RESUMO
Impaired neural plasticity may be a core pathophysiological process underlying the symptomatology of schizophrenia. Plasticity-enhancing interventions, including repetitive transcranial magnetic stimulation (rTMS), may improve difficult-to-treat symptoms; however, efficacy in large clinical trials appears limited. The high variability of rTMS-related treatment response may be related to a comparably large variation in the ability to generate plastic neural changes. The aim of the present study was to determine whether negative symptom improvement in schizophrenia patients receiving rTMS to the left dorsolateral prefrontal cortex (DLPFC) was related to rTMS-related brain volume changes. A total of 73 schizophrenia patients with predominant negative symptoms were randomized to an active (n=34) or sham (n=39) 10-Hz rTMS intervention applied 5 days per week for 3 weeks to the left DLPFC. Local brain volume changes measured by deformation-based morphometry were correlated with changes in negative symptom severity using a repeated-measures analysis of covariance design. Volume gains in the left hippocampal, parahippocampal and precuneal cortices predicted negative symptom improvement in the active rTMS group (all r⩽-0.441, all P⩽0.009), but not the sham rTMS group (all r⩽0.211, all P⩾0.198). Further analyses comparing negative symptom responders (⩾20% improvement) and non-responders supported the primary analysis, again only in the active rTMS group (F(9, 207)=2.72, P=0.005, partial η 2=0.106). Heterogeneity in clinical response of negative symptoms in schizophrenia to prefrontal high-frequency rTMS may be related to variability in capacity for structural plasticity, particularly in the left hippocampal region and the precuneus.
Assuntos
Córtex Pré-Frontal/fisiopatologia , Esquizofrenia/terapia , Estimulação Magnética Transcraniana/métodos , Adulto , Encéfalo/fisiopatologia , Método Duplo-Cego , Feminino , Humanos , Masculino , Plasticidade Neuronal/fisiologia , Córtex Pré-Frontal/diagnóstico por imagem , Escalas de Graduação Psiquiátrica , Esquizofrenia/complicações , Estimulação Magnética Transcraniana/psicologia , Resultado do TratamentoRESUMO
In the present study, we used single- and dual-task conditions to investigate the nature of topographical working memory to better understand what type of task can hamper performance during navigation. During dual-task conditions, we considered four different sources of interference: motor (M), spatial motor (SM), verbal (i.e. articulatory suppression AS) and spatial environmental (SE). In order to assess the nature of topographical working memory, we used the Walking Corsi Test, asking the participants to perform two tasks simultaneously (M, SM, AS and SE). Our results showed that only spatial-environmental interference hampers the execution of a topographical working memory task, suggesting a task-domain-specific effect. We also found general gender differences in the topographical working memory capabilities: men were more proficient than women, regardless of the type of interferences. However, like men, women performed worse when a spatial-environmental interference was present.
Assuntos
Memória de Curto Prazo/fisiologia , Desempenho Psicomotor/fisiologia , Memória Espacial/fisiologia , Navegação Espacial/fisiologia , Adulto , Feminino , Humanos , Masculino , Fatores Sexuais , Adulto JovemRESUMO
OBJECTIVES: To evaluate the efficacy of two alcohol-free antimicrobial mouthrinses in reducing plaque and gingivitis compared to an alcohol-containing rinse and toothbrushing alone. METHODS: One hundred and sixty healthy volunteers were enrolled in the randomized controlled trial. Participants were randomly and equally assigned to four groups: (i) toothbrushing + rinsing (0.06% CHX + 0.025% NaF, alcohol-containing rinse, positive control); (ii) toothbrushing + rinsing (0.06% CHX + 0.025% NaF, alcohol-free experimental rinse); (iii) toothbrushing + rinsing (0.06% CHX + 0.03% CPC + 0.025% NaF, alcohol-free experimental rinse); (iv) toothbrushing alone (negative control). At baseline, Quigley-Hein plaque index (QHI), modified proximal plaque index (MPPI), and papillary bleeding index (PBI) were recorded. All subjects brushed their teeth as usual during the study. Additionally, groups 1-3 rinsed twice daily. Eight weeks after baseline, indices were recorded again. anova with Bonferroni adjustment served for statistical analysis. RESULTS: One hundred and fifty-five participants were included into final analysis (i: n = 39, 2: n = 39, 3: n = 37, 4: n = 40). Experimental rinses (ii, iii) reduced QHI and MPPI to a higher extent than the negative control (iv), whereas no significant difference to the positive control was found. QHI: (i) 36.6%, (ii) 32.3%, (iii) 36.8%, (iv) 21.6%; MPPI: (i) 11.9%, (ii) 12.2%, (iii) 13.6%, (iv) 3.5%. For PBI, no statistically significant difference was found between groups: (i) 80.2%, (ii) 77.8%, (iii) 76.5% and (iv) 78.8%. CONCLUSIONS: With respect to QHI and MPPI, toothbrushing in combination with any rinse was more effective than toothbrushing alone. No statistically significant differences were found between the alcohol-free and the alcohol-containing control rinses.
Assuntos
Anti-Infecciosos Locais/uso terapêutico , Clorexidina/uso terapêutico , Placa Dentária/prevenção & controle , Gengivite/prevenção & controle , Antissépticos Bucais/uso terapêutico , Adulto , Cariostáticos/uso terapêutico , Cetilpiridínio/uso terapêutico , Índice de Placa Dentária , Etanol/uso terapêutico , Seguimentos , Humanos , Índice Periodontal , Método Simples-Cego , Fluoreto de Sódio/uso terapêutico , Escovação Dentária/métodos , Resultado do TratamentoRESUMO
Here we tested the prognostic impact of genomic alterations in operable localized pancreatic ductal adenocarcinoma (PDAC). Fifty-two formalin-fixed and paraffin-embedded primary PDAC were laser micro-dissected and were investigated by comparative genomic hybridization after whole genome amplification using an adapter-linker PCR. Chromosomal gains and losses were correlated to clinico-pathological parameters and clinical follow-up data. The most frequent aberration was loss on chromosome 17p (65%) while the most frequent gains were detected at 2q (41%) and 8q (41%), respectively. The concomitant occurrence of losses at 9p and 17p was found to be statistically significant. Higher rates of chromosomal losses were associated with a more advanced primary tumor stage and losses at 9p and 18q were significantly associated with presence of lymphatic metastasis (chi-square: p = 0.03, p = 0.05, respectively). Deletions on chromosome 4 were of prognostic significance for overall survival and tumor recurrence (Cox-multivariate analysis: p = 0.026 and p = 0.021, respectively). In conclusion our data suggest the common alterations at chromosome 8q, 9p, 17p and 18q as well as the prognostic relevant deletions on chromosome 4q as relevant for PDAC progression. Our comprehensive data from 52 PDAC should provide a basis for future studies with a higher resolution to discover the relevant genes located within the chromosomal aberrations identified.
Assuntos
Carcinoma Ductal Pancreático/genética , Deleção Cromossômica , Cromossomos Humanos Par 4 , Neoplasias Pancreáticas/genética , Adenocarcinoma/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Ductal Pancreático/patologia , Aberrações Cromossômicas , Cromossomos Humanos Par 17 , Cromossomos Humanos Par 18 , Cromossomos Humanos Par 8 , Cromossomos Humanos Par 9 , Hibridização Genômica Comparativa , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/patologia , Prognóstico , Análise de SobrevidaRESUMO
Civil aviation pilots who develop insulin-treated diabetes and want to renew a Commercial Pilot License (CPL) represent a medical, social and regulatory problem. This depends on justified concerns about hypoglycemia, the most threatening event for people who carry out jobs requiring a high level of concentration and reliability. This negatively affects social and working aspects of pilots' lives, who have a high profile and a high-cost professional qualification. It could be possible now to revise this attitude thanks to the availability of Continuous Glucose Monitoring (CGM) devices. CGM clearly showed to prevent hypoglycemic events in insulin-treated diabetic patients by allowing strict monitoring and trend prediction of glucose levels. By systematizing available data on such devices and present regulations in CPL issuance worldwide, our review can be used as handy tool for a fruitful discussion among the scientific community, national and international civil aviation regulators, stakeholders and pilots, aimed at evaluating the evidence-based opportunity to revise CPL issuance criteria for insulin-treated diabetic pilots. For the above-mentioned reasons, there are, among the regulatory administrations of Civil Aviation around the globe, several different approaches and limitations set for the subjects with insulin-treated diabetes who want to obtain, or renew, a CPL.
Assuntos
Aviação/normas , Automonitorização da Glicemia/instrumentação , Diabetes Mellitus Tipo 1/epidemiologia , Diabetes Mellitus/epidemiologia , Pilotos/normas , Automonitorização da Glicemia/métodos , Feminino , Humanos , Masculino , Reprodutibilidade dos Testes , Participação dos InteressadosRESUMO
The effect of tumor promoters and growth factors on the synthesis of urokinase and urokinase mRNA in human carcinoma cells has been investigated. In urokinase-producing human carcinoma cells (A1251), a 20-40-fold increase in urokinase mRNA level is obtained after treatment with 10 nM phorbol myristate acetate (PMA), a smaller effect (two- to fourfold) with 2 ng/ml platelet-derived growth factor (PDGF) and no effect with epidermal growth factor (EGF) (up to 50 nM). After treatment with PMA, urokinase mRNA level increases already at 30 min peaking 2-4 h thereafter. Cell line A431, which has an abnormally high number of EGF receptors, shows the same response to PMA, but also responds to EGF (two- to fourfold increase in mRNA). The kinetics are similar to those of A1251. Nuclear transcription experiments show that the PMA-induced increase in urokinase mRNA is due to increased synthesis. The protein synthesis inhibitor, cycloheximide (10 micrograms/ml), also increases the level of urokinase mRNA. When both cycloheximide and PMA are used, super-induction is observed. This result may indicate that a short-lived protein negatively regulates the level of urokinase. The different efficiency of the effectors (PMA and PDGF better than EGF) and their kinetics, as well as the effect of cycloheximide on urokinase mRNA synthesis, (a) are reminiscent of the effect of PDGF and PMA on competence phase genes (Kelly, K., B.H. Cochran, C.D. Stiles, and P. Leder, 1983, Cell, 35: 603-610), (b) demonstrate that the synthesis of urokinase is part of the early cellular response to these factors, and (c) provide a preliminary insight in the overproduction of urokinase by primary malignant tumors and transformed cells in culture.
Assuntos
Forbóis/farmacologia , RNA Mensageiro/biossíntese , Acetato de Tetradecanoilforbol/farmacologia , Transcrição Gênica/efeitos dos fármacos , Ativador de Plasminogênio Tipo Uroquinase/genética , Carcinoma de Células Escamosas , Divisão Celular , Linhagem Celular , Cicloeximida/farmacologia , Replicação do DNA/efeitos dos fármacos , Genes , Humanos , Cinética , Fator de Crescimento Derivado de Plaquetas/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/isolamento & purificaçãoRESUMO
OBJECTIVES: Traffic-related pollution is associated with cardiovascular disease in general, but previous studies suggested that low socioeconomic status (SES) groups might be more susceptible towards a negative impact. We examined whether the association between long-term exposure to high traffic and early signs of coronary artery disease is modified by SES. METHODS: Individual-level medical and social data from a population-based study were linked with census information on neighbourhood socioeconomic characteristics. Residential exposure to traffic was defined as proximity to major roads using a geographical information system. We studied associations between high traffic and coronary artery calcification (CAC) within strata of SES to examine effect modification. Data stem from an epidemiological study in Germany including 2264 women and 2037 men (45-75 years). RESULTS: High traffic and low SES were both associated with higher amounts of calcification (>or=75th age-specific percentile). More participants with low SES lived close to major roads while stratified analyses did not indicate higher susceptibility in low SES groups. Participants with low SES and simultaneous exposure to high traffic had highest levels of CAC. For example, the prevalence of high calcification was 23.9% in better-educated men with low traffic exposure but 37.7% in lower-educated men with high traffic exposure (women: 22.0% vs 28.1%). CONCLUSIONS: High traffic exposure was associated with coronary calcification in all social groups, but as low SES individuals had higher calcification in general and were also more often exposed to traffic, existing inequalities could be further shaped by traffic exposure.
Assuntos
Doenças Cardiovasculares/etiologia , Veículos Automotores/estatística & dados numéricos , Classe Social , Saúde da População Urbana/estatística & dados numéricos , Emissões de Veículos/análise , Idoso , Calcinose/epidemiologia , Calcinose/etiologia , Doenças Cardiovasculares/epidemiologia , Estudos Transversais , Exposição Ambiental/efeitos adversos , Exposição Ambiental/análise , Feminino , Alemanha/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Áreas de Pobreza , Prevalência , Características de Residência/estatística & dados numéricos , Fatores Socioeconômicos , Desemprego/estatística & dados numéricos , Emissões de Veículos/toxicidadeRESUMO
In addition to its role in invasion and metastasis of several tumors, the multifunctional urokinase receptor uPAR (urokinase plasminogen activator receptor) is directly involved in the growth of several cancer cells in vitro and in vivo. We have compared growth rate and oncogenic transformation in wild-type (wt) or uPAR-/- mouse embryonic fibroblasts (MEFs). Surprisingly, uPAR-/- MEFs grew faster than wt MEFs. This agreed with elevated levels of cell cycle mediators like extracellular signal-regulated protein kinase, p38, AP1 and Cyclin D1. Infection with a uPAR retrovirus reverted the effect, decreasing the growth rate. When MEFs were transformed with H-Ras(V12) and E1A oncogenes, the efficiency of transformation in uPAR-/- MEFs was higher than in wt. UPAR-/- MEFs grew faster at low serum, produced more colonies in agar and produced tumors in vivo in nude mice with a lower latency period. The properties of the heterozygous uPAR+/- MEFs were always intermediate. We conclude therefore that in MEFs uPAR concentration controls cell proliferation and the transforming activity of some oncogenes.
Assuntos
Proliferação de Células , Transformação Celular Neoplásica , Embrião de Mamíferos/metabolismo , Fibroblastos/metabolismo , Receptores de Superfície Celular/metabolismo , Animais , Apoptose , Células Cultivadas , Inibidor p16 de Quinase Dependente de Ciclina/genética , Inibidor p16 de Quinase Dependente de Ciclina/fisiologia , Embrião de Mamíferos/citologia , Fibroblastos/citologia , Regulação da Expressão Gênica , Homozigoto , Camundongos , Camundongos Knockout , Camundongos Nus , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Invasividade Neoplásica , Proteína Oncogênica p21(ras)/genética , Proteína Oncogênica p21(ras)/metabolismo , Receptores de Superfície Celular/genética , Receptores de Ativador de Plasminogênio Tipo Uroquinase , Fator de Transcrição AP-1/metabolismo , Transdução Genética , Transfecção , Vitronectina/metabolismoRESUMO
We have investigated the in vivo and in vitro regulation of the human urokinase-type plasminogen activator (uPA) gene by interleukin-1 (IL-1) and analyzed the transcription factors and signalling pathways involved in the response of the -2.0-kb uPA enhancer to IL-1 induction and to tetradecanoyl phorbol acetate (TPA) induction. Mutational analysis showed the cooperative activity of the Ets-binding site (EBS) and the two AP-1 elements of the enhancer. The results reveal that the EBS is required for the response to both inducers mediated by Ets-2, which is regulated at a level subsequent to DNA binding, by an IL-1- and phorbol ester-inducible transactivation domain. Both the IL-1 and the TPA-mediated induction result in a drastic increase of AP-1 binding to the downstream site of the enhancer (uPA 3' TPA-responsive element), while a mostly qualitative change, resulting from the interplay between ATF-2 homodimers and c-Jun-ATF-2 heterodimers, takes place at the upstream AP-1 element. The analysis of two distinct mitogen-activated protein kinase pathways shows that stress-activated protein kinase-Jun N-terminal kinase activation, resulting in the phosphorylation of ATF-2, c-Jun, and JunD, is required not only for the IL-1- but also for the TPA-dependent induction, while the extracellular signal-related kinase 1 (ERK-1) and ERK-2 activation is involved in the TPA- but not in the IL-1-dependent stimulation of the uPA enhancer.
Assuntos
Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Proteínas de Ligação a DNA , Proteínas Repressoras , Ativador de Plasminogênio Tipo Uroquinase/genética , Fator 2 Ativador da Transcrição , Sequência de Bases , Sítios de Ligação/genética , Linhagem Celular , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , DNA/genética , Primers do DNA/genética , Elementos Facilitadores Genéticos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-1/farmacologia , Dados de Sequência Molecular , Proteína Proto-Oncogênica c-ets-2 , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-jun/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Acetato de Tetradecanoilforbol/farmacologia , Transativadores/metabolismo , Fator de Transcrição AP-1/metabolismo , Fatores de Transcrição/metabolismo , Ativação TranscricionalRESUMO
The proteins of the Ets family are transcription factors involved in signal transduction, cell cycle progression, and differentiation. In this study, we report that thyroid cell neoplastic transformation is associated with a dramatic increase in ETS transcriptional activity, which is dependent on the accumulation of Ets-1, Ets-2, and other Ets-related proteins. Inhibition of ETS transactivation activity by the Ets-dominant negative construct (Ets-Z) induced programmed cell death in human thyroid carcinoma cell lines but not in normal thyroid cells. Apoptotic cell death induced by Ets-Z was dependent on the reduction of c-MYC protein levels, because it was prevented by overexpression of c-myc. Taken together, these data indicate that the induction of Ets-1 and Ets-2 transcription factors plays a pivotal role in thyroid cell neoplastic transformation.
Assuntos
Transformação Celular Neoplásica/metabolismo , Proteínas de Ligação a DNA , Proteínas Proto-Oncogênicas/biossíntese , Proteínas Repressoras , Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/metabolismo , Transativadores/biossíntese , Fatores de Transcrição/biossíntese , Apoptose/fisiologia , Sequência Consenso , DNA/metabolismo , Genes myc/genética , Humanos , Proteína Proto-Oncogênica c-ets-1 , Proteína Proto-Oncogênica c-ets-2 , Proteínas Proto-Oncogênicas/fisiologia , Proteínas Proto-Oncogênicas c-ets , Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/patologia , Transativadores/fisiologia , Fatores de Transcrição/fisiologia , Ativação Transcricional/fisiologia , Células Tumorais CultivadasRESUMO
Dimerization plays a pivotal role in modulating the activity of the c-Jun proto-oncogene product. Heterodimerization with activating transcription factor-2 (ATF-2) alters the DNA-binding specificity of c-Jun, allowing its targeting to several cAMP responsive element (CRE)-related sequences, which control a subset of AP-1-responsive genes. Here we show that a c-Jun/ATF-2 heterodimer binds to the AP-1 site (uPA 5'-TRE) essential for the activity of the human urokinase enhancer, conferring on this element several distinctive regulatory properties. The c-Jun/ATF-2 heterodimer was identified by binding competition assays, u.v. cross linking, and monospecific antibodies. In vitro binding studies revealed that the uPA 5'-TRE sequence is recognized by the cyclic AMP-unresponsive ATF-2 factor, but not by the cyclic AMP-inducible CREB. In addition, in vivo studies suggest that ATF-2 can mediate, at the same time, the activation of the c-Jun/ATF-2 site and the repression of the canonical collagenase AP-1 site. We report that heterodimerization with c-Fos does not increase the binding of c-Jun to the uPA 5'-TRE, in contrast to the increased binding at a consensus AP-1 site. Our data further suggest that c-Fos can act as a repressor of the c-Jun/ATF-2 binding site, revealing an important functional difference, with respect to canonical AP-1 elements.
Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/química , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Elementos Facilitadores Genéticos/fisiologia , Proteínas Proto-Oncogênicas c-fos/química , Proteínas Proto-Oncogênicas c-fos/metabolismo , Proteínas Proto-Oncogênicas c-jun/química , Proteínas Proto-Oncogênicas c-jun/metabolismo , Fatores de Transcrição/metabolismo , Ativador de Plasminogênio Tipo Uroquinase/genética , Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Fator 2 Ativador da Transcrição , Sequência de Bases , Sítios de Ligação , Humanos , Zíper de Leucina/fisiologia , Dados de Sequência Molecular , Ésteres de Forbol/farmacologia , Conformação Proteica/efeitos dos fármacos , Proto-Oncogene Mas , Transcrição Gênica/fisiologiaRESUMO
We have recently reported that neoplastic transformation of two rat thyroid epithelial cell lines by retroviruses carrying the v-mos and v-ras Ki oncogenes is associated with a drastic increase of AP-1 activity. The most important effects were represented by the dramatic junB and fra-1 gene induction, which was abolished by the block of the transformation-induced HMGI-C protein synthesis. Here, we have further characterized the transformation-dependent AP-1 activity, by analysing the expression of different jun- and fos-related components, in rat thyroid cell lines transformed by several oncogenes, in human thyroid carcinoma cell lines, and in naturally occurring human thyroid tumours. A significant increase of Fra-1 and JunB protein levels was detected in all oncogene transformed rat thyroid cell lines. Fra-1 gene induction was demonstrated to occur also in human thyroid carcinoma cell lines and tissues. Conversely, c-Jun and JunD proteins, rather than JunB, accumulated in human thyroid carcinoma cell lines. An induction of AP-1 target genes was also detected both in rat and human thyroid transformed cell lines. Therefore, in vivo and in vitro thyroid cell transformation is associated with important compositional changes in the AP-1 complex and an increased transcriptional activity.
Assuntos
Transformação Celular Neoplásica , Oncogenes , Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/genética , Fator de Transcrição AP-1/metabolismo , Animais , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Genes jun , Proteína HMGA2 , Proteínas de Grupo de Alta Mobilidade/biossíntese , Humanos , Proteínas de Neoplasias/biossíntese , Reação em Cadeia da Polimerase , Proteínas Proto-Oncogênicas c-fos/biossíntese , Proteínas Proto-Oncogênicas c-jun/biossíntese , Ratos , Tireoglobulina/biossíntese , Glândula Tireoide/patologia , Tireotropina/farmacologiaRESUMO
Glucose-6-phosphate dehydrogenase-deleted embryonic stem (ES) cells (G6pd Delta) proliferate in vitro without special requirements, but when challenged with oxidants fail to sustain glutathione disulphide reconversion to reduced glutathione (GSH), entering a condition of oxidative stress. Here, we investigate the signalling events downstream of GSH oxidation in G6pd Delta and wild-type (wt) ES cells. We found that G6pd Delta ES cells are very sensitive to oxidants, activating an apoptotic pathway at oxidant concentrations otherwise sublethal for wt ES cells. We show that the apoptotic pathway activated by low oxidant concentrations is accompanied by mitochondria dysfunction, and it is therefore blocked by the overexpression of Bcl-X(L). Bcl-X(L) does not inhibit the decrease in cellular GSH and reactive oxygen species formation following oxidant treatment. We also found that oxidant treatment in ES cells is followed by the activation of the MEK/extracellular signal-regulated kinase (ERK) pathway. Interestingly, ERK activation has opposite outcomes in G6pd Delta ES cells compared to wt, which has a proapoptotic function in the first and a prosurvival function in the latter. We show that this phenomenon can be regulated by the cellular GSH level.
Assuntos
Apoptose/fisiologia , Glucosefosfato Desidrogenase/metabolismo , Glutationa/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Células-Tronco/citologia , Animais , Apoptose/efeitos dos fármacos , Caspases , Diamida/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , MAP Quinase Quinase 4 , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Camundongos , Mitocôndrias/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Células-Tronco/efeitos dos fármacos , Reagentes de Sulfidrila/farmacologia , Proteína bcl-XRESUMO
fra-1 gene overexpression has been shown to represent a general event in thyroid cell transformation in vitro and in vivo. Moreover, inhibition of FRA-1 protein synthesis by stable transfection with a fra-1 antisense construct significantly reduces the malignant phenotype of the transformed thyroid cells, indicating a pivotal role of the fra-1 gene product in the process of cellular transformation. In the attempt to define the potential use of FRA-1 protein detection in the diagnosis of thyroid diseases, we analyzed Fra-1 expression by a combination of immunohistochemistry and reverse transcription-PCR (RT-PCR) assay in 174 samples of thyroid nodules (22 nodular hyperplasias, 102 follicular adenomas, 34 papillary carcinomas, 12 follicular carcinomas, and 4 anaplastic carcinomas) representative of the spectrum of thyroid tumor pathology. FRA-1 protein was abundant in all of the carcinoma samples (50/50, 100%), with an intense staining in the nucleus and the cytoplasm. Positive staining was also found in most of the adenomas (90 of 102; 88%), but in this case, the staining was restricted to the nucleus. Similar results were obtained from the analysis of thyroid goiters; however, the number of positive cases is lower than adenomas (8 of 22; 36%); moreover, the staining was not observed in all of the cells. Conversely, no FRA-1 protein was detectable in 12 normal thyroid tissue samples used as controls. RT-PCR analysis confirmed a higher fra-1 expression in papillary and follicular carcinomas compared with goiters and adenomas. fra-1 expression was also analyzed on 10 fine needle aspiration biopsy (FNAB) samples by RT-PCR. fra-1-specific mRNA was detected in seven of the eight FNABs corresponding to thyroid nodules that were eventually diagnosed as adenomas (three of four) and carcinomas (four of four) after surgery. Conversely, no fra-1 gene expression was observed in two FNABs derived from normal thyroid. Further studies are required before suggesting FRA-1 protein detection as a useful tool for the diagnosis of hyperplastic and neoplastic disorders of the thyroid gland.
Assuntos
Proteínas Proto-Oncogênicas c-fos/análise , Proteínas Proto-Oncogênicas c-fos/genética , Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/metabolismo , Nódulo da Glândula Tireoide/metabolismo , Biópsia por Agulha , Proteína HMGA1a , Proteínas de Grupo de Alta Mobilidade/análise , Proteínas de Grupo de Alta Mobilidade/genética , Humanos , Hiperplasia , Imuno-Histoquímica , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias da Glândula Tireoide/patologia , Nódulo da Glândula Tireoide/patologia , Fatores de Transcrição/análise , Fatores de Transcrição/genética , Células Tumorais CultivadasRESUMO
The hormonal regulation of the human urokinase type plasminogen activator (uPA) gene has been studied by introducing into mouse and rat Sertoli cell primary cultures a recombinant plasmid, in which the transcription regulatory elements of the cloned human uPA gene drive the expression of the bacterial chloramphenicol-acetyl-transferase gene. It was found to be expressed and regulated by FSH and (Bu)2cAMP in the mouse cells only, in agreement with data on the expression of the endogenous gene in rat and mouse gonads. The stimulation of transcription by FSH was evident in cultures from 13-day-old but not from 18-day-old mice, even though (Bu)2cAMP induction could be observed at both ages. Phorbol-myristate acetate was found to activate the human uPA promoter in Sertoli cell cultures from mice of both ages, even though the effect was less evident in cultures of 18-day-old animals. Deletion analysis of the human uPA 5'-flanking region showed that the distal enhancer element is not needed for (Bu)2cAMP induction, and that at least two promoter regions are involved in (Bu)2cAMP induced transcription. One of these cAMP responsive regions lies between nucleotides -72 and -29 from the CAP site. The sequence of this region would suggest the binding of transcription factor AP-2, a cell-specific mediator of both cAMP and phorbol esters action on gene expression. However, these sequences do not mediate phorbol ester activation of human uPA promoter in mouse Sertoli cells.
Assuntos
AMP Cíclico/farmacologia , Hormônio Foliculoestimulante/farmacologia , Ativadores de Plasminogênio/genética , Regiões Promotoras Genéticas/genética , Células de Sertoli/metabolismo , Transcrição Gênica/efeitos dos fármacos , Ativador de Plasminogênio Tipo Uroquinase/genética , Animais , Sequência de Bases , Células Cultivadas , AMP Cíclico/análogos & derivados , Fibrinolíticos/metabolismo , Masculino , Camundongos , Dados de Sequência Molecular , Ésteres de Forbol/farmacologia , Plasmídeos , Ativadores de Plasminogênio/metabolismo , Ativadores de Plasminogênio/fisiologia , Células de Sertoli/efeitos dos fármacos , Transfecção , Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Ativador de Plasminogênio Tipo Uroquinase/fisiologiaRESUMO
MicroRNAs (miRNAs) control cell cycle progression by targeting the transcripts encoding for cyclins, CDKs and CDK inhibitors, such as p27(KIP1) (p27). p27 expression is controlled by multiple transcriptional and posttranscriptional mechanisms, including translational inhibition by miR-221/222 and posttranslational regulation by the SCF(SKP2) complex. The oncosuppressor activity of miR-340 has been recently characterized in breast, colorectal and osteosarcoma tumor cells. However, the mechanisms underlying miR-340-induced cell growth arrest have not been elucidated. Here, we describe miR-340 as a novel tumor suppressor in non-small cell lung cancer (NSCLC). Starting from the observation that the growth-inhibitory and proapoptotic effects of miR-340 correlate with the accumulation of p27 in lung adenocarcinoma and glioblastoma cells, we have analyzed the functional relationship between miR-340 and p27 expression. miR-340 targets three key negative regulators of p27. The miR-340-mediated inhibition of both Pumilio family RNA-binding proteins (PUM1 and PUM2), required for the miR-221/222 interaction with the p27 3'-UTR, antagonizes the miRNA-dependent downregulation of p27. At the same time, miR-340 induces the stabilization of p27 by targeting SKP2, the key posttranslational regulator of p27. Therefore, miR-340 controls p27 at both translational and posttranslational levels. Accordingly, the inhibition of either PUM1 or SKP2 partially recapitulates the miR-340 effect on cell proliferation and apoptosis. In addition to the effect on tumor cell proliferation, miR-340 also inhibits intercellular adhesion and motility in lung cancer cells. These changes correlate with the miR-340-mediated inhibition of previously validated (MET and ROCK1) and potentially novel (RHOA and CDH1) miR-340 target transcripts. Finally, we show that in a small cohort of NSCLC patients (n=23), representative of all four stages of lung cancer, miR-340 expression inversely correlates with clinical staging, thus suggesting that miR-340 downregulation contributes to the disease progression.
Assuntos
Adenocarcinoma/genética , Adenocarcinoma/patologia , Apoptose/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Inibidor de Quinase Dependente de Ciclina p27/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , MicroRNAs/genética , Adenocarcinoma de Pulmão , Ciclo Celular/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Humanos , Proteínas de Ligação a RNA/genética , Regulação para Cima/genéticaRESUMO
Rats with unilateral dopamine denervation exhibit turning behaviour in response to the selective D1 agonist SKF 38393 only after a previous exposure to dopamine agonists. We demonstrate here that this 'priming' phenomenon is related to both an increased expression of the pre-existing AP-1 complex and the occurrence of novel AP-1 complexes which are formed by FosB- and JunD-related proteins. While the former protein is expressed as a consequence of the dopamine denervation, the latter is related to the first exposure to a dopamine agonist. Pre-treatment with MK-801, an antagonist for glutamatergic receptors, prevents both the priming development and the AP-1 compositional changes. Rotational behaviour induced by SKF 38393 closely correlates with the presence of the priming AP-1 complexes, regardless of the capability of the D1 agonist to induce the immediate-early gene cFos.
Assuntos
2,3,4,5-Tetra-Hidro-7,8-Di-Hidroxi-1-Fenil-1H-3-Benzazepina/farmacologia , Benserazida/farmacologia , Corpo Estriado/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Levodopa/farmacologia , Atividade Motora/fisiologia , Neurônios/metabolismo , Proteínas Proto-Oncogênicas c-fos/biossíntese , Proteínas Proto-Oncogênicas c-jun/biossíntese , Fator de Transcrição AP-1/metabolismo , Animais , Núcleo Celular/metabolismo , Corpo Estriado/efeitos dos fármacos , Denervação , Maleato de Dizocilpina/farmacologia , Masculino , Atividade Motora/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Oxidopamina , Ratos , Ratos Sprague-Dawley , Rotação , Comportamento Estereotipado/efeitos dos fármacosRESUMO
The authors describe the method used in Argentina for quantification of risk in controls of the potency and innocuity of foot and mouth disease vaccine. Quantitative risk analysis is a relatively new tool in the animal health field, and is in line with the principles of transparency and equivalency of the Sanitary and Phytosanitary Agreement of the Uruguay Round of the General Agreement on Tariffs and Trade (GATT: now World Trade Organisation [WTO]). The risk assessment is presented through a description of the steps involved in manufacturing the vaccine, and the controls performed by the manufacturer and by the National Health Animal Service (Servicio Nacional de Sanidad Animal: SENASA). The adverse situation is considered as the lack of potency or innocuity of the vaccine, and the risk is estimated using a combination of the Monte Carlo simulation and the application of a Bayesian model.
Assuntos
Aphthovirus/imunologia , Vacinas Virais/normas , Adjuvantes Imunológicos/normas , Animais , Argentina , Teorema de Bayes , Método de Monte Carlo , Probabilidade , Medição de Risco , Vacinas Virais/efeitos adversosRESUMO
Anomalies in the vascular structures of the neck, cranial base, temporal bone, and intracranial circulation may give rise to pulsatile tinnitus. If the anomalous sound is perceived also by others, then it is defined as objective tinnitus. Herein, the case is reported of right pulsatile tinnitus of one year's duration, which appeared after cranial trauma. Magnetic resonance angiography showed that the jugular bulb was dominant on the same side as the tinnitus. The tinnitus was recorded with a high-sensitivity microphone, while otoacoustic emissions were measured by distortion products during follow-up.