum). GldN was PCR-amplified, cloned into pET102/D-TOPO, and expressed in Escherichia coli. Bacteria expressing recombinant GldN (rGldN) were formalin-inactivated and injected intraperitoneally (i.p.) into rainbow trout with Freund's complete adjuvant (FCA) in four separate studies that used two different immunization protocols followed by challenge evaluations. Fish injected with E. coli only in FCA served as the control. Antibody responses to F. psychrophilum whole-cell lysates measured by ELISA were low in all four studies. Protection against F. psychrophilum challenge was observed in the first study, but not in the three following studies. The discrepancies in results obtained in the later studies are unclear but may relate to formalin treatment of the antigen preparations. Overall, it appeared that rGldN delivered i.p. as a crude formalin-killed preparation is not a consistent vaccine candidate, and more work is required. Additionally, this study illustrates the importance of conducting multiple in vivo evaluations on potential vaccine(s) before any conclusions are drawn.


Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Doenças dos Peixes/prevenção & controle , Infecções por Flavobacteriaceae/veterinária , Flavobacterium/imunologia , Oncorhynchus mykiss/imunologia , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/genética , Clonagem Molecular , Ensaio de Imunoadsorção Enzimática , Escherichia coli/genética , Doenças dos Peixes/imunologia , Infecções por Flavobacteriaceae/imunologia , Infecções por Flavobacteriaceae/prevenção & controle , Adjuvante de Freund/administração & dosagem , Expressão Gênica , Injeções Intraperitoneais , Análise de Sobrevida , Resultado do Tratamento , Vacinação/métodos , Vacinas de Produtos Inativados/administração & dosagem , Vacinas de Produtos Inativados/genética , Vacinas de Produtos Inativados/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
8.
PLoS One ; 9(3): e91855, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24651119

RESUMO

We have previously shown that the respiratory syncytial virus [RSV] can productively infect monocyte derived dendritic cells [MoDC] and remain dormant within the same cells for prolonged periods. It is therefore possible that infected dendritic cells act as a reservoir within the airways of individuals between annual epidemics. In the present study we explored the possibility that sub-epithelial DCs can be infected with RSV from differentiated bronchial epithelium and that in turn RSV from DCs can infect the epithelium. A dual co-culture model was established in which a differentiated primary airway epithelium on an Air Liquid Interface (ALI) was cultured on a transwell insert and MoDCs were subsequently added to the basolateral membrane of the insert. Further experiments were undertaken using a triple co-culture model in which in which macrophages were added to the apical surface of the differentiated epithelium. A modified RSV [rr-RSV] expressing a red fluorescent protein marker of replication was used to infect either the MoDCs or the differentiated epithelium and infection of the reciprocal cell type was assessed using confocal microscopy. Our data shows that primary epithelium became infected when rr-RSV infected MoDCs were introduced onto the basal surface of the transwell insert. MoDCs located beneath the epithelium did not become infected with virus from infected epithelial cells in the dual co-culture model. However when macrophages were present on the apical surface of the primary epithelium infection of the basal MoDCs occurred. Our data suggests that RSV infected dendritic cells readily transmit infection to epithelial cells even when they are located beneath the basal layer. However macrophages appear to be necessary for the transmission of infection from epithelial cells to basal dendritic cells.


Assuntos
Células Dendríticas/virologia , Epitélio/virologia , Macrófagos/virologia , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sinciciais Respiratórios/fisiologia , Adulto , Diferenciação Celular , Técnicas de Cocultura , Células Dendríticas/patologia , Células Epiteliais/patologia , Células Epiteliais/virologia , Epitélio/patologia , Fluorescência , Humanos , Macrófagos/patologia , Monócitos/patologia , Reprodutibilidade dos Testes , Infecções por Vírus Respiratório Sincicial/patologia , Replicação Viral
9.
Arterioscler Thromb Vasc Biol ; 33(6): 1257-63, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23559631

RESUMO

OBJECTIVE: Coarctation of the aorta is rarely associated with known gene defects. Blomstrand chondrodysplasia, caused by mutations in the parathyroid hormone receptor 1 (PTHR1) is associated with coarctation of the aorta in some cases, although it is unclear whether PTHR1 deficiency causes coarctation of the aorta directly. The zebrafish allows the study of vascular development using approaches not possible in other models. We therefore examined the effect of loss of function of PTHR1 or its ligand parathyroid hormone-related peptide (PTHrP) on aortic formation in zebrafish. APPROACH AND RESULTS: Morpholino antisense oligonucleotide knockdown of either PTHR1 or PTHrP led to a localized occlusion of the mid-aorta in developing zebrafish. Confocal imaging of transgenic embryos showed that these defects were caused by loss of endothelium, rather than failure to lumenize. Using a Notch reporter transgenic ([CSL:Venus]qmc61), we found both PTHR1 and PTHrP knockdown-induced defective Notch signaling in the hypochord at the site of the aortic defect before onset of circulation, and the aortic occlusion was rescued by inducible Notch upregulation. CONCLUSIONS: Loss of function of either PTHR1 or PTHrP leads to a localized aortic defect that is Notch dependent. These findings may underlie the aortic defect seen in Blomstrand chondrodysplasia, and reveal a link between parathyroid hormone and Notch signaling during aortic development.


Assuntos
Aorta/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Proteínas do Tecido Nervoso/genética , Receptor Notch1/genética , Receptor Tipo 1 de Hormônio Paratireóideo/genética , Transdução de Sinais/genética , Proteínas de Peixe-Zebra/genética , Animais , Coartação Aórtica/genética , Coartação Aórtica/fisiopatologia , Feminino , Masculino , Modelos Animais , Mutação/genética , Neovascularização Fisiológica/genética , Valores de Referência , Regulação para Cima , Peixe-Zebra
10.
Open Virol J ; 5: 114-23, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22046209

RESUMO

Respiratory Syncytial Virus (RSV) causes annual epidemics of respiratory disease particularly affecting infants. The associated airway inflammation is characterized by an intense neutrophilia. This neutrophilic inflammation appears to be responsible for much of the pathology and symptoms. Previous work from our group had shown that there are factors within the airways of infants with RSV bronchiolitis that inhibit neutrophil apoptosis. This study was undertaken to determine if RSV can directly affect neutrophil survival.NEUTROPHILS WERE ISOLATED FROM CITRATED VENOUS BLOOD (COLLECTED FROM HEALTHY ADULT VOLUNTEERS) BY DISCONTINUOUS PLASMA: Percoll gradient centrifugation and, in some experiments, further purified by negative immunomagnetic bead selection. The effect of RSV on neutrophil survival was measured by Annexin V-PE /To-Pro-3 staining and by morphological changes, using Dif-Quick staining of cytospins.Inhibition of neutrophil apoptosis was observed in neutrophils isolated by standard plasma:Percoll gradient when exposed to RSV but not in ultra pure neutrophil preparations. Adding monocytes back to ultra purified preparations restored the effect. The inhibition of apoptosis was observed with both active and UV inactivated virus. The effect is dependent on a soluble factor and appears to be dependent on CD14 receptors on the monocytes.

11.
Dev Comp Immunol ; 35(12): 1256-62, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21414351

RESUMO

Disease prevention is essential to the continued development of aquaculture around the world. Vaccination is the most effective method of combating disease and currently there are a number of vaccines commercially available for use in fish. The majority of aquatic vaccines are delivered by injection, which is by far the most effective method when compared to oral or immersion deliveries. However it is labor intensive, costly and not feasible for large numbers of fish under 20 g. Attempts to develop novel oral and immersion delivery methods have resulted in varying degrees of success but may have great potential for the future.


Assuntos
Imunidade Adaptativa , Infecções Bacterianas/imunologia , Vacinas Bacterianas/administração & dosagem , Doenças dos Peixes/prevenção & controle , Peixes/imunologia , Vacinação , Vacinas Virais/administração & dosagem , Viroses/imunologia , Administração Oral , Animais , Artemia/microbiologia , Artemia/virologia , Bactérias/imunologia , Infecções Bacterianas/microbiologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Doenças dos Peixes/virologia , Peixes/microbiologia , Peixes/virologia , Imersão , Injeções Intramusculares , Injeções Intraperitoneais , Nanopartículas/administração & dosagem , Concentração Osmolar , Plantas Geneticamente Modificadas/microbiologia , Plantas Geneticamente Modificadas/virologia , Som , Viroses/virologia , Vírus/imunologia
13.
Mol Cancer Ther ; 7(5): 1013-24, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18445660

RESUMO

Histopathologic grading of astrocytic tumors based on current WHO criteria offers a valuable but simplified representation of oncologic reality and is often insufficient to predict clinical outcome. In this study, we report a new astrocytic tumor microarray gene expression data set (n = 65). We have used a simple artificial neural network algorithm to address grading of human astrocytic tumors, derive specific transcriptional signatures from histopathologic subtypes of astrocytic tumors, and asses whether these molecular signatures define survival prognostic subclasses. Fifty-nine classifier genes were identified and found to fall within three distinct functional classes, that is, angiogenesis, cell differentiation, and lower-grade astrocytic tumor discrimination. These gene classes were found to characterize three molecular tumor subtypes denoted ANGIO, INTER, and LOWER. Grading of samples using these subtypes agreed with prior histopathologic grading for both our data set (96.15%) and an independent data set. Six tumors were particularly challenging to diagnose histopathologically. We present an artificial neural network grading for these samples and offer an evidence-based interpretation of grading results using clinical metadata to substantiate findings. The prognostic value of the three identified tumor subtypes was found to outperform histopathologic grading as well as tumor subtypes reported in other studies, indicating a high survival prognostic potential for the 59 gene classifiers. Finally, 11 gene classifiers that differentiate between primary and secondary glioblastomas were also identified.


Assuntos
Astrocitoma/diagnóstico , Astrocitoma/mortalidade , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/mortalidade , Perfilação da Expressão Gênica/métodos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Algoritmos , Proteínas Reguladoras de Apoptose , Astrocitoma/classificação , Astrocitoma/genética , Biomarcadores Tumorais/genética , Neoplasias Encefálicas/classificação , Neoplasias Encefálicas/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Fosfoproteínas/genética , Prognóstico , Reprodutibilidade dos Testes , Taxa de Sobrevida
14.
Fish Shellfish Immunol ; 24(5): 575-83, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18321729

RESUMO

In order to further characterise channel catfish (Ictalurus punctatus) Mx1, studies were initiated to amplify and clone the Mx1 promoter into a reporter vector, pGL3basic. Initially the Mx1 gene was amplified from genomic DNA and was found to have 12 exons and 11 introns, spanning a region over 6 kilobases (kb) in length. The Mx1 promoter was amplified using genome walking and during this process four additional Mx promoters were identified, suggesting the presence of five Mx genes in the channel catfish. All five promoters possess an interferon stimulated response element (ISRE) and the Mx1 promoter possessed two potential NF-kappabeta transcription sites. Following cloning each construct was transiently transfected into COS-7 and EPC cells for 24h and treated with 5 microg/ml poly I:C for 24h. An increase in expression of the reporter gene in response to poly I:C was noted in both cell lines in the pGL3Mx1 construct only. However, the reporter gene was also constitutively expressed in these cells. Constitutive expression was also observed in channel catfish ovary cells transiently transfected with pGL3Mx1 only. Treatment with 5 microg/ml poly I:C did not increase this expression, which may be due to high levels of cell death in this difficult to transfect cell line. The constitutive expression observed implies that a repressor element is missing in the 390 base pair sequence of the Mx1 promoter used in this study. These results suggest that only channel catfish Mx1 is involved in the type I interferon pathway and that the presence of an ISRE in a regulatory region is not necessarily indicative of a role in the type I interferon response.


Assuntos
Proteínas de Peixes/genética , Proteínas de Ligação ao GTP/genética , Genoma/genética , Ictaluridae/genética , Regiões Promotoras Genéticas/genética , Animais , Sequência de Bases , Células COS , Linhagem Celular , Regulação da Expressão Gênica/efeitos dos fármacos , Ordem dos Genes , Luciferases/metabolismo , Dados de Sequência Molecular , Proteínas de Resistência a Myxovirus , Poli I-C/farmacologia , Regiões Promotoras Genéticas/efeitos dos fármacos
15.
Res Dev Disabil ; 28(4): 362-85, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-16781115

RESUMO

This study evaluated two variants of a behavioral parent training program known as Stepping Stones Triple P (SSTP) using 74 preschool-aged children with developmental disabilities. Families were randomly allocated to an enhanced parent training intervention that combined parenting skills and care-giving coping skills (SSTP-E), standard parent training intervention alone (SSTP-S) or waitlist control (WL) condition. At post-intervention, both programs were associated with lower levels of observed negative child behavior, reductions in the number of care-giving settings where children displayed problem behavior, and improved parental competence and satisfaction in the parenting role as compared with the waitlist condition. Gains attained at post-intervention were maintained at 1-year follow-up. Both interventions produced significant reductions in child problem behavior, with 67% of children in the SSTP-E and 77% of children in the SSTP-S showing clinically reliable change from pre-intervention to follow-up. Parents reported a high level of satisfaction with both interventions.


Assuntos
Terapia Comportamental , Cuidadores/psicologia , Transtornos do Comportamento Infantil/terapia , Deficiências do Desenvolvimento/terapia , Educação , Deficiência Intelectual/terapia , Adaptação Psicológica , Adulto , Criança , Transtornos do Comportamento Infantil/psicologia , Pré-Escolar , Comportamento do Consumidor , Efeitos Psicossociais da Doença , Deficiências do Desenvolvimento/psicologia , Feminino , Seguimentos , Humanos , Masculino , Meio Social
16.
J Neuropathol Exp Neurol ; 65(11): 1049-58, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17086101

RESUMO

Brain tumors are the most common solid tumors of childhood, accounting for over 20% of cancers in children under 15 years of age. Pilocytic astrocytomas (PAs), World Health Organization grade I, are one of the most frequently occurring childhood brain tumors, yet little is known about genetic changes characterizing this entity. We have used microarray comparative genomic hybridization at 0.97 Mb resolution to study a series of PAs (n = 44). No copy number abnormality was seen in 64% of cases at this resolution. However, whole chromosomal gain (median 5 chromosomes affected) occurred in 32% of tumors. The most frequently affected chromosomes were 5 and 7 (11 of 44 cases each) followed by 6, 11, 15, and 20 (greater than 10% of cases each). Findings were confirmed by fluorescence in situ hybridization and microsatellite analysis in a subset of tumors. Chromosomal gain was significantly more frequent in PAs from patients over 15 years old (p = 0.03, Fisher exact test). The number of chromosomes involved was also significantly greater in the older group (p = 0.02, Mann-Whitney U test). One case (2%) showed a region of gain on chromosome 3 and one (2%) a deletion on 6q as their sole abnormalities. This is the first genomewide study to show this nonrandom pattern of genetic alteration in pilocytic astrocytomas.


Assuntos
Astrocitoma/genética , Neoplasias Encefálicas/genética , Dosagem de Genes , Análise de Sequência com Séries de Oligonucleotídeos , Adolescente , Adulto , Fatores Etários , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Astrocitoma/patologia , Neoplasias Encefálicas/patologia , Criança , Pré-Escolar , Feminino , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Lactente , Antígenos Comuns de Leucócito/metabolismo , Masculino , Microglia/metabolismo , Repetições de Microssatélites
17.
J Neuropathol Exp Neurol ; 65(6): 549-61, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16783165

RESUMO

Medulloblastomas and supratentorial primitive neuroectodermal tumors are aggressive childhood tumors. We report our findings using array comparative genomic hybridization (CGH) on a whole-genome BAC/PAC/cosmid array with a median clone separation of 0.97 Mb to study 34 medulloblastomas and 7 supratentorial primitive neuroectodermal tumors. Array CGH allowed identification and mapping of numerous novel, small regions of copy number change to genomic sequence in addition to the large regions already known from previous studies. Novel amplifications were identified, some encompassing oncogenes MYCL1, PDGFRA, KIT, and MYB not previously reported to show amplification in these tumors. In addition, one supratentorial primitive neuroectodermal tumor had lost both copies of the tumor-suppressor genes CDKN2A and CDKN2B. Ten medulloblastomas had findings suggestive of isochromosome 17q. In contrast to previous reports using conventional CGH, array CGH identified 3 distinct breakpoints in these cases: Ch 17: 17940393-19251679 (17p11.2, n = 6), Ch 17: 20111990-23308272 (17p11.2-17q11.2, n = 4), and Ch 17: 38425359-39091575 (17q21.31, n = 1). Significant differences were found in the patterns of copy number change between medulloblastomas and supratentorial primitive neuroectodermal tumors, providing further evidence that these tumors are genetically distinct despite their morphologic and behavioral similarities.


Assuntos
Neoplasias Encefálicas/genética , Meduloblastoma/genética , Tumores Neuroectodérmicos Primitivos/genética , Hibridização de Ácido Nucleico/métodos , Neoplasias Supratentoriais/genética , Adolescente , Adulto , Criança , Pré-Escolar , Aberrações Cromossômicas , Perfilação da Expressão Gênica , Humanos , Hibridização in Situ Fluorescente/métodos , Lactente , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos
18.
Nat Neurosci ; 9(5): 602-4, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16582904

RESUMO

Postnatal glutamatergic principal neuron synapses are typically presumed to express only calcium-impermeable (CI), GluR2-containing AMPARs under physiological conditions. Here, however, we demonstrate that long-term potentiation (LTP) in CA1 hippocampal pyramidal neurons causes rapid incorporation of GluR2-lacking calcium-permeable (CP)-AMPARs: CP-AMPARs are present transiently, being replaced by GluR2-containing AMPARs approximately 25 min after LTP induction. Thus, CP-AMPARs are physiologically expressed at CA1 pyramidal cell synapses during LTP, and may be required for LTP consolidation.


Assuntos
Hipocampo/citologia , Potenciação de Longa Duração/fisiologia , Células Piramidais/fisiologia , Receptores de AMPA/fisiologia , Animais , Animais Recém-Nascidos , Cálcio/metabolismo , Relação Dose-Resposta à Radiação , Estimulação Elétrica/métodos , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/fisiologia , Potenciais Pós-Sinápticos Excitadores/efeitos da radiação , Técnicas In Vitro , Potenciação de Longa Duração/efeitos dos fármacos , Potenciação de Longa Duração/efeitos da radiação , Camundongos , Antagonistas Nicotínicos/farmacologia , Técnicas de Patch-Clamp/métodos , Poliaminas/farmacologia , Células Piramidais/efeitos dos fármacos , Células Piramidais/efeitos da radiação , Ratos , Receptores de AMPA/química , Receptores de AMPA/deficiência , Fatores de Tempo
19.
Dev Comp Immunol ; 29(7): 627-35, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15784293

RESUMO

In vivo studies were carried out to investigate the protective effect of the interferon inducer poly I:C against channel catfish virus (CCV). Channel catfish were stimulated by intraperitoneal injection of 50 microg of poly I:C or PBS at various days prior to immersion challenge with CCV. Mortality in the poly I:C group was significantly reduced from 70% to 3% at day 1 compared to the PBS controls. Mortality increased at day 3 but was still significantly different from the PBS controls. Mx1 transcription was significantly higher only at day 1. In an additional study Mx1 transcription was monitored in the liver, kidney, gills, spleen, and intestine at various time points post-stimulation with either poly I:C or CCV. Mx1 mRNA was significantly elevated in all organs only at day 1 post-injection with poly I:C. In response to CCV, Mx1 transcription was not significantly elevated until day 3 post-challenge, but remained elevated in certain organs until day 7.


Assuntos
Proteínas de Ligação ao GTP/genética , Herpesviridae/imunologia , Ictaluridae/imunologia , Indutores de Interferon/farmacologia , Poli I-C/farmacologia , Transcrição Genética/efeitos dos fármacos , Animais , Proteínas de Ligação ao GTP/imunologia , Proteínas de Ligação ao GTP/metabolismo , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/mortalidade , Proteínas de Resistência a Myxovirus , Especificidade de Órgãos , RNA Mensageiro/metabolismo
20.
Fish Shellfish Immunol ; 16(3): 391-405, 2004 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15123306

RESUMO

A 2.5 kb full-length cDNA clone of a channel catfish (Ictalurus punctatus) Mx gene was obtained using RACE (rapid amplification of cDNA ends) polymerase chain reaction (PCR) from RNA extracted from the liver of poly I:C stimulated channel catfish. The gene consists of an open reading frame of 1905 nucleotides encoding a 635 amino acid protein. The predicted protein is 72.5 kDa and contains the dynamin family signature, a tripartite GTP binding motif and a leucine zipper, characteristic of all known Mx proteins. The catfish Mx protein exhibited 79% identity with perch Mx and between 71% and 74% identity with the three Atlantic salmon and the three rainbow trout Mx proteins. Mx mRNA was constitutively expressed in channel catfish ovary (CCO) cells, but in higher quantities in response to poly I:C treatment. Mx was induced in channel catfish following injection with channel catfish virus (CCV) and poly I:C.


Assuntos
Proteínas de Ligação ao GTP/genética , Expressão Gênica , Ictaluridae/genética , Ovário/metabolismo , Filogenia , Motivos de Aminoácidos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Southern Blotting , Primers do DNA , DNA Complementar/genética , Feminino , Proteínas de Ligação ao GTP/metabolismo , Herpesviridae/imunologia , Ictaluridae/imunologia , Ictaluridae/virologia , Dados de Sequência Molecular , Proteínas de Resistência a Myxovirus , Poli I-C/imunologia , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA