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1.
Eur J Pain ; 21(10): 1723-1731, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28722339

RESUMO

BACKGROUND: The goal was to test the effectiveness of a structured pain management programme after invasive electrophysiological interventions in cardiology including ablation of atrial fibrillation (AF) or ventricular tachycardia (VT) and implantation, or explantation, of pacemakers or implantable cardioverter defibrillators. METHODS: This was a prospective study with a pre-/post-design where a post-intervention group (116 consecutive patients) was compared to a pre-intervention group (102 consecutive patients) after implementation of a structured pain-management programme using the numeric rating scale (NRS 0-10) and classified as moderate-to-severe if NRS > 3. Measurements were recorded every two hours during the first 24 h post-operatively. The location of the pain and the amount of analgesic used were also recorded. RESULTS: The proportion of patients who experienced moderate-to-severe pain after the procedure decreased after initiation of the pain-management program: 47% versus 61%; p = 0.048. This difference was driven primarily by reduced pain late (8-24 h) after the procedure; 16% versus 39%; p < 0.001. The risk to develop late (8-24 h) post-procedural pain was reduced approximately three-fold after implementation of the pain-management programme (OR = 0.32, 95% CI 0.16-0.64, p = 0.001). Multivariate analysis indicated chronic pain, early pain (0-6 h), and type of intervention were associated with late post-interventional pain. In contrast, age, diabetes mellitus, BMI, gender and procedure time were not related. CONCLUSION: The findings illustrate the potential value of a structured pain-management programme. The proportion of patients who experienced moderate-to-severe pain after these electrophysiological procedures decreased significantly. SIGNIFICANCE: This is the first exploratory study that evaluates the impact of a multidisciplinary pain-management programme after cardiac electrophysiological interventions. It demonstrates that significant quality improvement is achievable following simple rules together with patient and staff education. The programme reduces the proportion of patients with moderate-to-severe pain after electrophysiological procedures significantly.


Assuntos
Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Ablação por Cateter/efeitos adversos , Manejo da Dor , Dor Pós-Operatória/terapia , Idoso , Idoso de 80 Anos ou mais , Analgésicos/uso terapêutico , Estudos Controlados Antes e Depois , Desfibriladores Implantáveis , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dor Pós-Operatória/etiologia , Estudos Prospectivos , Resultado do Tratamento
2.
Pathologe ; 38(2): 112-116, 2017 Mar.
Artigo em Alemão | MEDLINE | ID: mdl-28120026

RESUMO

We report a case of ALK1-positive anaplastic large cell lymphoma with expression of placental alkaline phosphatase (PLAP) in many tumor cells. Initially, due to the positivity of tumor cells for CD30 and PLAP, lymph node metastasis of a germ cell neoplasm was discussed. Anaplastic large cell lymphomas of T­cell lineage form a group of rare non-Hodgkin lymphomas with heterogeneous morphological and immunohistochemical appearance. They may imitate other neoplasms, such as large cell B­cell lymphomas, metastasis of a carcinoma, melanoma, embryonal carcinoma or seminoma, rhabdomyosarcoma and inflammatory myofibroblastic tumor. Only an extended immunohistochemistry panel leads to an accurate diagnosis.


Assuntos
Fosfatase Alcalina/metabolismo , Isoenzimas/metabolismo , Metástase Linfática/diagnóstico , Linfoma Anaplásico de Células Grandes/diagnóstico , Receptores de Activinas Tipo II/genética , Adulto , Biomarcadores Tumorais , Proteínas Ligadas por GPI/metabolismo , Humanos , Imuno-Histoquímica , Linfonodos/patologia , Linfoma Anaplásico de Células Grandes/genética , Masculino , Pescoço , Neoplasias Embrionárias de Células Germinativas/patologia
3.
J Atr Fibrillation ; 6(3): 934, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-28496900

RESUMO

Since the pulmonary veins (PVs) were identified as a major source of AF triggers, ablation strategies targeting the PVs have evolved from focal ablation inside the PVs to wide area circumferential PV isolation (PVI) which at this juncture is the standard approach. Despite the widespread popularity of PVI, a universal definition is lacking. While "entrance block" is a generally accepted endpoint for PVI, the role of "exit block" has yet to be determined. Inexcitability of the circular ablation line has been introduced as a promising additional endpoint for PVI and was associated with an improved clinical outcome in a randomized trial. Correct interpretation of PV electrograms during an ablation procedure is critical in terms of efficacy and safety. A variety of electrophysiological techniques help to correctly differentiate components of complex PV electrograms. Resumption of PV conduction after initially successful PVI leading to AF recurrence remains a major problem and confirmation of bi-directional conduction block does not exclude reversible tissue damage along the ablation line. Prolongation of post-PVI monitoring and application of provocative procedures such as the administration of adenosine after initial PVI to unmask dormant PV conduction may improve clinical outcome although there is lack of valid data supporting these strategies. This article aims on clarifying the electrophysiological criteria for complete pulmonary vein isolation and the explain the importance of this cornerstone in almost all atrial fibrillation ablation procedures.

4.
Curr Cardiol Rev ; 8(4): 354-61, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22920479

RESUMO

Atrial fibrillation (AF) is the most prevalent sustained arrhythmia in clinical practice. It is associated with significant morbidity and mortality and has been identified as an independent risk factor for ischemic stroke and thromboembolic events. Catheter ablation has become an established rhythm control therapy in patients with highly symptomatic drug-refractory AF. The definition of ablation success remains controversial since current symptom-based or intermittent electrocardiogram monitoring strategies fail to sufficiently disclose rhythm outcome. This failure is mainly related to the high incidence of asymptomatic AF recurrences, the unpredictable nature of arrhythmia relapses, and the poor correlation of symptoms and AF episodes. There is a clear correlation between the intensity of the monitoring strategy and the sensitivity for it to detect arrhythmia recurrences. Furthermore, several clinical studies assessing the long-term efficacy of catheter ablation procedures have reported late AF recurrences in patients who were initially considered responders to catheter ablation. In certain subsets of patients, precise long-term monitoring may help to guide therapy, e.g. patients in whom withdrawal of antithrombotic therapy may be considered if they are free of arrhythmia recurrences. Recently, subcutaneous implantable cardiac monitors (ICM) have been introduced for prolonged and continuous rhythm monitoring. The performance of a leadless ICM equipped with a dedicated AF detection algorithm has recently been assessed in a clinical trial demonstrating a high sensitivity and overall accuracy for identifying patients with AF. The clinical impact of ICM-based follow-up strategies, however, has to be evaluated in prospective clinical trials.


Assuntos
Fibrilação Atrial/cirurgia , Ablação por Cateter/métodos , Fibrilação Atrial/diagnóstico , Ablação por Cateter/tendências , Desfibriladores Implantáveis , Eletrocardiografia , Previsões , Humanos , Cuidados Pós-Operatórios/métodos , Prevenção Secundária , Resultado do Tratamento
6.
Colorectal Dis ; 14(10): 1276-86, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22309286

RESUMO

AIM: Immunosuppression and steroid medication have been identified as risk factors for complicated sigmoid diverticulitis. The underlying molecular mechanisms have not yet been elucidated. We hypothesized that glucocorticoid-induced tumour necrosis factor receptor (GITR) and matrix metalloproteinase-9 (MMP-9) might play a role. METHOD: GITR and MMP-9 were analysed at protein [immunohistochemistry/immunofluorescence (IF)] and messenger RNA level (real-time polymerase chain reaction) in surgical specimens with complicated and non-complicated diverticulitis (n=101). IF double staining and regression analysis were performed for both markers. GITR expression was correlated with clinical data and its usefulness as a diagnostic test was investigated. RESULTS: High GITR expression (≥41%) was observed in the inflammatory infiltrate in complicated diverticulitis, in contrast to non-complicated diverticulitis where GITR expression was low (P<0.001). High GITR expression was significantly associated with steroid use and pulmonary diseases (both P<0.001). MMP-9 expression correlated with GITR expression (R(2) =0.7268, P<0.0001, r=0.85) as demonstrated with IF double-staining experiments. Co-labelling of GITR with CD68, but not CD15, suggested that GITR-expressing cells in diverticulitis are macrophages. GITR expression was superior to C-reactive protein (CRP), white cell count and temperature in distinguishing complicated and non-complicated diverticulitis. CONCLUSIONS: Our results suggest that GITR expression in inflammatory cells might potentially indicate a molecular link between steroid use and complicated forms of acute sigmoid diverticulitis. Increased MMP-9 expression by GITR signalling might explain the morphological changes in the colonic wall of perforated and phlegmonous diverticulitis. Analysis of soluble GITR might be a promising strategy for future research.


Assuntos
Doença Diverticular do Colo/metabolismo , Proteína Relacionada a TNFR Induzida por Glucocorticoide/metabolismo , Imunossupressores/efeitos adversos , Metaloproteinase 9 da Matriz/metabolismo , Doenças do Colo Sigmoide/metabolismo , Esteroides/efeitos adversos , Idoso , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Biomarcadores/metabolismo , Estudos de Casos e Controles , Doença Diverticular do Colo/induzido quimicamente , Doença Diverticular do Colo/complicações , Doença Diverticular do Colo/diagnóstico , Feminino , Fucosiltransferases/metabolismo , Humanos , Imuno-Histoquímica , Antígenos CD15/metabolismo , Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Razão de Chances , Estudos Prospectivos , Curva ROC , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Doenças do Colo Sigmoide/induzido quimicamente , Doenças do Colo Sigmoide/complicações , Doenças do Colo Sigmoide/diagnóstico
7.
Cell Oncol (Dordr) ; 34(4): 315-26, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21573932

RESUMO

INTRODUCTION: The progressive growth of malignancies is accompanied by a decline in the immune response through mechanisms which are poorly understood. Apoptosis and induction of inflammation by tumor released cytokines as tumor escape mechanisms have been proposed to play an important role in colorectal carcinogenesis. METHODS: Expression of Tumor necrosis factor-alpha (TNF-α) was analyzed in colorectal cancer specimen and the cancer cell line HT-29 by immunohistochemistry and RT-PCR. TNF-α expression on protein and mRNA level were correlated with clinical characteristics and impact on survival. TNFR-1 was co-labelled with TNF-α and CD8+ cytotoxic T cells in immunofluorescence double staining experiments. RESULTS: 94% (n = 98/104) of the patients with CRC expressed TNF-α. High TNF-α expression was significantly associated with positive lymph node stage and recurrence of the tumor. Multivariate analysis revealed high TNF-α expression as an independent prognostic factor. Immunohistochemistry was correlated with RT-PCR results (т = 0.794). Immunofluorescence double staining experiments revealed increased TNFR-1 expression by CD8+ cells. CONCLUSIONS: TNF-α expression by tumor cells may be an efficient immunological escape mechanism by inflammation-enhanced metastases and probably by induction of apoptosis in tumor-infiltrating CD8+ immune cells resulting in a down regulation of the tumoral immune response. Our data support the role of tumor-derived TNF-α expression as an important promoter of tumoral immune escape mechanisms and malignant progression, and suggest that analysis on either protein (immunohistochemistry) or RNA level (RT-PCR) can be used effectively in this respect. Targeting TNF-α may be a promising option, especially in cases with high TNF-α expression and positive lymph node metastases.


Assuntos
Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/patologia , Linfonodos/patologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/patologia , Idoso , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/patologia , Neoplasias Colorretais/genética , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Linfonodos/efeitos dos fármacos , Linfonodos/metabolismo , Metástase Linfática/patologia , Linfócitos do Interstício Tumoral/efeitos dos fármacos , Linfócitos do Interstício Tumoral/patologia , Masculino , Pessoa de Meia-Idade , Variações Dependentes do Observador , Prognóstico , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Recidiva , Análise de Sobrevida , Fator de Necrose Tumoral alfa/genética
8.
Anal Cell Pathol (Amst) ; 33(3): 151-63, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20978325

RESUMO

INTRODUCTION: The progressive growth of malignancies is accompanied by a decline in the immune response through mechanisms which are poorly understood. Apoptosis and induction of inflammation by tumor released cytokines as tumor escape mechanisms have been proposed to play an important role in colorectal carcinogenesis. METHODS: Expression of Tumor necrosis factor-alpha (TNF-α) was analyzed in colorectal cancer specimen and the cancer cell line HT-29 by immunohistochemistry and RT-PCR. TNF-α expression on protein and mRNA level were correlated with clinical characteristics and impact on survival. TNFR-1 was co-labelled with TNF-α and CD8+ cytotoxic T cells in immunofluorescence double staining experiments. RESULTS: 94% (n=98/104) of the patients with CRC expressed TNF-α. High TNF-α expression was significantly associated with positive lymph node stage and recurrence of the tumor. Multivariate analysis revealed high TNF-α expression as an independent prognostic factor. Immunohistochemistry was correlated with RT-PCR results (τ=0.794). Immunofluorescence double staining experiments revealed increased TNFR-1 expression by CD8+ cells. CONCLUSIONS: TNF-α expression by tumor cells may be an efficient immunological escape mechanism by inflammation-enhanced metastases and probably by induction of apoptosis in tumor-infiltrating CD8+ immune cells resulting in a down regulation of the tumoral immune response. Our data support the role of tumor-derived TNF-α expression as an important promoter of tumoral immune escape mechanisms and malignant progression, and suggest that analysis on either protein (immunohistochemistry) or RNA level (RT-PCR) can be used effectively in this respect. Targeting TNF-α may be a promising option, especially in cases with high TNF-α expression and positive lymph node metastases.


Assuntos
Neoplasias Colorretais/metabolismo , Metástase Linfática/fisiopatologia , Recidiva Local de Neoplasia/fisiopatologia , Fator de Necrose Tumoral alfa/metabolismo , Neoplasias Colorretais/genética , Células HT29 , Humanos , Imuno-Histoquímica , Recidiva Local de Neoplasia/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Necrose Tumoral alfa/genética
10.
Cancer Biomark ; 7(6): 285-94, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21694467

RESUMO

BACKGROUND: Esophageal adenocarcinomas (EACs) arise due to gastroesophageal reflux, with Barrett's esophagus (BE) regarded as precancerous lesion. Glucocorticoid-induced TNFR family-related Receptor (GITR)-mediated inflammation of tumor infiltrating leucocytes (TILs) in the tumor microenvironment might play a role during the multistep carcinogenetic process as either tumor promoting factor according to an inflammatory microenvironment or as a feature of anti-tumor activity. METHODS: Immunohistochemical analysis of GITR expression was analyzed in esophageal cancer (n=70: 41 EAC with BE, 19 EAC without BE, and n=10 esophageal squamous-cell carcinomas, ESCC), the adenocarcinoma cell line OE-33, and peripheral blood leucocytes (PBLs) of EAC patients, furthermore in biopsies of BE without intraepithelial neoplasia (IN) (n=18). Results were correlated with clinicopathological parameters and five-year survival rates. Immunohistochemical GITR expression results were confirmed on mRNA level (RT-PCR). RESULTS: Quantification showed a significant increase of 25% GITR positive TILs in EAC with BE (p< 0.05) compared to 13% in adjacent BE, 24% in EAC without BE, 14% in ESCC, and 1% in BE without IN. High GITR levels were not significantly associated with clinicopathologic features which may predict worse clinical outcome and had no impact on survival (p= 0.7878). Increased GITR expression of peripheral blood leucocytes (PBLs) in EAC patients was shown on protein level (32%) and confirmed by RT-PCR (3.7-fold difference compared to normal tissue). CONCLUSIONS: This study provides for the first time evidence that GITR expression of TILs is associated in the pathogenesis of Barrett's esophagus. Our findings suggest that GITR-expression of TILs is associated with cancer progression. Its role as either tumor promoting factor %according to an in the inflammatory microenvironment or as a feature of anti-tumor activity and promising target for molecular therapies needs to be substantiated in further investigations.


Assuntos
Adenocarcinoma/metabolismo , Esôfago de Barrett/metabolismo , Neoplasias Esofágicas/metabolismo , Proteína Relacionada a TNFR Induzida por Glucocorticoide/biossíntese , Leucócitos/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/patologia , Idoso , Esôfago de Barrett/genética , Esôfago de Barrett/patologia , Linhagem Celular Tumoral , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Proteína Relacionada a TNFR Induzida por Glucocorticoide/genética , Humanos , Imuno-Histoquímica , Leucócitos/patologia , Masculino , Mucosa/metabolismo , Mucosa/patologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sobrevida
11.
Protoplasma ; 226(3-4): 137-46, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16333572

RESUMO

The determination of protein-protein interactions is becoming more and more important in the molecular analysis of signal transduction chains. To this purpose the application of a manageable and simple assay in an appropriate biological system is of major concern. Bimolecular fluorescence complementation (BiFC) is a novel method to analyze protein-protein interactions in vivo. The assay is based on the observation that N- and C-terminal subfragments of the yellow-fluorescent protein (YFP) can only reconstitute a functional fluorophore when they are brought into tight contact. Thus, proteins can be fused to the YFP subfragments and the interaction of the fusion proteins can be monitored by epifluorescence microscopy. Pairs of interacting proteins were tested after transient cotransfection in etiolated mustard seedlings, which is a well characterized plant model system for light signal transduction. BiFC could be demonstrated with the F-box protein EID1 (empfindlicher im dunkelroten Licht 1) and the Arabidopsis S-phase kinase-related protein 1 (ASK1). The interaction of both proteins was specific and strictly dependent on the presence of an intact F-box domain. Our studies also demonstrate that etiolated mustard seedlings provide a versatile transient assay system to study light-induced subcellular localization events.


Assuntos
Proteínas F-Box/metabolismo , Luz , Plantas/metabolismo , Sequência de Bases , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Proteínas Luminescentes , Dados de Sequência Molecular , Mostardeira/genética , Petroselinum/genética , Fitocromo A/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Plasmídeos , Protoplastos/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Plântula/genética , Transdução de Sinais , Espectrometria de Fluorescência/métodos , Transfecção , Técnicas do Sistema de Duplo-Híbrido
12.
J Inherit Metab Dis ; 28(5): 743-57, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16151906

RESUMO

Glycosaminoglycans are accumulated in both mucopolysaccharidoses (MPS) and mucolipidoses (ML). MPS I, II, III and VII and ML II and ML III patients cannot properly degrade heparan sulphate (HS). In spite of the importance of HS storage in the metabolic pathway in these diseases, blood and urine HS levels have not been determined systematically using a simple and economical method. Using a new ELISA method using anti-HS antibodies, HS concentrations in blood and urine were determined in MPS and ML II and ML III patients. HS concentrations were determined in 156 plasma samples from MPS I (n = 23), MPS II (n = 26), MPS III (n = 24), MPS IV (n = 62), MPS VI (n = 5), MPS VII (n = 5), ML II (n = 8) and ML III (n = 3), and 205 urine samples from MPS I (n = 33), MPS II (n = 33), MPS III (n = 30), MPS IV (n = 82), MPS VI (n = 7), MPS VII (n = 9), ML II (n = 8) and ML III (n = 3). The ELISA method used monoclonal antibodies against HS. MPS I, II, III and VII and ML II and III patients had significant elevation in plasma HS, compared to the age-matched controls (p < 0.0001). Eighty-three out of 89 (93.3%) of individual values in the above MPS types and ML were above the mean +2SD of the controls. In urine samples, 75% of individual values in patients with those types were above the mean +2SD of the controls. In contrast to the previous understanding of the HS metabolic pathway, plasma HS levels in all five MPS VI and 15% of MPS IV patients were elevated above the mean +2SD of the controls. These findings suggest that HS concentration determined by ELISA, especially in plasma, could be a helpful marker for detection of the most severe MPS I, II, III, VI and VII and ML II, distinguishing them from normal populations.


Assuntos
Ensaio de Imunoadsorção Enzimática/métodos , Heparitina Sulfato/química , Mucolipidoses/diagnóstico , Mucopolissacaridoses/diagnóstico , Adolescente , Biomarcadores/metabolismo , Química Clínica/métodos , Criança , Pré-Escolar , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Glicosaminoglicanos/química , Heparina/química , Heparitina Sulfato/sangue , Heparitina Sulfato/urina , Humanos , Lactente , Recém-Nascido , Mucolipidoses/sangue , Mucolipidoses/urina , Mucopolissacaridoses/sangue , Mucopolissacaridoses/urina
13.
J Inherit Metab Dis ; 28(2): 187-202, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15877208

RESUMO

The mucopolysaccharidoses (MPS) is characterized by accumulation of glycosaminoglycans (GAGs), and mucolipidosis (ML) by accumulation of GAGs and sphingolipids. Each type of MPS accumulates specific GAGs. The lysosomal enzymes N-acetylgalactosamine-6-sulphate sulphatase and beta-galactosidase involve the stepwise degradation of keratan sulphate (KS). Deficiency of these enzymes results in elevation of KS levels in the body fluids and in tissues, leading to MPS IV disease. In this study, we evaluated blood and urine KS levels in types of MPS and ML other than MPS IV. Eighty-five plasma samples came from MPS I (n = 18), MPS II (n = 28), MPS III (n = 20), MPS VI (n = 3), MPS VII (n = 5) and ML (n = 11) patients while 127 urine samples came from MPS I (n = 34), MPS II (n = 34), MPS III (n = 32), MPS VI (n = 7), MPS VII (n = 9) and ML (n = 11) patients. KS levels were determined using the ELISA method. Plasma KS levels varied with age in both control and patient populations. In all age groups, the mean values of plasma KS in MPS and ML patients were significantly higher than those in the age-matched controls. Plasma KS values in four newborn patients were above the mean + 2SD of the age-matched controls (mean, 41 ng/ml). Overall, 85.9% of individual values in non-type IV MPS and ML patients were above the mean + 2SD of the age-matched controls. For urine KS levels, 24.4% of individual values in patients were above the mean + 2SD of the age-matched controls. In conclusion, KS in blood is elevated in each type of non-type IV MPS examined, in contrast to the conventional understanding. This finding suggests that measurement of KS level provides a new diagnostic biomarker in a wide variety of mucopolysaccharidoses and mucolipidoses in addition to MPS IV.


Assuntos
Sulfato de Queratano/sangue , Sulfato de Queratano/urina , Mucolipidoses/metabolismo , Mucopolissacaridoses/metabolismo , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Especificidade de Anticorpos , Biomarcadores , Criança , Pré-Escolar , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática , Humanos , Lactente , Recém-Nascido , Sulfato de Queratano/imunologia , Pessoa de Meia-Idade , Mucolipidoses/diagnóstico , Mucopolissacaridoses/diagnóstico , Sensibilidade e Especificidade
15.
Mol Genet Genomics ; 265(6): 964-76, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11523788

RESUMO

The common plant regulatory factors (CPRFs) from parsley are transcription factors with a basic-leucine-zipper motif that bind to cis-regulatory elements frequently found in promoters of light-regulated genes. Proposed to function in concert with members of other transcription factor families, CPRFs regulate the transcriptional activity of many target genes. Here, we report that, in contrast to CPRF2, which operates as a transcriptional activator, CPRF1 functions as repressor in vivo. Two-hybrid screens using CPRF1 and CPRF2 as "baits" resulted in the isolation of four novel parsley proteins which interact with either CPRF1 or CPRF2 in vivo. Three of these factors represent new parsley bZIP factors, designated CPRF5-CPRF7, whereas the fourth, named CPRF1-interacting protein (CIP), shows no homology to any other known protein. CPRF5 and CIP specifically interact with CPRF1, whilst CPRF6 and CPRF7 exclusively form heterodimers with CPRF2. CPRF5, CPRF6 and CPRF7 are transcription factors that exhibit sequence-specific DNA-binding as well as transactivation abilities, whereas the function of CIP remains elusive. The newly isolated CPRFs and CIP are constitutively localized in the nucleus in parsley protoplasts. Furthermore, mRNA accumulation studies revealed that the expression of these novel bZIP genes and CIP is not altered by exposure to light. We discuss the possible roles of the newly identified proteins in CPRF1- and CPRF2-dependent target gene expression.


Assuntos
Apiaceae/genética , Apiaceae/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Sequência de Aminoácidos , Apiaceae/efeitos da radiação , Fatores de Transcrição de Zíper de Leucina Básica , Células Cultivadas , Clonagem Molecular , Sequência Conservada , Proteínas de Ligação a DNA/química , Fatores de Ligação G-Box , Zíper de Leucina , Luz , Dados de Sequência Molecular , Proteínas de Plantas/química , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes/metabolismo , Proteínas Repressoras/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo , Raios Ultravioleta
16.
J Cell Sci ; 114(Pt 3): 475-80, 2001 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11171317

RESUMO

Plants monitor changes in the ambient light environment by highly specialised photoreceptors, which include the red/far-red photoreversible phytochromes, the blue-light-absorbing cryptochromes and phototropin and the so-far-unidentified UVB photoreceptor(s). Light easily penetrates plant organs/tissues and reaches even the subcellular compartments of various cell types. Therefore, it is not surprising that the determination of the intracellular localisation of photoreceptors has been, for many years, a major, and often controversial, subject of plant photobiology and cell biology research. Phototropin, one of the blue-light photoreceptors of higher plants, controls phototropism by monitoring the direction of light, and it is localised in or at the plasmalemma. In contrast, the subcellular localisation of phytochromes changes dynamically and exhibits a very complex pattern. These photoreceptors are localised in the cytosol in dark- grown tissues. Irradiation, however, induces import of phytochromes into the nucleus. The import occurs in a light-quality- and light-quantity-dependent fashion and, as such, seems to be unique to higher plants. Light-induced accumulation of phytochromes in the nuclei correlates well with various physiological responses mediated by these photoreceptors. These observations indicate that light-dependent intracellular redistribution of phytochrome photoreceptors is one of the major regulatory steps in photomorphogenesis.


Assuntos
Luz , Complexo de Proteínas do Centro de Reação Fotossintética/metabolismo , Transdução de Sinais/efeitos da radiação , Escuridão , Plantas/metabolismo , Plantas/ultraestrutura , Transporte Proteico
17.
Plant J ; 22(2): 125-33, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10792828

RESUMO

Phytochromes (phy) are a family of photoreceptors that control various aspects of light-dependent plant development. Phytochrome A (phyA) is responsible for the very low fluence response (VLFR) under inductive light conditions and for the high irradiance response (HIR) under continuous far-red light. We have recently shown that nuclear import of rice phyA:GFP is regulated by VLFR in transgenic tobacco. The import is preceded by very fast, light-induced formation of sequestered areas of phyA:GFP in the cytosol. Here we report that expression of the Arabidopsis phyA:GFP fusion protein in phyA-deficient Arabidopsis plants complements the mutant phenotype. In these transgenic Arabidopsis lines, both light-dependent cytosolic formation of sequestered areas of the phyA:GFP as well as VLFR or HIR-mediated nuclear import of the fusion protein was observed. By contrast, light-dependent nuclear import of the same fusion protein was induced only by continuous far-red light (HIR) but not by pulses of far-red light (VLFR) in transgenic tobacco. These results demonstrate that photoregulation of intracellular partitioning of the Arabidopsis phyA:GFP differs significantly in different genetic backgrounds.


Assuntos
Arabidopsis/fisiologia , Núcleo Celular/metabolismo , Luz , Nicotiana/fisiologia , Fitocromo/metabolismo , Plantas Tóxicas , Proteínas Recombinantes de Fusão/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis , Compartimento Celular/fisiologia , Núcleo Celular/fisiologia , Proteínas de Fluorescência Verde , Proteínas Luminescentes/genética , Microscopia de Fluorescência , Fitocromo/genética , Fitocromo/fisiologia , Fitocromo A , Plantas Geneticamente Modificadas , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/fisiologia , Nicotiana/metabolismo
18.
Plant J ; 22(2): 135-45, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10792829

RESUMO

Photomorphogenesis of higher plants is regulated by photoreceptors including the red/far-red light-absorbing phytochromes, blue-UV/A sensing cryptochromes and as yet uncharacterized UV/B receptors. Specific phototransduction pathways that are controlled by either individual or interacting photoreceptors mediate regulation. Phytochrome B (phyB) is the major red light-sensing photoreceptor. Phototransduction mediated by this light sensor has been shown to include light-dependent nuclear import and interaction of phyB with transcription factor-like proteins in the nucleus. Here we report that nuclear import of phyB and physiological responses regulated by this photoreceptor exhibit very similar wavelength- and fluence rate-dependence. Nuclear import of phyB is insensitive to single red, blue and far-red light pulses. It is induced by continuous red light and to a lesser extent by continuous blue light, whereas far-red light is completely ineffective. The data presented indicate that light-dependent partitioning of phyB exhibits features characteristic of blue light responsiveness amplification, a phenomenon that is thought to be mediated by interaction of phyB with CRY1.


Assuntos
Núcleo Celular/metabolismo , Luz , Nicotiana/fisiologia , Células Fotorreceptoras , Fitocromo/metabolismo , Plantas Tóxicas , Proteínas Recombinantes de Fusão/metabolismo , Fatores de Transcrição , Compartimento Celular , Núcleo Celular/fisiologia , Proteínas de Fluorescência Verde , Proteínas Luminescentes/genética , Microscopia de Fluorescência , Fitocromo/genética , Fitocromo/fisiologia , Fitocromo B , Plantas , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/fisiologia , Frações Subcelulares/metabolismo , Nicotiana/metabolismo
19.
Exp Neurol ; 161(1): 174-82, 2000 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10683283

RESUMO

Ribosomes are integral constitutens of the protein synthesis machinery. Polymerase I (POL I) is located in the nucleolus and transcribes the large ribosomal genes. POL I activity is decreased in ischemia but nothing is known so far on POL I in perinatal asphyxia. We investigated the involvement of POL I in a well-documented model of graded systemic asphyxia at the level of activity, mRNA, protein, and morphology. Caeserean section was performed at the 21st day of gestation. Rat pups still in the uterus horns were immerged in a water bath for asphyctic periods from 5-20 min. Brain was taken for measurement of pH, nuclear POL I activity, and mRNA steady state, and protein levels of RPA40, an essential subunit of POL I and III. Silver staining and transmission electron microscopy with morphometry when appropriate were used to examine the nucleolus. Brain pH and nuclear POL I activity decreased with the length of the asphyctic period while POL-I mRNA and protein levels were unchanged. Accompanying the decrease in brain pH we found significant changes of nucleolar structure in the course of perinatal asphyxia at the light and electron microscopic level. As early as ten min following the asphyctic insult, morphological disintegration of the nucleolus was observed. The changes became more dramatic with longer duration of perinatal asphyxia. We conclude that severe acidosis may be responsible for decreased POL activity and for disintegration of nucleoli in neurons. This condition may lower the ribosome content in neonatal neurons and impair protein synthesis.


Assuntos
Asfixia Neonatal/metabolismo , Nucléolo Celular/enzimologia , Lobo Frontal/enzimologia , RNA Polimerase I/metabolismo , Animais , Animais Recém-Nascidos , Northern Blotting , Nucléolo Celular/ultraestrutura , Modelos Animais de Doenças , Feminino , Regulação Enzimológica da Expressão Gênica , Humanos , Concentração de Íons de Hidrogênio , Recém-Nascido , Microscopia Eletrônica , Gravidez , RNA Polimerase I/análise , RNA Polimerase I/genética , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Coloração pela Prata , Transcrição Gênica/fisiologia
20.
Semin Cell Dev Biol ; 11(6): 505-10, 2000 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11145880

RESUMO

Phytochromes in harmony with blue light photoreceptors play a major role in controlling plant growth and development from germination to seed maturation. Light absorption by phytochromes triggers a signaling cascade, phototransduction, which culminates in regulated gene expression. A major regulatory step at the cellular level, which affects specificities of light-induced physiological responses, seems to be the light-quality and light-quantity dependent nuclear import of the phytochromes themselves. The correlations found between the nuclear import of phytochromes (phyA and phyB) and various physiological responses regulated by these photoreceptors provides strong support for this hypothesis.


Assuntos
Núcleo Celular/metabolismo , Citoplasma/metabolismo , Células Fotorreceptoras , Fitocromo/metabolismo , Plantas/metabolismo , Fatores de Transcrição , Transporte Ativo do Núcleo Celular , Escuridão , Luz , Fitocromo A , Fitocromo B , Células Vegetais , Plantas/enzimologia , Proteínas Quinases/metabolismo , Transdução de Sinais
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