RESUMO
The aim of this study was to investigate the effects of acute glyphosate (active ingredient) exposure on the oxidative stress biomarkers and antioxidant defenses of a hybrid surubim (Pseudoplatystoma sp). The fish were exposed to different herbicide concentrations for 96 h. The thiobarbituric acid-reactive substances (TBARS), protein carbonyls and antioxidant responses were verified. The 15 mg a.pL(-1) of herbicide resulted in the death of 50% of the fish after 96 h. An increase in liver and muscle TBARS levels was observed when fish were exposed to the herbicide. The protein carbonyl content was also increased in the liver (4.5mg a.pL(-1) concentration) and brain (2.25 mg a.pL(-1) concentration). The antioxidant activities decreased in the liver and brain after exposure to herbicide. Levels of ascorbic acid in the liver (2.25 mg a.pL(-1) and 4.5 mg a.pL(-1) concentrations) and brain (2.25 mg a.pL(-1) concentration) were increased post-treatment. Levels of total thiols were increased in the liver and brain (2.25 mg L(-1) and 7.5mg a.pL(-1), respectively). Glyphosate exposure, at the tested concentrations affects surubim health by promoting changes that can affect their survival in natural environment. Some parameters as TBARS and protein carbonyl could be early biomarkers for Roundup exposure in this fish species.
Assuntos
Peixes-Gato/fisiologia , Glicina/análogos & derivados , Herbicidas/toxicidade , Fígado/efeitos dos fármacos , Músculos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Ácido Ascórbico/análise , Biomarcadores/metabolismo , Glicina/toxicidade , Fígado/química , Fígado/metabolismo , Músculos/química , Músculos/metabolismo , Oxirredução , Carbonilação Proteica , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , GlifosatoRESUMO
JUSTIFICATIVA E OBJETIVOS: A constante de equilíbrio entre o plasma e o sítio efetor (ke0) é utilizada pelos modelos farmacocinéticos para prever a concentração do fármaco em seu local de ação (Ce). Seria interessante que a Ce de propofol fosse semelhante na perda e na recuperação da consciência. O objetivo deste estudo foi avaliar o desempenho clínico de duas diferentes ke0 (rápida = 1,21 min-1 e lenta = 0,26 min-1) com relação à Ce durante a perda e a recuperação da consciência, usando o modelo farmacocinético de Marsh. MÉTODO: Participaram deste estudo 20 voluntários adultos sadios do sexo masculino. Em todos os voluntários, administrou-se propofol em regime de infusão alvo-controlada, modelo farmacocinético de Marsh ke0 rápida e, em outra oportunidade, usou-se o mesmo modelo farmacocinético com a ke0 lenta. Inicialmente, o propofol foi infundido em concentração-alvo plasmática de 3,0µg.mL-1. A perda de consciência e a recuperação de consciência basearam-se na resposta ao estímulo verbal. A Ce foi anotada no momento da perda e da recuperação da consciência. RESULTADOS: Na perda e na recuperação da consciência a Ce pela ke0 rápida foi diferente (3,64 ± 0,78 e 1,47 ± 0,29µg.mL-1, respectivamente, p < 0,0001), enquanto com a ke0 lenta a Ce foi semelhante (2,20 ± 0,70 e 2,13 ± 0,43µg.mL-1, respectivamente, p = 0,5425). CONCLUSÕES: Do ponto de vista clínico, a ke0 lenta (0,26 min-1) incorporada ao modelo farmacocinético de Marsh apresentou melhor desempenho que a ke0 rápida (1,21 min-1), uma vez que a concentração de propofol prevista em seu local de ação na perda e recuperação da consciência foi semelhante.
BACKGROUND AND OBJECTIVE: The constant equilibrium between the plasma and effect site (ke0) is used by pharmacokinetic models to calculate a drug concentration in its site of action (Ce). It would be interesting if Ce of propofol was similar at loss and recovery of consciousness. The objective of this study was to evaluate the clinical performance of two different ke0 (fast = 1.21 min-1, and slow = 0.26 min-1) in relation to Ce during loss and recovery of consciousness using Marsh pharmacokinetic model. METHODS: Twenty healthy adult male volunteers participated in this study. In all volunteers propofol was administered as target-controlled infusion, Marsh pharmacokinetic model for fast ke0 and, at a different time, the same pharmacokinetic model with slow ke0 was used. Initially, propofol was infused with a serum target-controlled infusion of 3.0 µg.mL-1. Loss of consciousness and recovery of consciousness were based on response to verbal stimulus. Ce was recorded at the moment of loss and recovery of consciousness. RESULTS: On loss and recovery of consciousness, the Ce for fast ke0 was different (3.64 ± 0.78 and 1.47 ± 0.29 µg.mL-1, respectively, p < 0.0001), while with slow ke0 the Ce was similar (2.20 ± 0.70 and 2.14 ± 0.43 µg.mL-1, respectively, p = 0.5425). CONCLUSIONS: Clinically, the slow ke0 (0.26 min-1) incorporated in the Marsh pharmacokinetic model showed better performance than the fast ke0 (1.21 min-1), since the calculated concentration of propofol at the effect site on loss and recovery of consciousness was similar.
JUSTIFICATIVA Y OBJETIVOS: La constante de equilibrio entre el plasma y el sitio efector (ke0), se usa por los modelos farmacocinéticos para prever la concentración del fármaco en su región de acción (Ce). Sería interesante que el Ce de propofol fuese similar en la pérdida y en la recuperación de la conciencia. El objetivo de este estudio, fue evaluar el desempeño clínico de dos diferentes ke0 (rápida = 1,21 min-1 y lenta = 0,26 min-1), con relación a la Ce durante la pérdida y la recuperación de la conciencia, usando el modelo farmacocinético de Marsh. MéTODO: Participaron en este estudio, 20 voluntarios adultos sanos del sexo masculino. A todos los voluntarios se les administró propofol en régimen de infusión objeto controlada, modelo farmacocinético de Marsh ke0 rápida y en otro momento, se usó el mismo modelo farmacocinético con a ke0 lenta. Inicialmente, el propofol se infundió en concentración-objeto plasmática de 3,0 µg.mL-1. La pérdida de la conciencia y la recuperación de la conciencia estuvieron basadas en la respuesta al estímulo verbal. La Ce fue anotada en el momento de la pérdida y de la recuperación de la conciencia. RESULTADOS: En la pérdida y en la recuperación de la conciencia, la Ce por la ke0 rápida, fue diferente (3,64 ± 0,78 y 1,47 ± 0,29 µg.mL-1, respectivamente, p < 0,0001), mientras que con la ke0 lenta la Ce fue parecida (2,20 ± 0,70 y 2,13 ± 0,43 µg.mL-1, respectivamente, p = 0,5425). CONCLUSIONES: Desde el punto de vista clínico, la ke0 lenta (0,26 min-1) incorporada al modelo farmacocinético de Marsh, presentó un mejor desempeño que la ke0 rápida (1,21 min-1), pues la concentración de propofol prevista en su región de acción en la pérdida y en la recuperación de la conciencia fue similar.
Assuntos
Adulto , Humanos , Masculino , Estado de Consciência/efeitos dos fármacos , Hipnóticos e Sedativos/farmacocinética , Propofol/farmacocinética , Modelos BiológicosRESUMO
BACKGROUND AND OBJECTIVE: The constant equilibrium between the plasma and effect site (ke0) is used by pharmacokinetic models to calculate a drug concentration in its site of action (Ce). It would be interesting if Ce of propofol was similar at loss and recovery of consciousness. The objective of this study was to evaluate the clinical performance of two different ke0 (fast = 1.21 min(-1), and slow = 0.26 min(-1)) in relation to Ce during loss and recovery of consciousness using Marsh pharmacokinetic model. METHODS: Twenty healthy adult male volunteers participated in this study. In all volunteers propofol was administered as target-controlled infusion, Marsh pharmacokinetic model for fast ke0 and, at a different time, the same pharmacokinetic model with slow ke0 was used. Initially, propofol was infused with a serum target-controlled infusion of 3.0 µg.mL(-1). Loss of consciousness and recovery of consciousness were based on response to verbal stimulus. Ce was recorded at the moment of loss and recovery of consciousness. RESULTS: On loss and recovery of consciousness, the Ce for fast ke0 was different (3.64 ± 0.78 and 1.47 ± 0.29 µg.mL(-1), respectively, p < 0.0001), while with slow ke0 the Ce was similar (2.20 ± 0.70 and 2.14 ± 0.43 µg.mL(-1), respectively, p = 0.5425). CONCLUSIONS: Clinically, the slow ke0 (0.26 min(-1)) incorporated in the Marsh pharmacokinetic model showed better performance than the fast ke0 (1.21 min(-1)), since the calculated concentration of propofol at the effect site on loss and recovery of consciousness was similar.
Assuntos
Estado de Consciência/efeitos dos fármacos , Hipnóticos e Sedativos/farmacocinética , Propofol/farmacocinética , Adulto , Humanos , Masculino , Modelos BiológicosRESUMO
Three rusts, Puccinia nassellae, Uromyces pencanus, and Puccinia graminella, are being studied as potential biological control agents for Nassella neesiana (Chilean needle grass) in Australia and New Zealand. An understanding of the life cycle of a pathogen is desirable before its release as a biocontrol agent is considered, to enable the assessment of the risks involved in such a release. Field observations and experiments have been carried out to elucidate the life cycles of these three pathogens. Puccinia nassellae cycles as urediniospores and produces dormant teliospores. Dormancy of teliospores has been broken through manipulation in the laboratory, but resulting basidiospores have failed to infect N. neesiana plants under the conditions tested. Uromyces pencanus cycles as urediniospores and its telia appear to have lost the capacity to produce basidiospores. Aecia have been reported for this rust in the literature. However, evidence is provided that these aecia in fact belong to the life cycle of P. graminella. Puccinia graminella produces only aecia and telia. The aeciospores have been shown to be repetitive (aecidioid urediniospores). Teliospores germinate directly without a dormant phase, but resulting basidiospores failed to infect N. neesiana plants under the conditions tested. The role of teliospores in the life cycle of all three rusts remains unknown. Although the autoecious nature of their life cycles has not been proven experimentally, neither is there any evidence that they are heteroecious. Practical and theoretical implications of these findings are discussed.
Assuntos
Basidiomycota/crescimento & desenvolvimento , Poaceae/microbiologia , Esporos Fúngicos/crescimento & desenvolvimento , Australásia , Basidiomycota/patogenicidade , Doenças das Plantas/microbiologia , Esporos Fúngicos/patogenicidadeRESUMO
The use of conditioned medium (CM) from human cardiac explants (HCEs) as a potential source of paracrine factors for adult stem cell signaling has never been evaluated. We hypothesized that HCEs might provide a source of soluble factors triggering the differentiation of mesenchymal stem cells (MSCs) into cardiomyocyte-like cells. By using two-dimensional electrophoresis (2-DE) gels/mass spectrometry and antibody macroarray assays, we found that HCEs release macromolecules, including cytokines, growth factors and myocardial and metabolism-related proteins into the culture medium. We identified a total of 20 proteins in the HCE-CM. However, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 2-DE, these 20 proteins account for only a fraction of the total number of proteins present in the HCE-CM. We also found that CM increased the proliferation of bone marrow-derived-MSCs (BM-MSCs) in vitro. Unlike the other effects, this effect was most evident after 48 h of culture. Moreover, we examined the effect of HCE-CM on levels of mRNA and protein for specific cardiac markers. We showed that a surprisingly big fraction of BM-MSCs (3.4-5.0%) treated in vitro with HCE-CM became elongated and began to express cardiac markers, consistent with their possible differentiation into cardiomyocyte-like cells. Our in vitro model may be useful not only per se, but also for studies of the mechanisms of action of soluble factors involved in cell differentiation, paving the way for possible new protein-based treatments in the future.
Assuntos
Células da Medula Óssea/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Meios de Cultivo Condicionados/química , Meios de Cultivo Condicionados/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Miócitos Cardíacos/efeitos dos fármacos , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Células Cultivadas , Técnicas de Cultura , Citocinas/química , Citocinas/farmacologia , Eletroforese em Gel Bidimensional , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/química , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Espectrometria de Massas , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Análise em Microsséries , Miócitos Cardíacos/citologia , Miócitos Cardíacos/metabolismo , Proteínas/química , Proteínas/farmacologia , RNA Mensageiro/metabolismo , Solubilidade , Fatores de Tempo , Técnicas de Cultura de TecidosRESUMO
Endothelial progenitor cells (EPCs), which express the CD133 marker, can differentiate into mature endothelial cells (ECs) and create new blood vessels. Normal angiogenesis is unable to repair the injured tissues that result from myocardial infarction (MI). Patients who have high cardiovascular risks have fewer EPCs and their EPCs exhibit greater in vitro senescence. Human umbilical cord blood (HUCB)-derived EPCs could be an alternative to rescue impaired stem cell function in the sick and elderly. The aim of this study was to purify HUCB-derived CD133(+) cells, expand them in vitro and evaluate the efficacy of the purified and expanded cells in treating MI in rats. CD133(+) cells were selected for using CD133-coupled magnetic microbeads. Purified cells stained positive for EPC markers. The cells were expanded and differentiated in media supplemented with fetal calf serum and basic fibroblast growth factor, insulin-like growth factor-I and vascular endothelial growth factor (VEGF). Differentiation was confirmed by lack of staining for EPC markers. These expanded cells exhibited increased expression of mature EC markers and formed tubule-like structures in vitro. Only the expanded cells expressed VEGF mRNA. Cells were expanded up to 70-fold during 60 days of culture, and they retained their functional activity. Finally, we evaluated the therapeutic potential of purified and expanded CD133(+) cells in treating MI by intramyocardially injecting them into a rat model of MI. Rats were divided into three groups: A (purified CD133(+) cells-injected); B (expanded CD133(+) cells-injected) and C (saline buffer-injected). We observed a significant improvement in left ventricular ejection fraction for groups A and B. In summary, CD133(+) cells can be purified from HUCB, expanded in vitro without loosing their biological activity, and both purified and expanded cells show promising results for use in cellular cardiomyoplasty. However, further pre-clinical testing should be performed to determine whether expanded CD133(+) cells have any clinical advantages over purified CD133(+) cells.
Assuntos
Antígenos CD/metabolismo , Sangue Fetal/citologia , Glicoproteínas/metabolismo , Infarto do Miocárdio/terapia , Peptídeos/metabolismo , Transplante de Células-Tronco , Antígeno AC133 , Animais , Sequência de Bases , Capilares/crescimento & desenvolvimento , Diferenciação Celular , Proliferação de Células , Primers do DNA/genética , Células Endoteliais/citologia , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Sangue Fetal/imunologia , Sangue Fetal/metabolismo , Humanos , Separação Imunomagnética , Técnicas In Vitro , Recém-Nascido , Masculino , Infarto do Miocárdio/fisiopatologia , Neovascularização Fisiológica , Fenótipo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Transplante Heterólogo , Fator A de Crescimento do Endotélio Vascular/genética , Função Ventricular EsquerdaRESUMO
Mesenchymal stem cells (MSCs) have received special attention for cardiomyoplasty because several studies have shown that they differentiate into cardiomyocytes both in vitro and in vivo. Nitric oxide (NO) is a free radical signaling molecule that regulates several differentiation processes including cardiomyogenesis. Here, we report an investigation of the effects of two NO agents (SNAP and DEA/NO), able to activate both cGMP-dependent and -independent pathways, on the cardiomyogenic potential of bone marrow-derived mesenchymal stem cells (BM-MSCs) and adipose tissue-derived stem cells (ADSCs). The cells were isolated, cultured and treated with NO agents. Cardiac- and muscle-specific gene expression was analyzed by indirect immunofluorescence, flow cytometry, RT-PCR and real-time PCR. We found that untreated (control) ADSCs and BM-MSCs expressed some muscle markers and NO-derived intermediates induce an increased expression of some cardiac function genes in BM-MSCs and ADSCs. Moreover, NO agents considerably increased the pro-angiogenic potential mostly of BM-MSCs as determined by VEGF mRNA levels.
Assuntos
Células-Tronco Adultas/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Hidrazinas/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Miócitos Cardíacos/citologia , Doadores de Óxido Nítrico/farmacologia , Penicilamina/análogos & derivados , Adulto , Células-Tronco Adultas/citologia , Células-Tronco Adultas/metabolismo , Idoso , Antígenos CD/genética , Cardiomioplastia , Diferenciação Celular/genética , Células Cultivadas , Conexina 43/genética , Expressão Gênica , Marcadores Genéticos , Coração/fisiologia , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Pessoa de Meia-Idade , Células-Tronco Multipotentes/citologia , Células-Tronco Multipotentes/efeitos dos fármacos , Células-Tronco Multipotentes/metabolismo , Proteínas Musculares/genética , Óxido Nítrico/metabolismo , Penicilamina/farmacologia , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
OBJETIVO: As células progenitoras endoteliais (CPE), caracterizadas pelo marcador CD133+, contribuem para a neovascularização, e o aumento no número dessas células pode ser uma ferramenta terapêutica promissora. O sangue de cordão umbilical humano contém um número significante de CPE, sugerindo a possibilidade do uso destas células para a revascularização de tecidos isquêmicos. O objetivo desse trabalho foi analisar a funcionalidade das células CD133+ diferenciadas in vitro. MÉTODOS: As células diferenciadas foram caracterizadas por citometria de fluxo; a expressão do mRNA de VEGF foi avaliada por RT-PCR e a funcionalidade, por meio de ensaios de formação de túbulos capilares. RESULTADOS: As células diferenciadas perderam os marcadores de CPE, mantiveram em níveis baixos os marcadores das linhagens hematopoética e monocíticas e aumentaram a expressão dos marcadores de células endoteliais adultas. As células diferenciadas apresentaram transcritos no mRNA de VEGF e mostraram-se capazes de formar túbulos capilares in vitro. CONCLUSÃO: As células CD133+ diferenciadas in vitro em células endoteliais demonstraram serem funcionalmente ativas, abrindo perspectiva para seu uso futuro em aplicações terapêuticas.
OBJECTIVE: Endothelial progenitor cells (EPC) caracterized by the CD133+ marker, contribute to the neovascularization. Increasing EPC number in vitro could be a promising therapeutic tool. Human umbilical cord blood maintains a significant number of EPC, suggesting the possibility to use these cells to induce the revascularization of ischemic tissues. The aim of this study was to analize the in vitro function of differentiated CD133+ cells. METHODS: Cells were characterized by flow cytometry, VEGF mRNA expression was evaluated by the RT-PCR analysis and the functionally by essays of capillary tubes formation. RESULTS: Differentiated cells lost EPC markers, maintained low levels of markers for hematopoietic and monocytic cell lines and increased the expression of adult endothelial cell markers. Differentiated cells expressed VEGF mRNA and were capable to induce in vitro capillary tubules formation. CONCLUSION: CD133+ cells differentiated into endothelial cells in vitro are functionally active initiating the possibility of their use in future therapeutic applications.
Assuntos
Adolescente , Adulto , Feminino , Humanos , Adulto Jovem , Antígenos CD , Diferenciação Celular/fisiologia , Células Endoteliais/fisiologia , Sangue Fetal/citologia , Glicoproteínas , Neovascularização Fisiológica , Peptídeos , Células-Tronco/fisiologia , Capilares , Células Cultivadas , Citometria de Fluxo , Parto , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Mensageiro/metabolismo , Células-Tronco/citologia , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto JovemRESUMO
OBJECTIVE: Endothelial progenitor cells (EPC) characterized by the CD133+ marker, contribute to the neovascularization. Increasing EPC number in vitro could be a promising therapeutic tool. Human umbilical cord blood maintains a significant number of EPC, suggesting the possibility to use these cells to induce the revascularization of ischemic tissues. The aim of this study was to analyze the in vitro function of differentiated CD133+ cells. METHODS: Cells were characterized by flow cytometry, VEGF mRNA expression was evaluated by the RT-PCR analysis and the functionally by essays of capillary tubes formation. RESULTS: Differentiated cells lost EPC markers, maintained low levels of markers for hematopoietic and monocytic cell lines and increased the expression of adult endothelial cell markers. Differentiated cells expressed VEGF mRNA and were capable to induce in vitro capillary tubules formation. CONCLUSION: CD133+ cells differentiated into endothelial cells in vitro are functionally active initiating the possibility of their use in future therapeutic applications.
Assuntos
Antígenos CD , Diferenciação Celular/fisiologia , Células Endoteliais/fisiologia , Sangue Fetal/citologia , Glicoproteínas , Neovascularização Fisiológica , Peptídeos , Células-Tronco/fisiologia , Antígeno AC133 , Adolescente , Adulto , Capilares , Células Cultivadas , Feminino , Citometria de Fluxo , Humanos , Parto , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células-Tronco/citologia , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto JovemRESUMO
Taphrina entomospora is one of the few species of the genus described on native plants of the Southern Hemisphere and also one of the few leaf pathogens known on Nothofagus species. The anatomical changes it produces on N. pumilio leaves, and its morphology, cytology, and sporogenesis were studied. The fungus is a perennial species that overwinters as mycelium in the foliar buds and infects the developing leaves, so the whole blade develops the disease symptoms. Interveinal areas of the leaves become chlorotic, thickened and rounded. Palisade parenchyma fails to develop, with spongy parenchyma developing as packed, rounded, isodiametric cells with little intercellular space. The mycelium is subcuticular, dikaryotic, and produces ascogenous hyphae, asci, and ascospores as described for other species in the genus. Before ascus discharge, ascospores bud in a regular, unique way. The life-cycle of T. entomospora is compared with other representative taxa in the genus and the distribution of this pathogen is discussed.